518 Transactions British Mycological Societythat he described the species. It was presumably published later than1900, as otherwise one would have expected Gran (1900) to mention it inhis comments on O. lauderiae.

Meunier provided no written description, but published a number ofdrawings of the fungus on different hosts, showing the exit tube, but notthe flagellation of the zoospores, so that the exact identity of the speciesremains in doubt. Petersen (1905) referred it to Eurychasma Magn. andScherffel (1925) to Ectrogella Zopf.

Nomenclaturally Meunier was in error in providing a superfluous nomennooum for the species, so the name Olpidium phycophagum should be placed insynonomy with O. lauderiae, whatever that organism turns out to be.

I am grateful to Mr R. Ross, Keeper of Botany, British Museum (Nat.Hist.), for the provision of Xerox copies of the relevant parts of Gran'sand Meunier's papers, and to Professor T. Braarud of Oslo for searchingthrough Gran's publications for O. dityli.

REFERENCES

CUPP, E. E. (1943). Marine plankton diatoms of the west coast of North America.Bulletin. Scripps Institution ofOceanography. Technical Series 5, no. I.

GRAN, H. H. (1900). Bemerkungen tiber einiger Plankton-diatomeen. Nyt Magazin forNaturuidenskabeme 38, 105-128.

JOHNSON, T. W. & SPARROW, F. K. (1961). Fungi in oceans and estuaries. Weinheim:J. Cramer.

MEUNIER, A. (1910). Microplankton des Mers de Barents et de Kara, Due d'Orleans,Campagne Arctique de 1907. Brussels.

PETERSEN, H. E. (1905). Contributions a la connaissance des Phycomycetes marins(Chytridinae Fischer). Oversigt K. Danske Videnskabemes Selskabs Forhandlinger, pp.439-488.

SCHERFFEL, A. (1925). Endophytische Phycomyceten-Parasiten der BaciIIariaceen undeinige neue Monadinen. Ein Beitrag zur Phylogenie der Ooomyceten (Schroter),Archivfiir Protistenkunde 51, 1-141.

RESERVES IN SPORES OF AQUATIC HYPHOMYCETES

N. SINGH*

Department of Biological Sciences, University of Exeter

Although chemical makeup of spores of a few fungi has been partiallyelucidated (Foster, 1949; Shu, Tanner & Ledingham, 1954; Owens,Novotny & Michels, 1958; Lingappa & Sussman, 1959; Tokunaga &Bartnicki-Garcia, 1971; Mills & Cantino, 1974), nothing has been reportedpreviously in this regard about the spores of aquatic hyphomycetes. Inthe present contribution results of an investigation of the chemicalcomposition of spores of two aquatic hyphomycetes are reported.

* Present address: Fourah Bay College, University of Sierra Leone, Freetown,Sierra Leone.

Trans. Br. mycol. Soc. 64 (3), (1975). Printed in Great Britain

Notes and Brief Articles

Table I. Composition of spores ofLemonniera aquaticaand Articulospora tetracladia

Percentage of dry weightA

L. aquatica A, tetracladia

Glucose 0'75 traceReducing sugars 1'1 0'94Trehalose 2'7 5'1Unaccounted for· 3'27 6'4 2

Total alcohol solublecarbohydrates 7,82 12'46

Glycogen 1'17 1'9Total acid-soluble

carbohydrates 60'34 62'67Total lipids 23'47 15'28

• By difference,

Lemonniera aquatica de Wildman and Articulospora tetracladia Ingold wereused. For the production of spores, cultures on 2 %malt-extract agar atleast 10 days old were cut into disks with a sterile I em cork borer and20 disks put in a sterile screw-top filter bottle containing 200 ml steriledistilled water. There were ten replicates. The bottles were aerated at15° by means of compressed air forced through water by hypodermicneedles (Webster & Towfik, 1972). The air was filtered through sterilecotton wool. After 24 h bubbling, water was replaced with fresh steriledistilled water. After a further 4-day period spore suspensions were pooledand filtered through four layers of muslin to separate mycelial fragmentsfrom spores. The clean spore suspension was then filtered through an81£m Millipore filter. The spores were thoroughly washed with distilledwater and then scraped off the filter.

