恒温和变温培养对羊肚菌菌丝生长及菌核形成影响的比较研究

为了探究恒温和变温培养对羊肚菌菌丝生长和菌核形成的影响,设置-9℃-30℃的7个恒温处理,3个变温处理,通过观察和记录在两种不同培养方式下菌丝的长势、生长速度、菌落干重、产核时间、菌核数量等内容,从而比较出恒温和变温培养对羊肚菌菌丝生长和菌核形成的影响效果。结果表明,在恒温条件下羊肚菌菌丝生长和菌核发育比变温好,20℃最适宜菌丝生长,菌丝生长速度较快,菌落干重最大;25℃最适宜菌核生长发育,产核最早。菌核数量最多。     

The effect of temperature on the infection of onions by Sclerotium cepivorum

Studies on Sclerotium cepivorum infection of salad onions in artificially infested soil showed that under field conditions infection was greatest in late spring and summer and declined to a low level in the late autumn and winter months. Change of infection levels was found to be correlated with the effect of soil temperature in the field on the germination of sclerotia. This was substantiated in laboratory studies which demonstrated that sclerotium germination, mycelial growth and seedling infection were all markedly influenced by temperature.     

Factors influencing the production of sclerotia in the wild rice (Zizania aquatica) pathogenSclerotium hydrophilum

Sclerotium hydrophilum was shown to be auxoheterotrophic for thiamine, with the addition of this vitamin being required for the induction of sclerotia on defined media, but riboflavin and pyridoxine also have a positive effect. In the absence of thiamine, an increase in glucose concentration lead to a decrease in the yield of sclerotia; however, the addition of thiamine negate this inhibition and, instead, as the glucose concentration increased a higher proportion of sclerotial initials matured. Overall it was found that thiamine, specifically the pyrimidine component of thiamine, is crucial for initiating sclerotium production, while glucose stimulates maturation. The effect of light on sclerotium production was found to be complex and dependent on the growth medium. Light is not required for either the induction or maturation of sclerotia, but continuous irradiation of developing cultures with either white light or black light induces an endogenous rhythm whereby sclerotia are formed every 48h. When exposed to alternating light/dark regimes mycelium that formed in the light does not mature sclerotia, but dark-formed mycelium does, even if it is subsequently exposed to light.     

Factors Influencing Formation of Sclerotia in Grifola umbellate （Pers.） Pilaet Under Artificial Conditions

The effects of substrate composition and temperature on myceilal growth and sclerotium production in Grlfola umbellate （Pers.） Pilaet were Investigated In the present study. The Induction of sclerotla of G. umbellate was affected greatly by the type of medium, as well as the type of carbon source. Malt-extract agar was able to induce the production of sclerotia. The production of sclerotia was also observed when the carbon source in the GPC agar medium （glucose 20 g/L, peptone 6 g/L, corn steep liquor 10 g/L, and agar 15 g/L） was replaced with glycerol or mannitol. Altering the composition of the GPC medium with milk powder, thiamine hydrochlorlde, extract of Armlllarla mellea, active clay, dlatomite, kaolin, or arginlne did not induce the production of sclerotla. A temperature range of 18-25 ℃ was suitable for both mycellai growth and sclerotium formation. Glycerol significantly Induced slerotium formation on nutrient supplemented with sawdust substrates In bottle culture. 24S-Polyporusterone A and polyporusterone B were assayed In samples of natural and cultured sclerotla. Both natural and cultured sclerotla contained 24S- polyporusterone A and polyporusterone B.     

Growth, sclerotia and aflatoxin production by Aspergillus parasiticus: influence of food preservatives

The influence of six food preservatives on control of aflatoxin production by Aspergillus parasiticus was tested in SMKY and defined media at three concentrations, viz., 0.1, 0.5 and 1.0%. Propionic acid completely inhibited the yield of mycelia and sclerotia, and aflatoxin production in culture medium, mycelia and sclerotia of A. parasiticus at all concentrations, whereas citric acid showed inhibition only at 0.5 and 1.0% concentrations. Sodium metabisulphite did not permit mycelial growth and aflatoxin biosynthesis in SMKY liquid medium but allowed production of sclerotia and aflatoxin on solid media, while the rest of the food preservatives had only marginal inhibitory effects.     

