稻瘟病菌生理小种酯酶同工酶特性的研究

 运用聚丙烯酰胺凝胶等电聚焦法对来自江、浙两省经全国统一鉴别寄主鉴定的稻瘟病菌7群12个生理小种的16个菌株菌体酯酶同工酶进行了分析比较,结果表明:稻瘟菌菌体酯酶同工酶带有13-18条,等电点分布范围为3.00-7.20。在等电点4.15附近有各生理小种共同的酶带。同一生理小种群内不同生理小种间酯酶同工酶的相似系数一般大于0.50,不同生理小种群的生理小种间酶谱的相似系数一般为0.30-0.40,也存在少数不是同一生理小种群的小种间酶谱相似系数大于0.50的情况。供试各生理小种间酶谱均存在不同程度的差异,表现为酶带数目的多少和等电点分布的不同。来自不同省份经鉴别寄主鉴定为同一生理小种的菌株间酶谱的相似系数亦仅0.05-0.60。研究结果初步反映出供试大多数稻瘟菌菌株间菌体酯酶同工酶的相似性与致病性有关,但也受菌株来源地生态环境影响。本文还讨论了稻瘟病菌致病性容易变异但在许多情况下又具有相对稳定性的可能机制,探讨在一定范围内聚丙烯酰胺凝胶等电聚焦技术用于稻瘟菌生理分化和致病性变异监测的可能性及尚待进一步研究解决的穗题等。     

黑龙江省稻瘟病菌生理小种的鉴定

应用３个籼稻、４个粳稻组成的中国鉴别品种将黑龙江省１１０个稻瘟病菌菌株划分为４群７个生理小种。用日本清泽的１２个单基因抗性鉴别品种将上述菌株划分为７７个生理小种。对＋、ｔｉ－ａ、Ｐｉ－ｋ、Ｐｉ－ｚ基因致病频率高（７０．４％～８９％）的生理小种遍布全省各稻区；对Ｐｉ－ｔ、Ｐｉ－ｂ、Ｐｉ－ｚｔ基因致病频率低（０～１９．３％）的生理小种分布窄，这３个基因可用于抗病育种。试验表明应用日本清泽的１２个粳稻单基因抗性鉴别品种更适宜黑龙江省稻瘟病菌生理小种的鉴定     

稻瘟病菌生理小种菌体可溶性蛋白特性初探

 采用聚丙烯酰胺凝胶等电聚焦法对来自江苏、浙江两省经全国统一鉴别寄主鉴定的稻瘟病菌7个生理小种群、12个生理小种的16个菌株菌体可溶性蛋白进行了分析比较,结果表明:供试菌株菌体可溶性蛋白电泳谱带有37~43条,谱带等电点分布范围为3.00~8.15。除个别菌株外,同一生理小种群内各生理小种问菌体可溶性蛋白电泳图谱的相似系数大于0.70,不同生理小种群的生理小种间菌体可溶性蛋白电泳图谱的相似系数一般为0.50~0.60。供试各生理小种群、生理小种乃至同一生理小种的不同菌株间,其可溶性蛋白电泳图谱存在不同程度的差异,表现为谱带数目的多少和部分谱带等电点(谱带位置)分布的不同。     

稻纹枯菌酯酶同工酶、可溶性蛋白及致病力的研究初报

 对稻纹枯菌25个菌株分别进行了致病力测定、酯酶同工酶和可溶性蛋白分析.结果表明:不同菌株存在明显的致病力分化,而酯酶同工酶图谱却有较强的一致性,它们的主酶带基本相同,但在副酶带上反映出一定的同工酶表型异质性,表现为带的数目及着色深浅的不同;可溶性蛋白图谱比酯酶同工酶图谱表现出更多的多样性,大多数致病力较强的菌株都有l~2条特征谱带,而且它们的谱带数目比弱毒力菌株的多出3~5条.     

