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THE
PROGEEDINGS
OF THB
LINNEAN SOCIETY
OF
New SoutH WALES VAS
FOR THE YEAR
IL)
VOL. LXXXVII.
WITH SIXTEEN PLATES. 306 Text-figures.
SYDNEY: PRINTED AND PUBLISHED FOR THE SOCIETY BY
AUSTRALASIAN MEDICAL PUBLISHING CO. LTD., Seamer Street, Glebe, Sydney,
and SOLD BY THE SOCIETY. 1963.
CONTENTS.
CONTENTS" OF “PROCEEDINGS, 1962
PART 1 (No. 398). (Issued 30th July, 1962.)
Pages
Presidential Address, Bighty-seventh Annual General Meeting, 28th March, 1962
(the President, Professor J. M. Vincent, absent overseas) :
Summary of Year’s Activities Nol Shae Pens eee aarp eae nor geste we ley Australian Studies of the Root-nodule Bacteria. A Review .. .. .. 8-88 Elections .. Big What bia ere ae ate ne ev aia A REN Ol ee GN ba 4 Balance Sheets for the Year ending 28th February, 1962 .. .. .. .. .. .. 5-7
Selecting for Virulence on Wheat while Inbreeding Puccinia graminis var. secalis. By I. A. Watson and N. H. Luig. (Plate i.) .. .. .. .. .. .. «. «. 39-44
A Study of some Smuts of Sorghum spp. By R. F. N. Langdon. (One Text-figure.) 45-50
Studies in Australian Loranthaceae. I. Nomenclature and New Additions. By B. A.-Barlow. (One Text-figure))’ J “SSP Rie Sehr. 5. 8. Ue ew) le DIE 6
The Reproduction and Early Life Histories of the Gastropods Bembicium auratum (Quoy and Gaimard) (Fam. Littorinidae), Cellana tramoserica (Sower.) (Fam. Patellidae) and Melanerita melanotragus (Smith) (Fam. Neritidae).
By D. T. Anderson. (Twenty-two Text-figures.) .. .. .. .. .. .. .. 62-68
The Host Plant Relationship of an Australian Swallowtail, Papilio aegeus, and its Significance in the Evolution of Host Plant Selection. By George O. Stride and R. Straatman. (Communicated by Dr. D. F. Waterhouse.) (Four Text- figures. ) PEURR cetos Wahi Se Doin | | el ge ae te emer. 00 ys iets alae fr eee eRe ee ce TS
A Revised Classification of the Australian Amphiuridae (Ophiuroidea). My H. Barraclough Fell. (Communicated by Elezabeth O. Pope.) .. .. .. .. .. 79-838
Galls of Agromyzidae (Dipt.) on Pittosporum undulatum Andr. By HBrich M. Hering. (Communicated by C. E. Chadwick.) (Plate ii; four Text-figures. ) 84-91
A New Species of Trigonalid Wasp parasitic on the Sawfly Perga affinis Kirby (Hymenoptera). By E. F. Riek. (Five Text-figures.).. .. .. .. .. «.. 92-95
A New Genus of Australian Stoneflies (Plecoptera, Gripopterygidae). By EH. F. Riek. (Six Text-figures. ) .. 96-98
CHG of
CONTENTS.
PART 2 (No. 399). (Issued 10th January, 1963.)
Asexual Intercrosses between Somatic Recombinants of Puccinia graminis. By I. A. Watson and N. H. Luig ..
The Genus Walchiella (Acarina, Trombiculidae). By Robert Domrow. (Sixty- three Text-figures. )
Observations on some Australian Forest Insects. 12. The Taxonomy of Zenarge turneri Rohwer (1918) (Hymenoptera: Argidae), the Cypress Pine Sawfly. By K. M. Moore. (Six Text-figures.)
Observations on some Australian: (Forest Insects. 13. A Comparison of the Biology of the Cypress Pine‘Sawfly Subspecies. By K. M. Moore. (One Text-figure. ) SCTE PN Eee). RES daca Cheap PS Gey Bloat dees Po oe
Hyla phyllochrous Gunther (Amphibia) as an Addition to the Fauna of Victoria, with the Description of a New Race and a Note on the Name of the Genus. By Stephen J. Copland. (One Map.)
