核盘菌弱毒株Ep-1PN培养性状的有性遗传特性

观察和测定了核盘菌(Sclerotinia sclerotiorum (Lib.) de Bary)弱毒株Ep-1PN单孢分离物的生长、菌落扩展和菌落形态等培养性状.结果发现来自7个子囊盘的1574个单孢分离物中有1560个分离物的培养性状与正常菌株的没有显著差异,与亲本相似的分离物只有6个,其它8个分离物介于弱毒株Ep-1PN与正常菌株之间.弱毒株Ep-IPN培养性状的有性遗传不遵循孟德尔核遗传规律.     

核盘菌弱毒株Ep—IPN培养性状的有性遗传特性

观察和测定了核盘菌（Ｓｃｌｅｒｏｔｉｎｉａ ｓｃｌｅｒｏｔｉｉｏｒｕｍ（Ｌｉｂ．）ｄｅｅ Ｂａｒｙ）弱毒株Ｅｐ－ＩＰＮ单孢分离物的生长、菌落扩展和菌落形态等培养性状。结果表明：来自７个子囊盘的１５７４个单孢分离物中有１５６０个分离物的培养性状与正常菌株的没有显著差异,与亲本相似的分离物只有６个,其它８个分离物介于弱毒株Ｅｐ－ＩＰＮ与正常菌株之间。弱毒株Ｅｐ－ＩＰＮ培养性状的有性遗传不遵循孟德尔遗传     

苎麻疫霉有性生殖导致菌落形态和生长速率在无性后代持续变异的研究*

研究了苎麻疫霉（PhytophthoraboehmeriaeSawada）菌株JS-5自交S1代单卵孢株的菌落形态和生长速率在其单游动孢子无性系后代的遗传与变异。结果表明,亲本菌株JS-5的菌落形态和生长速率在连续4代单游动孢子无性系后代稳定遗传,而该菌株的自交S1代的约2／3单卵孢株的上述性状在其单拘无性后代中发生分离,且上述性状的分离在无性单孢后代中至少可连续保持3～4代。本研究结果提示,苎麻疫霉有性生殖导致上述性状的变异,除有性生殖过程中的基因重组外,有性生殖还诱导产生了一种新的机制参与上述性状遗传与变异的调控。本研究还观察到供试菌株及其后代所产生的游动孢子绝大多数细胞内只含有1个细胞核,表明上述机制与异核现象无关。有性生殖导致生物学性状在随后的无性后代中发生持续变异可能是苎麻疫霉生物学性状遗传多样性的重要机制之一。     

两株毒力不同的核盘菌产草酸、果胶酶的比较

对强弱毒力不同的核盘菌Ep-1PNA5和Ep-1PN的主要致病因子草酸和果胶酶的产生进行了比较研究。结果发现强毒力的Ep-1PNA5和弱毒力Ep-1PN都可产生草酸,并且在发病油菜活体组织上,弱毒力Ep-1PN的病组织中的草酸含量高于Ep-1PNA5病组织;两个菌株在发病油菜活体组织上的果胶酶产量没有差异,但在诱导培养基中Ep-1PN菌株比Ep-1PNA5的果胶酶产量高。这一结果表明弱毒力Ep-1PN菌株毒力的衰退并不是因为其所携带的dsRNA因子抑制了草酸和果胶酶所产生的。     

