水稻白叶枯病成株抗性与过氧化氢含量及几种酶活性变化的关系

 分析了水稻白叶枯病成株抗性与过氧化氢(H₂O₂)含量及过氧化物酶(POD)、多酚氧化酶(PPO)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性变化的关系。试验结果表明:接种白叶枯病菌T7133后,苗期与成株期植株体内H₂O₂含量上升以及POD、PPO和SOD活性增加。与苗期叶片相比,成株期叶片H₂O₂含量高,且PPO和SOD的酶活性增强,而POD的酶活性则降低。苗期和成株期CAT的酶活性均低于对照,成株期比苗期CAT活性更低。这些结果表明,H₂O₂、PPO、SOD和CAT可能与水稻白叶枯病成株抗性之间存在一定的关联,而POD则没有直接关系。     

中生菌素对水稻悬浮细胞过氧化物酶基因转录表达的影响

用白叶枯菌（HB84—17）、10μg／ml中生菌素和10μg／ml中生菌素＋白叶枯菌处理水稻抗、感白叶枯病近等位基因系CBB4和沈农1033悬浮细胞，采用斑点杂交的方法研究了中生菌素对水稻悬浮细胞过氧化物酶基因转录表达的影响。结果表明，白叶枯菌、中生菌素和中生菌素＋白叶枯菌处理都能诱导悬浮细胞中过氧化物酶基因的转录表达。对于CBB4，白叶枯菌处理3h时过氧化物酶基因开始诱导转录表达，6h达到高峰，诱导表达时间持续至24h；中生菌素处理3h时开始大量表达，6h达到，高峰，大量表达持续至72h。对于沈农1033，白叶枯菌处理48h时过氧化物酶基因才开始诱导转录表达；中生菌素处理，3h就开始大量转录表达，并持续至48h。中生菌素＋白叶枯菌处理的感、抗病品种过氧化物酶基因的诱导转录表达强度和模式与单用中生菌素处理相同。     

OsHsp18.0-CI调控水稻对白叶枯病的抗性

 热激蛋白广泛存在于各种生物中,响应多种生物胁迫和非生物胁迫。前期研究结果显示,在水稻中超量表达小热激蛋白OsHsp18.0-CI基因,能通过增强水稻的基础防卫反应提高对水稻细菌性条斑病的抗性。本研究发现,OsHsp18.0-CI基因也能够受到白叶枯病菌的诱导表达,超量表达OsHsp18.0-CI的转基因水稻也增强了对多个白叶枯病致病菌株的抗性。转录组测序分析发现,OsHsp18.0-CI基因介导的水稻对白叶枯病的抗病信号途径有部分类似于其介导的对细菌性条斑病的抗性路径,同时也有大量新的未知功能基因的参与。以上结果表明,OsHsp18.0-CI基因受到病原菌的侵染时,也能通过增强水稻的基础防卫反应来提高对白叶枯病的抗性。     

菠萝泛菌与水稻白叶枯病菌双重PCR检测方法的建立与应用

近年来水稻发生了一种由菠萝泛菌Pantoea ananatis引起的新型细菌病害，其发生时期与白叶枯病相近，病症与白叶枯病类似。为了实现该病害与白叶枯病的快速检测，本研究基于泛菌属看家基因acnA,通过序列比对，设计了22对特异性引物，分别与已知的白叶枯病菌检测引物XOO80进行配对并筛选，建立了一种双重PCR检测方法，可从菠萝泛菌和白叶枯病菌中分别扩增出910 bp和162 bp的特异性条带，而其他10种非目标细菌均未有扩增条带，25μL体系中可稳定地从至少1 pg/μL的基因组DNA模板中扩增出特异性条带。本研究建立的菠萝泛菌与水稻白叶枯病菌双重PCR检测方法具有良好的特异性和灵敏度，为水稻细菌病害病原鉴定及防治提供了技术支撑。     

