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V

JANUARY, 1951

PACIFIC

SCIENCE

A QUARTERLY DEVOTED TO THE BIOLOGICAL AND PHYSICAL SCIENCES OF THE PACIFIC REGION

IN THIS ISSUE: Collins — The Cayenne Pineapple • Schwabe — Life History of Oxyspirura mansoni • Aka- mine — Viability of Forest Tree Seeds • Marples — Pacific Symphytognathid Spiders • Webster — Poly- nesian Species of Myoporum • Hubbs^ — Carcharhinus longimanus in the East-Central Pacific • Neal — Flowers of Cassia fistula • Schwartz and Schwartz^ — The Lace-necked Dove in Hawaii • NOTES

Published by

THE UNIVERSITY OF HAWAII HONOLULU, HAWAII

BOARD OF EDITORS

Leonard D. Tuthill, Editor-in-Chief (on leave) Department of Zoology and Entomology, University of Hawaii

O. A. Bushnell, Assistant Editor; Acting Editor-in-Chief Department of Bacteriology, University of Hawaii

Elbert G. Smith, Acting Assistant Editor Department of Chemistry, University of Hawaii

D. Elmo Hardy

Associate Entomologist, University of Hawaii Agricultural Experiment Station

Robert W Hiatt Department of Zoology and Entomology University of Hawaii

Carl L. Hubbs

Scripps Institution of Oceanography La Jolla, California

G. L. Kesteven

Office of the Commissioner General for the United Kingdom in South East Asia Singapore

Luna B. Leopold Hydraulic Engineer

U.S. Geological Survey, Washington 25, D.C.

Joseph E Martin

Plant Pathologist, Experiment Station Hawaiian Sugar Planters’ Association PO. Box 2450, Honolulu 4, Hawaii

Marie C. Neal Botanist

Bishop Museum, Honolulu 35, Hawaii

Harold S. Palmer Department of Geology, University of Hawaii

Harold St. John

Department of Botany, University of Hawaii

Oscar E Sette U.S, Fish and Wildlife Service EO. Box 3830, Honolulu, Hawaii

G. Donald Sherman

Chemist, University of Hawaii Agricultural Experiment Station

Thomas Nickerson, Managing Editor Juliette Wentworth, Assistant Managing Editor Office of Publications and Information, University of Hawaii

SUGGESTIONS TO AUTHORS

Contributions to Pacific biological and physi- cal science will be welcomed from authors in all parts of the world. Manuscripts may be ad- dressed to the Editor-in-Chief, PACIFIC SCIENCE, University of Hawaii, Honolulu 14, Hawaii, or to individual members of the Board of Editors. Use of air mail for sending correspondence and brief manuscripts from distant points is recom- mended.

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script. Authors should not overlook the need for good brief papers, presenting results of studies, notes and queries, communications to the editor, or other commentary.

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Manuscript form. Manuscripts should be typed on one side of standard-size, white bond paper and double-spaced throughout. Pages should be consecutively numbered in upper right-hand corner. Sheets should not be fastened together in any way, and should be mailed flat. Inserts should be either typed on separate sheets or {Continued on inside back cover)

PACIFIC SCIENCE

A QUARTERLY DEVOTED TO THE BIOLOGICAL AND PHYSICAL SCIENCES OF THE PACIFIC REGION

VOL. V JANUARY, 1951 NO. 1

Previous is: ' ublished October 25, 1950

CONTENTS

PAGE

Notes on the Origin, History, and Genetic Nature of the Cayenne Pineapple.

J. L. Collins . 3

Studies on Oxyspirura mansoni, the Tropical Eyeworm of Poultry. II. Life History.

Calvin W Schwabe 18

Viability of Hawaiian Forest Tree Seeds in Storage at Various T emperatures and

Relative Humidities. Ernest K. Akamine 36

Pacific Symphytognathid Spiders. B. J. Marples 47

The Polynesian Species of Myoporum. Grady L. Webster 52

Record of the Shark Carcharhinus longimanus, Accompanied by Naucrates and

Remora, from the East-Central Pacific. Carl L. Hubbs 78

Common and Aberrant Flowers of Cassia fistula. Marie C. Neal 82

A Survey of the Lace-necked Dove in Hawaii. Charles W Schwartz and

Elizabeth Reeder Schwartz 90

NOTES:

Tiuo Juvenile Pointed-tailed Ocean Sunfish, Masturus lanceolatus, from

Hawaiian Waters. Joseph E. King 108

News Notes 110

Pacific Science, a quarterly publication of the University of Hawaii, appears in Janu- ary, April, July, and October. Subscription price is three dollars a year; single copies are one dollar. Check or money order payable to University of Hawaii should be sent to Pacific Science, Office of Publications, University of Hawaii, Honolulu 14, Hawaii. Reprints of articles from Volumes I and II are available to subscribers.

Notes on the Origin, History, and Genetic Nature of the Cayenne Pineapple^

J. L. Collins^

Although plants and fruits of the pine- apple had been carried from America to Europe a number of times following their dis- covery by Columbus in 1493, it was almost 100 years before they were successfully grown in Europe. They were first grown to fruiting in Holland about 1690 and in England some 30 years later, following the development of artificial heating of glasshouses.

Thereafter, a great rivalry and enthusiasm developed in the growing of pineapples by the gardeners of the large estates in England and on the continent. They imported varie- ties from America, exchanged varieties among themselves, and in some instances developed new varieties by growing the seeds which were occasionally found in their fruits or were ob- tained from the West Indies. As a conse- quence of this general interest in the growing of pineapples, the horticultural journals of the times frequently carried articles or notices concerning the culture of this fruit.

Like a number of important agricultural crops, the Cayenne variety of the pineapple, Ananas comosus (L.) Merr., appeared upon the horticultural scene with no definite record of the manner or the place of its origin. The first reference in the literature to the Cayenne variety appears to be the short notice carried in the Gardeners Chronicle (England) of March 6, 1841, under the column heading of "'For- eign Correspondence,” as follows:

^Published with the approval of the Director as Miscellaneous Paper No. 48 of the Pineapple Research Institute of Hawaii, Honolulu, Hawaii. Manuscript received August 23, 1950.

^Department of Genetics, Pineapple Research Insti- tute of Hawaii.

Paris, Feb. 4, 1841. As information is con- tinually demanded by horticulturists regard- ing pines, perhaps a few words as to the varieties in request in Paris and the mode of culture may not be unacceptable. The Pot- ager, or Royal Kitchen Garden Versailles has the merit of producing this and other fruits in the greatest perfection. . . . The pinery con- sists of a vast number of pits, succession and fruiting houses, the whole of which are heated with hot water; it contains about 2000 plants among which are fine specimens of Queens, Cayenne Lisse and Espineux, Black Jamaica . . . in all about 40 varieties. Only four kinds are considered desirable for general cultiva- tion; of these, however, more than 1000 plants are annually fruited, namely 700 Queens and 300 Cayennes, Endville, and Providence. The Cayenne, both smooth and prickly, is of an excellent flavor and weight on the average from 9 to 12 pounds.

This article continues to record the names of the three principal growers, or nurserymen, for the French market, one each in Paris, Versailles, and Meridon, who list Cayenne plants along with some other varieties.

From this date, 1841, we trace the history of the Cayenne variety and map its distribu- tion in geography and time.

In this first reference to the Cayenne variety, mention is made of the smooth-leaved and spiny-leaved forms. Genetic studies in Ha- waii (Collins, 1936: 467) have shown that the smooth-leaved type infrequently produces spiny leaves; if we can judge from this modern behavior, then we can assume that the original Cayenne variety had smooth-edged leaves and also produced mutations giving rise to the spiny-leaved form as bud sports.

C3}

4

PACIFIC SCIENCE, Vol. V, January, 1951

In the Gardeners Chronicle for January 6, 1844, we find another reference to Cayenne in the section on ''Notices to Correspondents,” as follows: "The Cayenne pine is from French Guiana; cannot state its price, that is the af- fair of the nurseryman. ...” A reader of this journal had obviously written to the editor inquiring about the place of origin of Cayenne and the price of plants, and the editor’s reply indicates that it had been imported from French Guiana.

In a description of the Cayenne variety in "The Pineapple Manual” (Anonymous, ca. 1870), we find the statement that "This fine variety was imported from Cayenne many years ago and is now cultivated in many places,”

These scattered and brief references, to- gether with whatever weight may be given because of the name it bears, point rather ■clearly to Cayenne in French Guiana as the place from which this variety was introduced into France some time before 1840.

The fact that three French nurserymen in 1841 had sufficient material to offer Cayenne plants in their catalogues indicates its pres- ence in France for a number of years previous to that date. In the "Pineapple Manual” men- tioned above, the statement is made that "Cayenne is not very free in producing suckers.” At the present time we consider that healthy plants will produce an average of about four reproductive shoots each in a 2- year growth period. The rate of increase from a small beginning would be very slow and could well require 15 or more years to pro- duce enough plants so that three nurserymen could ojffer them for sale. Accordingly we may assume that a few plants may have reached France around 1820 to 1825.

MIGRATIONS OF THE CAYENNE VARIETY

With the distribution of plants to nursery- men in France, Cayenne soon became a favored variety in Europe because of its large, well-shaped fruits, good color, and fine flavor, although the Queen variety was also widely

grown and well received. The earlier distribu- tion of Cayenne to other countries appears to have stemmed from England and not directly from France. This early distribution from England is also emphasized by its early appearance in some of the British colonies, it having reached Australia by 1858 and Jamaica in 1870, although it came to Jamaica via Florida. In Australia its development was fairly rapid and from 1890 to 1895 that coun- try was able to furnish many slips and suckers for expanding the pineapple industry in the Hawaiian Islands, although the first Cayennes came to Hawaii from Florida in 1885 and Jamaica in 1886.

The decade from 1885 to 1895 was a period of accumulation of the Cayenne variety in the Hawaiian Islands. Introductions of pineapple plants were made from 11 different tropical countries (Florida, England, Jamaica, Baha- mas, Trinidad, Puerto Rico, Mexico, Aus- tralia, Singapore, Samoa, and Algeria) four of which were known to have included the Cayenne variety. It is possible that it may have been included in shipments from other countries as well, but the records do not list the varieties in some importations (Collins, 1934: 129).

Figure 1 shows two major centers of distri- bution of the Cayenne variety to other areas. England played this role during the last half of the nineteenth century, although the first European propagation was accomplished in France. Hawaii has been a major source of distribution during the first half of the twen- tieth century. It was doubtless sent to still other countries from these three major centers of distribution, but at present we do not have complete records. For instance, it was im- ported from Ceylon into South Africa, but we do not know how it reached Ceylon.

THE ORIGIN OF CAYENNE

The discussion of the Cayenne variety up to this point is based upon documentary evi- dence. In trying to trace the origin of the Cayenne pineapple beyond the realm of

The Cayenne Pineapple — COLLINS

5

1820

1863

Azores

I860

Florida 1870

Jamaica

1886

France 5 plants

Holland

^and

about

1835

England

Belgium

1858

Ceylon

ca. 1895

I

South Africa

1897

Australia

Haiti

Cuba

Mexico

Philippines

Formosa

Africa

(Kenya)

Fig. 1. Distribution of Cayenne following its introduction into France in 1820, Two major centers for dis- tribution appear: first England and then Hawaii.

6

PACIFIC SCIENCE, Vol. V, January, 1951

definitely known or documented history, cer- tain additional conclusions regarding its still earlier history have been based upon circum- stantial evidence.

As has already been stated, some of the early horticultural journals in England men- tioned that the Cayenne variety had been im- ported from French Guiana. The name it bears is also circumstantial evidence for that con- clusion. This latter statement is based upon the frequency with which plants, particularly the pineapple, are given names according to the place from which the varieties are ob- tained, even though they already possess local names. Among pineapples there are a number of varieties named after geographical areas in the West Indies and South America. Among these may be mentioned the follow- ing:

WEST INDIES SOUTH AMERICA

Antigua

Providence

Jamaica

St. Kitts

Cuban

Havannah

Puerto Rico

Bahia

Esmeralda

Pernambuco

Taboga

Surinam

Trinidad

Demerara

In the hope that further knowledge regard- ing the history of the Cayenne variety might be found in French Guiana, a search was made of the botanical literature for reports of plants and plant collections in the northern part of South America. This search disclosed the in- formation that in 1819 the French govern- ment sent an expedition to the French coloni- al possessions in America and the Pacific Ocean area to collect plants and seeds for the French botanical gardens at Paris and Ver- sailles. Samuel Perrottet, the botanist on this expedition, filed a report of the plants col- lected, in which he records finding a new variety of pineapple growing at Cayenne, in French Guiana, having spineless leaves (the others in the region having spiny leaves) and delicately flavored fruits averaging about 20 pounds in weight (Perrottet, 1825: 103).

He collected and sent to Versailles five

plants of this variety in 1820. This date fits in well with that which we have suggested above as being about the time the Cayenne variety is presumed to have been introduced into France.

The description, as it appeared in Perrot- tet’s report, is included here as a part of the history of this variety, with a free translation following it.

Bromelia mai-pouri, Perr. Cette nouvelle. espece ananas provident de Cayenne; cinq plantes ont ete, comme je Fai dit deposes au jardin des primeurs a Versailles. Le mai-pouri n’a point les feuilles armees de dents comme ses congeneres; ses fruits, d’un manger fort delicat, pesent d’ordinaire 10 kilogrammes (20 livres), et sont tres-beaux. M.

This new species of Ananas was procured from Cayenne; there were five plants as I have said, deposited in the garden of new varieties at Versailles. The mai-pouri does not have spiny leaves like its relatives; its fruits, of a very delicate flavor, weigh on an average 10 kilograms (20 pounds) and are very fine.

The description given by Perrottet is in it- self not sufficient to establish the identity of Bromelia mai-pouri’ d.n6. the Cayenne variety. In fact, the 20-pound average weight of fruit mentioned by Perrottet is difficult to believe of any variety of pineapple and must repre- sent an error in the record. The varieties Trini- dad and Cabezona were reported to have fruits sometimes reaching 20 to 24 pounds, but there is no other reference to a variety having an average fruit weight anywhere near 20 pounds.

In commenting upon this description of Bromelia mai-pouri^ Dr. L. B. Smith, of the National Museum in Washington, D. C., re- marked that it looked as if someone made a major error between collection and publica- tion. An inquiry concerning these plants sent to the Natural History Museum in Paris re- sulted in information that no herbarium speci- mens now existed of this Mai Pouri variety, but that they were considered the same as Smooth Cayenne in 1850 by Gautier, 'hhe famous grower of Parisian pineapples.” The

The Cayenne Pineapple—COLLINS

7

director of the museum stated that the dupli- cation of names had been published, al- though the term '*Mai Pouri” remains an un- supported name.

The supporting evidence of the date of its introduction into France and the postulated date based on the offering of plants for sale by nurserymen in 1841, together with the statement that Gautier believed the two to be identical in 1850, leads us to conclude that the five plants collected by Perrottet in 1820 represent the beginning of the Cayenne vari- ety in France.

How did this variety get started in French Guiana? This question cannot now and per- haps may never be answered with certainty. The collection of five slips sent to France in 1820 we believe to be of the variety now known as Cayenne. Some time after it reached France it was given the name it now bears.

The name used by the French botanist in his report is somewhat confusing, since he calls it a new species of Ananas^ yet listed it as Bromelia, a closely related genus in the pine- apple family. However, Perrottet probably used the name given to him at the time he collected it. The name ''Mai Pouri,” by which this variety may have been known in French Guiana, probably followed the pattern of indicating the place from which it origin- ated when it first appeared in French Guiana. The quest of the origin of the Cayenne was then continued in a search for a geographical area or locality having the name "Mai Pouri."

In Colombia, at the junction of the Tri- parro River with the Orinoco River, is a small village .called "Maipures." In this region, and in the watershed of the Venturari River in Venezuela, also a tributary of the Orinoco, lived the Maipure tribe of Indians. We now believe that the Cayenne pineapple originated in the interior region of Venezuela long occu- pied by the Maipure tribe of Indians, and that it was probably grown and used by them for a long period of time (Fig. 2). Velez (1946: 427), who recently traveled through the upper Orinoco River basin, states that the Piaroa

Indians have had under cultivation since time immemorial several pineapple varieties which yield large, well-flavored fruits.

Another circumstance which may also have a connection with Maipure pineapple con- cerns the Esmeralda pineapple variety, grown in Mexico. This variety is very similar, if not identical, to the Cayenne variety. Esmeraldas is the name of a small village at the mouth of the Esmeraldas River in northern Ecuador, in South America. The Maipure pineapple could have been carried from Venezuela or Colombia into the Esmeraldas region of Ecua- dor and thence into Mexico, where it was given the name of the place from which it had most recently come. We have no indication that it was known as Maipure while being grown in Ecuador, but there is good evidence that the Esmeralda and the Cayenne are the same variety.

How long had this variety been grown in the country of the Maipure Indians and why was it not found by Europeans at an earlier date? To the first question there is no definite answer. We suspect that the history of this variety may extend back into the antiquity of American civilizations along with such not- able plants as corn, tobacco, and potatoes.

To the second question a logical answer is apparent. That area of South America is even today largely unexplored, partly because it has long been inhabited by Indians who have had little contact with outside people and because of the difficulty of travel in this little- frequented and still primitive area. The Maipure Indians, who at one time roamed over this area, no longer exist as a distinct ethnic group.

Sir Walter Raleigh, who conducted an ex- pedition in 1595 up the Orinoco River for about 200 miles, reported having received quantities of pineapples from the Indians in the deep interior of the country. The village of Maipure on the Orinoco River is at the place now marked on maps as the head of navigation. This is some 200 miles farther up the Orinoco than the place reached by Ra-

8

PACIFIC SCIENCE, Vol. V, January, 1951

leigh. Could Sir Walter Raleigh and his men have been the first white men to find and eat the Cayenne pineapple?

If the Cayenne pineapple originated in the Maipure Indian country, the parent variety or species may still be present as a part of the native vegetation. Other members of the hy- brid population of which our Cayenne variety was a member may still be among those used by the Indians. Could some of the sibs of the Cayenne variety be better fitted for large-scale commercial production? Could some of the sibs have inherited resistance to diseases which Cayenne failed to obtain? Could some of the sisters of Cayenne have inherited two or more genes for yellow flesh color, where Cayenne obtained only one?

These are some of the questions which have been raised by this study of the origin of the Cayenne variety of pineapple, to which an- swers should sometime be obtained.

VARIATIONS WITHIN THE CAYENNE VARIETY

Two types of hereditary variations, which may be designated as strain differences and mutations, exist within the variety.

Strain differences

The term ' 'strain” is used here to indicate hereditary differences in fruits or plants with- in a variety when the manner of origin of these differences is not known. A long time ago growers of this variety recognized strain differences. The Journal of the Jamaica Agri- cultural Society (about 1900) carried a state- ment that there was a good and a poor kind of Smooth Cayenne and that the latter was also known as the Honolulu kind!

Captain John Kidwell, an early pioneer of the pineapple industry in Hawaii, stated (Kidwell, 1904) that there were two distinct types in Hawaii, and that he considered one much superior to the other. The poor type of Cayenne, according to his statement, had been imported from Queensland, Australia,

under the name of Smooth Cayenne. He de- scribed it as having a very large plant produc- ing fruits weighing from 7 to 15 pounds and numerous slips on the peduncle beneath the fruit. The fruits, while of good quality, were conical in shape and possessed a very large core. The other type, which he called the "true” Cayenne, was similar in appearance, but the plants were smaller and produced fruits from 5 to 7 pounds in weight. This type did not produce slips on the peduncle be- neath the fruit. As a consequence of this latter characteristic, this good strain was necessarily propagated from suckers and the crowns.

In 1887 a report of the United States De- partment of Agriculture on tropical and sub- tropical fruits mentions three kinds of Cayenne being grown in the Key West region of Florida. These were listed as Smooth Cayenne, Spine-leaved Cayenne, and Thomp- son’s Smooth Cayenne. No description of the third kind was given, but its listing as a separate strain indicates that it may have been different in some respect from the others.