不同寄主来源的串珠镰孢生物学性状及其在无性后代的遗传与变异

自安徽、山东、湖北、江苏等地多点采集棉花红腐病、水稻恶苗病、玉米穗腐病的病组织,经分离、鉴定和纯化,获得107个串珠镰孢（Fusarium monilifoFine）菌株。对上述来源于棉花、水稻、玉米的串珠镰孢的菌落形态、生长速率和产孢量等生物学性状及其在无性后代的遗传与变异进行了研究。结果表明,不同寄主来源的串珠镰孢菌株的菌丝生长温度范围和最适温度大致相同,但在菌落形态特别是色素方面存在明显差异,生长速率和产孢量也存在显著差异。棉花菌株的平均生长速率最大,玉米菌株生长速率最小,水稻菌株生长速率居中,相同群体的不同菌株间生长速率有极显著差异;玉米菌株产孢量最大,棉花菌株产孢量最小,水稻菌株产孢量居中。方差分析显示,不同寄主菌株群体间产孢量存在显著差异,而同一寄主群体的不同菌株间产孢量均无显著差异,说明菌株产孢量大小主要与其寄主种类有关,而与地区来源关系不大。遗传测定结果表明,分离自棉花、玉米和水稻的串珠镰孢的菌落形态和生长速率在单分生孢子后代均可稳定遗传;产孢量性状遗传有两种情况：分离自棉花和水稻的串珠镰孢菌株Fm1和Fm31的产孢量性状在单分生孢子后代均可稳定遗传,而分离白玉米的串珠镰孢菌株Fm19的产孢量性状在单分生孢子第一代（CG1）发生变异。     

苎麻疫霉有性生殖导致菌落形态和生长速率在无性后代持续变异的研究

研究了苎麻疫霉（PhytophthoraboehmeriaeSawada）菌株JS-5自交S1代单卵孢株的菌落形态和生长速率在其单游动孢子无性系后代的遗传与变异。结果表明,亲本菌株JS-5的菌落形态和生长速率在连续4代单游动孢子无性系后代稳定遗传,而该菌株的自交S1代的约2／3单卵孢株的上述性状在其单拘无性后代中发生分离,且上述性状的分离在无性单孢后代中至少可连续保持3～4代。本研究结果提示,苎麻疫霉有性生殖导致上述性状的变异,除有性生殖过程中的基因重组外,有性生殖还诱导产生了一种新的机制参与上述性状遗传与变异的调控。本研究还观察到供试菌株及其后代所产生的游动孢子绝大多数细胞内只含有1个细胞核,表明上述机制与异核现象无关。有性生殖导致生物学性状在随后的无性后代中发生持续变异可能是苎麻疫霉生物学性状遗传多样性的重要机制之一。     

两株致病力不同的核盘菌形态变化研究

比较了强弱致病型不同的核盘菌Ep-1PNA5和Ep-1PN的菌落、菌丝、细胞的形态变化。结果发现强致病型的Ep-1PNA5在平板上的菌落呈圆形均匀扩展,气生菌丝少,形成的菌核多,细胞的原生质均匀浓密,内部结构完整。弱致病型的Ep-1PN菌落呈扇形扩展,不均匀,气生菌丝发达,形成的菌核少,菌丝顶端分枝异常,有原生质外渗,且原生质不均匀,内部结构不完整。Ep-1PN致病力减弱和生长抑制可能与其菌丝结构和细胞形态变异有关。     

金针菇担孢子核相及遗传属性的研究

以3个不同的金针菇菌株为材料,研究了其担孢子的核相及遗传属性。荧光染色观察显示,担孢子核相以双核为主,双核孢子、单核孢子和无核孢子分别占80.2%、7.5%和12.3%。源于单孢分离物的菌丝为有隔膜、无锁状联合的多核菌丝。在交配试验中,源于不同菌株单孢分离物的菌丝原生质体的配对形成具锁状联合的菌落,而源于同一单孢分离物的菌丝原生质体的配对则形成无锁状联合的菌落,暗示担孢子中的两个核具有相同的交配型。RAPD分析显示,源于同一单孢分离物的菌丝原生质体为10个随机引物所扩增的图谱彼此完全相同,印证了担孢子中的双核是同质的。此外,观察表明,一个担子上着生有4个担孢子。因此,金针菇是一种具4个含同质双核担孢子的四极性蕈菌。     