中国稻瘟病菌的遗传多样性

 用美国普度大学M.Levy实验室提供的一个散布的重复序列探针(MGR-cPB586),对中国稻区田间分离的13个稻瘟病菌株DNA的EcoRI限制性片段长度多态性进行了研究。结果表明:稻瘟病菌不同生理小种间的RFLP普遍存在,MGR-DNA指纹能清楚地区分中国稻瘟病菌的主要生理小种,并能反映出同一生理小种内复杂的亲缘关系。     

我国水稻当家品种对稻瘟病、白叶枯病的兼抗反应初探

用多个致病力不同的白叶枯病菌株,和全国10省12个研究单位提供的稻瘟病混合菌以及10个生产上常见的生理小种,于1987、1988年对全国种植面积达10万亩以上的145个水稻品种进行了抗白叶枯病谱、抗稻瘟病混合菌和生理小种测定。结果表明国际稻26、金早6号、金早7号、扬稻1号(BG910)、青华矮6号等21个品种能抗多个白叶枯病菌株和抗多个稻瘟病小种混合菌。对10个生理小种的抗谱率相差较大。     

广东稻瘟病菌的遗传宗谱与致病性的关系

不同病圃同一套寄虫主的田间表现及其穗颈瘟分离菌株的遗传宗谱和致病性分析试验结果表明,不同生态稻区稻瘟病菌的小种分布类型不同,对品种的致病性也存在差异。将已经过ＲＦＬＰ和ＰＣＲ技术进行ＤＮＡ指纹分析,并划分为不同遗传宗谱的稻瘟病菌株进行致病性测定。结果表明,菌株的致病性与其遗传宗谱类型、生理小种类型、寄主及采集地点存在密切的关系：（１）同一遗传宗谱、分离自相同寄主、鉴定出来的生理小种类型又较接近的菌     

黑龙江省不同来源稻瘟病菌致病性分析

将从黑龙江‘龙粳17’、‘吉特639’和‘垦稻10’病田中分离的9、8、8个稻瘟病菌菌株喷雾接种到4套鉴别寄主上,结果表明,中国稻瘟病菌生理小种鉴别寄主将这3个来源的菌株分别划分成8、7和4个生理小种,日本清泽鉴别品种则将其分别划分成9、8和8个生理小种,以黑龙江本地水稻品种作为鉴别品种,则可将其分别划分成8、7和5个致病型,以24个水稻抗稻瘟病单基因系为鉴别品种,则将上述菌株分别划分为9、7和8个致病型。     

玉米弯孢叶斑病菌致病类型分化研究

 利用鉴别寄主和同工酶技术,对采自黑龙江、吉林、辽宁和河北4省玉米弯孢叶斑病菌菌株的致病类型进行分析。研究表明,鉴别寄主技术将26个菌株的致病类型划分为4种类型,其中强和中等致病类型主要分布在辽宁省瓦房店市、吉林省公主岭市和河北省保定市。玉米弯孢叶斑病菌在酯酶、苹果酸脱氢酶、过氧化物酶、多酚氧化酶、超氧化物歧化酶同工酶谱存在差异,不同菌株间的某些同工酶谱带数差异显著。     

黑龙江省稻瘟病菌生理小种鉴定与分析

2006年,应用日本清泽鉴别寄主共分离测定了黑龙江省稻瘟病菌单孢菌株178个,结果共划分为104个生理小种。其中77.7号小种为黑龙江省的优势小种,比例为4.49%;677.7、377.7号小种为强毒力小种,能侵染91.67%的鉴别品种,从基因角度来说,Pi-ta2、Pi-zt可在黑龙江省的优势小种区利用,Pi-ks,Pi-t不可在黑龙江省大规模利用,此外,地域较近地区的稻瘟病菌的生理小种群体也有较大的差异。     

辽宁省稻瘟病菌对稻瘟灵的敏感性监测

 利用菌丝生长速率法,测定了自辽宁省主产稻区2009-2010年分离的87株稻瘟病菌对稻瘟灵的敏感性。结果表明,稻瘟灵的抑制中浓度EC₅₀值为0.35~7.01 μg/mL,敏感性差异达20倍;且2009年稻瘟病菌对稻瘟灵的敏感性高于2010年。根据野生敏感型病原群体对药剂敏感性呈正态分布的原理,辽宁省稻瘟病菌对稻瘟灵的敏感性基线定为(1.77±0.80)μg/mL,辽宁省主要稻区均有抗稻瘟灵菌株出现,除2010年在铁岭稻产区监测到1株中抗菌株外,大部分稻瘟病菌株都表现为低抗水平。稻瘟病菌不同生理小种群对稻瘟灵的敏感性无明显差别。     