Ultrastructural basis of sclerotial survival in soil

The ultrastructure of developing and mature sclerotia ofSclerotium rolfsii was studied with a scanning electron microscope. The mature sclerotium is disconnected from the mycelium and well differentiated. The rind is composed of rather thick-walled empty cells. The cortex cells are large and almost completely filled with vesicles, whereas the medullar cells are smaller and some of them are very thickwalled.     

Variability in Indian isolates of Sclerotium rolfsii

Variability among 26 isolates of Sclerotium rolfsii collected from various hosts/soil samples and localities in India is reported. The isolates varied in colony morphology, mycelial growth rate, sclerotium formation, teleomorph production and sclerotial size and color. Out of 26 isolates, only 4 produced the teleomorph stage on Cyperus rotundus rhizome meal agar medium. Mycelial incompatibility among the isolates was also seen, and out of 325 combinations, only 29 combinations (8.9%) showed compatible reactions. Based on mycelial compatibility, 13 vegetative incompatibility groups (VCG) were identified among the isolates. HPLC analysis of the ethyl acetate fraction of culture filtrates of the isolates revealed 10-22 peaks. Six peaks were identified as gallic, oxalic, ferulic, indole-3-acetic acid (IAA), chlorogenic, and cinnamic acids. Oxalic, IAA, and cinnamic acids were present in the culture filtrates of all the isolates in varying amounts. The other three phenolic acids were not detected in some of the isolates. A comparative HPLC analysis of sclerotial exudate, sclerotia, mycelia, and culture filtrates of two S. rolfsii isolates (leaf spot- and collar rot-causing) producing different symptoms on their respective hosts revealed variation in the content of phenolic acids, IAA, and oxalic acid.     

Fungi isolated from Sclerotia ofSclerotium cepivorum and from soil and their effects upon the pathogen

Summary Data are presented on the antagonistic effects of the fungi isolated from sclerotia ofSclerotium cepivorum and from nonrhizosphere soil taken from around the roots of infected onions upon mycelial growth and sclerotial germination ofS. cepivorum. Most of the isolated fungi especiallyPenicillium species were antagonistic to mycelial growth. Sclerotial germination was slightly inhibited by diffusates of these fungal isolates. Testing the antifungal effect of someAllium extracts against the fungal isolates by the inhibition zone method showed that garlic extract has the greatest antifungal effects and onion extract is the least potent. However, spore germination tests indicated that onion extract completely inhibits the spore germination of all test fungi. The role of host-plant extracts in stimulating sclerotial germination is discussed.     

THE EFFECT OF SOME PHYSIOLOGICAL AND ENVIRONMENTAL FACTORS ON SCLEROTIAL ASPERGILLI

Rudolph , Emanuel D. (Ohio State U., Columbus.) The effect of some physiological and environmental factors on sclerotial Aspergilli. Amer. Jour. Bot. 49(1): 71–78. Illus. 1962.—The effect of varying conditions of carbon-nitrogen balance, temperature, pH, and light upon the formation of sclerotia by 6 species of Aspergillus (A. alliaceus, A. avenaceus, A. flavus, A. quercinus, A. sclerotiorum and A. wentii) was studied. On Czapek's agar, optimal growth as well as maximum production of sclerotia and conidia took place at high sucrose and nitrate concentrations. In general, fewer sclerotia were formed with glucose than with sucrose, and very poor growth took place with lactose. Sclerotia were formed best at temperatures that were optimal or below optimal for mycelial growth. The ranges of pH through which sclerotia were formed were narrower than those through which conidia and mycelia were formed. Light had no effect upon sclerotium formation. The formation of sclerotia in A. alliaceus was found to represent the strand-type development. A number of UV-induced strains and a spontaneous mutant strain of A. alliaceus showing varying amounts of sclerotium and conidium production are characterized. It is suggested that the sclerotia in Aspergillus are sterile stromata.     

The effect of glucose and lactose on beta-D-galactosidase activity and formation of sclerotia in Sclerotium rolfsii.