水稻转录因子OsBTF3对不同病原菌和信号分子的基因表达反应

 转录因子基因OsBTF3在水稻品种日本晴悬浮细胞中受白叶枯病菌(Xanthomonas oryzae pv.oryzae,Xoo)诱导表达。为了阐明OsBTF3在水稻叶组织中的表达特征,本研究利用RT-Q-PCR技术,对经3种亲和性病原菌[水稻白叶枯病菌(Xoo)、水稻条斑病菌(Xooc)和稻瘟病菌(Mg)]接种和4种信号分子[脱落酸(ABA)、水杨酸(SA)、茉莉酸甲酯(MeJA)、乙烯(ETH)]诱导处理的水稻叶片中OsBTF3的转录本进行了定量分析。结果表明,OsBTF3对Xoo、Xooc和Mg侵染的基因表达反应均显著地受到诱导,但反应速度和强度略有差异。而4种信号分子对OsBTF3表达的诱导作用差异较大,ABA诱导活性最强,MeJA和ETH次之,SA诱导作用不显著。因此,OsBTF3基因表达不仅具有病原菌Xoo、Xooc和Mg的诱导性,而且也具有信号分子MeJA、ETH和ABA的应答性。     

不同接种方法对甘蓝黑腐病菌胞外多糖突变体症状产生的影响

 1992~1994年采用中国水稻白叶枯病菌致病型研究方法,测定108个以江汉稻区为主的湖北水稻白叶枯病分离菌珠,在5个中国鉴别品种(IR26、爪哇14、南粳15、特特普和金则30)成株上的抗感反应,将病菌划分为O、Ⅰ、Ⅱ、Ⅲ、Ⅳ致病型。Ⅳ型菌31个,占参试菌株的28.7%,占采样县(市)的61.1%,出现频率最高,分布最广,是江汉稻区流行的优势致病型,其次是Ⅲ、Ⅱ型菌。此结果与1989~1991年测定湖北省白叶枯病致病型结果基本一致,流行优势致病型仍是Ⅳ型,但致病力指数由1989~1991年的72.3下降至59.3,菌株毒力有所减弱。白叶枯病是江汉稻区水稻生产的潜在威胁,而目前种植的水稻品种多不抗IV型菌,所以今后在育种过程中,要多考虑转育含Xa-4基因品种。在白叶枯病疫区建议种植抗性稳定的扬辐籼2号、扬稻4号、秀水664、威优64等品种。     

感染白叶枯病后水稻叶片光合同化物对病害的补偿作用

水稻抗白叶枯病品种ＩＲ２６和感病品种金刚３０在拔节期接种白叶枯病菌后，接种叶片净光合速率均下降，而与病叶相邻的健叶的净光合速率则明显增加，在穗期，ＩＲ２６接种叶片的＾１４Ｃ同化物累积率和运输到穗的百分率均高于金刚３０；ＩＲ２６与病叶相邻的健叶，＾１４Ｃ同化物输向接种病叶和穗的百分率高于金刚３０。这表明抗病品种ＩＲ２６接种病叶和健康叶片光合同化物对病菌侵染和产量损失的补偿作用较强，相反，金刚３０则不     

茉莉酸甲酯对水稻白叶枯病的诱导抗性及相关防御酶活性的影响

为探索茉莉酸甲酯(methyl jasmonate,MeJA)诱导水稻抗白叶枯病的效应,采用MeJA喷雾处理剪叶接种法,测定MeJA对水稻幼苗的白叶枯病病情指数、白叶枯病病菌Xanthomonas oryzae pv.oryzae的抑菌效果及对叶片过氧化物酶(peroridase,POD)、过氧化氢酶(catalase,CAT)、超氧化物歧化酶(superoxide dismutase,SOD)、多酚氧化酶(polyphenol oxidase,PPO)和苯丙氨酸解氨酶(phenylalnaine ammonialyase,PAL)等相关防御酶活性的影响.0.05 ～ 2.0 mmol/L的MeJA能降低水稻幼苗白叶枯病的病情指数,但对水稻白叶枯病菌无直接抑菌活性;0.1 mmol/L MeJA的诱导效果最好,处理48h后,感病品种温229和抗病品种嘉早312的诱导效果分别为73.18％和70.43％;0.05 ～2.0mmol/L的MeJA处理水稻叶片中POD、CAT、SOD、PPO和PAL活性呈上升趋势.研究表明MeJA能诱导水稻幼苗对白叶枯病的抗性,且诱导抗性的产生与MeJA提高水稻相关防御酶的活性有关.     