Studies on the Inheritance of Rust Resistance in Oats. 1. Inheritance of Stem Rust Resistance in Crosses involving the Varieties Burke, Laggan, White Tartar and Anthony. By Y. M. Upadhyaya and EH. P. Baker ..
A Trigonalid Wasp (Hymenoptera, Trigonalidae) from an Anthelid Cocoon (Lepidoptera, Anthelidae). By E. F. Riek. (Five Text-figures.)
A New Encyrtid Genus Parasitic on Bug Eggs. By EH. F. Riek. (Nine Text- ATR Sep) Ramer er ee PRE Mess caer 5 Cece een ga pee rn ea te ed ew
Zine Deficiency on the Darling Downs, Queensland. By B. R. Hewitt Notes on Plant Parasitic Fungi. I. By J. Walker. (Plate iii.)
The Actual Identity of Captain Cook’s Kangaroo. By Tom Iredale and Ellis Troughton. (Two Text-figures.)
The Development of the Polychaete Galeolaria caespitosa Lamarck (Fam. Serpulidae). By J. C. Andrews and D. T. Anderson. (Seven Text-figures.)
A New Species of Hchthroplexis, an Encyrtid Hyperparasite of Lerp-forming Psyllids on Eucalypts (Hymenoptera, Chalcidoidea). By E. F. Riek. (Seven Text-figures. )
Australian Liverworts. I. Haplomitrium intermedium, sp. nov. (Calobryales). By Geoffrey K. Berrie. (Two Text-figures. )
Some Insects and Terrestrial Arthropods from Heron Island, Queensland. By C. E. Chadwick
Studies on the Inheritance of Rust Resistance in Oats. II. The Mode of Inheritance of Crown Rust Resistance in the Varieties Landhafer, Santa Fe, Mutica Ukraine, Trispernia and Victoria in their Crosses with Susceptible Varieties. By Y. M. Upadhyaya and E. P. Baker
Alan Neville Colefax, 1908-1961. (Memorial Series, No. 20.) (With Portrait, Plate iv.)
Revision of the Thynnidae. Part V. A Contribution towards a Knowledge of the Thynnidae of the Philippines, Indonesia, New Guinea, The Solomons, New Caledonia and Lord Howe Island. By K. E. W. Salter. (Plates v-viii; thirty-four Text-figures. )
Pages 99-104
105-115
116-124
125-136
137-140
141-147 148-150
151-155 156-161 162-176
177-184
185-188
189-190 191-195
196-199
200-219
220-222
223-266
CONTENTS.
PART 3 (No. 400). (Issued 8th April, 1963.)
Sir William Macleay Memorial Lecture, 1962. Living Membranes—Frontiers of Research at the Boundaries of Life. By R. N. Robertson. (Plate ix; two Text-figures. ) ; ; :
Bat Ticks of the Genus Argas Gadasiies ana eee 5. Suece eae of Larvae
from Australian and New Guinea Carios-group Populations. By Harry Hoogstraal and Glen M. Kohls. (Communicated by Dr. Bruce McMillan.) (Twelve Text-figures. )
A New Genus of Gall-forming Biabhyscenaipnantie Pteromalidae Gatndasntera: Chalcidoidea) from Western Australia. By E. F. Riek. (Five Text-figures. )
A New Encyrtid (Hymenoptera, Chalcidoidea) Genus of Parasites of Lerp- forming Psyllids on Hucalyptus. By H. F. Riek. (Seven Text-figures.)
Gynodioecism in Leucopogon melaleucoides A. Cunn. By Alison McCusker ..
Amendments to the Disposal of Type Specimens of Species of Culex (Lophoceraomyia) from New Guinea. By Donald H. Colless
Notes on Australian Mosquitoes (Diptera, Culicidae). vi. Five New Victorian Species and a Description of the Larva of Aédes milsoni (Taylor). By N. V. Dobrotworsky. (Five Text-figures.) we a Feb Ee eceetat
New Species of Ohakunea Edwards and a Related New Genus with Notes on the Relationships of Heterotricha Loew. (Diptera). By Donald H. Colless. (Two Text-figures. ) SEEM ree ai A np ene ee re earn eem ne,
Notes on Australasian Tanyderidae, with Description of a New Species of Radinoderus Handl. (Diptera). By Donald H. Colless. (One Text-figure.)