Addition of 0.5% (w/v) lactose to a glucose-mineral mdeium (SM) induced formation of sclerotia and beta-D-galactosidase (beta-D-galactoside galactohydrolase)(EC 3.2.1.23) synthesis in Sclerotium rolfsii types A and R; These effects as well as lactose uptake were inversely related to glucose concentration within the tested range of 0.5 to 2.5% (w/v). Transfer of lactose-grown colonies to a glucose-supplemented medium nullified the inducible effect of lactose on formation of sclerotia, whereas transfer to water agar did not. It is concluded that glucose nullifies the effect of lactose on S. rolfsii by interfering with its active uptake.     

Broad spectrum action of phenazine against active and dormant structures of fungal pathogens and root knot nematode

Antifungal antibiotic from Pseudomonas chlororaphis isolate PA23 was identified as Phenazine using TLC and HPLC. Phenazine recorded the highest inhibition zone of 21?mm with 35.55% percent inhibition of mycelial growth of Pythium aphanidermatum over control. It had a significant effect on the hyphal morphology of P. aphanidermatum and on spore germination of Botryodiplodia theobromae and Alternaria solani. Disorganization of hyphal morphology of P. aphanidermatum includes vacuolization, cell content degeneration and hyphal lysis. Similarly interaction of phenazine with Rhizoctonia solani resulted in abnormal swelling of hyphal tips was noticed in the hyphal tips. Similarly the germination of sclerotia of Macrophomina phaseolina, R. solani and Sclerotium rolfsii were completely inhibited by phenazine at a concentration 50?μl. Incubation of the eggs of the root knot nematode Meloidogyne incognita in 30?μl concentration of phenazine, completely suppressed the hatching of juveniles.     

Inhibition of growth of Sclerotinia minor and other pathogens by citrinin in the filtrate of Penicillium citrinum

Penicillium citrinum Thom, isolated from the sclerotia of Sderotinia minor, was cultured in a broth of Czapek-Dox for 4 to 8 weeks. The filtrate obtained was incorporated into potato dextrose agar or Czapek-Dox agar at different concentrations (v/v). The amended media were tested for mycelial growth of S. minor and other pathogens. Mycelial growth of S. minor was completely inhibited on media amended with 20% (v/v) filtrate of P. citrinum, and considerable inhibition of S. minor occurred at 10 and 15% concentrations. Mycelial growth of S. major, Sclerotium rolfsii, Rhizoctonia solani (AG-4) was inhibited by similar concentrations of filtrate of P. citrinum. Inhibitor(s) in the filtrate were extracted with ethyl acetate and tentatively identified as citrinin. Citrinin was shown to be an active component in the filtrate against mycelial growth of S. minor, S. major and Sclerotium rolfsii.Cooperative investigation of U.S. Department of Agriculture, Agriculture Research Service and Oklahoma State University. Journal Article No. 4989, Oklahoma Agricultural Experiment Station, Oklahoma State University, Stillwater.Mention of a trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by USDA or by Oklahoma State University, or imply their approval to the exclusion of other products or vendors that may also be suitable.     

The occurrence of sclerotium rot on Momordica charantia caused by Sclerotium rolfsii in Korea

Sclerotium rolfsii is one of the most destructive pathogens and thought to affect a broad range of plant hosts. In July 2014, an occurrence of sclerotium rot was observed on bitter melon (Momordica charantia L.) in Jinju, South Korea. The rot symptoms were most developed on stems and fruit near the soil line, and infected bitter melon plants withered and eventually died. White mycelial mats with numerous sclerotia were produced on diseased stems and fruit near the soil surface. Based on the morphological characteristics, pathogenicity tests, and DNA sequencing and phylogenetic analysis of the internal transcribed spacer (ITS) ribosomal RNA (rRNA) gene region, the causal fungus was identified as S. rolfsii Saccardo. This is the first report of sclerotium rot caused by S. rolfsii on bitter melon in Korea. The recent occurrence of sclerotium rot on bitter melon poses a potential threat to its production in Korea.     