我国水稻白叶枯病菌致病力分化研究

综述了我国水稻白叶枯病菌致病力分化及其致病基因和致病机制的研究进展 ,查明稻白叶枯病菌致病力分化状况是进行水稻抗白叶枯病育种及其抗性品种利用的基础。     

植物促生细菌诱导水稻对白叶枯病抗性的初步研究

水稻白叶枯病是使水稻遭受重大损失的主要病害。曾有利用白叶枯病原菌胞外多糖（ＥＰＳ）和白叶枯病原菌弱毒株诱导水稻对白叶枯病害抗病性的报道。但有关应用植物促生细菌（ＰｌａｎｔＧｒｏｗｔｈＰｒｏｍｏｔｉｎｇＢａｃｔｅｒｉａ,ＰＧＰＢ）诱导水稻对白叶枯病害抗性的研究还未见报道。本文采用分离于水稻苗期根内、根面的具有固氮能力的植物促生细菌阴沟肠杆菌（Ｅｎｔｅｒｏｂａｃｔｅｒｃｌｏａｃａｅ）ＭＲ１２和争论产碱菌（Ａｌｃａｌｉｇｅｎｅｓｐａｒａｄｏｘｕｓ）Ｒ４作为诱导子处理孕穗期水稻叶片,研究了它们对水稻白…     

Expression profiling of rice genes in early defense responses to blast and bacterial blight pathogens using cDNA microarray

In order to understand the defense machinery in the model cereal crop rice, we performed a large-scale analysis of rice gene expression in response to rice blast Magnaporthe grisea (M. grisea) or Magnaporthe oryzae and bacterial blight Xanthomonas oryzae pv. oryzae (Xoo) during the early incompatible and compatible interactions. Using a gene chip containing 10 254 rice cDNAs representing 9240 unique genes, we identified 794 and 612 genes differentially expressed in the incompatible and compatible rice–M. grisea interactions, respectively, with 274 genes co-regulated during both interactions. In the rice–Xoo pathosystem, 454 and 498 differentially expressed genes were identified in the incompatible and compatible interactions, respectively, including 237 co-regulated genes in the both interactions. By clustering differentially regulated genes from all these interactions, we identified 29 co-regulated genes in the all four interactions, and 86 and 74 co-regulated genes in the two incompatible and two compatible interactions, respectively. These differentially expressed genes could be classified into three categories, including M. grisea- and Xoo-regulated, M. grisea-specific, and Xoo-specific. The expression patterns of representative defense-related genes were further confirmed by RT-PCR. The large-scale expression data from our microarray analysis indicated the existence of distinctive as well as shared defense pathways between the rice–M. grisea and rice–Xoo interactions.     

非亲和性稻瘟病菌对水稻的诱导抗性

为研究稻瘟病菌Magnaporthe oryzae不同菌株间的相互作用,选择与单抗性基因系水稻IRBL5-M （携带抗性基因Pi5）表现为亲和性的菌株HN52与非亲和性的菌株HN119为研究对象,将其单独或混合接种到单抗性基因系水稻IRBL5-M中,并通过荧光显微镜观察接种后水稻叶鞘的发病情况及病斑面积,测定接种后水稻内相关抗性基因OsWRKY45、OsNPR1、OsPR10、OsMAPK2的表达量以及活性氧的变化。结果显示,相较于单独接种亲和性菌株,混合接种后单抗性基因系水稻IRBL5-M病斑发病面积减少;混合接种中亲和性菌株HN52菌丝侵染能力降低,侵染菌丝细胞间扩展率显著降低73.13%;同时单抗性基因系水稻IRBL5-M中OsWRKY45、OsNPR1、OsPR10和OsMAPK2抗性基因表达量显著增加,水稻叶片中活性氧含量增加,表明在菌株混合侵染过程中,非亲和性菌株可通过激发水稻的抗性反应来降低亲和性菌株对水稻的侵染程度。     

水稻白叶枯病菌酸性还原酮加双氧酶的鉴定分析

 酸性还原酮加双氧酶(ARD)催化很多原核和真核生物中的甲硫氨酸急救途径(MSP)倒数第二步。本研究鉴定了水稻白叶枯病菌Xanthomonas oryzae pv. oryzae (Xoo)中的酸性还原酮加双氧酶, 命名为xard。Xoo 菌株PXO99^A、MAFF311018和KACC10331中的xard核苷酸序列完全相同。xard基因突变菌株在甲硫基腺苷(MTA)为唯一硫源时不能正常生长。这一结果证明Xard在MSP中起作用。xard突变体和野生型菌株PXO99^A 接种水稻IR24后病斑长度数据表明该基因突变对Xoo在水稻上的毒性没有影响。     