Notes on the Taxonomy of the Aédes scutellaris Group, and New Records of A. paullusi and A. albopictus es Culicidae). By Donald H. Colless. (One Text-figure. ) petals :
The Biology of Roeselia lugens (walte.), the Gum-leaf Skeletonizer Moth, with Particular Reference to the Hucalyptus camaldulensis Dehn. (River Red Gum) Forests of the Murray Valley Region. By K.-G. Campbell. (Plate x; seven Text-figures. ) OE TOA NET SEP REE Ce seed OS aay Mt
Australasian Ceratopogonidae (Diptera, Nematocera). Part ix. The Genus Macrurohelea. By David J. Lee. (Five Text-figures.)
On a Collection of Plants of Permian Age from Baralaba, Queensland. By J. F. Rigby. (Plates xi-xii.) Maree oak ska! ud) aes treme meats
Australasian Ceratopogonidae (Diptera, Nematocera). Part x. Additional Australian Species of Culicoides. By David J. Lee and Eric J. Reye. (Plate xiii; 55 Text-figures.) A ORE Ts hace eae ene Le
“Sandflies’”’ as Possible Vectors of Disease in Domesticated Animals in Australia. By D. J. Lee, E. J. Reye and A. L. Dyce .. PER dk Sestate sc
The Influence of the Tide Cycle on Certain Species of Culicoides (Diptera, Ceratopogonidae). By Eric J. Reye and David J. Lee. (Ten Text-figures.)
Chromosome Races in Goodenia bellidifolia Sm. By W. J. Peacock, Linnean Macleay Fellow in Botany. (Plates xiv-xv; two Text-figures. )
Structural Geology of Part of the Tamworth Trough. By Keith A. W. Crook (Plate xvi; twelve Text-figures. )
Abstract of Proceedings
List of Members
List of Plates crass ~ fete OOO) ee ee
List of New Genera, New Species and New Subspecies
Index
Pages
. 267-274
. 275-280
281-282
. 283-285
286-289
290
. 291-302
. 303-308
309-311
312-315
. 316-338
339-340
. 341-351
. 352-863
. 364-376
377-387
. 388-396
.. 397-409 .. 410-418 . 419-425
426 426
. 427-431
ANNUAL GHNERAL MEETING. 28TH MaArcH, 1962.
The Wighty-Seventh Annual General Meeting was held in the Society’s Rooms, Science House, Sydney, on Wednesday, 28th March, 1962.
Dr. I. V. Newman, Vice-President, occupied the chair.
The minutes of the Highty-Sixth Annual General Meeting, 29th March, 1961, were read and confirmed.
The Honorary Treasurer (Dr. A. B. Walkom) presented the balance sheets for the year ending 28th February, 1962, duly signed by the Auditor, Mr. 8. J. Rayment, F.C.A., and his motion that they be received and adopted was carried unanimously.
In the absence overseas of the President (Professor J. M. Vincent) the Honorary Secretary (Dr. W. R. Browne) read the first portion of the Presidential Address as follows:
I wish to thank the Society for the privilege of having filled the position ‘of President Guring the past year, and the Council for its support in conducting the Society’s affairs. Particularly once again, there must be recorded our deep gratitude to Dr. W. R. Browne and Dr. A. B. Walkom for their continued services as Honorary Secretary and Honorary Treasurer-HMditor. I should also like to express appreciation to Miss G. L. Allpress, our Assistant Secretary. Together with all members of Council, I am conscious of how much we owe to the efficient management of the regular affairs of the Society and the maintenance of its library.
REPORT ON THE AFFAIRS OF THE SOCIETY FOR THE YEAR.
The Society’s Proceedings for 1961, Vol. 86, Parts 1 and 2, were published in 1961 and Part 3 in February, 1962. Volume 86 consists of 322 pages, 11 plates and 215 text- figures. An increase in the charges for printing the Proceedings commencing with Vol. 87 (1962), Part 1, was made in November, 1961. Revised Library Regulations were adopted by the Council on 22nd March, 1961.
During the year twelve new members were added to the list, two members died, five members resigned and two were removed from the list of members. The numerical strength of the Society at 1st. March, 1962, was: Ordinary members, 241; Life Members, 31; Corresponding Member, 1; total, 2738.