Lipid extraction was carried out according to the method of Lingappa& Sussman (1959) and extracted lipids were chromatographed accordingto the procedure of Owens et al. (1958). For carbohydrate extraction themethod of Lingappa & Sussman (1959) was followed with slight modi­fication. Spores were extracted with boiling 80 % (v/v) ethanol. Totalcarbohydrates were determined by the anthrone method of Loewus (1952).Reducing sugars were determined by the method of Nelson (1944) usingthe modified reagent of Somogyi (1952) and glucose by the use of glucose­oxidase (Boehringer Mannheim, GMBH, Germany). Trehalose andglycogen were estimated by the methods ofNg, Smith & McIntosh (1974).

The results are presented in Table I. The amount of alcohol solublecarbohydrates varied considerably in both species though yields of acid­soluble carbohydrates were similar in both cases. The presence of aminoacids in the extracts may have contributed to the unaccounted for por­tions of the alcohol soluble carbohydrates. As compared with Neurosporatetrasperma Shear & B. O. Dodge (Lingappa & Sussman, 1959) the sporesof these fungi contain a much higher proportion of alcohol-insoluble

Trans. Br. mycol. Soc, 64 (3), (1975)' Printedin Great Britain

520 Transactions British Mycological Societycarbohydrates. Trehalose formed 35 %of the total alcohol soluble carbo­hydrates in both species. Total lipid yields were also different in bothspecies. In both cases, during the elution of columns, highest peakscorresponding to the carotenoid fractions were obtained on the recorder.Further, distinct orange-yellow bands were seen moving down the columnswhen eluants for carotenoids were added to the columns. The effluentswere also deep yellow in colour. It seems therefore that a large proportionof the lipid content in these species is made up of carotenoids.

Trehalose occurs widely in fungi and it is considered to be an endogenoussource of energy (Mandels, Vitols & Parrish, 1965; Hanks & Sussman,1969; Sussman & Lingappa, 1959; Horikoshi & Ikeda, 1966, cited by Nget al, 1974). For the germination of spores of these two fungi the stimulusof contact with a surface is important and 50 %germination can be ob­tained after 4 h of contact with a surface. However, spores kept suspendedin distilled water by bubbling air through the suspension for a similarperiod do not show germination. A supply of exogenous nutrients such asglucose (0'1 %) does not enhance the rate of germination (Singh &Webster, unpublished). Trehalose and glycogen therefore may be im­portant sources of energy for these spores. I t will be interesting to investigatethe fate of these reserves during the germination of spores in these species.Further work is continuing along these lines.

I wish to express my thanks to Professor John Webtser for allowing methe use oflaboratory facilities and for his most helpful suggestions. Thanksare also due to MrJohn Havell for his technical advice on various matters.

REFERENCES

FOSTER,]. W. (1949). Chemical activities offungi. New York: Academic Press.HANKS, D. L. & SUSSMAN, A. S. (1969)' The relation between growth, conidiation and

trehalase activity in Neurospora crassa, American Journal ofBotany56, 1152-1159.HORIKOSHI, K. & IKEDA, Y. (1966). Trehalose in conidia of Aspergillus oryzae. Journal of

Bacteriology gI, 1883-1887.LINGAPPA, B. T. & SUSSMAN, A. S. (1959). Endogenous substrates of dormant, activated

and germinating ascospores of Neurospora tetrasperma. Plant Physiology 34,466-472.LOEWUS, F. (1952). Improvement in anthrone method for determination of carbo­

hydrates. Analytical Chemistry 24,219.MANDELS, G. R., VITOLS, R. & PARRISH, F. W. (1965). Trehalose as an endogenous

reserve in spores of the fungus Myrothecium oerrucaria. Journal of Bacteriology go,1589-1598.

