Effect of salinity on spore germination of terrestrial and marine fungi. 1975 $$$$
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ቤተ መጻሕፍቲ ባይዱAND
E. B. GARETH JONES
Department of Biological Sciences, Portsmouth Polytechnic, Portsmouth, PO I 2D r
(With
2
Text-figures)
The combined effect of salinity and temperature on spore germination in I 1 terrestrial, freshwater and marine fungi was investigated. Spore germination in the terrestrial fungi decreased with increasing salinities and these inhibitory effects were more marked at low temperatures. The five marine fungi examined exhibited a broad tolerance to both salinity and temperature stresses.
32-2
Transactions British Mycological Society
Table
1.
Incubation times (hours) for spores at various temperatures
Fungi Temperature (0C)
A
Terrestrial Mucor hiemalis Wehimer. Chaetomium globosum Kunze Ex Fr. t Sordaria fimicola (Rob.) Ces. and de Not.] Penicillium notatum Westlingt Freshwater Heliscus lugdunensis Sacco and Therryt Tetracladium setigerum (Grove) Ingoldf Marine Corollospora maritima Werdermannt Torpedospora radiata Meyers'[ Asteromyces cruciaius (F. and Mme. Moreau) ex Hennbertj Dendryphiella salina (Suth.) Pugh and Nicotj Zalerion maritimum (Linder) Anastasiouf
MATERIALS AND METHODS
Seawater was collected at the mouth of Langstone harbour on a rising tide, filtered, and stored in carboys until required. Glass-distilled water was employed for seawater dilutions. The method employed for the measurement of spore germination was adapted from that described by Jackson (1958). Four agar disks (6 mm) were cut from petri dishes containing 10 ml non-nutrient 2 % agar, made up with a range of seawater dilutions at 10 % intervals. The agar disks at
each salinity were placed on sterile filter paper, thoroughly moistened with the water of the same salinity, in a sterile petri dish. Replicate sets were made up for each temperature (10-30°) and incubated for several hours before inoculation. After inoculation, the experimental petri dishes were incubated for varying periods of time depending on the fungi and temperature used (Table I). Spores were considered to have germinated when they had produced a germ-tube of one spore diameter or more in length. On completion of the experiment, the agar disks were placed on a glass slide and a glass coverslip bearing a drop of lactophenol cotton blue positioned on the upper surface of each disk. Germinated and ungerminated spores in each of four random microscope fields on each agar disk were counted, with the aid of an eye-piece grid graticule, and the results expressed as percentage spores germinated. The fungi tested are listed in Table I and the spore suspensions were prepared as follows: (I) M. hiemalis, P. notatum, A. cruciatus, D. salina and Z. maritimum. A stock agar plate bearing sporulating mycelium was flooded with 10 ml of a 0'02 % solution of Tween 80 in distilled water, several glass beads introduced and the plate shaken vigorously to obtain a uniform suspension of spores. The resultant suspension was centrifuged and the spores washed in fresh Tween 80 solution. The spore concentration was determined and adjusted, when necessary, by dilution to approximately 106 spores/ml. One drop of this spore suspension was placed on each agar disk and the petri dishes incubated as described above. (2) C. globosum, S. fimicola, C. maritima and T. radiata. Perithecia were removed aseptically from sporulating cultures and crushed in 3-4 ml of sterile Tween 80 solution using a fine rod. The larger fragments of peri-
Physiological studies of marine fungi have tended to concentrate on their salinity requirements (Meyers, 1968; Tubaki, 1969) and their ability to degrade wood (Jones, 1971). In the salinity studies, ion uptake and the effect of salinity on the vegetative growth of marine fungi only have been investigated. The effect of salinity on reproduction and spore germination has been much neglected. Meyers & Simms (1965) report that ascospores of Lindra thalassiae Orpurt, Meyers, Boral & Simms failed to germinate on a distilled-water medium. Borut and Johnson (1962) studied the effect of distilled water, 3·5 % NaCI, autoclaved, filtered or fresh untreated seawater on the spore germination of eight terrestrial fungi isolated from the Neuse-Newport estuary, U.S.A. Percentage germination in distilled water ranged from 45 (Gliocladium fimbriatum Gilman and Abott) to 100 (Curvularia pallescens Boedijn and Paecilomyces puntonii (Vuill.) Nann.). Spore germination of all the fungi, except C. pallescens, was inhibited in seawater, the spores of G.fimbriatum and Trichoderma lignorum (Tode) Harz failing to germinate in that medium. The present work was undertaken to investigate the effect of salinity and temperature on the spore germination of a range of terrestrial, freshwater and marine fungi.