Lecturettes were given at the following meetings: April, Collecting Insects in Tasmania, by Mr. D. K. McAlpine; June, Pollen and its Chronological Significance, by Dr. A. R. H. Martin; July, The New Zealand Glow-worm, by Dr. Aola Richards; September, Geographical Patterns of Chromosome Change in the Australian Flora, by Dr. S. Smith-White; October, The Argentine Ant Eradication Campaign in Sydney, by Mr. Gordon Pasfield; November, The Impact of New Techniques on Taxonomy, by Mr. R. C. Carolin. The discussions which followed the lecturettes gave an added interest to the proceedings of the meetings. Our thanks and appreciation are expressed to the lecturers. Owing to the holding of the A.N.Z.A.A.S. meeting in Brisbane from 29th May to 2nd June, 1961, no Ordinary Monthly Meeting of the Society was held in May.
Library accessions from scientific institutions and societies on the exchange list amounted to 1,962, compared with 1,912 in the previous year. Members and institutions continued to borrow books and periodicals from the library as previously, and many requests for microfilm and photo-printing copies of old and rare articles were granted by sending the required publication on loan to the Fisher Library, University of Sydney, for that purpose. The following new exchange relations were entered into by the Society: Academie des Sciences de Bulgarie, Sofia, Bulgaria; Botanical Institute, University of Zagreb, Zagreb, Yugoslavia; Instituto de Zoologia (Museu Bocage), Lisbon. Portugal. Council decided, for various reasons, to discontinue exchange of publications
PROCEEDINGS OF THE LINNEAN Society oF NEW SOUTH WALES, 1962, Vol. Ixxxvii, Part 1.
A
2 PRESIDENTIAL ADDRESS.
with the following: University of Pentisylvania, Philadelphia, U.S.A.; Indian Agri- cultural Research Institute, New Delhi, India; U.S. Department of Agriculture, Washington, U.S.A.; Institut Pasteur, Tunis, Tunisia, and New Zealand Oceanographic Institute, Wellington, New Zealand. A number of members took advantage of the Council’s offer of the Macleay Memorial Volume and Jubilee Brochuré,
Mr. G. P. Whitley, F.R.Z.S., and Miss Elizabeth C. Pope, M.Sc., C.M.Z.S., were elected members of Council in December, 1961, in place of Dr. J. W. Evans, who had resigned, and the late Mr. A. N. Colefax.
Mr. S. J. Copland and Dr. I. V. Newman, who were appointed the Society’s delegates to the Conservation Conference held on 18th November, 1961, presented a report to the Council.
Dr. I. V. Newman represented the Society at an Extraordinary General Meeting of the National Parks Association of N.S.W. (Central Region) on 28th November, 1961, to consider a merger between the Association and the Caloola Club.
The total net return from the Society’s one-third ownership of Science House for the year ended 31st August, 1961, was £1,274/18/10.
Linnean Macleay Fellowships.
Mr. W. J. Peacock, B.Sc., was reappointed to a Linnean Macleay Fellowship in Botany tenable for one year from 1st January, 1962. A summary of Mr. Peacock’s research during the past year is as follows:
During the year research was carried out in two main fields, one being cyto- evolutionary studies in the Goodeniaceae, the other concerning problems of chromosome structure. A polyploid series has been found in Goodenia bellidifolia, diploid, tetraploid, hexaploid and octoploid forms occurring. The fact that the ehromosome races have sharply defined, non-overlapping, but adjoining areas of distribution, indicates that the different forms have strict eco-physiological tolerances. When this is considered with the actual distribution pattern, a reasonable inference can be made that polyploidy has been of prime importance in migration of the species into the new habitats provided by the elevation of the Great Dividing Range in the late Pliocene. Another example of comparatively recent evolutionary change is found in Dampiera stricta which aiso contains intraspecific polyploids, ranging from diploids to decaploids. Of particular interest is the relationship between the diploid and tetraploid forms in the central and southern coastal regions of New South Wales. In contrast to the geographically separated occurrences of chromosome races in G. bellidifolia, the diploid and tetraploid of D. stricta occur in the one area in a complex mosaic. The tetraploid appears to be replacing the diploid which has a disjunct relic distribution. Brunonia australis presents two pan-Australian patterns of change, one in level of polyploidy, the other in the frequency of structural hybrids. That migration has been from west to east is indicated both by the occurrence of tetraploid and octoploid forms in the south-east of the continent and by the high frequency of interchanges in eastern populations. An interesting point is that there is strong evidence that Brunonia has entered Tasmania in two separate migrations, one probably being as recent as the Pleistocene. A number of other species in the Goodeniaceae show geographical patterns and are being investigated. Research has also been carried out on certain problems of chromosome structure. An analysis using X-ray breakage techniques has demonstrated that Brunonia australis and Vicia faba have subchromatids which have both structural and functional significance. The analysis has been carried another stage further in Brunonia in which a quadripartite structure of the chromatid has been shown. Vicia was chosen in the above experiments because recent studies on chromosome duplication using autoradio- graphic techniques indicated that the chromatid of Vicia faba was of a unineme structure. Since the results of the X-ray and autoradiographic analyses were not in agreement, the autoradiographic experiments (Taylor, Wood and Hughes) were repeatad. The results obtained did not parallel those obtained by Taylor et al. (other workers have also failed to repeat these experiments); however, the new results were compatible with the X-ray experiments. Further experiments are in progress.