Evidence of parasexual exchange of DNA in the rice blast fungus challenges its exclusive clonality

ABSTRACT We applied DNA markers to determine whether parasexual recombination may contribute to the extreme genetic diversity and variability observed in Magnaporthe grisea, the causal agent of rice blast disease. Dispersed repetitive elements and mapped, low-copy restriction fragment length polymorphism (RFLP) probes were used to detect transfers of DNA between cultured isolates of M. grisea. Low-copy RFLP probes also were used to detect putative recombinants among isolates from well-characterized field populations of the pathogen. Microscopic examination of tufted mycelium between cocultured isolates revealed frequent hyphal fusions. Hyphal tips and conidia were recovered without selection from tufted zones in two separate vegetative pairings involving isolates with dissimilar haplotypes, based on the repetitive element MGR586. Haplotypic changes were observed at a higher frequency in tuft derivatives than in subcultures of each isolate alone. From 136 tuft derivatives analyzed, 5 putative recombinant haplotypes were identified. Introgression was demonstrated with two independent repetitive elements, fosbury and MGR586, as probes on DNA digested with several restriction enzymes. Introgressions were characterized by addition of 1 to 10 MGR586 bands, and 1 to 3 fosbury bands from one parent into the background of the other. Polymorphic single-copy probes were used to analyze putative recombinants. One probe detected an introgression event as predicted by analysis with MGR586. To assess the possible role of parasexual recombination in field populations of the pathogen, isolates in the Philippines previously grouped based on DNA fingerprinting were analyzed with low-copy RFLP markers. Polymorphism in single-copy loci typically was seen between, but not within, putative pathogen lineages. One lineage (designated lineage 4), however, was polymorphic for several probes. For some isolates, alleles at these loci comigrated with alleles characteristic of other lineages, suggesting the transfer of DNA fragments between lineages. One isolate was apparently a merodiploid, carrying an allele typical of lineage 4 plus another allele characteristic of a different lineage. In a survey of isolates from the Indian Himalayas, a merodiploid also was found with single- or low-copy probes. Examination of MGR586 profiles of the putative recombinant and its putative donor strains showed the expected introgression of MGR586 bands. The detection of parasexual DNA exchanges in wild-type strains under unselected conditions and the existence of merodiploids in nature suggest that parasexual recombination occurs in field populations of M. grisea. This raises questions concerning exclusive clonality in the blast fungus.     

稻瘟病菌生理小种及其毒性

用我国一套稻瘟病菌生理小种鉴别品种对分离自云南省红河州个旧、石屏、建水不同种植模式的68个稻瘟病菌单孢菌株进行生理小种测定,同时以日本8个单基因鉴别品种对供试菌株进行基因分析。测定结果为B₉、B₁₃、B₁₅、B₂₇、B₂₉、B₃₁、C₉、C₁₁、C₁₅、D₁、E₃和G₁12个生理小种,其中粳型小种占60.29%,籼型小种占39.71%。此结果与以往云南省稻瘟病菌生理小种的变化趋势相吻合。供试的68个菌株进行毒性基因分析表明Av-K^s+和Av-i+频率毒性基因较高。     

Pyricularia grisea Isolates Causing Gray Leaf Spot on Perennial Ryegrass (Lolium perenne) in the United States: Relationship to P. grisea Isolates from Other Host Plants