In Hawaii we recognize three strains of Cayenne, all of which produce good canning fruits but differ in some plant and fruit char- acters. The one used most extensively throughout the Islands is known simply as Cayenne. The second strain, grown mostly on Kauai, is known as the "Hilo” variety, or as the Hilo Cayenne. The Hilo variety differs from Cayenne in at least three characters. The most prominent difference is in the absence of slips in the Hilo variety.. It also produces a smaller plant and fruit, with the fruit more cylindrical and of better average quality. It produces more suckers than Cayenne and the plants have a darker green leaf color. These differences are about the same as those Cap- tain Kidwell used to distinguish between Cayenne and what he called the Queensland type. The strain we now call Hilo appears to

Fig. 2. Location of the probable area of origin of the Cayenne pineapple, showing the location of the town of Maipures on the Orinoco River. The Venturari River (Ven. R.) area, for a long time the country of the Maipure Indians, is also shown.

Ceprn«l>t l«)7 by tb* Unrrtnitr ef Chk*90

10

PACIFIC SCIENCE, Vol. V, January, 1951

TABLE 1

Classification of Somatic Mutations in Cayenne

FRUIT MUTATIONS

I. Foliar proliferation of the ftuitlet

1. Crowning Beauty

2. Hour Glass

3. Slipping Beauty

II. Multiplication of floral organs

4. Flowering Beauty (increase in petal number)

5. Multiple sepals and bracts

III. Disappearance of floral structures

6. Dry fruit (only the floral bract subtending

the ftuitlet remains)

7. Bottle Neck (the upper one half to one third

of the fruit is like the dry fruit)

IV. Changes in fruit characters

8. Elongated bracts and sepals

9. Big eyes

10. Slender

11. Elongated fruit

12. Self seedy

13. Rough (pointed eyes)

14. Non-porous flesh

15. White flowers

16. Nubbin (dwarf and abnormal fruit — eyes

small and distorted, surface very rough) M226

be identical with the one Kidwell called the 'True” Cayenne.

Our third strain is known as the Collar-of- Slips, because of the excessive number of slips produced around the base of the fruit. It has a smaller fruit than the Hilo, is later in maturing its plant and ratoon crops, and is generally considered an undesirable type.

These Cayenne strains have been present as components of the variety for a long, long

Fig. 3. A plant of the Cayenne variety, showing slips, suckers, and a mature fruit.

PLANT MUTATIONS

V. Single character changes

17. Spiny leaves

18. Absence of anthocyanin

19. Linear anthocyanin

20. Blush anthocyanin

21. Intensified anthocyanin

22. Albino (no chlorophyll)

23. Streaked anthocyanin

24. Waxy (absence of trichomes)

25. Semi- waxy (reduction in trichomes)

26. Mealy (increased trichomes)

27. Few slips (573, M4W)

28. Increased number of slips (L69)

29. Multiple crowns

30. Increased wilt tolerance (resistance)

31. Increased chlorophyll (LH8)

VI. Multiple character changes

32. Paper Leaf (degenerate plant)

33. B.B. (gigas form)

34. Lanai (short leaf type)

35. Driver’s Dwarf (dwarf)

time. How they originated or which repre- sents the true or original Cayenne cannot now be determined with certainty.

Figure 3 shows a typical Cayenne plant with slips, suckers, and a mature fruit.

Mutations

The term '‘mutation” is applied to new hereditary variations which appear de novo in a population of plants or animals, caused by a change in the structure of a chromosome and thus in the hereditary constitution of the in- dividual.

Mutations have occurred in the Cayenne variety, giving rise to a series of new or changed forms; some are so profoundly changed that they no longer show the variety characters, others show only minute changes.

These mutant forms are treated separately from the strains already discussed, principally because we know something about the origin of these mutations and we do not know how the strains originated. It is quite possible that the latter also started as somatic mutations.

The mutations in Cayenne can be con- veniently divided into six classes on the basis

The Cayenne Pineapple — Collins

11

Fig. 5. A normal Cayenne plant in the center, with the short leaf Lanai mutation on the one on the right. These represent changes in opposite directions from the normal.

and the long leaf

of the characters which are altered. Four classes alFect the fruit and two classes affect both fruit and plant characters (Table 1).

The mutations listed in the first five classes of Table 1 are relatively slight or small char- acter alterations. They influence one or a few characters and do not, for the most part,

[v;, Fig. 4. A normal Cayenne fruit cut in longitudinal section on the right, showing internal structures of core, carpels, and vascular fibers. On the left is the mutant type Crowning Beauty, showing foliar pro- liferation of the fruitlets and absence of normal carpel structures.

represent profound genetic changes, even though the character alteration may be con- spicuous. The mutant Crowning Beauty (Fig. 4) is an example of this kind of mutation.

Those in Class VI, however, stand apart from the rest because they represent simul- taneous changes in a number of different morphological and physiological characters.

The Lanai type and B.B. mutations (Fig. 5) appear to be complementary to each other in their deviation from the normal variety. They vary from typical Cayenne in opposite direc- tions in at least six characters, shown in Table 2.

TABLE 2

Characters of Mutants B.B. and Lanai Compared WITH Those of the Normal Variety

CHARACTER

VARIATION FROM TYPICAL CAYENNE

In B.B.

In Lanai

Leaf number	decreased	increased
Leaf length ......	increased	decreased
Fruit size	larger	smaller
Fruit maturity ....	later	same as normal
Plant size	larger	smaller
Spines	more	normal

The Paper Leaf mutant departs so widely from the normal variety in a number of char-

flrfPrQ If nn 1r»no-f»r r/=»c<=‘mK1<=»c fL#=> C' oTr_

12

enne at all. It is considered a degenerate type, because of its small, worthless fruit and weak- ly growing plant. It probably could not sur- vive in field competition with normal plants.

Driver’s Dwarf (Fig. 6) has a diminutive form of plant and fruit, combined with collar of slips and an intensified purple anthocyanin pigment in the leaves. This may represent a change from the normal slip type of the Cay- enne variety to the collar-of-slip strain. On the other hand, the mutation giving rise to dwarf and increased anthocyanin may have occurred in a collar-of-slip strain plant which was al- ready in the field population. The dwarfing and anthocyanin intensification appear to have been simultaneous changes.

Fig. 6. A normal Cayenne fruit on the left, with a fruit of the mutant Driver’s Dwarf on the right.

Discussion of these more obvious mutant types naturally leads to the question of the possible occurrence of other mutations which are less conspicuous and less easily measured. Undoubtedly mutations of this kind take place, but the demonstration of the existence of mutations of slight phenotypic effect is much more difficult, because of overlapping variations of a non-hereditary nature.

Some examples of these small or elusive mutant effects are briefly described to show that at least a few are present in the variety.

PACIFIC SCIENCE, Vol. V, January, 1951

Two Cayenne clones were found to show a slightly darker green color, which could be recognized easily when a fairly large group of these were growing adjacent to a group of the normal variety. A clone may be thought of as one plant multiplied many times by vegeta- tive reproduction, and all plants of a clone have identical heredity. Chemical analyses of leaves revealed a higher chlorophyll content in these darker green leaves. A mutation of slight effect on chlorophyll production was probably the origin of these darker green clones.

The elongated fruit mutation listed in Table 1 develops a large number of multiple crowns and fasciated fruits when grown in warm, dry areas, but only normal plants ap- pear in the cool, more moist areas. In this case the additional character of multiple crowns is quite obvious under one environ- mental condition and disappears (or merges into the normal pattern) when grown under other conditions.

The number of slips produced per plant is a character subject to considerable variation due to differences in the environment during the growth period. This is well illustrated by the effect of relative crowding of plants in the field. Close planting reduces the number of slips, and, conversely, widely spaced plants permit a larger number of slips to develop. Table 3 shows the variation in number of slips resulting from an experimental planting of Cayenne with 10, 12, 14, and 16 inches be- tween plants (that is, with different degrees of crowding) .

TABLE 3

Effect of Different Plant Spacings on Slip Production

PLANT SPACING AVERAGE NUMBER

IN INCHES OF SLIPS

10 2.51

12 3.33

14 3.95

16 4.36

Odds — 19 to 1 0.43

Odds — 99 to 1 0.60

Because of the susceptibility of this char- acter to variation due to environmental dif-

The Cayenne Pineapple — ^COLLiNS

13

MEAN NUMBER OF SLIPS PER PLANT

Fig. 7. Ten Cayenne clones showing mean slip numbers for a period of 7 years, together with lowest and highest yearly means.

ferences, the discovery of genetic variations (somatic mutations) which have an effect on slip number is more difficult than locating mutations which are not subject to environ- mental modification. However, by repeated tests under similar environmental conditions it has been possible to demonstrate the oc- currence of mutations affecting number of slips in this variety.

Figure 7 shows the mean number of slips per plant for 10 clones over a period of seven plant crop harvests, together with lowest and highest limits of the yearly means for that period. Five of the clones had a range of annual mean slip number which did not over- lap the range of three other clones. Two clones, E and G, had a range of variation which overlapped all the other groups. There can be little doubt about a genetic difference existing between any one of the three high slip- producing clones, H, I, J, and any one of the five low-producing clones, A, B, C, D, and F.

The three high slip-producers, H, I, and J, do not appear^ to differ among themselves. Clone G, overlapping in its range with both the high and low slip producers, appears to be genetically different from both groups.

These mutant clones can be compared with each other, but should not be compared with the mean of the variety, first because they are a part of the variety and second because — the variety being a mixture of genetic types — random samples would not always include the same distribution of variant types. Fur- thermore, most Cayenne populations include some plants of the collar-of-slip strain, which produces a very high number of slips.

The somatic mutations in Cayenne include a range of types varying from those having

These conclusions are based upon another repli- cated plot test which showed significant differences as listed here, and we presume that the causes of the differences were genetic.

14

PACIFIC SCIENCE, Vol. V, January, 1951

marked, easily recognized characters to those differing so slightly that only statistical ana- lyses of quantitative data will identify them.

They are not frequent in occurrence, with the exception of a few which have been shown to be frequently mutating types (Collins, 1936). The mutations discussed here have been collected over a period of 20 years or more and from among millions of normal plants.

This variety is as stable genetically as are vegetatively propagated, highly heterozygous diploid varieties in general. Were this not true, the variety could not have been main- tained through this long period of time.

Cayenne is not the single clone it probably was at the beginning. Now it is a collection of clones, all having the same general char- acters but usually differing in one or a few characters or degrees of expression of char- acters. The more obvious mutations and those too poorly adapted to survive in the general population have been and are being reduced to small percentages or eliminated. The re- maining population heterozygosity consists of minor character alterations which are car- ried along by asexual propagation in the general mass of cultivated plants. As examples of these latter clonal types, reference can be made to some of those listed in Table 1, such as the clone with less porous fruits, self- seedy types, low and high slip-producing forms, and increased amount of chlorophyll.

As to their value to the organism (in a cultivated variety, positive value in horticul- ture) these somatic mutations follow the known pattern of randomly occurring muta- tions; the great majority are either detrimental or of no advantage to the organism. Only three of those which have been studied appear to have possible advantage in pineapple cul- ture, and only two of these, the wilt-resistant mutant and the high slip-producing type, are of possible importance.

TETRAPLOID CAYENNE

The pineapple normally has 50 chromo-

somes in its somatic cells. This is considered to be the diploid number for the genus Ananas. However, tetraploid Cayenne plants having 100 chromosomes were obtained after treating shoot growing points with colchicine solution (Kerns and Collins, 1947). The im- mediate results from these treatments were various kinds of chimeras of diploid and tetra- ploid tissues, together with plants which either died early in growth or reverted to nor- mal diploid tissue throughout.

By careful selection of buds from the tetra- ploid sectors of chimeras during several suc- cessive vegetative generations, constant new types which fall into the following three classes on the basis of the amount and loca- tion of tetraploid tissue were obtained.

Class 1 was completely tetraploid.

Class 2 was tetraploid except for a diploid epidermis.

Class 3 was diploid except for a tetraploid epidermis.

Classes 1 and 2 were alike in all visible characters. Class 3 was like the normal diploid in all visible characters.

The tetraploid has been compared in Table 4 with the diploid in a number of important characters. The fruit weight is less in the tetra- ploid and the fruit has fewer eyes than in the diploid. The average eye weight, however, is higher in the tetraploid, showing that the individual eyes of the tetraploid are larger. The Brix (dissolved solids including sugars) of the tetraploid fruit is lower than in the diploid fruit. The characters of fruit acidity, translucence, and vitamin C content are high- ly variable, so that no significant differences were obtained. These characters are readily altered by different environmental conditions.

Tetraploid plants are taller than diploids, but they have fewer leaves and produce fewer slips. They do not differ in average leaf length, although the tetraploids have wider leaves. The tetraploids also have a higher percentage of water in the leaves and, as a consequence, a lower percentage of dry matter per unit weight of green leaf tissue.

The Cayenne Pineapple — Collins

15

The fruits are seedless, as are the diploids. The chromosomes perform regularly in germ- cell formation, and functional gametes are produced.

Crosses between the Cayenne tetraploid and varietal hybrid tetraploids produce viable seed readily, but crossing with diploid forms produces very little seed; this is similar to the results obtained when varietal hybrid tetra- ploids are crossed with diploids.

TABLE 4

Comparison of the Fruit and Plant Characters IN THE Diploid and Tetraploid Cayenne

CHARACTERS	DIPLOID	TETRAPLOID
Fruit weight (lbs.)	5.8	4.0*
Eye number	140.4	105.6*
Eye weight (gm.)	16.3	16.8*
Brix	15.4	13.8*
Acidity (per cent)	0.71	0.86
Vitamin C	19.2	20.8
Translucence	2.7	2.3
Plant height (cm.)	28.1	30.5*
Number of active leaves. . .	57.6	44.2*
Leaf length (cm.)	65.0	65.2
Leaf width (cm.)	5.4	6.2*
Percentage water in leaves	81.8	83.5*
Percentage dry matter in leaves	18.2	16.5*
Pollen grain diameter (mi- crons)	47.0	64.0*
Stomata size (microns) ....	22.0	31.0*

*Significant differences.

In many plants, and particularly in orna- mentals, valuable horticultural characters are obtained by doubling of the chromosomes (Emsweller, 1948: 570). However, the tetra- ploid Cayenne is inferior to the diploid form in several important characters (Table 4).

GENETIC NATURE OF CAYENNE

The Cayenne pineapple normally has seed- less fruits, although it does produce normal germ cells. The seedlessness results from a condition known as self-incompatibility. Under these conditions the germ, or sex, cells of the same individual do not unite in ferti- lization to form embryos. Germ cells from two different varieties, Cayenne and Queen, for instance, are mutually compatible and

seeds are produced following cross pollination between the varieties.

Inasmuch as inbreeding is thus denied be- cause of self-incompatibility, the studies of the genetic nature of Cayenne have been con- ducted largely from the results of varietal crossings and from examination of the soma- tic mutations which have appeared in field populations.

Cayenne has a diploid number of 50 chro- mosomes; in the somatic cells they appear slightly elongated or almond-shaped (Collins and Kerns, 1931: 140); in the germ cells, they are almost round. While the process of germ- cell production is usually normal, some ab- normalities occur. The most frequent ab- normality is the formations of germ cells containing 50 chromosomes, which is double the ordinary number of chromosomes in gametes. These appear in the mature sex cells as giant pollen grains and ovules.

In crosses between Cayenne and other varieties, these giant germ cells give rise to occasional hybrids having 75 or 100 chromo- somes instead of the normal number of 50 for this species.

The number of these plants whose cells have 75 and 100 chromosomes is far below the expected number on the basis of the per- centage of 50-chromosome germ cells pro- duced. This shows that only a small propor- tion of the plant gametes function in ferti- lization and that selective fertilization in favor of the normal 25-chromosome gametes takes place.

Hybrid populations resulting from crossing Cayenne with other varieties provide some information regarding the genotype or heredi- tary constitution of the Cayenne variety.

The variety is heterozygous for many reces- sive and dominant genes. Table 5 gives some information on the genotype of Cayenne ob- tained from crossing with other varieties.

INBREEDING IN CAYENNE

In discussing the somatic mutations which have appeared, mention was made of the self-

16

PACIFIC SCIENCE, Vol. V, January, 1951

TABLE 5

The Genotype of Cayenne with Respect to Certain Characters

CAYENNE	CONTRASTING	CAYENNE	CAYENNE DOMINANCE
CHARACTER	CHARACTER	GENOTYPE	RELATION
Spiny tip leaves	Spiny leaves	Ss	dominant
Spiny tip leaves	piping leaves	PP	recessive
Anthocyanin in leaves	no anthocyanin	Aa	intermediate
Yellow flesh	white flesh	Yy	intermediate
Purple petals	white petals	WW	dominant
Chlorophyll	no chlorophyll	Cc	dominant
Normal fruit	proliferation	*	recessive
Seedless	seedy	*	recessive
Long leaves	short leaves	*	dominant

*Undetermined.

fertile mutation. In this form, the self- incompatibility characteristic of Cayenne is replaced by self-fertility as a result of a domi- nant mutation, but in all other respects the variety characters remain unchanged.

Inbreeding can now be carried out in the variety by using these self-fertile mutants. Inbreeding is a potent tool for dragging re- cessive skeletons out of hereditary closets and displaying them in the light of day for all to see. By the use of this tool we have pried out of Cayenne recessive secrets which it has long carried buried within its genotype.

About one fourth of the inbred progeny are semi-lethal and die in an early seedling stage. Another fourth are so weak that they grow very slowly and reach fruiting maturity long after the parent Cayenne. Most of the inbreds are low in vigor, with only a few ap- proaching that of the Cayenne variety. They exhibit a wide variation in fruit and plant types, including many fruit and crown fascia- tions. The inbred population supplies evi- dence that Cayenne is not only heterozygous for many recessive and some dominant char- acters, but that it exhibits heterosis or hybrid vigor.

A number of variations which had appeared in the variety as somatic mutations also ap- pear in the inbred population in numbers in- dicating Mendelian segregation in a hetero- zygous genotype.

The general characteristics of the inbred

populations supply some evidence regarding the parentage of Cayenne itself. The origin of this variety as a hybrid between any of the known species of Ananas can be ruled out, we believe. The parent or parents producing this variety through sexual reproduction must be considered at present as unknown varieties of A. comosus, or of such a variety of A. comosus and a species of pineapple not known at the present time.

SUMMARY

The Cayenne variety of pineapple was first mentioned in an English horticultural journal in 1841. Evidence is presented to show that it came from French Guiana in 1820. It is presumed to have been grown by the Maipure Indians in the upper Orinoco River valley long before it reached French Guiana. The manner and time of its origin are obscure.

After its introduction into France and Eng- land, it became established in many tropical countries during the 100 years between 1840 and 1940.

The genotype of the variety is highly heterozygous and it exhibits hybrid vigor in its growth. The variety is self-incompatible and must be propagated vegetatively. During its long period of vegetative propagation a number of somatic mutations have appeared, including one giving the mutant type self- compatibility. The present Cayenne is a mis- cellaneous collection of clones. The diploid chromosome number is 50, with 100-chro-

The Cayenne Pineapple — Collins

17

mosome tetraploids obtained by treatment of diploids with colchicine. The tetraploids are inferior to the diploids.

REFERENCES

Anonymous. Ca. 1870. The pineapple man- ual, being a guide to the successful cultiva- tion of the fruit and construction and management of the pinery by contributors to the Journal of Horticulture. London: Jour. Hort. and College Gardener.

Collins, J. L. 1934. Introduction of pine- apple plants into Hawaii and some brief accounts of pioneer pineapple growing. Pineapple Quart. 4: 119-120.

1936. A frequently mutating gene in

the pineapple. Ananas comosus (L.) Merr. Amer. Nat. 70: 467-476.

and Kenneth R. Kerns. 1931.

Genetic studies of the pineapple. I. A pre- liminary report upon the chromosome number and meiosis in seven pineapple

varieties (Ananas sativus Lindl.) and Bro- melia Pinguin L. Jour. Hered. 22: 139-142.

Emsweller, S. L. 1948. Breeding of orna- mental plants. Amer. Soc. Hort. Sci., Proc. 51: 565-574.

Kerns, K. R., and J. L. Collins. 1947. Chimeras in the pineapple. Hered. 38: 323-330.

Kidwell, Capt. John. 1904. The cultivation of pineapples in Hawaii. Hawaii. Forester and Agr. 1 (12).