PRESIDENTIAL ADDRESS. 3
Linnean Macleay Lectureship in Microbiology.
Dr. Y. T. Tchan, Linnean Macleay Lecturer in Microbiology, University of Sydney, submitted the following report on his work for the year ended 3lst December, 1961: “Since my return from study leave my activity has been mainly concerned with teaching. However, some research has been carried out during this period. The presence of Azotobacter in the Rhizospheres of wheat has been studied. Attempts to establish Azotobacter in the vicinity of the root system of Australian varieties of wheat have been unsuccessful, but some suggestions of stimulation could be noticed. This requires further study. Further work has been done to correlate the algal and field trial methods of stimulating the mineral requirement of soil. This work will probably be completed during 1962. Study of the Gram stain by Electron-Microscopy has been handicapped by the difficulty of marking ultra-thin sections. New embedding methods are under investigation, to overcome the difficulties.”
Obituaries.
It is with regret that the following deaths during the year are recorded:
Michel Francois (Michael Frank) Albert died at his home, “Boomerang”, Elizabeth Bay, Sydney, on 19th January, 1962, at the age of 87. Mr. Albert was educated at Fort Street High School and the University of Sydney. He entered the music publishing company of J. Albert & Son, Pty., Ltd., established by his father. In association with two other partners he formed the Australian Broadcasting Company, which established the Australian National Broadcasting Service, later taken over by the Australian Broad- casting Commission. At the time of his death Mr. Albert was chairman of the Common- wealth Broadcasting Corporation with stations in Sydney (2UW), Brisbane (4BC) and a country district of Queensland. He was a founder-director of the Australian Performing Rights Association. During his lifetime Mr. Albert made substantial gifts to many institutions, including the University of Sydney, where he helped to form the Chair of Music, St. Paul’s College and the Red Cross. He was a fellow of the Royal Geographical Society and a founder-director of the Royal Motor Yacht Club of N.S.W. He was aiso a founder-member of the Royal Automobile Club and a member of the University Ctub and the Royal Sydney Yacht Squadron. Mr. Albert joined this Society in 1927, taking up Life Membership on his election.
Allen Neville Colefax, B.Sc., died in Sydney on 7th December, 1961, at the age of 53. He was Senior Lecturer in Zoology at the University of Sydney, for fourteen years, regularly broadcast popular science talks in the A.B.C. Children’s Hour, and was known to thousands of radio listeners as ‘““‘Tom the Naturalist’. At the Council meeting of the Society on 13th December, 1961, the following motion was carried, members standing in silence: “That this Council desires to place on record its great sorrow and sense of loss in the death of Allen Neville Colefax. Mr. Colefax was elected a member of the Society in 1931, was President in 1951 and was a member of Council from 1943 till the time of his death. He served the Society wisely and well, and endeared himself to his colleagues on the Council by his modesty, sincerity and unaffected kindliness. The members of Council wish to express their deep sympathy with his wife and family.” Mr. Colefax contributed four papers to the Proceedings, also two with Professor W. J. Dakin and one with Mr. G. P. Whitley.
