2
Short Communications TIPPER, D. j. & BOSTIAN, K. A. (1984). Double-stranded ribonucleic acid killer systems in yeast. Microbiological Reviews 48, 125-156. VAN VUUREN, H. j.]. & WINGFIELD, B. D. (1986). Killer yeasts - cause of stuck fennentations in a wine cellar. South African Journal of Enology and Viticulture 7, 113-118. WESOLOWSKI, M. & WICKNER, R. B. (1984). Two new double- stranded RNA molecules showing non-mendelian inheritance and heat inducibility in Saccharomyces cerevisiae. Molecular and Cellular Biology 4, 181-187. WICKNER, R. B. (1979). The killer double-stranded RNA plasmids of yeast. Plasmid 2, 303-322. 367 WICKNER, R. B. (1981). Killer systems in Saccharomyces cerevisiae. In Molecular Biology ofthe Yeast Saccharomyces cerevisiae: Life Cycle and Inheritance (ed. J. N. Strathem, E. W. Jones & J. R. Broach), pp. 415-444. Cold Spring Harbor, N.Y. YOUNG, T. W. (1987). Killer yeasts. In The Yeasts 2 (2nd edn) (ed. A. H. Rose & J. J. Harrison), pp. 131-164. London, New York: Academic Press. YOUNG, T. W. & YAGIU, M. (1978). A comparison of the killer character in different yeasts and its classification. Antonie van Leeuwenhoek 44, 59-77. Two aquatic hyphomycetes as endophytes in Alnus glutinosa roots P. J. FISHER Department of Biological Sciences, University of Exeter, EX4 4PS, UK O. PETRINI Mikrobiologisches Institut, ETH-Zentrum, CH-8092 Zurich, Switzerland Two aquatic hyphomycetes as endophytes in Alnus glutinosa roots. Mycological Research 92 (3): 367-368 (1989). The aquatic hyphomycetes Tricladium splendens and Campylospora parvula have been isolated from surface-sterilized roots of Alnus glutinosa. Key words: Alnus glutinosa, Aquatic hyphomycetes, Endophytic fungi. Aquatic hyphomycetes have been found in abundance on decaying leaves and twigs of deciduous trees in unpolluted streams. The use of the term aquatic implies that such fungi live in water, but evidence is accumulating that they can also be found in terrestrial situations. Bandoni (1981) has summarized the occurrences of aquatic hyphomycetes from non-aquatic habitats. It is of interest that. of the 34 examples cited, only 3 refer to the roots of plants. Waid (1954) isolated Varicosporium e10deae Kegel from the root surfaces of beech seedlings grown in beechwood soil, and Nemec (1969) isolated Anguillospora longissima (Sacc. & Syd.) Ingold and Tetracladium marchalianum de Wild. from the roots of diseased strawberry plants. In a study of endophytic fungi in the bark and xylem of Alnus glutinosa (L.) Gaertner, we have demonstrated colon- ization of the bark of roots by the aquatic hyphomycetes Tricladium splendens Ingold and Campylospora parvula Kuzula (Fig. 1). The latter fungus may have once previously been recorded in Britain from a stream in Kent (Ingold, 1975), whilst T. spendens can be found in most streams containing suitable substrates. It is also one of the few aquatic hyphomycetes that will form spores when its mycelium is exposed to the atmosphere. Roots of Alnus glutinosa were sampled from 3 trees 26 standing on the bank of the Exeter Canal, Grid Reference SY 925915. The roots were scrubbed clean of all surface soil and subjected to surface-sterilization by the method of Fisher, Anson & Petrini (1986). Then the bark was stripped off with a sterile scalpel and cut into 1 em lengths before being placed in groups of 5 onto 1'5 % malt (MEA) extract. Plates were incubated at ambient temperature (ca 20°C) for 5-14 d depending on the growth rate of the fungi. Isolation was by transfer of mycelium to 2 % MEA. After 4 wk incubation at 20°, strips 1 em wide were cut from each culture and submerged in sterile distilled water and further incubated at 10° in diffuse daylight. After 4--0 d, spores of T. splendens and C. parvula were formed. This is the first record of aquatic hyphomycetes as endophytes in the roots of a plant. Only 1'6 % of the 300 pieces of bark sampled grew T. splendens and 0'7% C. parvula, compared with the most common root endophyte, Cylindrocarpon destructans (Zins.) Scholten, which colonized 19 % of all bark fragments. The infrequent occurrence of T. splendens and C. parvula suggests that these species may only be able to establish themselves as marginal endophytes in the roots of certain trees which grow near streams. Specimens (dried sporulating cultures and microscope slides) have been preserved as HME 4353 C. parvula, HME 4354 T. splendens. Cultures of C. parvula have been deposited at MYC 92

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Page 1: Two aquatic hyphomycetes as endophytes in Alnus glutinosa roots

Short Communications

TIPPER, D. j. & BOSTIAN, K. A. (1984). Double-stranded ribonucleicacid killer systems in yeast. Microbiological Reviews 48, 125-156.

VAN VUUREN, H. j.]. & WINGFIELD, B. D. (1986). Killer yeasts- cause of stuck fennentations in a wine cellar. South AfricanJournal of Enology and Viticulture 7, 113-118.

WESOLOWSKI, M. & WICKNER, R. B. (1984). Two new double­stranded RNA molecules showing non-mendelian inheritance andheat inducibility in Saccharomyces cerevisiae. Molecular and CellularBiology 4, 181-187.

WICKNER, R. B. (1979). The killer double-stranded RNA plasmids ofyeast. Plasmid 2, 303-322.