苎麻疫霉有性生殖导致菌落形态和生长速率在无性后代持续变异的研究

研究了苎麻疫霉菌株ＪＳ－５自交Ｓ１代单卵孢株的菌落形态和生长速率在其单游动孢子无性系后代的遗传与变异。结果表明,亲本菌株ＪＳ－５的菌落形态和生长速率在连续４代单游动孢子无性系后代稳定遗传,而该菌株的自交Ｓ１代的约２／３单卵孢株的上述性状在其单孢无性后代中发生分离,且上述性状的分离在无性单孢后代中至少可连续保持３￣４代。本研究结果提示,苎麻疫霉有性生殖导致上述性状的变异,除有性生殖过程中的基因重组外     

马铃薯晚疫病菌单孢分离物生物学特性的初步研究*

研究了马铃薯晚疫病菌单游动孢子分离的方法,并对其单游动孢子分离物菌落生长直径、产孢量以及对甲霜灵的敏感性进行了初步研究。结果表明同一菌株的不同单游动孢子菌落生长直径和产孢量有显著差异,但不同游动孢子分离物对甲霜灵敏感性没有显著差异。     

Oxalic acid production and its role in pathogenesis of Sclerotinia sclerotiorum

Abstract Sunflower plants were inoculated with a virulent isolate of Sclerotinia sclerotiorum and with the same isolate nutritionally conditioned to produce small amounts of oxalic acid. The preconditioned isolate behaved as hypovirulent. Tomato plants were inoculated with four S. sclerotiorum isolates of increasing virulence. A close correlation among disease severity, accumulation of oxalic acid, decrease in pH and inhibition of polyphenoloxidase in both infected host tissues was demonstrated. Oxalic acid production as an important factor of virulence in S. sclerotiorum is emphasized and its effect on the phenolic metabolism of the host via inhibition of polyphenoloxidase is suggested.     

A Novel Mycovirus That Is Related to the Human Pathogen Hepatitis E Virus and Rubi-Like Viruses

Previously, we reported that three double-stranded RNA (dsRNA) segments, designated L-, M-, and S-dsRNAs, were detected in Sclerotinia sclerotiorum strain Ep-1PN. Of these, the M-dsRNA segment was derived from the genomic RNA of a potexvirus-like positive-strand RNA virus, Sclerotinia sclerotiorum debilitation-associated RNA virus. Here, we present the complete nucleotide sequence of the L-dsRNA, which is 6,043 nucleotides in length, excluding the poly(A) tail. Sequence analysis revealed the presence of a single open reading frame (nucleotide positions 42 to 5936) that encodes a protein with significant similarity to the replicases of the “alphavirus-like” supergroup of positive-strand RNA viruses. A sequence comparison of the L-dsRNA-encoded putative replicase protein containing conserved methyltransferase, helicase, and RNA-dependent RNA polymerase motifs showed that it has significant sequence similarity to the replicase of Hepatitis E virus, a virus infecting humans. Furthermore, we present convincing evidence that the virus-like L-dsRNA could replicate independently with only a slight impact on growth and virulence of its host. Our results suggest that the L-dsRNA from strain Ep-1PN is derived from the genomic RNA of a positive-strand RNA virus, which we named Sclerotinia sclerotiorum RNA virus L (SsRV-L). As far as we know, this is the first report of a positive-strand RNA mycovirus that is related to a human virus. Phylogenetic and sequence analyses of the conserved motifs of the RNA replicase of SsRV-L showed that it clustered with the rubi-like viruses and that it is related to the plant clostero-, beny- and tobamoviruses and to the insect omegatetraviruses. Considering the fact that these related alphavirus-like positive-strand RNA viruses infect a wide variety of organisms, these findings suggest that the ancestral positive-strand RNA viruses might be of ancient origin and/or they might have radiated horizontally among vertebrates, insects, plants, and fungi.     