PRESIDENTIAL ADDRESS.
Australian Studies of the Root-nodule Bacteria. A Review. (For full text see pages 8-38.)
This review deals with Australian work with the root-nodule bacteria since the pioneering efforts of the Society’s first Bacteriologist, Dr. R. Greig-Smith. A short historical account tracing the establishment of research groups interested in the subject between then and the present day is followed by a more detailed treatment under tne headings: the bacterium, interaction between bacterium and host, and application of
4 PRESIDENTIAL ADDRESS.
knowledge. Studies of the bacterium itself deal with cytology and composition (which have been relatively neglected in the past), antigenic composition of the rhizobia, factors affecting their growth, antagonistic and mutually beneficial effects involving rhizobia themselves and in relation to other organisms, bacteriophage, variation (developed particularly in connection with a relatively well studied set of substrains). and taxonomy (in which fresh interest is developing). The interaction between bacterium and its host is treated first in relation to general considerations of the symbiosis; then in more detail in respect of the morphology and metabolism of the nodule, specificity in relation to invasion and fixation and the influence exercised by the environment. So far as it is possible to treat it separately, this knowledge is applied to a consideration of the need for seed inoculation in Australia, the supply of seed inoculants and requirements for their successful use.
No nominations of other candidates having been received, the Chairman declared the following elections for the ensuing year to be duly made: President: Professor B. J. F. Ralph, B.Se., Ph.D., A.A.C.1.
Members of Council: R. H. Anderson, B.Sc.Agr.; Elizabeth C. Pope, M.Sc., C.M.Z.S.; EH. Le G. Troughton, C.M.Z.S., F.R.Z.S.; T. G. Vallance, B.Se., Ph.D.; J. M. Vincent, D.Sc.Agr., Dip.Bact.; and G. P. Whitley, F.R.Z.S.
Auditor: S. J. Rayment, F.C.A. Following the declaration the Chairman installed Professor Ralph as President.
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PRESIDENTIAL ADDRESS. AUSTRALIAN STUDIES OF THE ROOT-NODULE BACTERIA. A REVIEW. By J. M. VINCENT. [28th March, 1962.]
Synopsis.
This review deals with Australian work with the root-nodule bacteria since the pioneering efforts of the Society’s first Bacteriologist, Dr. R. Greig-Smith. A short historical account tracing the establishment of research groups interested in the subject between then and the present day is followed by a more detailed treatment under the headings: the bacterin, interaction between bacterium and host, and application of knowledge.
Studies of the bacterium itself deal with cytology and composition (which have been relatively neglected in the past), antigenic composition of the rhizobia, factors affecting their growth, antagonistic and mutually beneficial effects involving rhizobia themselves and in relation to other organisms, bacteriophage, variation (developed particularly in connection witt. a relatively well studied set of substrains), and taxonomy (in which fresh interest is developing).
The interaction between bacterium and its host is treated first in relation to generai considerations of the symbiosis; then in more detail in respect of the morphology and metabolism of the nodule, specificity in relation to invasion and fixation and the influence exercised by the environment.
So far as it is possible to treat it separately, this knowledge is applied to a consideration of the need for seed inoculation in Australia, the supply of seed inoculants and requirements for their successful use.
INTRODUCTION.
It is now more than 60 years since this Society’s first Macleay Bacteriologist, Dr. R. Greig-Smith, published on the root-nodule bacteria (Greig-Smith, 1899, 1901, 1906a-c, 1907 and 1911). Whilst it is true that, in the absence of plant tests, some of Greig- Smith’s cultures could be of doubtful validity, and that most of his generalizations (Greig-Smith, 1906c and 1907) as to a relationship between gum production and nitrogen fixation can hardly be sustained today, the fact remains that he pioneered this field in Australia when knowledge of the organisms anywhere was in its beginnings.
Otherwise Australian work during the first 30 years of the century rested largely with State Departments of Agriculture and led to the issuing of cultures to farmers (New South Wales, 1914; Western Australia, 1926; and most other States by 1938).
Strong, while attached to the Council for Scientific and Industrial Research Division of Soils in Adelaide, which was at that time responsible for the issue of cultures in South Australia, published his important paper on specificity between Rhizobium and clover host (Strong, 1937). Since then the root-nodule bacteria have remained one of the interests of the C.S.I.R.O. Division of Soils.