367

WICKNER, R. B. (1981). Killer systems in Saccharomyces cerevisiae. InMolecular Biology ofthe Yeast Saccharomyces cerevisiae: Life Cycleand Inheritance (ed. J. N. Strathem, E. W. Jones & J. R. Broach),pp. 415-444. Cold Spring Harbor, N.Y.

YOUNG, T. W. (1987). Killer yeasts. In The Yeasts 2 (2nd edn) (ed.A. H. Rose & J. J. Harrison), pp. 131-164. London, New York:Academic Press.

YOUNG, T. W. & YAGIU, M. (1978). A comparison of the killercharacter in different yeasts and its classification. Antonie vanLeeuwenhoek 44, 59-77.

Two aquatic hyphomycetes as endophytes in Alnus glutinosaroots

P. J. FISHER

Department of Biological Sciences, University of Exeter, EX4 4PS, UK

O. PETRINI

Mikrobiologisches Institut, ETH-Zentrum, CH-8092 Zurich, Switzerland

Two aquatic hyphomycetes as endophytes in Alnus glutinosa roots. Mycological Research 92 (3): 367-368 (1989).

The aquatic hyphomycetes Tricladium splendens and Campylospora parvula have been isolated from surface-sterilized roots of Alnusglutinosa.

Key words: Alnus glutinosa, Aquatic hyphomycetes, Endophytic fungi.

Aquatic hyphomycetes have been found in abundance ondecaying leaves and twigs of deciduous trees in unpollutedstreams. The use of the term aquatic implies that such fungilive in water, but evidence is accumulating that they can alsobe found in terrestrial situations.

Bandoni (1981) has summarized the occurrences of aquatichyphomycetes from non-aquatic habitats. It is of interest that.of the 34 examples cited, only 3 refer to the roots of plants.Waid (1954) isolated Varicosporium e10deae Kegel from theroot surfaces of beech seedlings grown in beechwood soil, andNemec (1969) isolated Anguillospora longissima (Sacc. & Syd.)Ingold and Tetracladium marchalianum de Wild. from the rootsof diseased strawberry plants.

In a study of endophytic fungi in the bark and xylem ofAlnus glutinosa (L.) Gaertner, we have demonstrated colon­ization of the bark of roots by the aquatic hyphomycetesTricladium splendens Ingold and Campylospora parvula Kuzula(Fig. 1). The latter fungus may have once previously beenrecorded in Britain from a stream in Kent (Ingold, 1975),whilst T. spendens can be found in most streams containingsuitable substrates. It is also one of the few aquatichyphomycetes that will form spores when its mycelium isexposed to the atmosphere.

Roots of Alnus glutinosa were sampled from 3 trees

26

standing on the bank of the Exeter Canal, Grid ReferenceSY 925915. The roots were scrubbed clean of all surface soiland subjected to surface-sterilization by the method of Fisher,Anson & Petrini (1986). Then the bark was stripped off witha sterile scalpel and cut into 1 em lengths before being placedin groups of 5 onto 1'5 % malt (MEA) extract. Plates wereincubated at ambient temperature (ca 20°C) for 5-14 ddepending on the growth rate of the fungi. Isolation was bytransfer of mycelium to 2 % MEA. After 4 wk incubation at20°, strips 1 em wide were cut from each culture andsubmerged in sterile distilled water and further incubated at10° in diffuse daylight. After 4--0 d, spores of T. splendens andC. parvula were formed. This is the first record of aquatichyphomycetes as endophytes in the roots of a plant. Only1'6 % of the 300 pieces of bark sampled grew T. splendens and0'7% C. parvula, compared with the most common rootendophyte, Cylindrocarpon destructans (Zins.) Scholten, whichcolonized 19 % of all bark fragments. The infrequent occurrenceof T. splendens and C. parvula suggests that these species mayonly be able to establish themselves as marginal endophytesin the roots of certain trees which grow near streams.

Specimens (dried sporulating cultures and microscope slides)have been preserved as HME 4353 C. parvula, HME 4354T. splendens. Cultures of C. parvula have been deposited at

MYC 92

Page 2: Two aquatic hyphomycetes as endophytes in Alnus glutinosa roots

Short Communications 368

Fig. 1. A, Conidium of Tricladium splendens; B, conidium development of Campylospora parvula; C and 0, mature spores. Bar scale =

20 IJm.

CAB International Mycological Institute (1MI 327408), and atthe Czechoslovak Collection of Microorganisms, 662 43 Brno,Accession Number CCM 8044.

We are indebted to Dr Ludmila Marvanova for confirmingthe identification of C. parvu/a.

REFERENCES

BANDON!, R.]. (1981). Aquatic hyphomycetes from terrestriallitter. In The Fungal Community (ed. D. T. Wicklow & G. C. Carroll),pp. 693-708. New York: Marcel Dekker.

FISHER, P. ]., ANSON, A. E. & PETRINI, O. (1986). Fungalendophytes in Wex eUTopaeus and Wex gallii. Transactions of theBritish Mycological Society 86, 153-156.

INGOLD, C. T. (1975). An illustrated guide to aquatic and water­borne hyphomycetes with notes on their biology. FreshwaterBiological Association Scientific Publication 30, 24-25.

NEMEC S. (1969). Sporulation and identification of fungi isolatedfrom root-rot in diseased strawberry plants. Phytopathology 59,1552-1553.

WAID, ]. S. (1954). Occurrence of aquatic hyphomycetes upon theroot surfaces of beech grown in woodland soils. Transactions of theBritish Mycological Society 37, 420-421.