Perrottet, Samuel. 1825. Catalogue rais- sonne des plantes introduites don les colonies frangaise de Mascareigne et de Cayenne, et de celles rapportees vivantes des mers d’Asie et de la Guyane au Jardin des Plantes de Paris. Mem. Soc. Linn. 3(3): 89-151.

Velez, Ishmael. 1946. Wild pineapples in Venezuela. Science 104: 427-428.

Studies on Oxyspirura mansoni, the Tropical Eyeworm

of Poultry. II. Life History

Calvin W, Schwabe^

This paper, the second in a series, presents the results of a study made in Hawaii of the life history of Oxyspirura mansoni, the tropical eyeworm of poultry. This nematode has been shown by previous investigators to possess a digenetic life cycle in which it utilizes as the definitive host many species of domestic and wild birds and, as the secondary host, a single species of burrowing cockroach, Pycnoscelus surinamensis.

The parasite is widely distributed in the warmer parts of the world. It has been re- ported from Brazil (de Magalhaes, 1888; Almeida, 1933), China (Cobbold, 1880), Indo-China (Fielding, 1928^), Mauritius (Emmerez and Megnin, 1901), Reunion (Ozoux, 1910), British West Indies (Hutson, 1943), Formosa (Kobayashi, 1927), British and Dutch East Indies (Picard, 1929; Field- ing, 1928^), Florida and Jamaica (Ransom, 1904), Hawaii (Wilcox and McClelland, 1913), Guam (Fielding, 1928^), and Samoa (Alicata, personal communication). Its near relative (or synonym), Oxyspirura parvovum, has been reported from Australia by Dodd

(1909).

Apparently Manson’s eyeworm has ap- proximately the same circumtropical distri- bution as its intermediate host (reviewed in Schwabe, 1949). Neither parasite nor host is known to exist in Hawaii at altitudes exceed- ing 3,000 feet.

^Department of Zoology and Entomology, Univer- sity of Hawaii. Present address: Department of Bac- teriology, School of Veterinary Medicine, Alabama Polytechnic Institute, Auburn, Alabama. Manuscript received May 16, 1950.

The eyeworm is considered important eco- nomically in many tropical and subtropical areas. The roach is objectionable not only in that it serves as the vector for the eyeworm, but also because it reportedly damages both roots and bark of certain ornamental and crop plants (reviewed by Schwabe, 1949) • Previous to this study the development of the larval and adult stages of this parasite was not well known.

MATERIALS AND METHODS

Surinam roaches in all stages of develop- ment were easily collected from soil beneath chicken houses and in chicken yards. Because the incidence of natural infection with the parasite among such roaches approached 100 per cent, only laboratory-raised nymphs were employed in the experimental infections.

Adult Pycnoscelus surinamensis females were housed individually in 4-inch glass stacking dishes. A 3 -inch disk of filter paper was placed in each dish to provide a hiding place for the roaches during the daytime. Nymphs, iso- lated at birth, were housed together in the same manner as the adults. The roaches thrived on a diet of whole-wheat bread and water.

Embryonated eggs for infecting laboratory- raised roaches were obtained by macerating gravid female worms. This procedure proved more satisfactory than the involved method of concentration and separation of embryo- nated eggs from the feces of infected birds.

The gravid worms were taken from the eyes of infected chickens obtained from poultry farms in Manoa Valley and Waialae, Hono-

[18}

Oxyspirura mansoni — Schwabe

19

lulu. The parasites were removed from the eyes of the birds by holding the head rigid and inserting the tips of a pair of dull- pointed forceps beneath the nictitating mem- brane. Worms thus obtained usually survived for at least 24 hours in physiological saline at room temperature.

Young nymphs to be infected were first isolated without food or water in clean stack- ing dishes for at least 48 hours. Nymphs which were dead or enfeebled after this treat- ment were removed. Gravid female eye- worms were macerated in a drop of water to free the eggs from the vagina and uteri. The entire mass was then soaked up with a small bread crumb. The starved nymphs rapidly ate the moistened bread. It was later found that the nymphs would just as readily consume living female worms so this method of infec- tion was adopted.

A roach to be examined was pinned, dorsal side downward, on a paraffin block. The head was severed with a sharp scalpel and the pos- terior abdominal segment was teased from the remainder of the abdomen with fine dissect- ing needles. The rectum remained attached to this segment and the entire alimentary tract was withdrawn in this manner. The alimen- tary tract and the remainder of the roach were each placed in a few drops of physiological saline solution on separate clean glass slides. Free third-stage larvae, if present, could, with the unaided eye, then be seen wandering about on the slide. Encysted second- and third-stage larvae were readily observed en- tangled in the Malpighian tubules or at- tached elsewhere along the alimentary canal, particularly in the region of the rectum.

To observe the late first-stage larvae, the abdominal fat was dissected away from the body wall, placed in a drop of physiological saline solution, and examined beneath a cover slip with the aid of a compound microscope.

In examinations for early first- stage larvae, the crop and esophagus were severed from the remainder of the alimentary canal, teased apart, and examined as a wet mount with a

compound microscope. This procedure was repeated on separate slides for the midgut and hindgut.

First-stage larvae were examined as live wet mounts or were stained vitally with methylene blue or fixed in Bouin’s solution. Second- and third-stage larvae were examined alive by compressing the cyst beneath a cover slip or by freeing the larvae from the cysts and fixing them in Bouin’s solution. Fourth-stage larvae were killed in Bouin’s solution and examined as wet mounts.

When living larvae were too active for study they were anesthetized by a crystal of chloral hydrate introduced beneath the cover slip.

The chickens used in these experiments were all hybrid stock (Rhode Island Red X New Hampshire Red) obtained at the age of 2 weeks from the Department of Poultry Hus- bandry, University of Hawaii. They were housed on wire and fed commercial growing mash.

Third-stage larvae were first obtained in the laboratory by the dissection of large numbers of infected roaches. Inasmuch as this con- sumed so much time and the larvae were not obtained in the numbers desired, a more satis- factory method was devised. It was observed that when infected roaches were torn apart and placed in physiological saline solution, heated to approximately 37° C., the larvae immediately began to migrate from the tis- sues of the roaches and to settle to the bottom of the container. This behavior of the larvae suggested the use of the Baermann apparatus, with which an ample supply of infective lar- vae was readily obtained.

THE ADULT EYEWORM

The sexually mature worms are found be- neath the nictitating membranes and in the conjunctival sacs and naso-lacrimal ducts of domestic chickens, ducks, and a number of other wild and domesticated birds.

Anatomy of the adult eyeworm

The adult Oxyspirura mansoni is a slender.

20

PACIFIC SCIENCE, Vol. V, January, 1951

TABLE 1

Reported Measurements of Adult Female Eyeworms

MEASUREMENT	SCHWABE (Oahu, T.H.) 1948-49	RANSOM ' (Florida) 1904	MAGALHAES (Brazil) 1888	SWEET (Australia) 1910*	FIELDING (Australia) 1928*
Length f	15-17	12-18	I6.2t	13.5-20	15.5-17
Width (ant. end)	69.1-75	50
Width (max.)	369-538	400-430		270-390	270-420
Width (at anus)	92 1	90-100
Esophagus length t	1.55t	1.5
Esophagus width (post, end)	107t	80-100
Intestine width (at esophagus)	61. 5t
Intestine width (max.)	77-100	100
Anus to post, end	446-477	400-530	400-530	390-440	364-509
Protostom length	15.4-22	15-25
Protostom width	35.7t	25-30
Mesostom length	22-31	25-30
Mesostom width	17.3t	20-25
Excretory pore from ant. end	410t	350-400
Nerve ring from ant. end	232J	250		220-300	270-320
Vulva diameter	46-61	40-50
Vulva to post. endf.	1.23-1.31	1-1.4	1-1.33	0.78-1.07	0.91-1.55
Vagina length f	2
Vagina width (near uteri)	57.U
Vagina width (near vulva)	36t
Uteri width	123-146	100
Ovary width	22-30

^Oxyspirura parvovum.

fThese measurements in millimeters; all others in microns. JAverage measurements (10 specimens).

TABLE 2

Reported Measurements of Adult Male Eyeworms

MEASUREMENTS	SCHWABE (Oahu, T.H.) 1948-49*	RANSOM (Florida) 1904	SWEET (Australia) 1910t	FIELDING (Australia) 1928t
Length!	9.2-13	12-14	9.2-14.5	8.2-15.47
Width (ant. end)	75	50		74
Width (max.)	339	200-350	260-330	254-327
Width (at cloaca)	143	65-150		136
Esophagus length!	1.37
Esophagus width (post, end) ‘	107
Intestine width (max.)	111
Buccal capsule length	53.5
Excretory pore from ant. end	410
Nerve ring from ant. end	285
Cloaca to post, end	285	320-400		253-390
Long spicule length!	4.15			3.64-4.55
Short spicule length	232			214-235

*Average measurements (10 specimens).

\Oxyspirura parvovum.

JThese measurements in millimeters; all others in microns.

Oxyspirufa mansoni — Schwabe

21

I

Fig, 1. Photomicrograph showing the anterior end of an adult Oxyspirura mansoni.

white, thread-like worm attenuated at both the anterior and posterior ends. Adults studied in the laboratory measured from 9 to 17 mm. in length (Tables 1 and 2). The pos- terior end is more slender and in the male is curved ventrad. The thin, transparent, outer cuticle is smooth; neither transverse nor longitudinal striations were discernible on worms studied. Magalhaes (1888) reported fine transverse striations; these were not ap- parent to Emmerez and Megnin (1901) or to Ransom (1904).

The following papillae are common to both sexes: a small cervical papilla with a short hair-like process located on each side of the body approximately 440 ju from the anterior end; a pair of small latero-caudal papillae near the posterior extremity of the tail; six minute oral papillae; and four large sublateral cepha- lic amphids. In the male, six pairs of papillae

Fjg. 2. R'-Oph ’’Ik', rli-: ead

of :ia .)d(]|r rn di. Oxy'iJ'it .n - ".tL'.iyh

suriOLind tlie cl«j.iCjl opeijing. Iciur p.ars pre- ana 1 and twt'» pjit’s postan.il.

I'he rruisck arr.ingeirM^r,*- i-; of rlv;- poly- rnyarian type, witii a tan-blic M-Tangeirient of muscle fibers in the an.d iegi<-u of Lk.i.!! rhe male and the fem.de, .irucljin*r the lectunj to the dorsal wall.

Fig. 3. Photomicrograph showing the posterior end of an adult female Oxyspirura mansoni.

22

Fig. 4. Photomicrograph of median cross section through an adult female eyeworm.

The cuticular buccal capsule is six-lobed and, as is characteristic of the Thelaziidae, is divided into two chambers, the anterior pro- tostom and the posterior mesostom. The esophagus is of the typical spirurid type and is approximately 1.5 mm. in length. It is dis- tinctly divided into a short anterior muscular portion and a posterior glandular portion. The lumen is restricted and triradiate when viewed in cross section. A well- developed esophageal-intestinal valve separates the eso- phagus from the intestine.

The thin-walled intestine extends almost the entire length of the body, varying only slightly in diameter, and joins the esophagus anteriorly and the rectum posteriorly. The in- testinal wall is composed of a single layer of short, ciliated columnar cells. The rectum is thick-walled and muscular and terminates at the ventral anus in the case of the female. In the male, the ejaculatory duct joins the rectum ventrally to form the cloaca.

PACIFIC SCIENCE, Vol. V, January, 1951

The excretory system is of the simple H or oxyuroid type. From the ventral pore the common excretory duct extends dorsally through the large excretory cell and divides on the ventral side of the esophagus to join the two lateral excretory canals. These lie em- bedded in the lateral lines of the body wall. The lateral lines, which extend almost the en- tire length of the body, are, apparently, each a single large multi-nucleate cell. The lateral canals terminate posteriorly at the latero- caudal papillae.

A nerve ring surrounds the esophagus, cephalic of the excretory pore. In association with it are four large ganglia, two dorsal and two ventral, and a number of smaller cells. A dorsal and a ventral nerve cord extend pos- teriorly the length of the body, and several smaller nerve fibers extend into the cephalic region.

The mature male possesses two spicules of unequal length, both of which are cuticular in nature, transversely striated, and hollow. The shorter, which is "trough-shaped,” mea- ures about 200^ in length by 30^t in maxi- mum width, is only slightly protrusible, and acts primarily as a guide for the longer, more slender spicule. The long spicule is 3 to 4.2 mm. in length by 10^ in maximum width and is capable of being protruded from the cloaca almost its entire length. A short muscular ejaculatory duct extends anteriorly from the cloaca and terminates in a thin-walled seminal vesicle, an uncoiled organ extending over half the length of the body. The single long, coiled testis fills the remainder of the body cavity.

In the female the vulva, which measures 46 to 51m in diameter, is located ventrally in

30^

Fig. 5. The short, "trough-shaped” spicule of the adult male eyeworm.

Oxyspirura mansoni — SCHWABE

23

the posterior half of the body. It is surround- ed by a cuticular ring of highly refractile ma- terial. The vagina extends anteriorly from the vulva and is divisible into two parts, by rea- son of the thickness of the wall. The proxi- mal portion is nearly 150 in length and 35 ju in diameter. The distal thicker- walled portion is approximately 60 fx in diameter. The vagina branches to form the two thin- walled uteri. These extend forward to the posterior region of the esophagus, where they reflex and ex- tend back to the region of the vulva. The diameter of the uteri decreases to 22-30 /x to form the two ovaries which lie as a coiled mass in the posterior portion of the body cavity.

HOSTS OF THE EYEWORM

Nematodes inhabiting the eyes of birds have been reported from a number of species. Most of these parasites are members of the genera Thelazia and Ceratospira, or are species of the genus Oxyspirura other than 0. mansoni.

The hosts of only those eyeworms which have been definitely identified as Oxyspirura mansoni (or parvovum) are recorded here.

Definitive hosts

Manson’s eyeworm has been reported to occur naturally in the following birds: do- mestic chicken {Callus domesticus) (Cobbold,

1880); turkey {Meleagris gallopava) (Cram, 1927); peafowl {Favo cristatus) (Magalhaes, 1888); English sparrow {Passer domesticus) (Illingworth, 1931; Alicata, 1947); mynah bird {Acridotheris tristis) (Alicata, 1947); Chinese dove {Streptopelia chinensis) (Alicata, 1947; Schwartz and Schwartz, 1949); Japanese quail {Coturnix coturnix japonica) (Schwartz and Schwartz, 1949); pheasant {Phasianus torquatus torquatus and P. vesicolor vesicolor) (Schwartz and Schwartz, 1949).

Through the cooperation of Paul Breese, Director, the birds in the Honolulu Zoo are being examined for the presence of Oxyspirura mansoni. This survey is not complete as yet, but the following new hosts may be recorded: the great Argus pheasant (Argusianus argus argus) and the Siamese fireback pheasant {Diardigallus diardi) .

Role ofi the natural reservoir hosts imthe spread of the parasite in Hawaii

During the course of this investigation mynah birds, English sparrows, and Chinese doves have been trapped on the University campus and have been found to harbor Man- son’s eyeworm. Similar observations have been made by other investigators in other parts of Hawaii (Illingworth, 1931; Alicata, 1947; Schwartz and Schwartz, 1949; Tanada, personal communication) .

TABLE 3

Numbers of infective Larvae per Roach (100 Surinam Roaches Examined at Each Locality)

STAGE	RANGE	MIYATA POULTRY FARM, WAIALAE, OAHU	UNIVERSITY FARM, MANOA VALLEY, OAHU*	WAIKIKI, OAHUf
	Max.	36	3
Adult roaches	Min.	16	0	None
	Ave.	23
	Max.	34	2
Nymphs	Min.	1	0	None
(Final instar)	Ave.	3
	Max.	6	0
Young nymphs	Min.	0	0	None

*Roach population low (daily removal of manure, weekly chlordan spray). fApproximately 4 miles from nearest poultry farm.

24

PACIFIC SCIENCE, Vol. V, January, 1951

Since these commonly infected birds are widespread and numerous throughout the Hawaiian Islands, it seemed desirable to as- certain the role they play in the dissemination of the parasite. To determine this, the degree of infestation of Surinam roaches was estab- lished at (1) a heavily infested poultry farm in Waialae, Honolulu, Oahu; (2) a relatively clean poultry yard in Manoa Valley, Hono- lulu; and (3) a residential area in Waikiki, Honolulu. In all three places roaches were numerous.

As shown in Table 3 the roaches from (1) harbored a large number of infective eye- worm larvae per roach. At (2), where low in- festation of the chickens resulted from strict sanitation and frequent removal of the ma- nure, but where large numbers of wild birds gathered to feed, the number of larvae per roach was very low. At (3), an area at some distance from any poultry farm, but where wild birds were also very numerous, no larvae were found in the many roaches examined.

Inasmuch as domestic fowl and wild birds are the only sources of infection of the local roaches these data indicate that the local wild birds are of little importance as reservoir hosts in the dissemination of the eyeworm popu- lation. Apparently their feces are too scattered to be eaten to any great extent by the roaches.

Mammals as hosts of Manson s eyeworm

In no case has Oxyspirura mansoni been known to occur naturally in the eyes of a mammal. Fielding (1927) found that infec- tive eyeworm larvae placed in the eyes of guinea pigs would develop to maturity.

To check Fielding’s observations on an- other mammal, I obtained several white rats. Each of these was forcibly fed four infected roaches. The next day the eyes were anes- thetized with a 5 per cent solution of butyn and examined carefully for eyeworm larvae. No larvae were found.

Approximately 30 infective larvae were then introduced by pipette into the mouth of one

of the rats. On examination of the eyes 24 hours later, no larvae were found.

Two rats then received approximately five eyeworm larvae per eye. One was examined after 10 days and larvae were seen in both eyes. The rat was necropsied and the worms were removed. They had molted to the fourth stage and the reproductive organs were de- veloping normally. The second rat was killed 25 days after the larvae were placed in its eyes. Adult male and female worms were re- covered from both eyes.

LIFE HISTORY OF THE PARASITE

Apparently little effort had been made be- fore about 1927 to ascertain the life cycle of Oxyspirura mansoni. Previously Emmerez and Megnin (1901), and Emmerez (1918), Ran- som (1904), and Ozoux(1910) had attempted to transmit the infection directly from one chicken to another with embryonated eggs and/or first-stage larvae; but they carried the work no further when their efforts were un- successful.

Kobayashi (1927) in Formosa and Sanders (1928) in Florida found that an intermediate host, which they identified as the cockroach Pycnoscelus surinamensis Linn., was essential for the completion of the life cycle of the para- site. Previously Fielding (1926) had shown the same roach to be the intermediate host of the Australian eyeworm of poultry, Oxy- spirura parvovum Sweet. Fielding (1927 and 1928^) studied the developmental anatomy of the Australian eyeworm, but because of his uncertainty as to whether the species was 0. mansoni or 0. parvovum., he referred to it simply as the eyeworm of poultry in his latter paper. The validity of 0. parvovum as a species was questioned by Tryon (1926). This lack of clarity as to species, together with the fact that Fielding’s drawings were not identifiable with the material found in Hawaii, led me to investigate the life history of Manson ’s eye- worm under Hawaiian conditions.

Life cycle in general

The eggs of the parasite are laid by the

Oxyspkuramansoni — Schwabe 25

definitive host. They gain entrance when the host roach is consumed by a suitable bird. The larvae seldom pass farther than the crop of the definitive host (Fielding, 1926), where they migrate from the tissues of the roach under stimulation of heat and moisture.

The infective larvae then crawl up the eso- phagus, reach the roof of the mouth, and gain entrance to the eyes through the naso- lacrimal ducts. During this study worms have been observed in the eyes of a chicken within 5 minutes after the ingestion of an infected roach.