MILLS, G. L. & CANTINO, E. C. (1974). Lipid composition of the zoospores of Blasto­cladiella emersonii. Journal of Bacteriology uS, 192-201.

NELSON, N. (1944). A photometric adaptation of the Somogyi method for the deter­mination of glucose. Journal ofBiological Chemistry I53, 375-380.

NG, A. M. L., SMITH,]. E. & MciNTOSH, A. F. (1974). Changes in carbohydrate com­position and trehalase activity during conidiation of Aspergillus niger in continuousand batch culture. Transactions of the British Mycological Society 63, 57-66.

OWENS, R. G., NOVOTNY, H. M. & MICHELS, M. (1958). Oomposition of conidia ofNeurospora sitophila. Contributions riftheBoyce Thompson Institute Ig, 355-374·

SHU,P., TANNER, K. G. & LEDINGHAM, G. A. (1954). Studies on the respiration ofrestingand germinating uredospores ofwheat stem rust. Canadian JournalofBotany 32, 16-23.

Trans. Br. mycol. Soc. 64 (3), (1975). Printed in Great Britain

Notes and Brief Articles 52 1

SOMOGYI, M. (1952). Notes on sugar determination. Journal if Biological Chemistry 195,19-23·

SUSSMAN, A. S. & LINGAPPA, B. T. (1959)' Role of trehalose in ascospores of Neurosporatetrasperma. Science 130, 1343.

TOKUNAGA, J. & BARTNICKI-GARCIA, S. (1971). Cyst wall formation and endogenouscarbohydrate utilization during synchronous encystment of Phytophthora palmivorazoospores. ArchivjUr Mikrobiologie 88, 119-126.

WEBSTER, J. & TOWFIK, F. H. (1972). Sporulation of aquatic hyphomycetes in relationto aeration. Transactions if the British Mycological Society 59, 353-364.

RESPONSES OF FUNGAL ZOOSPORES TO IONICFORMS OF RESINS

N. S1NGH*

Department ofBiological Sciences, University of Exeter

(With Plate 43)

Responses of zoospores to amino acids and sugars (Royle & Hickman,1964-; Zentmyer, 1966; Rai & Strobel, 1966; Dukes & Apple, 1961;Bimpong & Clerk, 1970) and dilute acids (Katsura & Hosomi, 1963, citedby Ho & Hickman, 1967) are well known. However, it is extremelydifficult to distinguish the effects of whole compounds on zoospores fromthe effects due to ions if the compound ionizes in solution. Ho & Hickman(1967) adopted a new approach to this problem and studied the behaviourof zoospores of Phytophthora megasperma var. sojae Kaufman towards in­dividual ions adsorbed on resin particles. They showed that zoospores of thisfungus tended to accumulate and encyst in a ring a short distance from theHr-resin. Nearly all the cysts germinated. A similar pattern was observedwith OH--resins but germination was largely inhibited. Formate-,acetate-, chloride- and ammonium-resins did not elicit any detectableinfluence on zoospores. Responses of both uniflagellate and biflagellatefungal zoospores to various ionic resins are reported here. Zoospores wereobtained by usual procedures of growing the fungi on various solid mediaat their optimum temperatures and suspending disks cut from the marginof the colony or flooding the whole Petri dish with distilled water orpond water. In the case of Synchytrium taraxaci, de Bary & Woronin theinfected leaves were covered with a thin layer of distilled water and kept atroom temperature. Sporangia of Albugo candida (Hooke.) O. Kunge werescraped off infected leaves and kept suspended in distilled water at 4-0 for4 h to obtain zoospores.

Prepared resin particles were picked up on the tip of an inoculatingneedle and placed in the centre of a cover slip. A drop of zoospore sus­pension was then placed on the particles. The coverslip was inverted andplaced on a well made by sticking a metal ring to a microscope slide. The

* Present address: Fourah Bay College, University of Sierra Leone, Freetown,Sierra Leone.

Trans. Br. mycol. Soc. 64 (3), (1975). Printed in Great Britain