Morphological Transitions Governed by Density Dependence and Lipoxygenase Activity in Aspergillus flavus

Aspergillus flavus differentiates to produce asexual dispersing spores (conidia) or overwintering survival structures called sclerotia. Results described here show that these two processes are oppositely regulated by density-dependent mechanisms and that increasing the cell density (from 10¹ to 10⁷ cells/plate) results in the lowest numbers of sclerotial and the highest numbers of conidial. Extract from spent medium of low-cell-density cultures induced a high-sclerotium-number phenotype, whereas high-cell-density extract increased conidiation. Density-dependent development is also modified by changes in lipid availability. Exogenous linoleic acid increased sclerotial production at intermediate cell densities (10⁴ and 10⁵ cells/plate), whereas oleic and linolenic acids inhibited sclerotium formation. Deletion of Aflox encoding a lipoxygenase (LOX) greatly diminished density-dependent development of both sclerotia and conidia, resulting in an overall increase in the number of sclerotia and a decrease in the number of conidia at high cell densities (>10⁵ cells/plate). Aflox mutants showed decreased linoleic acid LOX activity. Taken together, these results suggest that there is a quorum-sensing mechanism in which a factor(s) produced in dense cultures, perhaps a LOX-derived metabolite, activates conidium formation, while a factor(s) produced in low-density cultures stimulates sclerotium formation.     

Antifungal Activity of Some Deoxyhypusine Synthase Inhibitors

Several inhibitors of deoxyhypusine synthase were examined for antifungal activity. All compounds, used at concentrations of 1.0 mM or less, reduced mycelial growth of the oat leaf stripe pathogen Pyrenophora avenae grown on solid media. Four of the compounds completely inhibited fungal growth, 4a doing so at a concentration of only 0.61 mM. Inhibition of fungal growth afforded by some of the compounds was more marked than was reflected by the radial growth measurements.     

Individual and Combined Effects of Certain Pesticides on Rhizoctonia solani, Sheath Blight Pathogen of Rice

The effect of different pesticides (fungicide, herbicide and insecticide) on the mycelial growth and formation and germination of sclerotia of sheath blight pathogen of rice, Rhizoctonia solani Kühn was studied in nutrient medium and in natural soil. Among the herbicides and insecticides, fenitrothion (insecticide) and thiobencarb (herbicide) were the most effective in inhibiting the mycelial growth and sclerotial formation. These pesticides effected significant inhibition of both mycelialgrowth and sclerotial formation at 7.5 to 10μg/ml concentration while fungicides carbendazim and tolclofosmethyl effected almost complete inhibition at 2.5 μg/ml. Interestingly, a synergistic increase in the toxicity to mycelial growth, sclerotial formation and sclerotial germination was noticed when the fungicide, mancozeb and the herbicide, thiobencarb were applied at subtoxic concentration in combination. Such synergistic interactions between pesticides in a combination leading to increased toxicity suggest that under the current practice of applying fungicides, herbicides and insecticides either simultaneously or in rotation, certain combinations may help in reducing the dose of a fungicide in disease control.     

Antifungal efficacy of chitosan and its thiourea derivatives upon the growth of some sugar-beet pathogens

Chitosan (CS) was modified by reaction with benzoyl thiocyanate to give a thiourea derivative (TUCS). The antifungal behavior of chitosan and its thiourea derivative was investigated in vitro on the mycelial growth, sporulation and germination of conidia or sclerotia of the following sugar-beet: Beta vulgaris pathogens isolated in Egypt, Rhizoctonia solani Kühn (AG(2-2)) Sclerotium rolfsii Sacc. and Fusarium solani (Mart.) Sacc. All the prepared thiourea derivatives had a significant inhibiting effect on the different stages of development on the germination of conidia or sclerotia of all the investigated fungi in the polymer concentration range of 5-1000 microg ml(-1). In the absence of chitosan and its derivative, R. solani exhibited the fastest growth of the fungi studied. However, growth tolerance of the modified chitosan was highest for F. solani and lowest for R. solani. The most sensitive to the modified chitosan stress with regard to their germination and number produced were the sclerotia of S. rolfsii. It has been found that the TUCS is a much better fungicidal agent (about 60 times more) than the pure CS against most of the fungal strains tested. The molecular weight and the degree of deacetylation were found to have an important effect on the growth activities of the pathogens.     