Genome‐wide identification of pathogenicity genes in Xanthomonas oryzae pv. oryzae by transposon mutagenesis

A transposon mutant library was constructed from the bacterial blight pathogen Xanthomonas oryzae pv. oryzae (Xoo) KACC10331 by Tn5 transposon mutagenesis. The susceptible rice cultivar Milyang 23 was inoculated with a total of 24 540 mutants resistant to kanamycin and 67 avirulent or reduced‐pathogenicity mutant strains were selected for study. Southern hybridization verified that 84 mutant strains had single‐copy insertions and their single‐transposon insertion sites were identified by sequencing analysis combined with thermal asymmetric interlaced (TAIL)‐PCR. The single‐transposon‐tagged sequences of 21 mutant strains belonged to pathogenicity‐related genes previously reported in Xanthomonas species, while the other 46 single‐transposon‐tagged sequences included diverse functional genes encoding, five cell‐wall‐degrading enzymes, three fimbrial and flagella assembly regulators, five regulatory proteins, 15 metabolic regulators and 18 hypothetical proteins, which were identified as novel pathogenicity genes of Xoo.     

Characterization of <Emphasis Type="Italic">Myoviridae</Emphasis> and <Emphasis Type="Italic">Podoviridae</Emphasis> family bacteriophages of <Emphasis Type="Italic">Erwinia amylovora</Emphasis> from Hungary - potential of application in biological control of fire blight

The virulence spectrum of 300 isolates of Xanthomonas oryzae pv. oryzae (Xoo), representing 17 districts of Punjab, Pakistan was elucidated through inoculation on a set of six rice IRRI-differentials. The virulence level was assessed by using principal component and cluster analysis. Among six principal components (PCs), PC-1 exhibited 59.3 % of the total variance. The highly virulent isolates clusters on the positive side of the ordination away from the point of intersection of PC1 and PC2 and classifies the Xoo isolates from slow disease to the highest disease causing entities. The 300 isolates were categorized into 29 pathotypes (Pt1-29) wherein the highly virulent pathotype (Pt-1), comprises of 39 Xoo isolates were widespread in 12 districts. The majority of Xoo isolates were moderately to least virulent (21.7–43 %) and average disease progress curves confirmed the field reactions of these pathotype clusters for an efficient recognition of Xoo isolates. Interaction of the pathogen with differentials harboring different resistant genes was well investigated in the current study for future management approaches for which the surveillance of the new Xoo pathotypes may expedite the disease resistant rice breeding programme in the country.     

Pathogenesis-related gene expression in rice is correlated with developmentally controlled Xa21-mediated resistance against Xanthomonas oryzae pv. oryzae

Disease resistance mediated by the resistance gene Xa21 is developmentally controlled in rice. We examined the relationship between Pathogenesis Related (PR) defense gene expression and Xa21-mediated developmental disease resistance induced by Xanthomonas oryzae pv. oryzae (Xoo). OsPR1a, OsPR1b, and OsPR1c genes were cloned and their induction was analyzed, in addition to the OsPR10a gene, at the juvenile and adult stages in response to a wildtype Xoo strain that induces a resistance response (incompatible interaction) and an isogenic mutant Xoo strain that does not (compatible interaction). We found that the adult stage leaves are more competent to express these OsPR1 genes and that the Xa21 locus is required for the highest levels of induction.     

EGFP-Rhm51 foci enable the visualization and enumeration of DNA double-strand breaks in <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis>

To detect and count DNA double-strand breaks (DSBs) during the life cycle of Magnaporthe oryzae, we constructed an expression vector for EGFP-Rhm51 fusion protein and introduced it into strain Ina86-137. EGFP-Rhm51 foci were detected, and the number of foci in mycelia treated with mitomycin C, which induces DSBs, was higher than in untreated samples, indicating that the foci enabled the visualization of DSBs during the life cycle. EGFP-Rhm51 foci were observed at all stages of the asexual cycle including when invasive hyphae formed in an intact rice leaf sheath, demonstrating that M. oryzae undergoes DSBs during vegetative and parasitic growth.