1,8-Dihydroxynaphthalene monoglucoside,a new metabolite of Sclerotinia sclerotiorum,and the effect of tricyclazole on its production

Isolate SS7 of Sclerotinia sclerotiorum was previously shown to produce and excrete into agar medium copious amounts of the melanin precursor 1,8-dihydroxynaphthalene. Much reduced quantities of this product were produced in the presence of tricyclazole, an inhibitor of pentaketide melanin biosynthesis. In this study, we demonstrate that young cultures of isolate SS7 produce 1,8-dihydroxynaphthalene monoglucoside, a new natural product not previously reported from fungi. When cultured in the presence of tricyclazole, such young cultures also accumulated two new monoglucosides of 1,3,8-trihydroxynaphthalene, which, as well as 1,8-dihydroxynaphthalene monoglucoside, were also obtained from cultures of two other isolates of S. sclerotiorum. It is proposed that rapid glucosylation of 1,3,8-trihydroxynaphthalene in young tricyclazole-inhibited S. sclerotiorum cultures accounts for the failure to observe 2-hydroxyjuglone or other metabolites usually associated with blockage of the pentaketide pathway to melanin in fungi.     

Pathogenicity of Sclerotinia sclerotiorum on Ranunculus acris in Dairy Pasture

Fifty-four isolates of Sclerotinia sclerotiorum from Ranunculus acris and other natural hosts were applied as mycelial infested kibbled wheat onto 6 month-old R. acris plants in two glasshouse screening experiments. Most isolates (90%) did not differ in their pathogenicity towards R. acris. One isolate, S. sclerotiorum G45, was selected based on its ability to cause severe disease and suppress regeneration of R. acris. A field experiment was conducted to determine the efficacy of S. sclerotiorum (G45) against R. acris in infested dairy pastures in the Takaka Valley, Golden Bay, New Zealand. Isolate G45 was formulated as a wettable powder and was applied as a slurry at 20 and 40 ml/plant in December 1995. After 10 weeks, regeneration from the crown of treated plants was apparent and a second application of S. sclerotiorum was made in February 1996. Best control of R. acris was obtained when the plants were inoculated in full flower in December. At the first time of treatment, the 40 ml application of S. sclerotiorum slurry reduced the total dry weight of R. acris by an average of 57%. The second application had no effect on total dry weight, possibly because moisture levels were not sufficient for S. sclerotiorum infection. This study confirmed S. sclerotiorum to be an aggressive pathogen of R. acris under both glasshouse and field conditions. As a result, this pathogen has potential as a mycoherbicide for R. acris. Further experiments are required to explore ways of enhancing the efficacy of S. sclerotiorum against R. acris by manipulation of the host, pathogen and environment.     

Cyclosporine A from a nonpathogenic Fusarium oxysporum suppressing Sclerotinia sclerotiorum

AIMS: To evaluate the antagonistic activity of Fusarium oxysporum nonpathogenic fungal strain S6 against the phytopathogenic fungus Sclerotinia sclerotiorum and to identify the antifungal compounds involved. METHODS AND RESULTS: The antagonistic activity of Fusarium oxysporum strain S6 was determined in vitro by dual cultures. The metabolite responsible for the activity was isolated by chromatographic techniques, purified and identified by spectroscopic methods as cyclosporine A. The antifungal activity against the pathogen was correlated with the presence of this metabolite by a dilution assay and then quantified. Cyclosporine A caused both growth inhibition and suppression of sclerotia formation. In a greenhouse assay, a significant increase in the number of surviving soybean (Glycine max) plants was observed when S. sclerotiorum and F. oxysporum (S6) were inoculated together when compared with plants inoculated with S. sclerotiorum alone. CONCLUSION: Fusarium oxysporum (S6) may be a good fungal biological control agent for S. sclerotiorum and cyclosporine A is the responsible metabolite involved in its antagonistic activity in vitro. SIGNIFICANCE AND IMPACT OF THE STUDY: Cyclosporine A has not been previously described as an inhibitor of S. sclerotiorum. Its minimum inhibitory concentration (MIC) of 0.1 microg disc(-1) makes it suitable to use as a biofungicide. In vivo experiments showed that F. oxysporum (S6) is a good candidate for the biocontrol of S. sclerotiorum in soybean.