江西稻瘟病菌稻巨座壳致病性分化年度动态变化

为明确江西水稻种植区稻巨座壳菌（稻瘟病病菌）的年际变化规律,本研究采用7个中国鉴别寄主与30个单基因鉴别品系两套鉴别寄主分别鉴定分析了2006-2018年间从江西37个水稻主要种植县市分离的1 161个稻瘟病单孢菌的生理小种、致病力、致病类型与无毒基因型等。研究结果表明,江西稻瘟病菌可以分成7群49个生理小种,其中ZA、ZB、ZC群为优势种群,ZB13为优势小种,出现频率为18.00%,以毒性较强的强致病力菌株为主;江西稻瘟病菌在生理小种构成、优势小种、致病力年际变化方面均具有3-5年的周期性;江西历年稻瘟病菌的致病类型较为丰富且存在年度差异,菌株致病类型占各年度总菌株数的82.79%-98.21%,优势致病型菌株占当年总菌株的3.57%-5.77%;历年稻瘟病菌的无毒基因个数为24-29个,其中Avr-Pizt、Avr-Piz5、Avr-Pik、Avr-Pik(C)在历年供试菌株中出现频率较高,说明与之相对应的抗瘟基因在江西抗病育种与抗性品种布局中具有较好的应用价值。     

稻瘟菌无毒基因研究进展

无毒基因编码的产物激发病原物与植物特异性相互作用。水稻与稻瘟菌之间的特异互作符合“基因对基因”关系。从研究稻瘟菌无毒基因的意义、已鉴定和克隆的稻瘟菌无毒基因、稻瘟菌无毒基因与其抗病基因的互作特点等几个方面,对稻瘟菌无毒基因研究进展作了简要评述 。     

致病疫霉超级生理小种遗传多样性分析

通过交配型和甲霜灵抗性以及线粒体DNA单倍型、SSR和AFLP基因型分析对40个超级生理小种菌株进行了遗传多样性分析。在被测菌株中发现了A1、A2和自育3种不同类型的交配型。其中,A1和自育型菌株数量多,分别为21株和14株,而A2交配型仅5株。甲霜灵抗性测定检测出高抗菌株26株,敏感菌株14株。线粒体DNA单倍型测定出Ia型和IIa型两种,比例接近1:1。基于5个基因座被测40个超级生理小种菌株共鉴定出了7种SSR基因型。利用6对荧光引物共检测到258条AFLP谱带,其中多态性谱带204条,多态性为79.1%。将供试的40个菌株划分为38个基因型,几乎每个菌株都为1个特有基因型。而且,我国南方和北方超级生理小种群体存在着明显的遗传差异。结果表明我国致病疫霉超级生理小种具有丰富的遗传多样性,可以推断致病疫霉中的任何小种都可在多个抗病基因的强大选择压力下,在短时间内通过与之对应的无毒基因快速突变而成为超级生理小种。当前对致病疫霉生理小种的鉴定及监测对生产上利用抗病品种防控晚疫病的指导意义不大。     

湖南桃江病圃稻瘟病菌的无毒基因及水稻抗瘟单基因联合抗性分析

【目的】鉴定湖南省桃江病圃稻瘟病菌无毒基因型,为合理搭配种植湖南省水稻抗瘟品种和抗病育种提供依据。【方法】在湖南桃江病圃采集水稻品种"丽江新团黑谷"(LTH)稻瘟菌病样,用单孢分离法分离稻瘟病菌单孢并纯化获得单孢菌株,用针刺离体法将菌株接种到以"LTH"为轮回亲本培育而成的24个含单抗瘟基因的水稻5叶期第5叶片上,对供试菌株进行无毒基因鉴定,并应用联合致病性系数和联合抗病性系数分析抗瘟基因组合间的互作。【结果】供试92个稻瘟病单孢菌株含有全部的24个无毒基因,对24个已知含单抗瘟基因的水稻材料表现出不同程度的毒力水平,含水稻抗瘟基因Pi-20对供试菌株抗菌频率最高,达54.35%;通过联合致病性系数和联合抗病性系数分析抗瘟基因组合间的互作,结果表明最佳搭配组合为Pi-20×Pi-k~s(RAC=0.28,PAC=0.23)。【结论】湖南省桃江病圃稻瘟病菌致病力较强,24个抗瘟基因多已感病化,含抗性基因Pi-20与Pi-k、Pi-k~s、Pi-3组合的水稻品种目前可在湖南省推广利用,但需研究引进新的抗瘟基因。     