In vitro inhibitory effects of rosemary and sage extracts on mycelial growth and sclerotial formation and germination of Sclerotinia sclerotiorum

The anti-fungal efficacy for two Labiate plants, rosemary (Rosmarinus officinalis L.) and Greek sage (Salvia fructicosa Mill.), against Sclerotinia sclerotiorum fungus (Lib.) de Bary has been investigated. The inhibitory effect of these plants as crude leaf ethanolic extract on the radial mycelial growth as well as on sclerotial production and germination was measured in vitro at various concentrations (stock?=?0.5?g dry leaf powder/ml ddH₂O) in the growth medium. In general, rosemary extract revealed a remarkable anti-fungal effect against the fungus, being more inhibitory than Greek sage in this respect. This was evident as total inhibition of radial mycelial growth by rosemary occurred at 10% extract concentration, while sage was half as potent producing such an effect at double the concentration (20%). Both rosemary and sage extracts were more inhibitory to sclerotial formation than to mycelial growth as the fungus ceased to produce any sclerotia at the lower concentrations of 5 and 5–10%, respectively. In addition, rosemary was highly effective in inhibiting sclerotia germination as total inhibition of germination occurred at 20% extract concentration at three?days and onward after incubation. Moreover, at this level, the survival of sclerotia was totally lost when examined after 12?days of incubation. For sage, inhibition of sclerotial germination/death was only 20% at 12th day of incubation. The results of this study indicate that the extracts of rosemary and Greek sage leaves could become natural alternatives to synthetic fungicides to manage diseases of S. sclerotiorum.     

Interactions between Sarocladium oryzae and stem attacking fungal pathogens of rice

In laboratory tests Sarocladium oryzae, the sheath rot pathogen of rice was found to inhibit the mycelial growth of other stem-attacking rice pathogens. Among those inhibited, Sclerotium oryzae and Gaeumannomyces graminis var. graminis were most sensitive while Pyricularia oryzae and Rhizoctonia solani were less sensitive. Tissue-based tests made with rice culm segments established that Sarocladium oryzae inhibits mycelial growth and delays sclerotium formation in R. solani. Cerulenin, the toxin produced by Sarocladium oryzae showed a toxicity pattern towards rice pathogens similar to that of Sarocladium oryzae. The stem rot pathogen, Sclerotium oryzae was most sensitive to cerulenin. In two greenhouse experiments, IR58 rice plants inoculated with Sarocladium oryzae alone or together with Sclerotium oryzae, G. graminis var. graminis or R. solani were found to have reduced plant height and increased tiller number. Sheath rot severity increased when Sarocladium oryzae was inoculated as a single pathogen or together with others. Sheath rot inoculation reduced stem rot in rice plants by 76 and 58%, respectively, in Experiment 1 and 2. By its known antagonistic interaction towards stem rot and crown sheath rot pathogens which are sensitive to it and by other unknown interactions, sheath rot emerges as the dominant disease.     

Influence of inocula and grains on sclerotia biomass and carotenoid yield of<Emphasis Type="Italic"> Penicillium</Emphasis> sp. PT95 during solid-state fermentation

Various inocula and grains were evaluated for carotenoid production by solid-state fermentation using Penicillium sp. PT95. Millet medium was more effective in both sclerotia growth and carotenoid production than other grain media. An inoculum in the form of sclerotia yielded higher sclerotia biomass compared to either a spore inoculum or a mycelial pellet inoculum. Adding wheat bran to grain medium favored the formation of sclerotia. However, neither the inoculum type nor addition of wheat bran resulted in a significant change in the carotenoid content of sclerotia. Among grain media supplemented with wheat bran (wheat bran:grain =1:4 w/w, dry basis), a medium consisting of rice and wheat bran gave the highest sclerotia biomass (15.10 g/100 g grain), a medium consisting of buckwheat and wheat bran gave the highest content of carotenoid in sclerotia (0.826 mg/g dry sclerotia), and a medium consisting of millet and wheat bran gave the highest carotenoid yield (11.457 mg/100 g grain).