The eyeworm eggs

During the course of this study several at- tempts were made to recover embryonated eggs from the eyes of infected birds. Fluid was removed from the eye with a pipette and examined beneath the microscope, but in no case were eggs observed. However, they were recovered in varying quantities from the crop, intestinal contents, and feces of infected birds. The eggs were elliptical in shape, had a shell thickness of 1 to 1.5 a-^d were approxi- mately the same size as those measured in the

TABLE 4

Reported Measurements of Eyeworm Eggs (All Measurements in Microns)

		SCHWABE	RANSOM	SWEET	FIELDING	FIELDING
STAGE	RANGE	(Oahu, T.H.)	(Florida)	(Australia)	(Australia)	(Australia)
		1948-49	1904	- 1910*	1928t	1928*
Embryonated eggs	Max.	60x45
from feces	Min.	42x23
	Ave.	53x40
Embryonated eggs from	Max.	50x28.5	65x45	45x30
vagina and uteri	Min.	35.7x17.8	50x40	33x25
	Ave.	41.6x23.7			43x31	41x30
Shelled, segmented eggs	Max.	42.9x32
from uteri	Min.	28.4x14.3
	Ave.	36.7x22.9
Unshelled, non-segmented	Max.	28.25x10.7				28x14
eggs from uteri	Min.	21.6x7.2				24x12
	Ave.	23.4x9.5	24x12

Fig. 6. Photomicrograph of the vagina of a gravid female eyeworm showing embryonated eggs.

adult females in the eyes of the definitive host. They are washed down the naso-lacrimal ducts with the eye fluid into the mouth, swallowed, and finally passed out with the excrement. The intermediate host is infected by eating embryonated eggs or first-stage lar- vae in the feces of infected birds. After a de- velopmental period of approximately 51 days in the roach, the larvae are infective to the

*0. parvovum,

fold fixed 0. mansoni eggs.

26

PACIFIC SCIENCE, VoL V, January, 1951

vagina of gravid female worms (Table 4). The shell had a smooth surface with no discerni- ble markings.

Ransom (1904) obtained embryonated eggs from the uteri of gravid worms. These were placed in ''salt solution'’ and were reported to have hatched in 3 days. The larvae obtained were dead, or were feeble and soon died. Sanders (1928) employed a similar technique, substituting distilled water for the saline solution, and found that embryonated eggs from gravid females hatched in 3 days.

Fielding (1927) succeeded in hatching liv- ing larvae in the following media: 0.9 per cent NaCl, 1.4 per cent NaCl, other physiological solutions, 1 per cent citrated fowl blood,, moistened earth, fowl feces (plus sand and saline), sterile fowl feces (plus saline and charcoal), moistened bread. Hutson (1943) was unsuccessful in hatching eggs in distilled water but obtained living first-stage larvae from cultures of distilled water plus fowl feces. Upon introduction into the body cav- ity of a roach these larvae developed normally.

It seemed desirable to determine, if possi- ble, whether eyeworm eggs hatch more readily in the digestive tract of the intermediate host or in fowl feces. Attempts were made to ob- tain mature eggs from the crops of infected chickens. (Some embryonated eggs obtained from the uteri of gravid worms may not be fully matured. Such eggs in other species of nematodes have been known to "hatch” ab- normally in artificial media.) Although eggs were usually present in the crop, they could not be separated in sufficient quantity for the experiment. This made it necessary to obtain embryonated eggs from macerated female worms. To avoid as far as possible the selec- tion of immature eggs, special care was taken to collect by use of a fine pipette only the largest eggs.

These were placed in three dishes contain- ing (1) physiological saline solution, (2) physiological saline solution plus sterilized fowl feces, and (3) physiological saline solu- tion plus the macerated alimentary tracts of

several Surinam roaches. None of the eggs had hatched by the end of the second day, but quiescent or feeble larvae were observed in each of the three cultures on the third day. None of the larvae survived. Although addi- tional work is indicated, apparently, under the conditions of this experiment, the medium had no appreciable effect upon the rapidity with which eyeworm eggs hatched.

Larvae observed both in the artificial media and in the alimentary tract of the intermedi- ate host apparently hatched in the following manner. (The terminology used is based on the description of the spirurid egg by Chris- tenson in Chitwood et al., 1940.) The chitin- ous middle layer of the shell separated from the inner vitelline membrane at either or both poles. The region of the poles then became thinner, producing polar operculations marked by ill-defined sub-terminal lines of fracture. One or both caps either broke off at the line of fracture or dissolved, leaving a barrel- shaped shell, open at either or both ends. The embryo then freed itself from the thin vitel- line membrane and squeezed through the polar opening.

Similar observations have been made by Ransom (1904), Sanders (1928), and Fielding (1927).

Development in the intermediate host

Eggs were obtained from gravid female worms and were fed to young laboratory- raised roach nymphs. One nymph was dis- sected 24 hours after feeding, and numerous embryonated and non-embryonated eggs were found in the crop. No eggs or egg shells were found in the feces of the nymphs ex- amined at this time.

At 48 hours, several first-stage larvae were found free in the lumen of the crop and an- terior midgut, and many others were observed in the process of hatching. The empty shells were barrel-shaped, which suggested that they hatched by splitting off one or both polar caps, as previously described. Several of the larvae were rather firmly attached to cellular

Oxyspirura mansoni — SCHWABE

27

debris by the tip of the tail. Embryonated eggs were found in the midgut until the seventh day, and numerous larvae were pres- ent until the ninth day. A single larva was found in the midgut on the eighteenth day. It was almost double the size of those seen there previously.

First-stage larvae were observed for the first time in the body cavity on the eighth day, and by the tenth day many were seen. Some were wandering freely in the cavity, but most were found in the adipose tissue lining the abdominal wall. Apparently they mi- grated through the wall of the midgut, and after wandering in the body cavity for a short time, burrowed into the abdominal fat. The larvae became lethargic at this stage and were extremely difficult to extricate. They grew only slightly in length, broadened consider- ably, lost their long, pointed tails, and as- sumed a more-or-less club-shaped appearance because of the enlargement of the several large rectal cells.

Incomplete encystment actually occurred in this stage in some instances, with the ill- defined cyst measuring approximately 0.3 mm. by 0.5 mm. Thereafter little change took place until about the seventeenth day when larvae measuring 250^^ to 320)U in length and with the cuticular sheath loosened at both the anterior and posterior ends were observed. Larvae in this pre-molting condition were seen until the twenty-fifth day, at which time a second-stage larva which measured 990)U in length was observed.

Complete encystment occurred in the se- cond stage of larval development. The cysts were nearly spherical in shape and measured approximately 0.8 mm. by 0.8 mm. They were found attached to the alimentary tract, particularly around the rectum and entangled in the Malpighian tubules. The cysts were thin-walled an-d transparent. The wall, ap- parently of loose connective tissue, consisted of a gelatinous matrix and a few scattered cells. The wall was richly tracheated, which constituted evidence that the cyst was formed

by the cockroach and not secreted by the parasite. The cysts were filled with fluid and the coiled larvae were able to move about freely in them.

On the thirty-third day encysted second- stage larvae measuring 1.8 mm. to 1.85 mm. were observed; those on the fortieth day had attained a length of 2.6 mm. to 4.5 mm. On the forty-fifth day a second-stage larva 6.62 mm. in length was observed in the process of molting. Ecdysis evidently takes place be- tween the forty-fifth and fiftieth days, at which time the last molting larvae were seen.

Many of the parasites were able to free themselves from their cysts through their in- creased activity during the period of ecdysis, and on the fifty-first day numerous third- stage larvae were found, some encysted, but most wandering freely in the body cavity. These early third-stage larvae measured from 7.4 mm. to 8.3 mm. in length and were ex- tremely active.

Whether or not those larvae which were unable to free themselves during the molting period eventually escape from the cyst could not be ascertained.

Upon the attainment of the third stage, the larvae were infective to the definitive host, as was shown by their appearance in the eyes of chickens within 5 minutes after they were in- troduced into the mouth by pipette.

Development in the definitive host

Large numbers of infective larvae were ob- tained from roaches by use of the Baermann apparatus. They were introduced by pipette into the mouths of 4- week-old chicks. The larvae migrated up the naso-lacrimal ducts and were observed to enter the eyes several minutes after they were introduced. Chicks were killed every 2 days, and the worms were removed from the eyes and examined micro- scopically.

On the second day the larvae appeared much the same as they did in the cockroach. On the fourth day several third-stage larvae in the process of molting were observed.

28

Fig. 7. a, First-stage larva; b, anterior end of late first-stage larva, c, first-stage larva in process of molting, showing loosened cuticle at anterior and posterior ends.

Fourth-Stage larvae were first seen on the fifth day, although some third-stage larvae were still present until the seventh day. The reproductive organs developed rapidly dur- ing this period: the vulva of the female was apparent on the seventh day and the develop- ing short spicule of the male on the ninth day.

A separate mesostom and protostom had begun to form by the thirteenth day. For most larvae the final molt occurred about the

PACIFIC SCIENCE, Vol. V, January, 1951

twenty-first day, and young adults were pres- ent on the twenty-third day. Embryonated eggs were first observed in the crop contents of a chicken on the thirty-second day. Egg laying apparently began between the thir- tieth day, when the previous negative ex- amination was made, and the thirty-second day.

DESCRIPTION OF THE LARVAL STAGES

First-stage larvae

Because of their small size and poor de- velopment the various structures of the first- stage larvae were discerned only with dif- ficulty. The newly hatched larvae were long and slender, being somewhat attenuated at the anterior and posterior ends. The anterior end was rather blunt and the relatively Jong tail was acute. In the later period of this stage, the tail shortened considerably, the relative growth in width exceeded the growth in length, and the body assumed a more-or-less club-shaped appearance because of the en- largement of the several large rectal cells. The maximum size observed in this stage was a length of 339m ^.nd a width of 24.9m- (See Table 5. Tables 5, 6, 7, and 8 show detailed measurements of a single typical larva for each of the days recorded. When size varia- tions among the larvae on a particular day were significant, detailed measurements of more than one larva are given. When the sex of the larvae could be readily determined, measurements of both male and female larvae for a particular day were made.)

The cuticle was very thin and transparent and bore fine transverse striations. No papillae were apparent in this stage.

The oral opening was surrounded by a ring of highly refractile material. The transparent esophagus extended approximately one-third the length of the body and terminated as a large spherical bulb, possessing what ap- peared to be an esophageal valve. The intes- tine consisted of a number of rather large, ill- defined cells containing numerous large vacu- oles of lipid-like material. The ventral anal

Oxyspimra mmsoni — Schwabe

29

TABLE 5

Measurements of First-stage Larvae (All Measurements in Microns)

measurements	DAYS AFTER INFECTION
3	5	8	17	18
Length	125	174	207	249	286
Width (ant. end)		7	7	7.5	7
Esophagus length		57.1		107	93
Anus to post. end.		49.9	46.4	39.2	32.2
Width (max.)	14.3	17.8	17.8	32	18

aperture was often seen with a posterior cuti- cular fold partially covering it, and a few large rectal cells (apparently three in number) were well developed.

A nerve ring was seen encircling the es- ophagus at approximately two-thirds of the distance from its anterior end.

Neither an excretory pore nor an excretory cell was apparent in this stage.

A minute genital primordium, approximate- ly 75 m from the posterior end, was visible on the ventral body wall in several living speci- mens.

Second-stage larvae

Rapid growth occurred during this stage of larval development. The length increased from 990 fj, in early second-stage larvae to 6.62 mm. in the latter portion of the stage. The larvae were long and slender with taper- ing tails, on the end of which four small

papillae were discernible in the later part of the stage.

The cuticle was relatively thin and bore marked transverse striations. A provisional buccal capsule had begun to form, but no cephalic papillae were visible.

The esophagus was long and slender and in the late second stage extended approxi- mately one-half the length of the body in some larvae. Two gland-like structures were apparent at its anterior end, and the esopha- geal-intestinal valve was well formed.

The intestine was a well-defined, thin- walled tube, varying only slightly in diameter and extending from the esophagus to the rectum. The cellular structure of its wall was apparent.

There were three large rectal cells and several smaller ones immediately anterior to the anus, which was located ventrally approx- imately 170-190 fx from the posterior end.

TABLE 6

Measurements of Second-stage Larvae

measurements	days after infection
27	33	40	40	45
Length	999.6 54 67.8	1.85*	2.6*	4.5*	6.62*
Width (ant. end)	53.5 89 24	38.5 107.8	62	52
Width (middle).'	170	172
Width at anus	92.4 400	77	85
Esophagus length		357	616	749
Esophagus width (post, end)		43
Anus to post, end	53.5	160	170	246	250
Excretory pore to ant. end	178	250	360.5
Nerve ring to ant. end.		89		160	185.6
Intestine width . .		62	112
Genital primordia to post, end			890

*These measurements in millimeters; all others in microns.

30

Fig. 8. Anterior and posterior ends of late second- stage larva.

An excretory pore and a large excretory cell were plainly visible on the ventral side of the esophagus, 250-300 ju from the anterior end.

The nerve ring and several small ganglion cells encircled the esophagus, about 100 n anterior to the excretory pore.

The genital primordium was visible in the living larvae ventrad of the intestine, approxi- mately 500 ju from the anus.

PACIFIC SCIENCE, Vol. V, January, 1951

Third-stage larvae

Third-stage larvae varied from 7.53 mm. to 8.31 mm. in length and had a maximum width of approximately 157 ju. Other than in this feature, their over-all appearance was much the same as that of the second-stage larvae.

The cuticle was smooth and no striations were discernible, either transverse or longi- tudinal. The anterior oral opening was six- lobed, and four sublateral papillae were plainly visible projecting at right angles to the surface. No caudal papillae were present in this stage.

The esophageal gland-like structures ap- parent in the second stage were not discerni- ble in the third-stage larvae, but a number of cells were seen to surround the esophagus at its anterior end. The esophagus was distinctly divided into an anterior muscular portion having a length of about 210 n and a pos- terior glandular portion approximately 600 /x in length. The remainder of the alimentary

TABLE 7

Measurements of Third-stage Larvae*

Days after infection 51*

Lengthf 7.53-8.31

Width (ant, end) 53.5

Width (max.) 157

Width (at anus) 92.3

Buccal capsule length. 35.7

Esophagus length. . 846

Esophagus width (post, end) . 71.5

Intestine width (max.). 107

Nerve ring from ant. end 178.5

Excretory pore from ant. end 303

Anus to post. end. 184

*In intermediate host. For measurements of third- stage in definitive host, see Table 7.

fThis measurement in millimeters; all others in mi- crons.

tract appeared much the same as that of the second-stage larvae.

In addition to the excretory pore and cell apparent in the second stage, two lateral ex- cretory canals were traced from the region anterior of the excretory pore to very near the tip of the tail. Each canal sent off an anterior branch which joined ventrad of the esophagus and entered the excretory cell as a common excretory duct. The lateral canals

Oxyspifura mansoni — SCHWABE

32

PACIFIC SCIENCE, Vol. V, January, 1951

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w

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'^These measurements in millimeters; all others in microns.

Oxyspirura mansoni — -Schwabe ■

were each embedded in a multinucleate lateral line 50 /I in width.

The nervous system appeared much the same_as in the second-stage larvae.

The sexes could be distinguished readily in the third stage. In the female the genital primordium could be seen attached to the ventral body wall approximately 650 /x from the posterior end. In the male the primordium was similarly situated but unattached. The primordium in both sexes measured approxi- mately 35 M by 14 /X in this stage and was composed of several cells.

Fourth-stage larvae

In general proportions the fourth-stage larvae differed little in appearance from those of the third stage. Early fourth-stage larvae measured about 10 mm. in length while those in the later portion of the stage measured from 12.8 to 14 mm.

Fine transverse striations were evident in the transparent cuticle. The four sublateral papillae each measured approximately 4.6 ju in length. In addition^ by the twenty-first day, the two latero-caudal papillae had begun to form, and the preanal and postanal papil- lae of the males were evident.

By the thirteenth day the separate pro- tostom and mesostom could be differentiated, and in the later part of the stage the buccal capsule appeared much the same as in the adult.

The remainder of the alimentary tract showed little change from that of the third- stage larvae, the intestine being displaced somewhat to allow room for the developing reproductive organs.

The lateral excretory canals were seen to terminate in the latero-caudal papillae in the late fourth-stage larvae. Other than this, the excretory system had apparently achieved its full development by the end of the third stage.

The only additional development noted in the nervous system in the fourth stage was the appearance of two large dorsal and two

33

large ventral ganglion cells near the nerve ring.

In the fourth-stage larvae the greatest de- velopment occurred in the reproductive sys- tem. By the fourth day the genital primord- ium in both sexes measured approximately 214 ju in length and in the case of the female was attached to the ventral body wall. The point of attachment, which later became the vulva, was approximately 894 ^ from the posterior end. In the male the genital pri- mordium became a U-shaped tube with two terminal bulbous growing tips developing toward the posterior end. By the ninth day the short spicule of the male had begun to form and the vulva of the female was ap- parent, although it remained closed externally during the entire fourth stage. The repro- ductive tract, at least in the female, appeared almost complete by the thirteenth day. By the sixteenth day the long spicule of the male could be recognized. The male repro- ductive system was well formed by the twenty-first day; the ejaculatory duct could be seen to enter the ventral side of the rectum at its posterior terminus, the anal papillae were evident, and both spicules were well developed.

Acknowledgments'. I wish to express my sincere appreciation to Dr. J. E. Alicata, Parasitolo- gist, Hawaii Agricultural Experiment Station, for suggesting the original problem and making certain facilities of the Department of Parasitology available to me; Mr. Paul Breese, Director of the Honolulu Zoo, for examining and permitting the examination of birds in the Honolulu Zoo; Dr. R. W. Hiatt, Chairman of the Department of Zoology and Entomology, University of Hawaii, for pro- viding such excellent facilities for my work; Mr. Hiromu Matsumoto, Junior Chemist, Hawaii Agricultural Experiment Station, for the white rats utilized in this work; Mr. Harry Miyata, Miyata Poultry Farm, for per- mitting the unrestricted use of his flocks and farm; and Dr. M. M. Rosenberg, Associate

34

PACIFIC SCIENCE, VoL V, January, 1951

Poultry Husbandman, Hawaii Agricultural Experiment Station, for providing the experi- mental chickens employed in this study.

REFERENCES

Alicata, J. E. 1947. Parasites and parasitic diseases of domestic animals in the Hawai- ian Islands. Pacific Set. 1(2): 69-84.

Almeida, [.^]. 1933. Arch. Esc. Sup. Agr. e Med. Vet. 10: 171. {fi^ide Reis, J., P. Nobrega, and A. S. Reis. 1936. Tratado De Doencas Das Aves. 327-328. Sao Paulo, Brazil.) [In Portuguese.]

Chitwood, B. C., et al. 1940. An introduction to nematology. Sect. I, Pt. Ill, and Sect. II, Pt. I: 125-240. M. B. Chitwood, Babylon,

N. Y.

CoBBOLD, T. S. 1880. (Prefatory note to P. Manson, "Further observations on microfilariae, with description of a new sptcits.”) Quekett Micros. Club, Jour. 6(44):

130-132.

Cram, E. B. 1927. Bird parasites of the nematode suborders Strongylata. Ascari- data and Spirurata. U. S. Natl. Mus., Bui. 140: 311-328.

Dodd, S. I909. Eyeworms of poultry. Queensland Dept. Agr., Rpt. 1-100. (Fide Fielding, 1926.)

Emmerez, C. I9I8. Poultry in Mauritius. Mauritius Dept. Agr., Bui. 12. (fide Field- ing, 1926.)

and P. Megnin. I90I. Un nouveau

parasite et une nouvelle maladie chez les poulets de File Maurice. \Paris\ Soc. de Biol. Compt. Rend. 53(33): 933-935. QPide Ransom, 1904.)

Fielding, J. W. 1926. Preliminary note on the transmission of the eyeworm of Australian poultry. Austral. Expt. Biol, and Med. Sci., Jour. 3: 225-232.

1927. Further observations on the

life history of the eyeworm of poultry. Austral. Expt. Biol, and Med. Sci., Jour. 4: 273-281.

1928^?. Additional observations on

the development of the eyeworm of poul- try. Austral. Expt. Biol, and Med. Sci., Jour. 5: 1-8.

1928^. Observations on eyeworms

of birds. Queensland Agr. Jour. July, 1928.

Hutson, L. R. 1943. Miscellaneous veterinary research in Antigua, B. W. 1. Studies on Manson s eyeworm of poultry. 1-32. (Un- published thesis: University of Toronto.)