基于多基因序列分析对尖孢镰孢菌古巴专化型(香蕉枯萎病菌)生理小种的鉴定

由尖孢镰孢菌古巴专化型Fusarium oxysporum f.sp.cubense,Foc引起的香蕉枯萎病是香蕉生产上的毁灭性病害,自1996年以来已对我国华南地区香蕉生产造成了严重危害。传统上香蕉枯萎病菌生理小种的鉴定主要采用人工接种鉴别寄主尔后测定病菌致病性的方法,但实验周期长,且受季节影响。以来自澳大利亚的香蕉枯萎病菌生理小种1号(BW1)、2号(Race 2)、3号(Race 3)以及亚热带4号(BW4)为对照,对分离自我国华南地区主要香蕉产区(广东、广西、海南、福建等省区)的14株香蕉枯萎病菌的单孢菌株进行致病性测定,并结合热带4号小种(TR4)和亚热带4号小种(ST4)的分子特异检测方法,确定其生理小种类型;同时,利用ITS、TEF‐1α、IGS、histone H3、β‐tubulin等5个主要用于镰孢菌系统发育学研究的基因,研究不同地区不同来源的Foc菌株之间的亲缘关系及其与非病原尖孢镰孢菌的关系,并评价这5个基因在香蕉枯萎病菌生理小种鉴定上的应用价值。研究结果表明:(1)来源于我国华南地区的4号小种主要为热带4号小种;(2)TEF‐1α、IGS、histone H3等3个基因片段能够将Foc中不同生理小种的菌株划分成不同的系统发育谱系,与致病性测定的结果具有对应关系,也能较好地反映尖孢镰孢菌种内菌株的亲缘关系,可用于香蕉枯萎病菌生理小种鉴定;(3)我国Foc 1号生理小种的遗传多样性高于4号生理小种,Foc 1号生理小种的菌系与来自香蕉果实上的非病原尖孢镰孢菌的亲缘关系比其与Foc 4号生理小种的菌系的亲缘关系更近。     

稻瘟菌无毒基因研究进展

稻瘟菌是引起水稻稻瘟病的病原物。水稻与稻瘟菌间存在广泛而特异的相互作用,是研究寄主与病原物互作的重要模式系统。本文对稻瘟菌与水稻互作最重要的激发子―无毒基因的研究现状进行了概括,讨论了无毒基因的定位、克隆方法以及已克隆无毒基因的功能及进化研究,同时对今后无毒基因研究的重要方向进行了探讨,为深入理解无毒基因的功能及与水稻可能的互作关系奠定了基础。     

2012–2013年中国禾柄锈菌小麦专化型小种动态及新毒力谱分析

监测具有潜在威胁的小麦秆锈病病原菌小种动态并分析其毒力谱变化,是培育抗病品种不可或缺的基础工作。在小麦秆锈病鲜有发生的情况下,于2012–2013年获得小麦田间秆锈病标样11份和经有性生殖过程的禾柄锈菌小麦专化型菌样22份。上述菌样经过分离、纯化,得到了53个单孢子堆菌株。利用国内最新采用的禾柄锈菌小麦专化型小种与毒力鉴别寄主体系进行分析,鉴定出13个生理小种,其中34C3RTGQM和34Oro II-MRGQM为优势小种,出现频率均为13.2%,首次发现了对Sr5+Sr11具有联合毒力的6个新小种,其出现频率处在1.9%–13.2%范围内。同时测定出Sr9e、Sr26、Sr31、Sr33、Sr37、Sr38、Sr47和Sr Tt3等8个抗秆锈病单基因对全部供试菌株表现抗病,余下40个单基因系则分别表现对1个以上至全部菌株感病。结论:(1)首次报道了6个对Sr5+Sr11具有危险性的联合毒力谱的新小种类型,这类毒力类型应当被密切关注;(2)明确了当前完全有效的8个抗秆锈病基因和其他部分有效的抗病基因;(3)初步说明了有性循环菌株与无性循环菌株在毒力结构上存在差异。     

四川省马铃薯晚疫病菌群体表型和遗传变异的分析

致病疫霉Phytophthora infestans引起的晚疫病是马铃薯的一种毁灭性病害。为了对四川省近年马铃薯晚疫病菌进行系统多样性分析,本研究从表现型和基因型两个方面鉴定了四川省马铃薯晚疫病菌的群体多态性。交配型、甲霜灵敏感性和生理小种的鉴定结果发现A1交配型菌株只有2份,A2型有29份,自育型12份;甲霜灵敏感菌株3株,中抗菌株22株,高抗菌株18株;共测定出11个生理小种,其中全毒力小种发生频率最高,占供试菌株的25.58%。基因型鉴定结果表明,43份材料共发现了10种SSR基因型,5个SSR标记在该种群上共产生了16个SSR位点,每个标记平均有3.2个位点,多态性信息含量平均值为0.46,SSR4是多态性最丰富的标记。本试验中A2菌株和自育型的基因型SSR相同,其中全毒力小种是当地的优势致病菌,该研究为今后四川马铃薯晚疫病的有效防控提供理论依据。     