Work at Sydney University was commenced independently by Jensen and myself at about the same time (1939). Jensen, again as the Society’s Bacteriologist, published a valuable series of papers on the root-nodule bacteria between 1941 and 1948. My own work is continuing and has had the cooperation of many students and post-graduate colleagues. Pate, during the time he was on the staff at the Botany School, continued in Sydney the work he had commenced in Northern Ireland and to which he has since returned.
The active interest of the C.S.I.R.O. Division of Plant Industry in Canberra goes back, on the one hand, to 1942 with the nutritional work initiated by Anderson, and, on the other, to the establishment by Nutman in 1953 of the group now headed by Bergersen.
State Departments of Agriculture, notably New South Wales, Western Australia, Tasmania and Queensland, have continued their earlier interest. In Queensland the particular problems of the tropical legume were tackled first by McKnight (1949) in the Department and with great vigour and initiative by Norris (1956) of the C.S.1.R.O.
PROCEEDINGS OF THE LINNEAN Society oF NEw SouTtH WALES, 1962, Vol. lxxxvii, Part 1.
BY J. M. VINCENT. 9
Most recently the Department of Agriculture in Papua—New Guinea has become actively interested in rhizobial problems.
The growth of interest and increase in the amount of Australian work is indicated by the dates of the papers on which this review is based. More than four times as many papers were published between the years 1941 and 1950 as in the previous ten years, and between 1951 and 1960 the number had again more than doubled.
It has been my pleasure to attempt this collation and presentation of what has become a considerable body of significant work.
I. THE BACTERIUM. 1. Cytology and Composition.
A start has recently been made in applying newer techniques of electron microscopy to the study of the fine structure of Rhizobium trifolii (Vincent, Humphrey and North, in preparation). In this way a classical double-layer cell wall and a double-layer cyto- plasmic membrane of similar dimensions have been demonstrated. A large part of the area inside the cytoplasmic membrane is occupied by granules about 50-80 mw in diameter. These have been demonstrated in thin sections and are liberated by rupturing the cell mechanically.
The well-known unevenness of staining of Rhizobium appears to be due to one to several large spherical aggregations of polymeric $-hydroxybutyrate that can amount to 40-50% of the cell dry weight. These bodies can be readily demonstrated with Sudan black and do not take up ordinary basic dyes. Extraction with chloroform removes the highly refractile polymer, and hence the strikingly granular appearance under the phase microscope, and leaves the corresponding parts of the cell weakly stainable with basophilic dyes. This suggests that the polymer is impregnated on a _ basophiiic (? protein-containing) material rather than simply displacing the cytoplasm. To some extent the abundance of polymer reflects the age of the cells; even more the abundance of carbon-containing nutrient.
The composition of the water-soluble extra-cellular gums has been studied for 17 strains, representing 4 species. The most striking result (Humphrey and Vincent, 1959) was that all of the 7 strains of Rh. meliloti produced polysaccharides that con- tained glucose but lacked glucuronic acid, whereas the latter was to be found in all of the remaining 10 strains that represented Rh. trifolii, Rh. leguminosarum and Ih. phaseoli. These ten could be grouped according to the proportion of glucuronic acid, but these groupings bore no relationship to species, as defined by invasive ability. Nor was there any apparent relationship between the nature of the gum and the somatic agglutinogens of the bacterial strains. In the course of this work, Humphrey (1959) also found that six out of seven of the strains of Rh. trifolii contained 4-O-methyl glucuronic acid. The seven strains of Rh. meliloti and the one of Rh. phaseoli lacked this component which was, however, found in one of the two strains of Rh. legumino- sarum. This appears to be the first report of this compound being found associated with a bacterium, although it is common enough in plant hemicelluloses.
From a systematics point of view it is relevant to note that gums of Azotobacter and Beijerinckia, though not studied in the same detail (Humphrey, unpublished), were certainly different from those of Rhizobium. Information as to the nature of the gum produced by Agrobacterium and rhizobia of the cowpea miscellany (if they can be induced to produce sufficient gum for study) would be of interest in relation to the taxonomy of the root-nodule bacteria generally.