Illingworth, J. F. 1931. Manson’s eyeworm distributed by English sparrows. Hawaii. Ent. Soc., Proc. 7: 461.

Kobayashi, H. 1927. On the life history of the Oxyspirura mansoni and pathological changes in the conjunctiva and ductus lacrymalis caused by the worm. Jap. Path. Soc., Trans. 17: 239-242.

DE Magalhaes, P. S. 1888. Notas helminth- ologicas. Brazil. Soc. de Med., Rev. 1(1): 5-20. (Fide Ransom, 1904.)

Ozoux, M. 1910. La filaire de I’oeil du dindon. [Paris] Soc. de Biol., Compt. Rend. 1: 974-975.

Picard, W. K. I929. In:Ned. Indie voorko- mende Pluimveezieten. [English summary.] Nederland. Indische Bl. v. Diergeneesk. 41(1): 42-48.

Ransom, B. H. 1904. Manson s eyeworm of chickens with a general review of nematodes parasitic in the eyes of birds. U. S. Dept. Agr., Bur. Anim. Indus., Bui. 60: 1-54.

Sanders, D. A. 1928. Manson’s eyeworm of poultry. Amer. Vet. Med. Assoc., Jour. 25(5): 568-584.

1929. Manson s eyeworm of poultry.

Fla. Agr. Expt. Sta., Bui. 206.

Oxyspirura mansoni — SCHWABE

35

ScHWABE, C, W. 1949. Observations on the life -history of Pycmscelus surmamensis Linn., the intermediate host of the chicken eye- worm in Hawaii. Hawaii. EnL Soc.^ Proc. 13(3): 433-436.

195 0^^. Studies on Oxyspirura mansoni.,

the tropical eyeworm of poultry. III. Pre- liminary observations on eyeworm patho- genicity. Vet. Res. 11(40): 286-

290. ■

1950^. Studies on Oxyspirura mansoni^

the tropical eyeworm of poultry. IV.

Methods for control. Hawaii. Ent. Soc., Proc. 14(1): 175-183.

Schwartz, C. W., and E. R. Schwartz. 1949. The game birds in Hawaii. 168 pp. Hawaii Board of Commissioners of Agri- culture and Forestry, Honolulu, T. H.

Tryon, H. 1926. [Validity of 0. parvovum.] Queensland^ Agr. Gaz. 25 (1926): 272. {Fide Fielding, 1926.)

Wilcox, E. V., and C. K. McClelland. 1913. Eyeworm of chickens. Hawaii Agr. Expt. Sta., Press Bui. 43: 1-14.

Viability of Hawaiian Forest Tree Seeds in Storage at Various Temperatures and Relative Humidities^

Ernest K. Akamine^

INTRODUCTION

In Hawaii the Territorial Board of Commis- sioners of Agriculture and Forestry is charged with the maintenance of forest reserves. Be- fore World War II forest tree seeds were im- ported from outside sources for reforestation purposes. During the war years, when seeds could not normally be imported because of the critical shipping situation, it was realized that some locally harvested tree seeds could not be kept under ordinary conditions for any extended period without impairment of their viability.

At the request of the Territorial Board of Commissioners of Agriculture and Forestry, studies on the viability of forest tree seeds were pursued at the University of Hawaii Agricultural Experiment Station between the years 1944 and 1948. Seeds of the following trees were tested: paper bark {Melaleuca Leu- cadendron Linn.), brush box {Tr/stanla con- ferva R. Br.), turpentine tree {Syncarpia lauri- folia Ten.), Norfolk Island pine {Araucaria excelsa R. Br.), mamani {Sophora chrysophylla Seem.), Monterey cypress {Cupressus macro- carpa Hartw.), and Indian sandalwood {San- talum album Linn.). It is the purpose of this paper to present the results of experiments designed to develop practical means of pro- longing the viability of seeds of these forest trees under storage.

^Published with the approval of the Director of the University of Hawaii Agricultural Experiment Station as Technical Paper 213. Manuscript received March 23, 1950.

^Department of Plant Physiology, University of Hawaii College of Agriculture, Agricultural Experi- ment Station.

It had previously been found that because the prevailing temperatures and relative hu- midities of the atmosphere in Hawaii are generally high, seeds of garden, field, and forage crops in ordinary storage deteriorated rapidly (Akamine, 1943). It was further de- termined that, in order to maintain their longevity, seeds should be stored in a medium in which either the temperature or the relative humidity is kept below that of the atmos- phere, or, better still, in a medium in which both the temperature and the relative hu- midity are kept below those of the air.

Germination and viability studies con- ducted on forest tree seeds have not been as extensive as those conducted on seeds of other species. Cold storage prolonged the life of Noble fir seed (Isaac, 1934). Moss (1938) found that seeds of three species of Populus produced a germination of 70 per cent after a storage period of 2 years under calcium chloride at 23° F. These seeds lost their via- bility in 2 to 4 weeks when they were stored at room temperature (70° F.). Coniferous seeds have been successfully stored for several years in sealed containers at 36° to 40° F., provided the moisture content of the seeds did not exceed 5 to 8 per cent (Heit and Eliason, 1940; Latour, 1942). The literature on viability studies of forest tree seeds has been reviewed by Tourney and Korstian (1931: 109-152) and by Baldwin (1942: 81- 94). More recently Crocker (1948: 28-66) and Porter (1949) also reviewed the work on storage studies of seeds, including those of forest trees.

136}

Viability of Seeds — Akamine

37

EXPERIMENTAL PROCEDURE

The seeds of the seven forest tree species used in these studies were harvested and col- lected from the various forest reserves throughout the Territory by the foresters of the Territorial Board of Commissioners of Agriculture and Forestry. The viability studies were conducted by the Department of Plant Physiology at the University of Hawaii Agri- cultural Experiment Station. Freshly harvested and cured seeds were used. Species and their storage periods are listed in Table 1,

Because of the dijSiculty encountered in most of the species in separating normal em- bryonated seed from empty seed by external appearance the ”per cent normal seed” for each species was determined from internal examination of lots of known numbers of seeds obtained at random (Table 1). Germina- tion tests were conducted on lots of seed counted out at random, and the percentage of germination was based on the "per cent normal seed.”

Seeds were stored at relative humidities of approximately 30, 45, 60, 75, and 90 per cent at temperatures of 45°, 59°, and 70°-80° F. (room temperature) The required humidities were maintained with solutions of sulphuric acid of various concentrations (Akamine, 1943) in large desiccators. Some seeds of each species were stored in airtight containers without humidity control at each temperature, and control lots of seeds stored in the open were also included under each temperature. At intervals during the storage period, the specific gravity of the sulphuric acid solutions was determined with a hydrometer, and any divergence from the required specific gravity was corrected by the addition of water or con- centrated sulphuric acid.

At the time of storage, an initial germina- tion test was conducted on the seeds of each species. Thereafter, germination tests were

^The cold storage facilities of the United States Department of Agriculture Bureau of Entomology and Plant Quarantine at Honolulu, T. H., were used during the early stages of this study.

conducted at intervals, once in approximately 1 to 2 months, at the beginning of the storage period, and once in approximately 6 months, during the latter part. Data on the germina- tion test period, germination condition, number of replications, and number of seeds per replication for seeds of each species are shown in Table 1. In all cases, tap water was used in the substratum. The sandalwood seed, with its seedcoat removed by hand, was treated with a fungicide ("Thiosan”). The seedcoat was removed to hasten germi- nation, and the fungicide was applied to pre- vent the contamination of the germination medium by mold organisms. Before testing them, the seeds of mamani were mechanically scarified for 2 to 3 minutes in a shaking machine (Akamine, 1942), using an equal amount of black sand and seed. The black sand was used to abrade the seedcoat, which in this seed is impervious to water and hence requires scarification in order for the seed to germinate. The few hard unswollen seeds re- maining at the end of the germination test were nicked on the seedcoat with a knife and left for an additional period to germinate. In all cases, all sound ungerminated seeds left at the end of the test were considered viable and included in calculation of the germina- tion percentage. Because of their enormous size, the seeds of Norfolk Island pine were germinated in a Minnesota seed germinator at room temperature instead of in the petri dish. The criterion of germination was the emergence of normal primary roots and shoots.

EXPERIMENTAL RESULTS

For the sake of brevity, the voluminous germination data on the stored seeds have been omitted in this paper, and, in their place, graphs^ constructed from these data are pre- sented in Figures 1 to 7. To present the trend of the germination more readily, the curves for these graphs were drawn from points ob-

^The author is indebted to Herbert Sakamoto for the construction of the graphs.

TABLE 1

Data on Stored Seeds of Forest Tree Species

H

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PACIFIC SCIENCE, Vol. V, January, 1951

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*Ail tests at room temperature (70°-80° F,).

Viability of Seeds — Akamine

39

tained by the use of * 'running averages” of the original data.

Paper bark seed

The germination behavior of this seed in storage at various temperatures and relative humidities is graphically presented in Figure 1. At each storage temperature, the relative humidity has influenced the viability of the stored seed (Fig. 1) . In general, the lower the humidity, the longer the seed is kept in a germinative condition. At room temperature, the 45 per cent relative humidity gave results slightly superior to the 30 per cent relative humidity. At 59° F., these two humidities were equally effective in prolonging the life of the seed. The 45 and 60 per cent relative humidities were slightly more effective than the 30 per cent relative humidity for main- taining the viability of the seed in storage at 45° F. At each storage temperature, the seed

tr

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45" F.

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59" F.

LJ

ol I \ ^ I

MAYS mays mays MAYS MAYS

1944 1945 1946 1947 1948

GERMINATION DATE

Fig. 1. The effect of temperature and relative humid- ity on the viability of stored paper bark seed.

kept in an airtight container maintained its viability just as well as that kept at 75 per cent relative humidity. The longevity curve for the seed stored open at room temperature lies approximately midway between the 60 and 75 per cent relative humidity curves. Since the relative humidity of the air is ap- proximately 68 per cent, it seems logical to assume that the curve for the open storage should be in this position. Although no at- tempt was made to maintain the humidity in the cold chamber and no humidity measure- ments were made, it may be surmised that seed stored open at 59° F. was subjected to relative humidities in the neighborhood of 30 and 45 per cent because the curves for these storage media lie in about the same plane (Fig. 1). At 45° F., the curve for the open storage lies in the neighborhood of the lower humidity storage curves after the first year of storage.

When the graphs in Figure 1 are superim- posed and a critical examination is made of them, it becomes evident that the effective- ness of the two lowest relative humidities (30 and 45 per cent) in preserving the viabil- ity of the seed was not influenced by the dif- ferent storage temperatures. At relative humidities of 60 and 75 per cent and under airtight conditions, 45° F. and room tempera- ture were each equally more effective than 59° F. in preserving the life of the seed. At a relative humidity of 90 per cent, 45° F. was more effective than 59° F., which in turn was more effective than room temperature in maintaining the longevity of the seed. In open storage, 59° F. maintained the viability of the seed more effectively than 45° F. and room temperature, which were equal to each other in effectiveness.

It can be seen from Figure 1 that the via- bility of the seed of paper bark could be maintained for about 4 years even in open storage at room temperature. The optimum storage condition at room temperature, how- ever, was one in which the relative humidity was kept below that of the air. Generally

40

PACIFIC SCIENCE, Vol. V, January, 1951

speaking, at temperatures of 59° and 45° F., open storage was about as effective as the low relative humidities in preserving the life of the seed.

Brush box seed

The germination behavior of the seed of brush box in storage under various conditions is depicted graphically in Figure 2. In general, regardless of the storage temperature, seeds stored at the low relative humidities main- tained their viability better than those stored at the high humidities (Fig. 2). At each tem- perature, the 45 per cent relative humidity was optimum, being superior even to the 30 per cent. The difference between these two hu- midities in their effectiveness in increasing the longevity of the seed, however, became smaller as the storage temperature was low- ered. At room temperature, the germination curve of the seed stored in airtight containers

h-

UJ

6 ERMINATI ON DATE

Fig. 2. The eflfect of temperature and relative humid- ity on the viability of stored brush box seed.

approached that of the seed in the 60 per cent relative humidity storage toward the latter part of the storage period. With prolonged storage at 59° the airtight storage curve was near the 30 per cent relative humidity storage curve. At 45° F., the airtight storage was surpassed only by the 45 per cent relative humidity storage in effectiveness in maintain- ing the viability of the seed. As in the case of the paper bark seed (Fig. 1), the germination curve of the seed stored open at room tem- perature lies approximately midway between the curves of the 60 per cent and 75 per cent relative humidity storage (Fig. 2). At 59° F., the open storage curve again lies between the 60 and 75 per cent realtive humidity curves, but at the end of the storage period, it had approached the curve of the higher one of these humidities. At 45° F., the open storage curve actually lies below that of the 75 per cent relative humidity curve.

When the graphs in Figure 2 are superim- posed, it is seen that when the seeds were stored at 30 per cent relative humidity, 45° F. was more effective than 59° F. and room tem- perature in maintaining the viability of the seeds. The two latter temperatures were about equally effective in this respect. The effective- ness of the 45 per cent relative humidity in prolonging the germinative power of the seed was not influenced by the different tempera- tures employed. At relative humidities of 60 and 90 per cent and in airtight storage, it was found that the lower the storage temperature, the longer the seeds remained viable. Tem- peratures of 45° and 59° F. were equally more effective than room temperature; in open storage seeds kept at 59° F. remained in a viable state longer than those kept at 45° F. Seeds stored open at the latter temperature in turn kept better than those stored at room temperature.

Since the longevity of the seed at 45 per cent relative humidity was the same at any storage temperature and since this humidity was the optimum for seed storage, it followed that even at room temperature, the seed of

Viability of Seeds — Akamine

41

brush box could be kept viable for several years provided it was stored at this humidity. Another effective method of prolonging the life of the seed was to store it in airtight con- tainers at 45° F. (Fig. 2).

Turpentine tree seed

The germination results of this seed in storage under various conditions are shown in Figure 3. Seeds stored at different relative humidities behaved differently at different temperatures. In general, at room tempera- ture, the lower the humidity the better the seed kept, although toward the latter part of the storage period, the 45 per cent relative humidity seemed somewhat superior to the 30 per cent humidity. After a prolonged stor- age period at room temperature, the viability of the seeds in airtight and open storage was approximately the same, and the germination

Fig. 3. The eflFect of temperature and relative humid- ity on the viability of stored turpentine tree seed.

curves for these seeds lie above the 75 per cent relative humidity curve and below the 60 per cent humidity curve (Fig. 3). Seeds stored at relative humidities of 30, 45, and 75 per cent maintained their original germination after 4 years of storage at 59° F. Storage at a relative humidity of 60 per cent and storage in air- tight and open containers were slightly less effective than the above humidities at this temperature. Storage at 90 per cent relative humidity at 59° F. was detrimental to the keeping quality of the seed. With prolonged storage at 45° F., relative humidities of 45 and 75 per cent seemed to be about the optimum media for maintaining the longevity of the seed; relative humidities of 30 and 60 per cent and airtight storage seemed to be the next best storage media. Open storage at this temperature was slightly inferior to the above storage conditions. Storage at 90 per cent relative humidity was inferior to all in retain- ing the viability of the seed.

When the graphs in Figure 3 are superim- posed, it is seen that at a relative humidity of 30 per cent, 59° F. was more effective than 45° F. or room temperature in retaining the viability of the seed. At relative humidities of 45, 60, and 75 per cent and in airtight storage, the seed kept in a viable condition longer at 45° and 59° F. than at room temperature. The two lower temperatures were equally effec- tive in this respect. The loss of viability of the seeds stored at a relative humidity of 90 per cent was proportional to the storage temperature; that is, the higher the tempera- ture, the more rapid was the loss in viability. With open storage, 59° F. was slightly superior to 45° F., which in turn was superior to room temperature in maintaining the life of the seed.

From the above considerations, it seemed that an optimum storage medium for the seed of turpentine tree was one in which the rela- tive humidity was maintained at either 30, 45, or 75 per cent at 59° F., or one in which the relative humidity was maintained at either 45 or 75 per cent at 45° F.

42

PACIFIC SCIENCE, Vol. V, January, 1951

Norfolk Island pine seed

The germination behavior of the seed of Norfolk Island pine in storage at various rela- tive humidities and temperatures is presented in Figure 4. At room temperature, no matter what the storage condition, the longevity of the stored seed was very short (Fig. 4). At this temperature, open storage seemed to be slightly superior to the other types of storage. At 59° F., the lower relative humidities, especially the 45 per cent, preserved the via- bility of the seed better than the higher hu- midities. At 45° F., the most striking occur- rence was the position of the 30 per cent relative humidity curve. As a storage medium, this humidity was inferior to all the other storage media. The 60 per cent relative hu- midity at 45° F. seemed about the optimum storage medium, with the 75 per cent hu- midity a close second. In general, up to a

GERMINATION DATE

Fig, 4. The effect of temperature and relative humid- ity on the viability of stored Norfolk Island pine seed.

relative humidity of 75 per cent, the higher the humidity, the better the seed kept in a viable state. At a relative humidity of 90 per cent, the seed did not keep any better than that in open storage. Though inferior to the 60 and 75 per cent relative humidities, the airtight storage was superior to the open stor- age for maintaining the longevity of the seed at 45° F.

When the graphs in Figure 4 are superim- posed, it will be seen that, in general, the germination curves of the seeds stored at 45° F. lie above those of the seeds stored at 59° F. and that the latter curves in turn lie above those of the seeds stored at room temperature. Thus it seems that regardless of the storage medium, the lower the temperature, the longer was the seed kept in a viable condition.

From the above considerations, it seemed that storing the seed of Norfolk Island pine at a relative humidity of 60 per cent at 45° F. extended the germinative life of this seed.

Mamani seed

In Figure 5 is presented graphically the germination behavior of the seed of mamani in storage under various conditions. Unfor- tunately, shortage of seed supply necessitated the termination of some germination tests before the experiment was concluded. As a result, in these instances, accurate compari- sons between the different storage media could not be made. Nevertheless, it seems that in open storage this seed remained viable for an extended period of time even at room temperature. At the lower temperatures of 45° and 59° F., open storage was probably superior to the other storage media.

When all the storage media are taken into consideration, it seems that as a whole the lower temperatures of 45° and 59° F. were slightly superior to room temperature in maintaining the longevity of mamani seed (Fig. 5). These low temperatures were equally effective in prolonging the viability of this seed.

From the above considerations, it followed

Viability of Seeds — ^Akamine

43

CD 50

SO CENT REL. HUM.

45 PER CENT REL. HUM.

€0 PER GENT REL. HUM.

75 PER CENT REL. HUM.

00 PER C£NT REL. HUM.

1 airticht

M OPEN

GERMINATION DATE

Fig. 5. The effect of temperature and relative humid- ity on the viability of stored mamani seed.

that Storage of mamani seed in the open at temperatures of 59° or 45° F. seemed to be the optimum storage condition for maintain- ing the viability of this seed over an extended period of time.

Monterey cypress seed

The germination behavior of Monterey cypress seed in storage under various condi- tions is presented in Figure 6. At room tem- perature, the lowest and the highest relative humidities were the least effective in main- taining the viability of this seed. The 45 and 75 per cent relative humidities were more ef- fective. The 60 per cent relative humidity was the most effective storage medium at room temperature. The airtight storage and open storage were inferior to only the 60 per cent relative humidity. At 59° F., the 30 per cent relative humidity was the least effective stor- age medium, and the 60 per cent relative

humidity was the most effective. The position of the germination curves of the 45, 75, and 90 per cent relative humidities was approxi- mately midway between the curves of these two humidities (Fig. 6). The curve of the air- tight storage approached that of the 30 per cent relative humidity, and the curve of the open storage approached that of the 60 per cent relative humidity. At 45° F., the most effective medium was the open storage, and the least effective was the airtight storage. The 30, 45, and 60 per cent relative humidities were equally less effective than the open storage. The highest relative humidities, 75 and 90 per cent, were somewhat superior to the airtight storage toward the latter part of the storage period in maintaining the viabil- ity of the seed at 45° F.

When the graphs in Figure 6 are superim- posed, it appears that at relative humidities of

GERMINATION DATE

Fig. 6. The effect of temperature and relative humid- ity on the viability of stored Monterey cypress seed.