海南省香蕉枯萎菌生理小种的RAPD分析

利用随机扩增多态性DNA(RAPD)分子标记方法对海南省香蕉枯萎病菌2个生理小种(小种1和小种4)进行遗传多样性分析,以筛选出的15个随机引物对采自海南省各市县发病蕉区的分别属于1号生理小种和4号生理小种的16个代表菌株及广东省2个1号和4号生理小种对照菌株进行RAPD-PCR扩增,结果产生97个RAPD分子标记,其中多态性的条带有76条,通过聚类分析探讨了供试小种间的亲缘关系,并寻找到了1、4号生理小种的特异性条带,为在分子水平上进行香蕉枯萎病菌生理小种鉴定提供更为便利的手段。     

水稻广谱抗稻瘟病基因研究进展

稻瘟病是水稻生产中的最严重病害之一,由于稻瘟菌小种的高度变异性,垂直抗性基因难以持续控制稻瘟病的危害,因此,克隆和利用广谱持久抗瘟基因被认为是解决稻瘟病问题最经济有效的策略。本文从广谱抗源的筛选与利用,广谱抗瘟基因的定位、克隆与应用等方面对水稻广谱抗稻瘟病基因研究取得的进展进行了概述,并介绍了广谱抗性分子机理的最新研究进展。基于国内外稻瘟病抗性基因研究的现状及趋势,以及我国丰富的抗瘟水稻种质资源,克隆越来越多的广谱抗瘟基因具有重要的理论与应用价值。     

植物病原物无毒基因及其功能

植物抗病基因与病原物无毒基因产物间直接或间接相互作用导致产生的基因对基因抗性是植物抗病性的重要形式。无毒基因已在多种植物病原物 ,包括真菌、细菌、病毒和卵菌等中得到克隆。绝大多数已克隆无毒基因之间 ,及其与已知蛋白之间 ,均无显著序列同源性。然而 ,多数已克隆植物抗病基因有较高序列一致性 ,产物往往具有相似的结构域。由序列一致性很高的抗病基因产物与没有明显序列同源性的无毒基因产物相互作用 ,介导产生的过敏性细胞坏死和抗病性 ,在产生速度、强度和组织特异性等方面均可能有显著差异。无毒基因具有双重功能 :在含互补抗病基因植物中表现无毒效应 ,而在不含互补抗病基因植物中显示小种、菌株、致病型、或种特异性毒性效应     

Fungal Avirulence Genes: Structure and Possible Functions

Avirulence (Avr) genes exist in many fungi that share a gene-for-gene relationship with their host plant. They represent unique genetic determinants that prevent fungi from causing disease on plants that possess matching resistance (R) genes. Interaction between elicitors (primary or secondary products ofAvrgenes) and host receptors in resistant plants causes induction of various defense responses often involving a hypersensitive response.Avrgenes have been successfully isolated by reverse genetics and positional cloning. Five cultivar-specificAvrgenes (Avr4,Avr9, andEcp2 fromCladosporium fulvum; nip1fromRhynchosporium secalis;andAvr2-YAMOfromMagnaporthe grisea) and three species-specificAvrgenes (PWL1andPWL2fromM. griseaandinf1fromPhytophthora infestans) have been cloned. Isolation of additionalAvrgenes from these fungi, but also from other fungi such asUromyces vignae,Melampsora lini, Phytophthora sojae,andLeptosphaeria maculans,is in progress. Molecular analyses of nonfunctionalAvrgene alleles show that these originate from deletions or mutations in the open reading frame or the promoter sequence of anAvrgene. Although intrinsic biological functions of mostAvrgene products are still unknown, recent studies have shown that twoAvrgenes,nip1andEcp2, encode products that are important pathogenicity factors. All fungalAvrgenes cloned so far have been demonstrated or predicted to encode extracellular proteins. Current studies focus on unraveling the mechanisms of perception of avirulence factors by plant receptors. The exploitation ofAvrgenes and the matchingRgenes in engineered resistance is also discussed.     

水稻稻瘟病抗性基因的克隆、育种利用及稻瘟菌无毒基因研究进展

稻瘟病是世界上影响水稻(Oryza sativa)粮食生产的主要病害之一, 抗病基因的发掘与利用是抗病育种的基础和核心。随着寄主水稻和病原菌稻瘟病菌(Magnaporthe oryzae)基因组测序和基因注释的完成, 水稻和稻瘟病菌的互作体系成为研究植物与真菌互作的模式系统。该文对稻瘟病抗病基因的遗传、定位、克隆及育种利用进行概述, 并通过生物信息学分析方法, 探讨了水稻全基因组中NBS-LRR类抗病基因在水稻12条染色体上的分布情况, 同时对稻瘟病菌无毒基因的鉴定及无毒蛋白与抗病蛋白的互作进行初步分析。最后对稻瘟病抗病基因研究存在的问题进行分析并展望了未来的研究方向, 以期为水稻抗稻瘟病育种发展和抗病机制的深入理解提供参考。     

