江汉平原水稻白叶枯病菌致病型的变异研究

 从水稻和稻田周围常见的10种杂草上来集梨孢属菌株,以涂沫法作交互接种,分离自牛筋草、狗尾草、法氏狗尾草、李氏禾、罗氏草、铺地黍、稗和马唐的梨孢菌与分离自水稻的梨孢菌可以交叉侵染其寄主,茭白和碎米莎草梨孢菌与稻梨孢菌不能交互侵染;自然诱发条件下来自牛筋草和狗尾草的梨孢菌与来自水稻的梨孢菌对其寄主同样发生交叉侵染。直接从杂草寄主上分离的梨孢菌株对水稻的致病性较弱,接种到水稻感病品种发病后再分离的菌株,对水稻致病性增强,对原寄主的致病性基本保持不变。推断稻瘟病区稻田周围牛筋草、狗尾草、法氏狗尾草、李氏禾、罗氏草、铺地黍、稗和马唐等杂草寄主上的梨孢菌在稻瘟病菌的积累和传播方面具有作用。     

芽胞杆菌CQBS03抑菌蛋白TasA基因的克隆及原核表达

为了探讨柑桔溃疡病生防菌芽胞杆菌Bacillus CQBS03菌株TasA基因的功能,采用PCR方法从CQBS03基因组DNA中扩增出编码TasA基因的全长DNA序列,并构建pEASY-E1/TasA原核表达载体,经大肠杆菌Escherichia coli表达获得TasA基因的融合表达蛋白,纸碟法检验融合蛋白对柑桔溃疡病菌Xanthomonas citri citri的抑制作用。结果显示,CQBS03菌株的TasA基因包含1个786 bp的完整开放阅读框（GenBank登录号为JQ309841）,编码261个氨基酸残基;该序列与来源于解淀粉芽胞杆菌B.amyloliquefaciens的1个已知同源TasA基因序列FJ713580的相似性达99.75%。原核表达产物经SDS-PAGE分析,检测到约31 kD的融合蛋白;纯化后的融合蛋白对柑桔溃疡病菌有明显的抑制作用,72 h后抑菌圈直径达11.5 mm。研究表明TasA基因是生防菌芽胞杆菌CQBS03抑制柑桔溃疡病菌的功能基因之一,并且该基因对原核表达宿主没有抑制作用,具有较好的开发利用前景。     

用免疫分离法检测稻种上的白叶枯病菌

 免疫分离法是一种新型的、高度灵敏和准确的种菌带菌检验法,它结合了利用专化性抗血清的选择法吸附作用和活细菌可形成菌落的两方面的优点,从种苗材料中分离到目标细菌,用纯菌测试它的回收率为50~70%,最低检出率为50~100个菌/ml,人工喷接种和病田收获的病种带菌率达87%,平均每粒自然病种上的细菌数在100个左右。免疫分离所得菌落经致病性测定,大多数为白叶枯菌。     

Genetical relationships between reactions to bacterial leaf spot,yellow mosaic and Cercospora leaf spot diseases in mungbean (Vigna radiata)

Summary Studies on the inheritance pattern of bacterial leaf spot (BLS), yellow mosaic (YM) and Cercospora leaf spot (CLS) reactions in crosses of BLS and YM resistant/tolerant but CLS susceptible × CLS resistant but BLS and YM susceptible parents indicated that resistances to BLS and CLS were governed by single dominant genes, whereas YM tolerance was a monogenic recessive character. The studies also indicated that these three genes were inherited independently. The simple inheritance pattern and independent assortment of the genes governing resistance/tolerance to these diseases suggest that the usual breeding methods will be adequate to develop multi-disease resistant mungbean cultivars.Paper XII in the series Studies on resistance in crops to bacterial diseases in India.     

Molecular characterization of the incitant of cowpea bacterial blight and pustule, Xanthomonas campestris pv. vignicola

Strains of Xanthomonas campestris pv. vignicola (Xcv), isolated from cowpea leaves with blight or minute pustules and collected from various geographic areas, were selected on the basis of pathological and physiological features. All strains were analyzed for genotypic markers by two methods: ribotyping with EcoRI endonuclease, and RFLP analysis with a plasmid probe (pthB) containing a gene required for pathogenicity from Xanthomonas campestris pv. manihotis. Ribotyping revealed a unique pattern for all the strains that corresponded to the previously described ribotype rRNA7. Based on polymorphism detected by pthB among Xcv strains, nine haplotypes were defined. The observed genetic variation was independent of the geographic origin of the strains and of pathogenic variation. Some haplotypes were widely distributed, whereas others were localized. In some cases, we could differentiate strains isolated from blight symptoms and pustules according to haplotypic composition. However, in most cases, no significant differences were observed. Our results and the previous pathogenic and biochemical characterizations suggest that the strains isolated from leaves with blight symptoms or minute pustules belong to the same pathovar. We provide information on pathogen diversity that can be used to identify and characterize resistant germplasm.     