44

PACIFIC SCIENCE, Vol. V, January, 1951

30 and 45 per cent and in open storage, the lower the temperature the longer the seed re- mains viable. At a relative humidity of 60 per cent and with airtight storage, the three stor- age temperatures were equally effective in preserving the viability of the seed. At 75 per cent relative humidity, temperatures of 45° and 59° F. were equally more effective than room temperature. At 90 per cent relative humidity, 59° F. was more effective than 45° F., which in turn was more effective than room temperature.

From the above considerations, it seemed that storing the seed of Monterey cypress in the open at 45° F. prolonged the life span of this seed better than storing under any other condition.

Indian sandalwood seed

Because of the limited supply of Indian sandalwood seed, only a few seeds were used in the germination tests (Table 1). Neverthe- less, the results obtained seemed reliable.

The germination status of this seed at vari- ous periods of storage under varying condi- tions is depicted graphically in Figure 7, which indicates that at room temperature, the low relative humidities of 30, 45, and 60 per cent seemed to be the optimum storage media. The high relative humidities of 75 and 90 per cent were the least effective in preserv- ing the life of the seed. The airtight storage and open storage were intermediate between these humidity groups in this respect. At 59° F., the lower the relative humidity the longer the seed kept viable. The airtight stor- age and open storage were less effective than the 45 per cent relative humidity but more effective than the 60 per cent humidity at this temperature. In general, the lower relative humidities were more effective than the higher humidities in maintaining the viability of the seed at 45° F. The optimum relative humidity was probably 45 per cent. Seeds stored in airtight containers and in the open retained their viability approximately to the

z

GERMINATION DATE

Fig. 7. The effect of temperature and relative humid- ity on the viability of stored Indian sandalwood seed.

same degree as those stored at the relative humidity of 60 per cent at this temperature.

When the germination curves of the seeds stored at the same humidities at each temper- ature are compared with each other, it will be seen that at relative humidities of 30, 45, and 60 per cent, 45° F. was more effective than 59° F., which in turn was more effective than room temperature in maintaining the via- bility of the seed (Fig. 7). At a relative hu- midity of 75 per cent, 45° and 59° F. were equally more effective than room tempera- ture. At a relative humidity of 90 per cent, the three temperatures were equal in effec- tiveness. In airtight storage and in open stor- age, the lower the temperature the longer the seed remained viable.

From the above considerations, it can be stated that the optimum storage condition for Indian sandalwood seed was one in which the

Viability of Seeds — Akamine

45

temperature and the relative humidity were maintained at 45° F. and 45 per cent, re- spectively.

DISCUSSION

If the longevity of the seeds of all species under all conditions of storage is considered as a whole, it will be seen that these seeds fall in one of the following groups: long-lived seed, medium-lived seed, and short-lived seed. The seed of mamani falls in the first group. In the second category fall the seeds of paper bark, turpentine tree, and brush box. The seeds of Monterey cypress, Indian sandal- wood, and Norfolk Island pine are short- lived.

The retention of viability by the seed of mamani was probably due to the impervious nature of its seedcoat, which interfered with the uptake of moisture and with respiration. It will be remembered that this seed required scarification for germination. Generally speak- ing, regardless of the condition of storage, this seed maintained its viability for 314 years (Fig- 5).

In general, among the medium-lived seeds, the seeds of paper bark and turpentine tree maintained their viability in storage for 4 years, slightly better than the seed of brush box (Figs. 1, 2, and 3).

In general, among the short-lived seeds, the seed of Monterey cypress retained its viability better than that of Indian sandalwood, which in turn maintained its viability better than the seed of Norfolk Island pine (Figs. 4, 6, and 7).

Although different species reacted some- what differently toward the various storage conditions, in general, the lower relative hu- midities were more conducive than the higher humidities to the preservation of viability. There were, however, two species (Monterey cypress and Norfolk Island pine) which were exceptions to this statement. Monterey cy- press seed stored at 30 per cent relative hu- midity at room temperature did not maintain its viability any better than that stored at 90

per cent relative humidity at this temperature (Fig. 6). The 30 per cent relative humidity at 59° F. was the least effective storage medium for this seed. At a temperature of 45° F., the 30 per cent relative humidity was also the least effective storage medium for the seed of Norfolk Island pine (Fig. 4).

SUMMARY

The effects of temperature and relative humidity on longevity of seeds of seven forest tree species in storage for several years were studied.

Although seeds of different species reacted somewhat differently toward the various stor- age conditions, in general it can be stated that the longevity of these seeds can be main- tained in Hawaii if the temperature of the storage medium is maintained at a lower level than that of the prevailing atmosphere (70°- 80° F.), if the relative humidity is maintained at a lower level than that of the prevailing at- mosphere (66-71 per cent), or if both the temperature and relative humidity are main- tained at lower levels than those of the air.

The following were found to be the opti- mum media for the storage of seeds of the individual species in Hawaii:

Paper bark— Relative humidity of 45 per cent at room temperature or open stor- age at 59° F.

Brush box — Relative humidity of 45 per cent at room temperature or airtight stor- age at 45° F.

Turpentine tree — Relative humidity of 45 per cent at 59° F.

Norfolk Island pine — Relative humidity of 60 per cent at 45° F.

Mamani — Open storage at 59° F.

Monterey cypress — Open storage at 45° F.

Indian sandalwood — Relative humidity of 45 per cent at 45° F.

REFERENCES

Akamine, E. K. 1942. Methods of increasing

the germination of koahaole seed. Hawaii Agr.

Expt. Sta. Cir. 21: 1-14.

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PACIFIC SCIENCE, Vol. V, January, 1951

1943. The effect of temperature and hu- midity on viability of stored seeds in Hawaii. Hawaii Agr. Expt. Sta. Bui. 90: 1-23.

Baldwin, H. I. 1942. Forest tree seed. xvi+ 240 pp., 28 figs. Chronica Botanica Co., Waltham, Mass.

Crocker, W. 1948. Growth of plants, v+459 pp., 171 figs. Reinhold Publishing Corp., New York.

Heit, C. E., and E. J. Eliason. 1940. Conifer- ous tree seed testing and factors affecting germination and seed quality. N. Y. State Agr. Expt. Sta. Tech. Bui. 255: 1-45.

Isaac, L. A. 1934. Cold storage prolongs the life of Noble fir seed and apparently in-

creases germinative power. Ecology 15: 216- 217.

Latour, H. J. 1942. Notes on the storage of red pine seed for 9 years. N. Y. State Con- servation Department Notes on Forest Investi- gations 38: 1. [Mimeographed.]

Moss, E. H. 1938. Longevity of seed and es- tablishment of seedlings in species of Populus. Bot. Gaz. 99: 529-542.

Porter, R. H. 1949. Recent developments in seed technology. Bot. Rev. 15: 221-344.

Toumey, j. W., and C. F. Korstian. 1931. Seeding and planting in the practice of forestry. 2d ed. xviii+507 pp., 162 figs. John Wiley and Sons, New York.

Pacific Symphytognathid Spiders

B. J. Marples^

The family Symphytognathidae was erect- ed by Hickman (1931) for a minute spider, Symphytognatha globosa, from Tasmania. The most characteristic feature is the absence of lung-books, the anterior spiracles leading into tracheal tubes which supply the cephalo- thorax as well as the abdomen, while the posterior spiracles are absent. In S. glohosa the female has no palps. Page (1937) and Gertsch (1941) have transferred several other genera into this family, or described new ones. In some of these only four eyes are present in- stead of the usual six, and the female palps may be merely reduced in size instead of ab- sent.

Some of the specimens described in the present paper were collected in 1946 in Upolu, Western Samoa, during a visit devoted to the collection of arachnids; a few hours of collecting in Suva, Fiji, on the return journey, yielded specimens of a closely related but quite distinct species. Because this is a pe- culiar and interesting family which has not previously been recorded from the Pacific is- lands, it seemed desirable to deal with both species together. The main Samoan collection is being dealt with as a whole.

In the following descriptions the leg index, given below the leg formula, is the length of the leg divided by the length of the carapace. The tibial index is the breadth of the proximal end of the patella expressed as a percentage of the combined lengths of the tibia and patella.

Vatu gen. nov.

Six eyes. Chelicerae fused for about half of

^Professor of Zoology, University of Otago, Dune- din, New Zealand. Manuscript received, March 28, 1950.

their length. Ventral end of the retromargin of the cheliceral groove with a large bifurcated process. Palp of female absent. Three claws, ventral one large. Abdomen pear-shaped with ventral spinnerets. Close to genus Symphytog- natha^ but different in web and coccoon and in a number of minor structural characters. Genotype: Vatu vitiensis.

Vatu vitiensis n. sp.

Three 9, Icf, 1 immature cf. From tree trunks, Suva, Fiji.

FEMALE: Length 1.20 mm. Carapace, ster- num, and mouth parts brown, legs lighter. Abdomen mottled greyish-brown above, darkening towards the posterior end. Ventral surface behind the spinnerets very much lighter, almost cream-coloured.

Carapace: Length 0.37 mm., breadth 0.32 mm. Highest near the eyes, which are near the anterior margin.

Eyes: Six, all pale. Posterior row recurved. Lateral eyes touching. PME separated from one another by one quarter of their diameter and from the PLE by rather more than their diameter. Ratio of eyes PLE : ALE: PME = 71:68:60. Eye group breadth 0.23 mm.

Chelicerae: No boss or stridulating organ. Apparently fused for rather less than half of their length. Fang stout with relatively large serrations, and longer than the groove. The ventral retromargin of the groove with a stout process bifurcated into two blunt teeth.

Maxillae: Converging but not meeting above the lip.

Lip: Broader than long.

Sternum: Length 0.23 mm., breadth 0.22 mm. Convex.

Valp: No vestige visible even in cleared

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PACIFIC SCIENCE, Vol. V, January, 1951

specimens, though a stout hair arises in the same spot.

, 14 2 3

Le^s: —

2.9 2.8 2.6 2.1

seen in a cleared specimen as in the figure. Pair of striated structures on the pedicel as in S. glohosa, where they were interpreted by Hickman (1931) as lyriform organs and by Petrunkevitch (1933) as stridulating organs.

			PATELLA
		FEMUR	AND TIBIA	METATARSUS	TARSUS	TOTAL
I		0.35	0.37	0.21	0.19	1.12
II		0.31	0.35	0.14	0.19	0.99
Ill		0.28	0.24	0.08	0.17	0.77
IV		0.35	0.35	0.14	0.21	1.05
Tibial Index I . . .	. . .13.1		Tibial Index IV.	13.6

Three claws, the median one being long, sharp and downcurved, and larger than the paired ones. The median claw on legs I and II has one tooth ; all the other claws are without pectinations. A few serrated bristles on the feet. No spines, but a large erect bristle towards the distal end of each patella, two smaller ones on tibia I, and one on tibia II. One trichobothrium on metatarsi I and II, three on tibiae I and II, two on III, and four on tibia IV.

Abdomen: Length 0.88 mm., breadth 0.56 mm. Pear-shaped with a rounded posterior end. The six spinnerets are ventrally placed, the anterior being the largest. Anal tubercle widened transversely. No colulus. Epigyne in the form of a transverse furrow overhung from in front by a rounded lip. Internal structure

MALE: Length 0.76 mm. Carapace, sternum, and mouth parts brown, legs light brown. Abdomen light yellowish-brown, posterior end black. Anterior part of the sides slightly darker, there being a very faint suggestion of a dorsal band joining these areas. More well marked are two triangular dark areas anterior to the posterior black cap. Together they form a procurved crescentic dark marking, interrupted in the mid-dorsal line by the yellowish-brown ground colour of the ab- domen.

Carapace: Length 0.30 mm., breadth 0.31 mm. Sloping up steeply to the conspicuous row of eyes. Anterior to the eyes is a sinuous ridge, concave above the chelicerae and curv- ing forward in the middle line. This has the appearance of being the anterior edge of the

Fig. 1. Patu vitiensis: a, female, ventral view; b, male, left side; c, female chelicera, dorso-retrolateral view; (7, female apparatus as seen in a cleared skin; e, male palp, retrolateral view; /, male .palp, ventral view.

Symphytognathid Spiders — Marples

49

carapace, but below it is a more or less verti- cal clypeus. The details are very difficult to make out, owing to the small size.

Eyes: Six, all pale. Posterior row slightly recurved. The PME are separated from one another by about half their diameter and from the PLE by almost their diameter. The lateral eyes are touching. Ratio of eyes, PLE: PME: ALE = 75:70:51. Breadth of eye group 0.23 mm. The PME are separated from the an- terior ridge by about half their diameter and from the anterior edge of the carapace by about two and a half times their diameter.

Cheltcerae: Small.

Maxillae and Lip: As in female.

Sternum: length 0.24 mm., breadth 0.19 mm.

Palp: Large and rounded palpal organ, de- tails as in figure.

, 12 4 3

Le^s:

3.8 3.3 2.7 2.4

Patu samoensis n. sp.

Eight 9 from minute sheet webs in the ir- regularities of tree trunks in forest, Apia, Upolu, Western Samoa.

female: Length 1.63 mm. Carapace, ster- num, and mouth parts dark brown. Legs lighter with dark annulations. Abdomen mottled greyish.

Carapace: Length 0.55 mm., breadth 0.43 mm. High and rounded.

Eyes: Six, all pale. Large and conspicuous, posterior row straight. PME separated from one another by less than a quarter of their diameter and from the PLE by less than half their diameter. Lateral eyes touching. Ratio of eyes, PME: PLE: ALE = 78: 68: 64! Eye groups breadth 0.30 mm. PME twice their diameter from the carapace edge.

Chelicerae: Apparently fused for about half their length and rather cylindrical in anterior view. The fang is longer than the groove and

I

II

III

IV

Tibial Index 1 11.3

PATELLA

FEMUR	AND TIBIA	METATARSUS	TARSUS	TOTAL
0.31	0.36	0.16	0.23	1.06
0.26	0.33	0.14	0.19	0.92
0.20	0.23	0.10	0.15	0.68
0.27	0.30	0.11	0.16	0.84

Tibial Index IV 12.5

One stout spine on the ventral surface of tibia II about one third of its length from the distal end.

Abdomen: Length 0.55 mm., breadth 0.33 mm. Pear-shaped with the narrower posterior end truncated. Spinnerets ventral and close to the petiolus.

Web and Coccoons: The webs were situated on tree trunks and consisted of fine horizontal sheets about 4 centimeters long. The silk was too fine for the arrangement of threads to be made out. No oblique thread was noticed as in the Samoan species, though a bunch of coccoons was hanging some way from the edge. Spiders were seen sitting in the centre instead of with the coccoons; possibly these may have been males while the females were with the coccoons.

has relatively large serrations. On the ventral retromargin of the groove is a stout process bifurcated into two sharp teeth. The cheli- cerae do not bulge forward in front of the edge of the carapace when seen in dorsal view.

Maxillae: Converging but not meeting above the lip.

Lip: Broader than long. Junction with ster- num distinct.

Sternum: Length 0.30 mm., breadth 0.29 mm. Convex.

Palp: No trace of a palp is to be seen even in a cleared specimen, though three or four hairs arise from the same spot. No palp was visible in the cleared skin of a first instar taken from a coccoon.

^ 12 4 3

2.6 2.4 2.4 1.9

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PACIFIC SCIENCE, Vol. V, January, 1951

I

II

III

IV..

Tibial Index 1 15.8

PATELLA

FEMUR	AND TIBIA	METATARSUS	TARSUS	TOTAL
0.43	0.46	0.26	0.30	1.45
0.38	0.43	0.22	0.30	1.33
0.31	0.31	0.16	0.22	1.00
0.42	0.42	0.24	0.25	1.33

Tibia! Index IV 14.3

Three claws, the third as large as the dorsal pair and sharply directed downwards. Claws slender and not pectinated, and pectinated bristles not present. No spines, but a large erect bristle at the distal end of each patella, two smaller ones on tibia I and one on tibia II. One trichobothrium on metatarsi I and II, three on tibiae I and II, two on tibia III, and four on tibia IV.

Abdomen: Length 1.16 mm., breadth 0.80 mm. Pear-shaped, projecting behind in a knob-ended process and provided with a pair of similar dorso-lateral processes. The six spinnerets are ventral, the anterior being the largest. No colulus. Anal tubercle large. The epigyne is visible only when the abdomen is removed. There is a transverse fold overhung by a backwardly directed lip whose edge is

dark and thickened. The appearance of a cleared specimen is shown in the figure.

In specimens cleared in potash the spiracles could be seen leading into an atrium from which numerous tracheal tubes supplied both the cephalothorax and abdomen. There was a transverse connecting tube between the atria. Sections showed five pairs of tracheal tubes, three large and two small, passing through the pedicel into the cephalothorax.

Web and Coccoon: The webs were spun in the irregularities of a fallen tree trunk. They con- sisted of a very fine horizontal sheet with very regular meshes. The sheet was an irregular polygon some 6 centimeters long, and an oblique thread extended upwards from the sheet between 1 and 2 centimeters from one end. The threads of the sheet radiated from

Fig. 2. Patu samoensis: a, female, dorsal view; b, female, left side; c, female chelicera, dorso-retrolateral view; d, female, eyes, dorsal view; e, female apparatus as seen in a cleared skin; /, female, anterior view of carapace and chelicerae.

Symphytognathid Spiders — ^Marples

51

the point of attachment of the oblique thread, but it was a sheet and not an orb web. The spider rested at this point together with a cluster of coccoons. The coccoons each con- tained only a single egg about 0.34 mm. in diameter. The coccoon consisted of a sphere some 0.66 mm. in diameter, woven loosely of fine threads so as to be transparent, the whole surface being covered with spirals of thick thread with the loops projecting from the sur- face. The whole structure was thus 0.83 mm. in diameter. All the silk was white. This is quite unlike the web and coccoon described by Hickman for S. glohosa. A first instar re- moved from a coccoon had a globular abdo- men with no trace of the projections.

SUMMARY

A new genus of spiders belonging to the family Symphytognathidae is described, with a species from Fiji and another from Samoa. Members of this family have been described

from Tasmania, New Caledonia, East Indies, and Central and South America, but apparent- ly not from the Pacific islands. The new genus seems to be close to Symphytognatha from Tasmania. •

REFERENCES

Face, L. 1937. A propos de quelques nouvel-

les araignees apneumones. Soc. Zool. Prance, Bui 62: 93-106.

Gertsch, W. J. 1941. Report on some arachnids from Barro Colorado Island, Canal Zone. Amer. Mus. Nat. Hist. Novi- tates 1146: 1-8.

Hickman, V. V. 1931. A new family of spi- ders. Zool Soc. London, Proc. 1321-1328.

Petrunkevitch, a. 1933. An inquiry into the natural classification of spiders, based on a study of their internal anatomy. Conn. Acad. Arts and Set., Trans. 31: 299-389.

The Polynesian Species of Myoporum^

Grady L. Webster^

INTRODUCTION

This revision was originally begun as an analysis of the notoriously polymorphic species Myoporum sandwicense of the Hawaiian Islands. After a considerable amount of work had been done, however, it was found that the forms of Myoporum from southern Poly- nesia were very similar. Because of the close- ness of relationship, it seemed both more practical and more worth while to treat all of the Polynesian species together.

In the Hawaiian Islands, Myoporum sandwi- cense is an evergreen shrub or tree which grows mostly in dry forests on leeward slopes from sea level to an altitude of 10,000 feet. Its plasticity in habit is remarkable, as it may become a tree 15 meters high in the dry forests (Rock, 1913: 427) or a creeping, fleshy, prostrate shrub on the beaches and low rocky islets. More rarely it may even invade the rain-forest, as in Waikolu Valley on Molokai.

The Hawaiians used the species, which they called naio, for timber in building houses (Brigham, 1908: 83). Apparently it was not cultivated, as it is in Rarotonga (Wilder, 1931: 100), for the perfume of the flowers. There are references (Bennett, 1832: 257; Hooker and Arnott, 1841: 93) concerning the attempts of traders of the 1820’s to substitute naio wood for sandalwood in the Chinese

iPart of a thesis submitted in partial fulfilment of re- quirements for the degree of Master of Arts in Botany at the University of Texas. Manuscript received Sep- tember 22, 1949.