Cell wall preparations of Rh. trifolii contained the rigid (“R’) layer components: glucosamine, muramic acid, glutamic acid alanine, and diaminopimelic acid, and the wide range of amino-acids generally found in the walls of Gram-negative bacteria (Humphrey and Vincent, in preparation). In this case such amino-acids comprised: lysine, aspartic acid, glycine, serine, valine, methionine, leucine and tryptophane. Glucose (or galactose) and rhamnose were also present. Humphrey and Vincent, seeking an explanation of the peculiar swollen shape of calcium-deficient cells (Vincent and Colburn, 1961), the walls of which still contain the “R” layer components, suggested
10 AUSTRALIAN STUDIES OF THE ROOT-NODULE BACTERIA,
that this element might provide rigidity by stabilizing areas of excess negative charge in the “R” layer components. It has recently been shown by the author that strontium (not barium) can replace calcium, though less efficiently, in providing normal rod- shaped cells.
2. Antigenic Constitution.
Earlier work from the author’s laboratory was largely concerned with the more detailed analysis of the antigenic composition of Rh. meliloti and Rh. trifolii, by separating the flagellar and somatic reactions, and using cross absorption tests (Vincent, 1941 and 1942; Hughes and Vincent, 1942; Purchase and Vincent, 1949; Purchase, Vincent and Ward, 195la; Vincent, 1953). This work has been dealt with in a con- solidated account on a previous occasion (Vincent, 1954a), which also discusses the several applications the method has been put to in the author’s laboratory.
Practically all the work done with Rhizobium has involved the agglutination reaction which remains the easiest technique to apply. Neither complement fixation nor precipitation has been used at all widely. Unpublished results recently obtained in this laboratory by Mrs. J. Dorman have shown that, with the exception of some non- reciprocal reactions to low titre, complement-fixation tests amongst rhizobia of clover, pea, lucerne, bean, cowpea, peanut, soybean, lupin and lotus have followed closely the situation as revealed by agglutination. Reciprocal cross reactions in both agglutination and complement fixation were found between clover and pea, cowpea and peanut, lupin and lotus. Otherwise no cross reaction was found.
Bloomfield (1959) has extended the author’s preliminary serological study of variants of Rh. trifolii, SU297 and SU298. His findings will be discussed under Variation.
3. Growth.
(a) Hydrogen-ion Concentration.
Jensen (1942a) has provided valuable information about the influence of pH on the growth of Rh. trifolii and Rh. meliloti and noted the greater acid tolerance of the former. He also found that Rh. trifolii had difficulty in initiating growth at a faintly alkaline reaction when the medium was well buffered. The critical acid reaction for the growth of Rh. trifolii was in the vicinity of pH 5-2, although one out of 24 strains was able to grow at pH 4:9 in buffered soil extract. The corresponding critical pH for Rh. meliloti was 5:4. These observations have fitted in very well with field experience, both so far as the natural occurrence of these two species of Rhizobium is concerned, and in relation to the degree of difficulty in establishing the Rh. trifolii in acid soil (Vincent and Waters, 1954). This species failed to grow at pH 4-8-5-0 and grew optimally in the vicinity of pH 7, there being some degree of inhibition under more alkaline conditions (comp. Jensen, above).
(b) Requirement of Divalent Cations.
The use of CaCO, or Ca(OH)., especially in experiments with soil, has caused some confusion as to whether the benefit was due to its effect on pH, or due to calcium as such. Norris has indeed been led to conclude that Rhizobium does not need calcium (Norris, 1958a@ and 1959a@). This is, however, contrary to our experience with Fh. trifolii and Rh. meliloti (Mullens and Vincent, unpublished) and that of Bergersen (1961c), the last with two strains of “cowpea” rhizobia as well. It is apparent that Norris’s failure to detect a calcium effect was due to restrictions imposed in his study by the desire to check a large collection of strains, necessarily in less detail. There is good reason to believe that the conditions Norris provided for growth were sub-optimal and that this, combined with the generally semi-quantitative nature cf his assessments, prevented a more positive result. The author’s detailed study of the response of total growth to concentration of calcium (Vincent, 1962) shows clearly why the undoubted stimulatory effects of low concentrations of this element are difficult to define. For example, the 0:016 mM calcium in Loneragan and Dowling’s basal medium (Loneragan and Dowling, 1958) is close to the amount this writer found sufficient