E. B. GARETH JONES
Department of Biological Sciences, Portsmouth Polytechnic, Portsmouth, PO I 2D r
(With
2
Text-figures)
The combined effect of salinity and temperature on spore germination in I 1 terrestrial, freshwater and marine fungi was investigated. Spore germination in the terrestrial fungi decreased with increasing salinities and these inhibitory effects were more marked at low temperatures. The five marine fungi examined exhibited a broad tolerance to both salinity and temperature stresses.
32-2
Transactions British Mycological Society
Table
1.
Incubation times (hours) for spores at various temperatures
Fungi Temperature (0C)
A
Terrestrial Mucor hiemalis Wehimer. Chaetomium globosum Kunze Ex Fr. t Sordaria fimicola (Rob.) Ces. and de Not.] Penicillium notatum Westlingt Freshwater Heliscus lugdunensis Sacco and Therryt Tetracladium setigerum (Grove) Ingoldf Marine Corollospora maritima Werdermannt Torpedospora radiata Meyers'[ Asteromyces cruciaius (F. and Mme. Moreau) ex Hennbertj Dendryphiella salina (Suth.) Pugh and Nicotj Zalerion maritimum (Linder) Anastasiouf
MATERIALS AND METHODS
Seawater was collected at the mouth of Langstone harbour on a rising tide, filtered, and stored in carboys until required. Glass-distilled water was employed for seawater dilutions. The method employed for the measurement of spore germination was adapted from that described by Jackson (1958). Four agar disks (6 mm) were cut from petri dishes containing 10 ml non-nutrient 2 % agar, made up with a range of seawater dilutions at 10 % intervals. The agar disks at
each salinity were placed on sterile filter paper, thoroughly moistened with the water of the same salinity, in a sterile petri dish. Replicate sets were made up for each temperature (10-30°) and incubated for several hours before inoculation. After inoculation, the experimental petri dishes were incubated for varying periods of time depending on the fungi and temperature used (Table I). Spores were considered to have germinated when they had produced a germ-tube of one spore diameter or more in length. On completion of the experiment, the agar disks were placed on a glass slide and a glass coverslip bearing a drop of lactophenol cotton blue positioned on the upper surface of each disk. Germinated and ungerminated spores in each of four random microscope fields on each agar disk were counted, with the aid of an eye-piece grid graticule, and the results expressed as percentage spores germinated. The fungi tested are listed in Table I and the spore suspensions were prepared as follows: (I) M. hiemalis, P. notatum, A. cruciatus, D. salina and Z. maritimum. A stock agar plate bearing sporulating mycelium was flooded with 10 ml of a 0'02 % solution of Tween 80 in distilled water, several glass beads introduced and the plate shaken vigorously to obtain a uniform suspension of spores. The resultant suspension was centrifuged and the spores washed in fresh Tween 80 solution. The spore concentration was determined and adjusted, when necessary, by dilution to approximately 106 spores/ml. One drop of this spore suspension was placed on each agar disk and the petri dishes incubated as described above. (2) C. globosum, S. fimicola, C. maritima and T. radiata. Perithecia were removed aseptically from sporulating cultures and crushed in 3-4 ml of sterile Tween 80 solution using a fine rod. The larger fragments of peri-
Physiological studies of marine fungi have tended to concentrate on their salinity requirements (Meyers, 1968; Tubaki, 1969) and their ability to degrade wood (Jones, 1971). In the salinity studies, ion uptake and the effect of salinity on the vegetative growth of marine fungi only have been investigated. The effect of salinity on reproduction and spore germination has been much neglected. Meyers & Simms (1965) report that ascospores of Lindra thalassiae Orpurt, Meyers, Boral & Simms failed to germinate on a distilled-water medium. Borut and Johnson (1962) studied the effect of distilled water, 3·5 % NaCI, autoclaved, filtered or fresh untreated seawater on the spore germination of eight terrestrial fungi isolated from the Neuse-Newport estuary, U.S.A. Percentage germination in distilled water ranged from 45 (Gliocladium fimbriatum Gilman and Abott) to 100 (Curvularia pallescens Boedijn and Paecilomyces puntonii (Vuill.) Nann.). Spore germination of all the fungi, except C. pallescens, was inhibited in seawater, the spores of G.fimbriatum and Trichoderma lignorum (Tode) Harz failing to germinate in that medium. The present work was undertaken to investigate the effect of salinity and temperature on the spore germination of a range of terrestrial, freshwater and marine fungi.