^Former graduate assistant in Botany, University of Hawaii, and Teaching Assistant in Biology, University of Texas; now at the Herbarium, University Museum, University of Michigan.

trade, but neither then nor since has the naio attained any real commercial value. Brown (1935: 279) reports that on the island of Rapa (in southern Polynesia) naio wood is used for building canoes and houses.

The real value of Myoporum to the Hawaiian Islands resides in its role in the formation of a dry forest cover and in the consequent checking of soil erosion. On most of the is- lands the dry forest region has been partially or completely denuded, with serious conse- quent erosion. Reforestation work has been carried out chiefly with quick-growing exotic trees such as Casuarina and Eucalyptus, but the appearance of the resulting vegetation is dis- appointing from an esthetic point of view. It is to be hoped that eventually the original dry forest trees, such as native species of Dio- spyros, Acacia, Sophora, and Myoporum, will regain a part of their lost dominance. Egler (1947: 425), studying the communities of the dry southeastern section of the Koolau Range on Oahu, predicted that although Myoporum was rare at that time it might in the future as- sume an important place in the Prosopis com- munity.

Acknowledgments: Dr. Harold St.John of the University of Hawaii originally suggested this study, made it possible to collect and observe the plants in the field, and has been of great assistance in criticizing and offering sugges- tions for the manuscript. Mr. Otto Degener loaned his large and important collections for study and discussed with me some of the forms which I was unable to see in the field. Dr. B. C. Tharp readily consented to having the study continue at the University of Texas, and he and Dr. W. Gordon Whaley kindly

[52}

Polynesian Species of Myopomm — Webster

read and criticized the manuscript. I also wish to thank Miss Marie Neal for making avail- able the facilities of the Bishop Museum Herbarium, Miss Mary Lou Jeffrey and Mr. William L. Brudon for drawing the illustra- tions, and the curators of the herbaria listed below for the loan of specimens:

Bishop Museum, Honolulu (BISH) ; Bois- ser Herbarium, Geneva (G-BOIS); Otto Degener, personal herbarium (DEG) ; Deles - sett Herbarium, Geneva (G-DEL) ; Gray Herbarium (GH); Royal Botanic Gardens, Kew (K) ; New York Botanical Garden (NY) ; Laboratoire de Phanerogamie, Paris (P) ; Riks- museum, Stockholm (S) ; University of Texas, Austin (T) ; Naturhistorisches Museum, Vien- na (W).

TAXONOMIC POSITION

The genus Myoporum consists of about 30 species scattered over a wide area including Mauritius, Australia, New Zealand, New Guinea, China, Japan, Micronesia, and Poly- nesia. It is characterized by its relatively actinomorphic corolla and its fleshy drupe with usually solitary ovules in the cells. Like other genera in the Myoporaceae, Myoporum has axillary flowers, stamens with confluent anther cells, pendent anatropous ovules, and gland-dotted, alternate leaves. The Verbe- naceae is probably the most closely related family, but it is well distinguished by its ovules which are not apically attached and by its opposite leaves which lack internal secretory tissue (Solereder, 1899: 711).

The Polynesian species of Myoporum be- long to section Fentacoelium of Gray (1866: 51), which has priority over the more ap- propriate name Eumyoporum of Bentham (1870: 2). Wettstein (1895: 360) distinguish- ed three sections which contained Pacific insular species — Pentacoelium, Polycoelium, and Eumyoporum — but there is no justification for placing these insular species in more than one section. Kraenzlin’s sections (1929: 15) are even more poorly founded, so it seems

53

that so far no really satisfactory subdivision of the genus has been made.

The Hawaiian species M. sandwicense was apparently first collected by Archibald Men- zies on Vancouver’s expedition and was at first referred to the New Caledonian M. tenuifolium by Hooker and Arnott (1841: 93). It was described as a new species, Poly- coelium sandwicense^ by De Candolle (1847: 705) and was redescribed in the genus Myoporum by Asa Gray (1866: 52), No other name was officially proposed for any of the Hawaiian plants until Leveille (1912: 63) published M. Fauriei based on a Faurie specimen from the island of Hawaii. Kraenz- lin (1929: 21)reduced the species to a variety but appears to have misapplied the name to specimens of var. sandwicense.

Hugh Cuming had collected Myoporum in the Austral Islands (Tubuai) in 1828, but over a hundred years went by before any new species were described from southern Poly- nesia. Cheeseman (1903: 291) listed as Myoporum sp. a Mangaian plant cultivated on Rarotonga. Wilder (1931: 100) referred the Rarotongan plant to M. sandwicense, but Skottsberg (1933: 165) described it as a new species — M. Wilderi—'a.n^. discussed its prob- able relationships to most of the other in- sular species. Two years later Brown (1935: 277) published three species from the Austral Islands and Rapa.

Variation in the Hawaiian myoporums has been discussed by Skottsberg in his paper on M. Wilderi and by Degener (1930: 261), but neither has published any new names although Degener illustrated a pubescent- leaved form and announced his intention of describing it in his Flora Hawaiiensis.

MORPHOLOGICAL CRITERIA

Because of the extreme polymorphism of many Hawaiian populations of Myoporum it is difficult to find morphological characters which do not vary as much between indi- viduals as between varieties. An accurate concept of the range of variation — arrived at

54

PACIFIC SCIENCE, VoL V, January, 1951

by observation of a great many specimens— is essential in defining the taxonomic charac- ters of the various groups.

Growth habit, as has been indicated, is so greatly modified by environment that it is probably of no taxonomic significance. The stem is significant only in distinguishing var. stellatum from var. Degeneri, the former hav- ing pubescent branchlets.

There are many leaf shapes but few of them are constant in any one population. Serrate margins are found sporadically among nearly all the Hawaiian varieties and must be used with extreme caution as a taxonomic character. On the other hand, this serrature is constant in the southern Polynesian forms and may be used dependably to separate M. Stokesii from M. rapense. Conspicuous dif- ferences in leaf texture occur among the Hawaiian plants, but in the main these appear to reflect environmental differences. The relative conspicuousness of the nervation depends in turn on leaf texture and is thus obviously unreliable.

The leaves of all the Polynesian species have subepidermal pellucid-punctate tissue, and the distribution and type of these pel- lucid spots appear to be good specific cri- teria. M. Stokesii and M. rapense are difficult to separate from the New Zealand M. laetum except for the fact that the glandular spots on the leaves of the latter are far more conspicuous. In M. laetum the pellucid spots are visible by reflected light (on the upper leaf surface) and give a distinct "polka dot” effect in transmitted light. The spots of the two Polynesian species are smaller, less con- spicuous in transmitted light, and almost invisible in reflected light.

A readily recognizable and often valuable leaf character in Hawaiian Myoporum is the presence of pubescence. Unfortunately, while pubescent leaves are constant features of varieties stellatum and Degeneri they may appear sporadically in populations of other varieties which ordinarily have glabrous leaves. These sporadic occurrences seem

often to be a response to wounding caused by grazing animals, although in some cases the lower "sucker” shoots from an uninjured plant may bear pubescent leaves. In fact, even if the leaves on basal or wound shoots are not pubescent, they are often serrate and larger than normal leaves. Furthermore, there is evidence that at least in some groups of Myoporum sandwkense the "juvenile” leaves of a young plant are of the abnormal serrate- pubescent type, while those formed later are entire and glabrous. Further investigations on this subject are much to be desired.

The number of flowers in the axillary clusters varies not only between plants but even on the same branch and thus does not appear to be of any great taxonomic value. The same can be said of the flower pedicels, which vary widely, but inconsistently, in length and thickness.

The number and length of the calyx lobes, within certain ranges of variation, are sig- nificant, while their shape (on account of its plasticity) is usually less so. The size and shape of the corolla have not been used for distinguishing varieties; although there are readily apparent differences, they are over- lapping and inconstant. The amount of pubescence on the corolla throat is usually quite variable and may or may not have significance.

The ovary is generally uniform in shape and constant in size, within any one variety. It is glabrous and has a somewhat expanded, dif- ferently colored basal region which seems to be nectariferous. At anthesis the flowers may be noticeably fragrant, although I have personally found the flowers of M. sand- wkense to be almost odorless. In marked contrast to the Hawaiian species in this regard are M. Stokesii and M. rapense, whose flowers are credited in collectors’ notes as ranging in scent from "strong sweet” to "fetid.”

The style furnishes some of the best tax- onomic criteria, especially in its length and in the presence or absence of pubescence.

Polynesian Species of My op orum— Webster

This last character is undoubtedly valuable in delimiting species, although it may oc- casionally be inconstant (on the islands of Hawaii and Raivavae). The abrupt basal curvature of the style observed in some of the specimens from southern Polynesia is an inconstant character and its importance is hard to assess on the basis of the small amount of material studied.

The number of stamens is one of the classical key characters for separating M. sandwicense from the other Polynesian species, but this distinction breaks down when a large number of Hawaiian specimens is ex- amined. The length and height of insertion of the stamens were found to be quite inconstant, and the degree of divergence of the anther cells is dependent on age and is of no taxonomic value at all.

The drupes of most of the Hawaiian varieties differ from the other Polynesian species in having a greenish- white exocarp. Some plants on the island of Hawaii (var. Fauriei), however, may have either whitish, pink, or purplish drupes. The drupes of M. Stokesii and M. rapense are pink or red and brick-red or purplish-red, respectively, while those of M. laetum are said to be purple. Judging from the variability of color in M. sandwicense var. Fauriei, however, it would be unwise to consider differences in shade of color as especially meaningful. The presence of color in the drupes of var. Fauriei and its absence in the other Hawaiian varieties do seem to be significant, however.

The bony endocarp is usually subglobose or top-shaped and often ridged at the top or along the sides. Its shape, although extremely variable, is a distinguishing character in some varieties and species. The number of cells in the drupe is inconstant, but a knowledge of the range of variation will assist in placing a specimen in the proper category. Counting the number of cells is not always easy as some of the abortive ones may be quite small. There is often a considerable degree of ster- ility, for many drupes will be found to

55

contain only one or two cells with well- developed seeds.

TAXONOMIC CATEGORIES

The species is here considered to be a population whose individuals have in com- mon an ensemble of fundamental morpho- logical characters different from that present in other species. It is reasonable to suppose that this morphological distinctiveness is due to various types of barriers (such as geographical, physiological, and genetical) which prevent a free genetic exchange with other species.

The categories subspecies and variety, as here used, apply to groups which occupy a discrete region within the total area of the species and which are characterized by a lack of sharp morphological distinctness from other groups within the species. A sub- species differs from a variety in its greater morphological differentiation, but does not necessarily occupy a greater area. Both are considered to lack genetic isolation — that is, varieties of the same species would be ex- pected to interbreed freely when brought together — and no doubt often owe their existence to geographical barriers.

In recent years there has been considerable discussion among taxonomists as to whether the geographically delimited groups within a species should be given the rank of sub- species or of variety. However, in this work it has seemed better to use both categories in order better to accommodate the great sub -specific range of differentiation.

The form, in the present intrepretation, is a group of plants which occur within a variety and differ by very minor or inconstant char- acters ; they may or may not be geographically localized. Specimens may also be referred to forms when they are inadequate to give an accurate concept of the population. Inasmuch as the forms listed in this study are not con- veniently separated morphologically and to some extent are designations based on in- complete knowledge, it seems unwise to

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PACIFIC SCIENCE, Vol. V, January, 1951

dignify them with Latin names. For con- venience they are listed by number, each under the island where it occurs.

DISTRIBUTION

So far species of Myoporum have not been discovered on any of the islands occurring in the 3,000-mile expanse of sea between Hawaii and the Austral Islands. It is not surprising that they are missing from low coral atolls such as Palmyra and Fanning, but their absence from the Society Islands, the Mar- quesas, and Makatea is unexpected, as is their absence from other large Pacific islands such as Samoa and Fiji.

The disjunct distribution of M. sandwi- cense — with two subspecies in the Hawaiian Islands and one in the Cook Islands (Man- gaia) — is not unique, however. For instance, Isachne distichophylla has been found only on Rarotonga and in the Hawaiian Islands. Hedy Otis section Bikkiocarpa has two species, one from West Maui and one from Rapa (see Fosberg, 1943: 25). The genus Phyllostegia has 23 species in the Hawaiian Islands and one in Tahiti. Our present state of knowledge is insufficient to explain these facts of dis- tribution. However, the theory of Zimmer- man (1948: 49-52) that high volcanic islands, now reduced to reefs or atolls by erosion, once existed as stepping-stones in the mid- Pacific indicates how these cases of bicentric distribution might have arisen.

At the time of Captain Cook’s visit Myoporum undoubtedly existed on all seven of the main Hawaiian Islands. It is now absent from the denuded island of Kahoolawe, but Forbes (1913: 86) learned from old visitors to the island that the naio had been seen there before the dry forest was wiped out. The relative abundance of the species at those localities where it still persists has no doubt undergone great changes. In some areas, as at Kaena Point on Oahu and the Puuwaawaa district of Hawaii, it may be as common as it was before the dry forests were disturbed; on the other hand it is rare or absent from

areas such as the Lualualei region on Oahu, western Molokai, and the Kohala peninsula of Hawaii, where probably it was once very abundant.

In the Austral Islands the destruction of the native forests has been so complete that a clear picture of the distribution of Myo- porum there may never be obtained. St. John tells me that he made a special effort to col- lect Myoporum on these islands during the Mangarevan Expedition of 1934 and that few plants of the genus could have been overlooked in the vestiges of the native for- est. Judging from his findings, M. rapense var. Skottsbergii and the form described by Brown as M. rimatarense may well be extinct.

On the basis of our present knowledge, however, it seems significant that M. rapense is found on the three lower islands of the Austral group and that its derivative M. Stokesii is restricted to Raivavae.

One of the chief difficulties in under- standing the distributional pattern of Poly- nesian Myoporum is that the means of long- range dispersal of the genus are still a matter of conjecture. It does not appear likely that the fruits are spread by ocean currents, since my own observations show that they will float for no more than a few days. Dispersal by birds is a possibility, since some endemic Hawaiian birds were known to have eaten the seeds; but there is no evidence that any migratory birds are fond of the fruits. It must be admitted that the dispersal mechanism of Myoporum still remains unknown, and that the accidental transport by hurricanes is as likely a possibility as any.

RELATIONSHIPS OF THE SPECIES

At present, and in spite of a fairly recent monograph (Kraenzlin, 1929), the genus Myoporum is in great need of critical revision. The seat of the greatest systematic difficulty is the complex of chiefly insular species which includes the three Polynesian species. Almost nowhere within this insular complex are the species well defined, and it is very difficult to

Polynesian Species of Myoporum — ^Webster

find morphological characters sufficiently constant to distinguish them.

Because of its great range of morphological variation, M. sandwkense poses a particularly difficult problem in species definition. In its most typical form the species is well charac- terized by its flowers with five stamens and a glabrous style, and by its whitish-colored drupes with five or more cells. To some ex- tent, however, every one of these characters proves to be inconstant when the complete range of variation of the species is ascertained. The other Polynesian species typically differ in having flowers with four stamens and pubescent styles as well as reddish-tinted drupes with fewer cells, but most of these characters are inconstant. However, M. sand- wkense not only approaches the other Poly- nesian species very closely, but also is hard to separate from M. tenui folium, a species lying to the west and southwest in Micronesia and Melanesia. This latter species is charac- terized by the small size of its flowers and fruits; but there are forms of M. sandwkense whose reproductive organs are of about the same dimensions. Of course, in all these cases species distinctions can be maintained by using rather complicated combinations of characters; for instance, Af. tenuifolium can be separated from M. sandwkense by the fact that the forms of the latter which have small flowers and fruit do not have pink-tinted drupes. Such contrived species differences are admittedly unsatisfactory, but they will have to serve for the present. The temptation to combine various of the insular species should be resisted until the entire group is surveyed, because only then will it be possible to define the exact limits of M. sandwkense and its nearest congeners.

Because the classification of the insular myoporums is still in a very imperfect state, one hesitates to speculate as to the phylogeny of the Polynesian groups. However, the fol- lowing observations on the possible relation- ships of the Hawaiian groups seem worthy of statement.

57

It is pretty clear that the group of M. sandwkense showing the greatest resemblance to the Austral Island species is ssp. Wilderi, although it is not possible to say whether or not the latter originated directly from or coordinately with the Austral Island species. Among the groups of Af. sandwkense from the Hawaiian Islands proper, var. Fauriei shows the greatest resemblance to ssp. Wilderi. Because var. Fauriei is restricted to the island of Hawaii, it seems likely that this island was the point of immigration of Myoporum. Degener (1930: 266), on the basis of the occurrence of pubescent, serrate leaves on certain wounded plants from Molokai, has suggested that the direct ancestor of the Hawaiian forms had leaves of this type. How- ever, this phenomenon does not seem to be a case of a reversion to an immediate ances- tral type, but rather the expression of a potentiality which is widespread, inasmuch as independent occurrences of pubescent leaves are known from other Hawaiian islands as well as from Australia, New Zealand, and New Caledonia.

It seems evident that var. sandwkense, the most widely distributed of the Hawaiian groups, is derived from var. Fauriei, while it in turn has given rise to varieties lanatense, Degeneri, and stellatum. The origin of ssp. St.-Johnii is more obscure. It could have come directly from the adjacent population of var. Fauriei, but the rather great morphological differences between the two entities make this rather questionable. The history of Myoporum on the island of Hawaii is made even more mysterious by the fact that var. sandwkense is absent from this island, even though it is presumed to have originated there. This suggests that the original popu- lation of var. sandwkense on Hawaii has become gradually modified into the group now known as ssp. St.-Johnii. This suggestion is of course hypothetical, but it would explain the curious anomaly of the absence of var. sandwkense from its presumed place of origin, as well as the occurrence on Hawaii of forms

58

PACIFIC SCIENCE, Vol. V, January, 1951

of ssp. St.-Johnii very similar to var. sand- wicense.

On each island of the Hawaiian group independent modifications, some of which are superficially similar and have resulted in a sort of parallel evolution, have arisen in the populations. This curious phenomenon, which is characteristic of populations on some other Pacific islands as well, has been remarked on by Degener (1930: 266), who realized the independent origins of the hairy- leaved and ''peach-leaved” forms on the various Hawaiian islands. Even more per- plexing from the taxonomic point of view is the occurrence of reversions of characters. Thus, in tracing the character of style pubescence in the Polynesian species, we find that it is present in M. rapense, beginning to disappear in M. Stokesd, absent in ssp. Wilderi, but reappearing in ssp. St.-Johnii. Naturally this sort of behavior of morpho- logical characters introduces great hazards in classifying and interpreting the phylogeny of the Polynesian species.

It is interesting that there seems to be little evidence of inter-varietal hybridization in the Hawaiian Islands. No indubitably hybrid specimens have been seen except possibly from the island of Hawaii, and here additional field work will be necessary to establish the actual hybrid nature of such specimens. The conclusion seems inevitable that evolution in Hawaiian Myoporum has been most strongly conditioned by geo- graphical isolation, while inter-varietal hy- bridization has not played an important part. The fast rate of differentiation and slow dis- persal rate of the populations have been acting together to produce an intense micro- evolution. The taxonomic difficulty of char- acterizing M. sandwicense is a reflection of the rapidity and recentness of this process. Within this one species can be found groups at all stages of differentiation — from individual variants to populations of almost specific distinctness.

Owing to the ruination of much of the

flora of the Austral Islands, our knowledge of the groups there is rather incomplete. Never- theless, it is clear that M. rapense is very closely related to M. laetum of New Zealand and is no doubt derived from it or a similar type. On Rapa and Tubuai it has changed little from the original immigrant, but on Raivavae it has given rise to and been almost supplanted by the polymorphic M. Stokesii, which itself is undergoing incipient speciation in the manner of M. sandwicense.

SYSTEMATIC TREATMENT

It is not pretended that the taxonomic ar- rangement which follows is definitive. Fur- ther collecting on some of the Hawaiian Islands may necessitate a reappraisal of some of the unnamed forms listed herein and should result in the discovery of additional forms and perhaps even new varieties. Ade- quate understanding of a highly polymorphic genus such as Myoporum requires accurate field observations, and careful notes by col- lectors concerning flower color and scent, fruit color, and leaf variation will be of great assistance in future studies.