ABSTRACT Gray leaf spot of perennial ryegrass (prg) (Lolium perenne), caused by the fungus Pyricularia grisea (teleomorph = Magnaporthe grisea), has rapidly become the most destructive of all turf grass diseases in the United States. Fungal isolates from infected prg were analyzed with several molecular markers to investigate their relationship to P. grisea strains found on other hosts. All of the molecular markers used in this study revealed that isolates from prg are very distantly related to those found on crabgrass. Fingerprinting with MGR586 (Pot3) revealed zero to three copies of this transposon in the prg pathogens, distinguishing them from isolates pathogenic to rice, which typically have more than 50 copies of this element. RETRO5, a newly identified retroelement in P. grisea, was present at a copy number of >50 in isolates from rice and Setaria spp. but only six to eight copies were found in the isolates from prg. The MAGGY retrotransposon was unevenly distributed in the prg pathogens, with some isolates lacking this element, some possessing six to eight copies, and others having 10 to 30 copies. These results indicated that the P. grisea isolates causing gray leaf spot are distinct from those found on crabgrass, rice, or Setaria spp. This conclusion was supported by an unweighted pair-group method with arithmetic average cluster analysis of single-copy restriction fragment length polymorphism haplo-types. Fingerprints obtained with probes from the Pot2 and MGR583 transposons revealed that the prg pathogens are very closely related to isolates from tall fescue, and that they share similarity with isolates from wheat. However, the wheat pathogens had fewer copies of these elements than those found on prg. Therefore, I conclude that P. grisea isolates commonly found on other host plant species did not cause gray leaf spot epidemics on prg. Instead, the disease appears to be caused by a P. grisea population that is specific to prg and tall fescue.     

中国部分地区稻曲病菌培养特性及其遗传多样性分析

采用生物学方法和RAPD-PCR技术,对来自11个省(市)84个菌株的菌丝生长速率、分生孢子数量、孢子萌发率及其遗传多样性进行分析,以明确中国部分地区稻曲病菌株的培养特性和遗传多样性。依据菌丝生长速率,菌株可被划分为快和慢2种类型,分别占58.33%和41.67%。依据产孢力和分生孢子萌发力,菌株可划分为强、中和弱3种类型。采用12条RAPD引物共扩增出323条带,多态性条带比率为98.14%,遗传距离变化范围为0.02～1.00。在遗传距离0.725水平上,所有菌株被划分成7个遗传聚类组,聚类组R4和R5为优势聚类组,并存在一些亚组。不同地区之间和同一地区内的菌株表现出不同程度的变异,内陆地区的菌株群体变异程度明显高于沿海地区。从采用相同接种体接种的水稻品种上分离的稻曲病菌具有紧密的亲缘关系。     

粤、桂、苏、皖四省稻瘟病菌有性态的研究

应用国际稻瘟病菌标准交配型菌株KA3和THl6对2000年从广东、广西、江苏和安徽4省采集的145个稻瘟病菌菌株的育性和交配型测定结果表明,广东、广西和安徽3省稻瘟病菌群体中存在着Matl.1和Matl.2两种交配型,江苏省的23个可交配菌株均为Matl.1。各省间稻瘟病菌群体的可交配率差异不大,总的可交配率为48.3%。有9个菌株与标准菌株交配产生了子囊孢子,可育率为6.2%。检测到2个两性菌株,不同地区稻瘟病菌群体雌雄性别的比例有显著性差异。用不同交配型的可育菌株进行互交,14个组合中只有1个组合产生子囊壳,但没有形成成熟的子囊孢子。     

西南地区稻瘟病菌对戊唑醇的敏感性基线建立及抗性监测

于2013—2014年分别从重庆市及四川、贵州省的16个区县采集分离获得902株稻瘟病菌,采用菌丝生长速率法测定了其中150个菌株对戊唑醇的敏感性。结果表明:其EC_(50)值分布在0.047 5~0.599 6μg/m L之间,平均EC50值为(0.215 4±0.149 7)μg/m L。菌株的不同敏感性频率呈连续单峰曲线分布,因此其平均EC_(50)值0.215 4μg/m L可作为田间稻瘟病菌对戊唑醇敏感性检测的参考标准。戊唑醇对菌株的最低抑制浓度(MIC)值≤4μg/m L,以4μg/m L作为抗性菌株监测的区分剂量,测定了各地病原菌对戊唑醇的田间抗性,发现自各地采集分离的902株稻瘟病菌多数对戊唑醇仍表现为敏感,仅2株为低水平抗性。