Association Genetics Reveals Three Novel Avirulence Genes from the Rice Blast Fungal Pathogen Magnaporthe oryzae

To subvert rice (Oryza sativa) host defenses, the devastating ascomycete fungus pathogen Magnaporthe oryzae produces a battery of effector molecules, including some with avirulence (AVR) activity, which are recognized by host resistance (R) proteins resulting in rapid and effective activation of innate immunity. To isolate novel avirulence genes from M. oryzae, we examined DNA polymorphisms of secreted protein genes predicted from the genome sequence of isolate 70-15 and looked for an association with AVR activity. This large-scale study found significantly more presence/absence polymorphisms than nucleotide polymorphisms among 1032 putative secreted protein genes. Nucleotide diversity of M. oryzae among 46 isolates of a worldwide collection was extremely low (θ = 8.2 × 10⁻⁵), suggestive of recent pathogen dispersal. However, no association between DNA polymorphism and AVR was identified. Therefore, we used genome resequencing of Ina168, an M. oryzae isolate that contains nine AVR genes. Remarkably, a total of 1.68 Mb regions, comprising 316 candidate effector genes, were present in Ina168 but absent in the assembled sequence of isolate 70-15. Association analyses of these 316 genes revealed three novel AVR genes, AVR-Pia, AVR-Pii, and AVR-Pik/km/kp, corresponding to five previously known AVR genes, whose products are recognized inside rice cells possessing the cognate R genes. AVR-Pia and AVR-Pii have evolved by gene gain/loss processes, whereas AVR-Pik/km/kp has evolved by nucleotide substitutions and gene gain/loss.     

Novel Mutations Detected in Avirulence Genes Overcoming Tomato Cf Resistance Genes in Isolates of a Japanese Population of Cladosporium fulvum

Leaf mold of tomato is caused by the biotrophic fungus Cladosporium fulvum which complies with the gene-for-gene system. The disease was first reported in Japan in the 1920s and has since been frequently observed. Initially only race 0 isolates were reported, but since the consecutive introduction of resistance genes Cf-2, Cf-4, Cf-5 and Cf-9 new races have evolved. Here we first determined the virulence spectrum of 133 C. fulvum isolates collected from 22 prefectures in Japan, and subsequently sequenced the avirulence (Avr) genes Avr2, Avr4, Avr4E, Avr5 and Avr9 to determine the molecular basis of overcoming Cf genes. Twelve races of C. fulvum with a different virulence spectrum were identified, of which races 9, 2.9, 4.9, 4.5.9 and 4.9.11 occur only in Japan. The Avr genes in many of these races contain unique mutations not observed in races identified elsewhere in the world including (i) frameshift mutations and (ii) transposon insertions in Avr2, (iii) point mutations in Avr4 and Avr4E, and (iv) deletions of Avr4E, Avr5 and Avr9. New races have developed by selection pressure imposed by consecutive introductions of Cf-2, Cf-4, Cf-5 and Cf-9 genes in commercially grown tomato cultivars. Our study shows that molecular variations to adapt to different Cf genes in an isolated C. fulvum population in Japan are novel but overall follow similar patterns as those observed in populations from other parts of the world. Implications for breeding of more durable C. fulvum resistant varieties are discussed.     

Soybean Resistance Genes Specific for Different Pseudomonas Syringae Avirulence Genes Are Allelic, or Closely Linked, at the Rpg1 Locus

RPG1 and RPM1 are disease resistance genes in soybean and Arabidopsis, respectively, that confer resistance to Pseudomonas syringae strains expressing the avirulence gene avrB. RPM1 has recently been demonstrated to have a second specificity, also conferring resistance to P. syringae strains expressing avrRpm1. Here we show that alleles, or closely linked genes, exist at the RPG1 locus in soybean that are specific for either avrB or avrRpm1 and thus can distinguish between these two avirulence genes.     

Avirulence proteins from haustoria-forming pathogens

A major insight that has emerged in the study of haustoria-forming plant pathogens over the last few years is that these eukaryotic biotrophs deliver suites of secreted proteins into host cells during infection. This insight has largely derived from successful efforts to identify avirulence (Avr) genes and their products from these pathogens. These Avr genes, identified from a rust and a powdery mildew fungus and three oomycete species, encode small proteins that are recognized by resistance proteins in the host plant cytoplasm, suggesting that they are transported inside plant cells during infection. These Avr proteins probably represent examples of fungal and oomycete effector proteins with important roles in subverting host cell biology during infection. In this respect, they represent a new opportunity to understand the basis of disease caused by these biotrophic pathogens. Elucidating how these pathogen proteins gain entry into plant cells and their biological function will be key questions for future research.