水稻白叶枯病菌鞭毛基体基因fliExoo缺失突变分析

 为了阐明水稻白叶枯病菌(Xanthomonas oryzae pv. oryzae,简称Xoo)鞭毛基体组分蛋白基因fliExoo的生物学功能,本研究用Gm^R抗性基因标记交换法成功构建了基因缺失突变体△fliExoo。与野生型菌株PXO99^A相比,△fliExoo鞭毛缺失,菌体易沉降,在0.3%半固体培养基上运动能力明显降低;尽管生长速率和胞外纤维素酶活性无明显变化,但胞外多糖产生和生物膜形成能力明显下降;对水稻品种日本晴的致病性和对非寄主烟草的致敏性反应明显减弱。基因互补可以使上述突变表型恢复。因此,鞭毛基因fliExoo突变不仅影响了细菌鞭毛依赖的运动性,而且对病原菌毒性相关的表型也产生了影响。     

甘蔗白条病及其致病菌Xanthomonas albilineans研究进展

由白条黄单胞杆菌Xanthomonas albilineans引起的甘蔗白条病是一种寄生在植物维管组织的系统性细菌病害,在全球多数种植甘蔗的国家或地区普遍发生,对甘蔗产业的发展构成潜在威胁。本研究综述了甘蔗白条病的发生和分布、传播与流行规律以及病原菌生物学与基因组特性、鉴定与检测、遗传多样性和致病机理等方面内容;提出加强抗病种质挖掘与新品种选育推广、加快抗病分子育种进程、切断病害传播途径、加强隔离检验检疫等病害防控策略;此外,评估了该病害在我国蔗区流行暴发的风险,展望今后甘蔗抗病分子育种、病原菌致病机制及其与寄主互作机理等方面的分子基础研究重点与应用前景。     

水稻品种与水稻细菌性条斑病菌的互作机制

水稻叶片接种3d后，水稻细菌性条斑病菌开始在叶片上大量增殖；接种10d后，病菌数量开始保持相对的稳定，但感病品种上的菌量比抗病品种上的菌量在10倍左右，病斑在接种后第3d开始出现，接种10d以后，病斑在感病品种上能继续快速发展，而在抗病品种上开始受到抑制，通过对抗病和感病品种叶片上的气孔观测发现，感病品种的气孔密度和气孔长度一般都较抗病品种大，相关分析表明，气孔长度理发师品种抗性相关。接种后对PAL酶的活性测定结果表明，PAL酶的活性与品种抗性呈负相关关系。     

Multilocus sequence typing analysis of Italian Xanthomonas campestris pv. campestris strains suggests the evolution of local endemic populations of the pathogen and does not correlate with race distribution

Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot in Brassicaceae. It is widespread in Italy and severe outbreaks occur under conditions that favour disease development. In this study a multilocus sequence typing approach (MLST) based on the partial sequence of seven loci was applied to a selection of strains representative of the main areas of cultivation and hosts. The aim was to investigate whether the long tradition of brassica crops in Italy has influenced the evolution of different Xcc populations. All loci were polymorphic; 14 allelic profiles were identified of which 13 were unique to Italian strains. Based on the seven loci, the most common genotype within the Italian Xcc strains (AP1) was also the most representative genotype found in worldwide Xcc strains. This genotype was included in a new clonal complex in addition to three other clonal complexes already identified in Xcc populations. The phylogenetic reconstruction using a concatenated dataset of four conserved protein-coding genes, dnaK, fuyA, gyrB and rpoD, showed that the Italian strains belonged to two genetic groups. Physiological races were also investigated for the first time in Italy. The race structure of Xcc was determined by inoculating eight differential Brassica lines belonging to five species and showed that, in Italy, race 4 is the most widespread, followed by races 1 and 6. No correlation was found between allelic profiles, host of isolation, geographical origin and races, although a prevalent race was identified within the same clonal complex.     

寄主植物中抑制十字花科黑腐病菌Ⅲ型分泌系统小分子化合物的鉴定

 寻找抑制植物病原菌III型分泌系统的植物源活性小分子化合物,是研发生物安全农药的重要途径之一。本研究采用水煮提取法从十字花科黑腐病菌寄主植物满身红萝卜中提取分离活性小分子化合物,利用高效液相-质谱联用解析出活性物质的单体结构。然后用荧光素酶基因luxAB构建融合报告系统以及定量PCR检测活性物质对十字花科黑腐病菌III型分泌系统的抑制效果,最后采用剪叶接种和压渗接种的方法研究活性小分子物质对十字花科黑腐病菌的生防作用。研究表明,植物鞘氨醇和二氢鞘氨醇对十字花科黑腐病菌III型分泌系统基因的转录表达有一定程度的抑制作用,但是对I、II、IV型分泌系统基因的表达没有明显的抑制作用。植物鞘氨醇在XCM1上影响菌的生长,而二氢鞘氨醇不影响菌的生长。同时,还发现这两种物质能显著降低Xcc在寄主植株满身红萝卜上的病害症状以及能够使Xcc在非寄主植物辣椒上引起过敏反应的能力丧失。该研究结果为深入研究小分子化合物对十字花科黑腐病菌III型分泌系统的作用机制及后续开发植物源抑制剂提供了一定的理论依据。