MYOPORUM Soland. ex Forst. f.. Prodr. 44. 1786.

Generic synonyms pertinent to the Poly- nesian species are:

Pentacoelium Sieb. & Zucc., Abh. Akad.

Muench. 4(3): 151. 1846.

Polycoelium A. DC., Prodr. 11: 705. 1847. Prinastrum Nutt, ex Gray, Am. Acad. Sci.

6: 53. 1866 (as synonym).

Low or erect shrubs, rarely tall trees. Leaves alternate (rarely opposite), sessile or with a winged petiole, linear to lanceolate or el- liptic, entire or serrate, glabrous or rarely pubescent, often viscid when young. Flowers axillary, solitary or clustered, ebracteolate- pedicellate; calyx usually deeply 5-lobed, green and herbaceous, gland-dotted. Corolla usually 5-lobed, imbricate in the bud, sub- actinomorphic, campanulate to funnelform.

Polynesian Species of My op or um — Webster

59

glabrous without, glabrous or pubescent within, usually glandular-punctate. Stamens usually 4, rarely more, alternate with the corolla lobes; filaments subulate or terete, glabrous or pubescent, adnate to the corolla; anther sacs confluent. Ovary of various shapes, often conic-cylindric, usually 2-5- celled (rarely the cells more numerous) ; style simple, usually as long as or longer than the ovary; stigma convex, entire or obscurely 2-5 -grooved (rarely bifid); ovules solitary or rarely paired in each cell of the ovary, anatropous, pendent from the apex of the cell. Fruit a fleshy drupe; endocarp bony, usually more or less subglobose but sometimes de- pressed or laterally compressed; seeds spindle- shaped, with scanty endosperm, the radicle superior.

TYPE SPECIES: Myoporum laetum Forst. f., here chosen as lectotype for the genus. It should be noted that Forster attributed the authorship of the genus Myoporum to Solan- der alone, and not to Banks and Solander, as subsequent writers have incorrectly done.

Section Pentacoelium (Sieb. Zucc.) A. Gray, Proc. Am. Acad. Sci. 6: 51-52. 1866.

'Pentacoelium Sieb. & Zucc., Abh. Akad. Muench. 4(3): 151. 1846 (as a genus).

Polycoelium (A. DC.) A. Gray, op. cit. (as a section).

Eumyoporum Bentham, Flora Australiensis 5: 2. 1870 (as a section).

Insularia Kraenzlin, Fedde Repert. Sp. Nov., Beih. 54: 15-16. 1929 (as a section).

Ovary mostly 2-4-celled; ovule, one in each cell. Drupe mostly subglobose, not strongly laterally compressed.

KEY TO THE POLYNESIAN SPECIES

This key is intended for use in determining specimens in flower and fruit. Sterile twigs from the base of a plant or from a wounded branch may sometimes have serrate and/or densely pubescent leaves very unlike the normal ones. Specimens of this unusual type

should be accompanied by normal flowering branches if the correct determination is to be made.

A. Style glabrous, or if hirsutulous then 2-3.5 mm. long and fruit not red-tinged. . . 1. M. sandwicense

B. Style glabrous.

C. Stamens usually 5 (less commonly 4 or 6); corolla glabrous or pubescent; leaves various,

not elliptic-spatulate and abruptly acute

2. ssp. sandwicense

D. Leaves glabrous.

E. Endocarp strongly depressed, not or scarcely ridged, the thick wall exceeding the cells in diameter. ... 5. var. lanaiense

E. Endocarp of various shapes, often ridged, rarely strongly depressed, and then the wall thinner than the cells in diameter.

F, Calyx lobes mostly 1-3 mm, long; corol- la often pubescent within; style mostly 1.5-3 mm. long; drupe always greenish- white; endocarp 2-6 mm. long, 4-10-

celled 3. var. sandwicense

F. Calyx lobes mostly 3-5 mm. long; corol- la glabrous (or rarely sparsely pubescent) within; style mostly 3-5 mm. long; drupe sometimes pink or purplish; endo- carp 5-10 mm. long, 4-6-celled

4. var, Fduriei

D. Leaves pubescent.

G. Hairs branched; branchlets pubescent; leaves lanceolate or elliptic-lanceolate . . 6. var. stellatum

G. Hairs unbranched.

H. Branchlets glabrous; leaves often broad- ly elliptic, entire or remotely serrate. .

7. var. Degeneri

H. Branchlets usually pubescent; leaves lanceolate.

I. Corolla not over 6 mm. long; style

less than 3 mm. long

var, sandwicense, pubescent form

I. Corolla over 6 mm. long; style 3 mm.

long or more var. Fauriei,

pubescent form

C. Stamens constantly 4; corolla densely pubes- cent; leaves elliptic-spatulate, abrubtly acute . .

9. ssp. Wilder i

B. Style hirsutulous; flower parts mostly 6-8; endo- carp usually 7-12-celled, often sharply ridged. .

8. ssp. St.-Johnii

A. Style hirsutulous (rarely glabrate in M. Stoke sii), 4-6 mm. long; stamens 4; fruit pink or red; endo- carp 3-6-celled.

J. Leaves entire; endocarp angled. . . 10. M. Stokesii

J. Leaves serrate; endocarp not or scarcely angled.

11. M. rapense

K, Calyx lobes not ciliate . 12. var. rapense

K. Calyx lobes ciliate 13. var, Skottshergii

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PACIFIC SCIENCE, Vol. V, January, 1951

1. Myoporum sandwicense A. Gray, Proc.

Am. Acad. Sci. 6: 52-53. 1866.

Polycoelium sandwicense A. DC., Prodr. 11: 706. 1847 (illegitimate name).

Usually much-branched shrubs 1-3 m. high, but stature varying from prostrate (littoral form) to trees 15 m. tall. Stem usually glabrous with smooth green or olive-brown bark when young, gray and cracking into chunks when older. Leaves alternate, often lanceolate but varying from linear-lanceolate to ovate, acute to long-acuminate, entire or often crenate or serrate, decurrent-petiolate,

3.5- 22 cm. long, 0.5-4 cm. wide; usually somewhat fleshy but varying from submem- branous to stiff-coriaceous when dry; usually glabrous or more rarely pubescent. Flowers axillary, in clusters of 2-10, often 1 or more abortive; pedicels terete or flattened, 0.5- 1.7 cm. long. Calyx lobes 4-9, mostly 5,

1- 6 mm. long, broadly ovate to oblong or lanceolate, green and herbaceous or slightly scarious on the minutely glandular- denticulate margins, glandular-punctate, usu- ally glabrous. Corolla 4-9- (mostly 5-) lobed,

4.5- 12 mm. long, campanulate to funnelform, usually glandular-punctate, entirely glabrous to densely pubescent within; color pure white, white with pink-purplish splotches, or entirely pink; corolla lobes ovate- deltoid to oblong, emarginate, obtuse, or apiculate,

2- 7 mm. long. Stamens inserted on the corolla, the same number as and alternate with the corolla lobes or occasionally 1 or 2 fewer; filaments flattened-subulate, 1.5-5 mm. long; anthers 0.5-1. 5 mm. long, the cells confluent by a common suture and thus changing from parallel to widely divergent at an thesis. Ovary cylindric-conic to de- pressed-conic, 1.5-4 mm. long, glabrous, green, 4-12-celled, the base with a yellow or purple apparently nectariferous ring; style terete or flattened, 1.5-6 mm. long; stigma convex, obscurely 2-5-grooved. Drupe fleshy, the exocarp greenish-white or creamy-white, rarely pinkish or purplish; endocarp 2-10

mm. long, 2-9 mm. broad, smooth or ribbed, subglobose or variously flattened, hard and bony when mature. Seeds one in each cell, spindle-shaped, papery when dry, 2-2.5 mm.

long.

The name Myoporum sandwicense dates from its use by Asa Gray in 1866, the origi- nal combination Polycoelium sandwicense A. DC. being illegitimate under the International Rules of Botanical Nomenclature, Article 60 (1). The generic name Polycoelium was a substitution for the earlier Pentacoelium of Siebold and Zuccarini, and hence was super- fluous when published. According to Article 69 of the rules, Gray’s adoption of the epithet sandwicense is treated as a new name and not as a new combination.

As here construed M. sandwicense em- braces three rather diverse subspecies, each of which could well be treated as a species if their differences were constant. I have been reluctant to reduce M. Wilderi to a subspecies, but if this were not done consistency would demand the elevation of ssp. St.-Johnii to specific rank, and this action does not seem warranted.

2. Myoporum sandwicense ssp. sand- wicense.

Myoporum sandwicense A. Gray, Proc. Am. Acad. Sci. 6: 52-53. 1866 (in part, excluding reference to Douglas collection).

Prostrate shrubs to tall trees. Leaves mostly lanceolate, less commonly almost linear or ovate or elliptic, never spatulate, 3.5-22 cm. long, 0.5-4 cm. broad, glabrous or pubescent. Flowers in clusters of 2-10 (mostly 3-5). Calyx mostly 5 -lobed, lobes broadly ovate to oblong or lanceolate, 1-5 mm. long. Corolla mostly 5 -lobed, glandular-punctate, glab- rous or pubescent within, 4.5-12 mm. long. Stamens isomerous with the corolla lobes, the fifth one occasionally reduced or absent. Ovary 4-10-celled, 1.5-4 mm. long; style glabrous, 1.5-6 mm. long. Drupe creamy- or greenish-white or rarely pinkish or purplish;

Polynesian Species of Myoporum — ^Webster

61

endocarp 2-10 mm. long, 2-9 mm. broad, subglobose to depressed.

This extremely variable and complex sub- species is restricted to the Hawaiian Islands.

3. Myoporum sandwicense ssp. sand-

wicense var. sandwicense.

Pis. I, 2-6; II, 16-17, 27-30; III, 38-40

Myoporum sandwicense A. Gray, Proc. Am. Acad. Sci. 6: 52-53. 1866 (in part, excluding reference to Douglas and Nuttall collections).

Leaves mostly lanceolate to oblong or ovate lanceolate, rarely sublinear or ovate,

4- 22 cm. long, 0.5-3. 8 cm. broad, glabrous (except in one form). Flowers in clusters of 2-9 (mostly 3-6); pedicels 0.3-1. 5 cm. long. Calyx mostly 5-lobed (rarely the lobes 4, 6, or 7); lobes broadly to narrowly ovate or oblong, 1-3 (rarely 4) mm. long. Corolla

5- lobed (rarely the lobes 6-7), glandular- punctate, glabrous or pubescent within, 4.5-8 (rarely to 9-5) mm. long. Stamens usually 5, occasionally one reduced (more rarely the number 4 or 6-8). Ovary 4-10-celled but mostly 5-7-celled; style 1.5-3 (rarely 4) mm. long. Drupe creamy-white, never pink or purplish; endocarp 2-6 (rarely 7) mm. long, mostly subglobose or top-shaped, usually ribbed.

LECTOTYPE: Isles Sandwich, Oahu, Remy 462 in the Gray Herbarium.

This extremely polymorphic variety is found at present on all the main islands except Hawaii and Kahoolawe. It is the heterogeneous residuum which remains after the more well-marked populations have been removed, and some of the forms enumerated here may warrant reappraisal when better known.

It has seemed best to arrange the various forms in a geographic sequence, first for the sake of convenience and also because forms from different islands which appear morpho- logically similar are most likely related not to each other but to adjacent forms on the same island. To give an idea of the corres- pondence and relationships of the forms,

however, the following key is submitted with the caution that a clear differentiation of the forms is not possible.

Key to forms of var. sandwicense

A. Leaves ali serrate, glabrous; style mostly 3-4 mm.

long Molokai Form 3

A. Leaves entire or occasionally serrate or pubescent and sharply serrate; style usually not over 3 mm, long.

B. Leaves glabrous.

C. Stamens 4; style mostly 2.5-4 mm. long. . . .

Oahu Form 5, Kauai Form 4

C. Stamens mostly 5, rarely more.

D. Style 2.5-4 mm. long; corolla glabrous, broadly campanulate, the lobes 5-7 .......

East Maui Form

D. Style mostly 1-3 mm. long; corolla often pubescent, the lobes mostly 5.

E, Corolla lobes 6-8, corolla pubescent. . . .

Oahu Form 4

E. Corolla lobes mostly 5, rarely 6.

F. Leaves large, 8-22 cm, long, often ser- rate; style 2-3 mm. long; corolla pubes- cent Kauai Form 1

F. Leaves smaller, 5-14 cm. long, mostly entire; corolla glabrous or pubescent.

G. Endocarp strongly depressed.

H. Calyx less than 2 mm. long

Oahu Form 3

H. Calyx over 2 mm. long

West Maui Form 2

, G. Endocarp more or less isodiametric, or at least not depressed.

I. Leaves linear-lanceolate, 8-10 times

as long as broad; calyx 2-3 mm. long. Molokai Form 2

I. Leaves mostly less than 8 times as long as broad.

J. Leaves broadly elliptic.

K. Leaves rather membranous . . . Kauai Form 3

K. Leaves fleshy, leathery when

dry Oahu Form 1

J. Leaves mostly lanceolate.

L. Leaves long-attenuate

Kauai Form 4

L. Leaves acute to acuminate.

M, Cells of drupe 4-5; corolla mostly about 5 mm. long

Molokai Form 1

M. Cells of drupe mostly 5-7, less commonly 8 or more, rarely 4; corolla often long- er than 5 mm

.... Niihau Form, Kauai Form 2, Oahu Form 2, Lanai Form, West Maui Form 1

B. Leaves mostly densely pubescent and serrate, sometimes glabrous entire leaves on the same plant Molokai Form 4

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PACIFIC SCIENCE, Vol. V, January, 1951

15

Polynesian Species of Myopomm — Webster

63

Specimens examined

HAWAIIAN ISLANDS (without Specific lo- cality) :

Bennett (K); Gaudichaud 39 (G-DEL, P); 1839, Gaudichaud (G-DEL); Macrae (K); Menzies (K); Wilkes Expedition (NY); ” Ha- waii,” Wilkes Expedition (NY).

NIIHAU:

At foot of plateau, South East, Stokes (BISH, NY); south corner of plateau, Stokes (BISH); Kalaalaau Valley, alt. 50 ft., St. John 22747 (BISH) ; Mokouia Valley, alt. 500 ft., St. John 23392 (BISH); Paniau, 1 mile S. of, crest of windward cliff, alt. 1,200 ft., St.John 23603 (BISH).

KAUAI:

Eorm 1. Leaves large, 8-22 cm. long, tend- ing to be serrate. Stamens 5.

Waimea, seacoast, Mann & Brigham 383 (in part) (G-BOIS, GH, NY); Waimea, Polihale, flat back of beach, St. John et al. 22936, 22937, 22961 (BISH, T).

Eorm 2. Leaves mostly smaller and entire, 5.5-14 cm. long. Stamens 5.

Without locality, Menzies (GH); Wawra (G-DEL); Wawra 2072, 2407 (W); Oct. 1916, Rock (BISH). Waimea, seacoast, Mann & Brigham 383 (in part) (BISH, GH); Napali Coast, at top of beach west of Kalalau Valley, St. John et al. 23208 (BISH, T); barren ridge, Waimea Drainage Basin, West Side, Forbes 982. K (BISH); Waimea, Milolii Ridge, on bare windswept headland, St. John & Fosberg 13711 (BISH); Waimea, Polihale, rocky lowland, alt. 30 ft., St.John et al. 22933 (BISH, T); Haeleele Valley, Degener 9736 (DEG) ; northwest side of Waimea Canyon, along ditch trail at 3,500 ft., Degener 20313

(DEG) ; Waimea, Kumuwela [Kumuweia] Ridge, bottom of gulch near stream, alt. 3,500 ft., St. John, Fosberg, & Oliveira 13832 (BISH); southeast side Kumuwela Ridge, moist open forest, alt. 1,100 m., Fosberg 12630 (BISH); Kaholuamanu, Forbes 319 (BISH); same locality [Kaholuamano], Rock? 92 (BISH), Rock 2478 (BISH, GH, NY), 3238 (BISH), 5259 (GH).

Form 3. Leaves elliptic, rather abruptly contracted to the tip.

Without specific locality, Remy 463 (GH).

Form 4. Like Form 2 but stamens 4.

Without specific locality, Wawra 2299 (W).

Form 3. Leaves attenuate-acuminate, entire. Stamens 5. Drupe 6-7 mm. long.

Without locality, Brodie (BISH); ridge west of the Hanapepe River, Heller 2432 (BISH, G-BOIS, G-DEL, GH, NY, P).

OAHU:

Form 1. Leaves fleshy, elliptic, up to 3 cm. wide (probably a mere ecological form) .

Without locality, Remy 461 (GH, P) ; Mokuleia, near sea, Degener 9730 (DEG, NY); Moku Manu 1. (islet off Mokapu Point), Munro (BISH).

Form 2. Leaves lanceolate. Stamens 5 or rarely 4 or 6. Drupe subglobose, 5-8-celled (up to 10-celled in specimens from KooJau Range). Styles mostly 1. 5-2.0 mm. long.

Without locality: Beechey Expedition (G- DEL, K), Remy 462 (GH), Wilkes Expedi- tion (GH).

Waianae Range: Kaena Pt., Bryan (BISH); Kaena Pt., Kuaokala, Cowan 726 (BISH); Kaena, in front of beach, alt. 15 ft., Cowan 732, 737 (BISH); Kaena Pt., near sea.

PLATE I

Leaf outlines of the Polynesian species of Myoporum.

EXPLANATION OF PLATE

M. sandwicense var. stellatum: (1) from Webster 1245, Oahu. M. sandwicense var. sandwicense: (2) from Forbes 148. Mo, Molokai; (3) from Degener 9730, Oahu; (4) from Fosberg 15155, Oahu; (5) from Degener 12184, Molokai; (6) from Heller 2452, Kauai. M. sandwicense var. Fauriei: (7) from Webster & Wilbur 1775, Hawaii; (8) from Rock 8535, Hawaii. M. sandwicense ssp. Wilderi: (9) from Wilder 781, Rarotonga. M. sandwicense var. Degeneri: (10) and (11) from Degener 12185, East Maui. M. Stokesii: (12) from St.John & Fosberg 15956, Raivavae; (13) from St.John & Fosberg 15964, Raivavae. M. rapense var. Skottsbergii: (14) from Cuming 1450, Tubuai. M. rapense var. rapense: (15) from Fosberg 11525, Rapa.

64

PACIFIC SCIENCE, VoL V, January, 1951

HAWAIIAN ISLANDS ASSEMBLED

Fig. 1, Distribution of iW. sandwicense var. sandwkense. Solid dots indicate exact locality, open dots approximate locality. (Base map, courtesy of Bernice P. Bishop Museum.)

Degener, Park, & Nitta 9731 (DEG); Kaena Pt., Forbes 1632.0 (BISH, NY); blufF above Kaena Pt., alt. 60 m., Fosberg 13133 (BISH); by railroad, Kaena Pt., alt. 15-20 ft., Neal (BISH); Kaena Pt., Russ (BISH), Topping (DEG); Nihoa Gulch, Mokuleia, alt. 150 ft., Hume 307 (BISH); same locality, alt. 400 ft., Liu (DEG); Nihoa Gulch, Kaena, base of cliffs, alt. 400-500 ft., St.John 11140 (BISH); near beach 1 mile east of Kaena Pt., Webster 1077, 1079, 1080 (BISH, T), 1078 (BISH); cliffs 2 miles east of Kaena Pt., alt. 250-350 ft., Webster 1086, 1091 (BISH, T), 1090 (BISH); at the top of the first gulch east of Kaena Pt., alt. 400 ft., Webster 1092. A (BISH, T), 1092. B (near to Form 3) (BISH); cliffs 1 mile east of Kaena Pt., alt. 400 ft., Webster & Cowan 1097 (BISH, T); near road 1 mile east of Kaena Pt., Webster & Krauss 1096 (BISH); Kewaula Valley, arid rocky valley, Degener, Park, & Nitta 9732 (DEG, NY); arid ravine near Kawaihapai, Degener 2197 (DEG, NY); steep brushy wall of gulch, south side Makua Valley, alt. 500 m.,

Fosberg 12330 (BISH); Mt. Kaala, forest at 2,500 ft., Degener, Murashige, & Kerr 19736 (tra