A New Cell Wall Located N-rich Protein is Strongly Induced During the Hypersensitive Response in Glycine Max L.

Soybean (Glycine max (L.) Merill, cv. Williams 82) plants and cell cultures respond to avirulent pathogens with a hypersensitive reaction. After inoculation of soybean with Pseudomonas syringae pv. glycinea, carrying the avirulence gene avrA, or zoospores from the fungus Phytophthora sojae Race 1, a resistance-gene-dependent cell death programme is activated. A new gene was identified by differential display of mRNAs that is specifically activated during the early phase of incompatible pathogen-soybean interactions but does not respond to compatible pathogens. The gene is strongly induced within 2h after addition of P. sojae zoospores. A similar kinetic pattern was observed for P. syringae (avrA) inoculated soybean cell cultures. The gene encodes a deduced protein of 368 amino acids with a very high content of asparagine and was therefore termed N-rich protein (NRP). The protein is composed of two distinct domains, of which only the C-terminal domain has striking homology to proteins of unknown function from other plants. An antibody raised against the recombinant NRP recognizes a protein of 42kDa. The protein is located in the cell wall as indicated by cell fractionation studies. Comparison of the genomic DNA-sequence with the cDNA, identified two introns within the open reading frame. The NRP-gene is not directly induced by salicylic acid or hydrogen peroxide, indicating a distinct and specific signal transduction pathway which is only activated during programmed cell death. The NRP-gene appears to be a new marker in soybean activated early in plant disease resistance.     

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。     

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.     

Activation of systemic disease resistance in pea by an avirulent bacterium or a benzothiadiazole, but not by a fungal leaf spot pathogen

Inoculation of first expanded leaves of pea seedlings with an avirulent strain of Pseudomonas syringae pv. pisi , or treatment with sprays of a benzothiadiazole (20 or 100  μ g a.i. mL⁻¹), decreased the susceptibility of subsequent leaves 7 or 14 days later to challenge inoculation with Mycosphaerella pinodes . Inoculation of first leaves with a virulent strain of P. syringae pv. pisi or with M. pinodes did not decrease the susceptibility of plants to M. pinodes . Treatments effective in decreasing susceptibility to M. pinodes were similarly active against Uromyces viciae-fabae and virulent P. syringae pv. pisi . Effective treatments also enhanced the activities of the enzymes β-1,3-glucanase and chitinase in untreated upper leaves 6 days later. Ineffective treatments for decreased susceptibility had no effect on the activity of the enzymes. None of the treatments enhanced peroxidase activities. The results are discussed in relation to the reported signalling effects of the benzothiadiazole and in relation to a suggested high activity of the avirulent P. syringae pv. pisi strain and inactivity of M. pinodes in enhancing natural signalling.     

Analysis of the gacS-gacA regulatory genes of spontaneous mutants of Pseudomonas fluorescens biocontrol strain PfG32R

Pseudomonas fluorescens strain PfG32R, a potential biocontrol agent against soilborne pathogens, frequently loses its antifungal activity and ability to produce enzymes. To characterize genetically the instability of these bacterial functions, we analyzed gacS and gacA genes of PfG32R and three spontaneous mutants of PfG32R (NR1, NR9, and ASW6), which had lost their ability to produce proteases and antifungal activities. The gacS and gacA sequences of PfG32R had 77%–89% and 78%–87% homology, respectively, with several known gacS and gacA homologues. All three spontaneous mutants were subjected to complementation analysis. Introduction of clones containing an intact gacS of PfG32R and another P. fluorescens strain Pf-5 as well as strain tea632 of P. syringae pv. theae complemented all three mutants restoring protease and antifungal activities, indicating a mutation in gacS. In sequencing analysis, the mutants had a deletion or change in amino acids in the conserved sensor kinase domains of GacS. The three mutants maintained both their antibacterial activity against Ralstonia solanacearum and Clavibacter michiganensis ssp. michiganensis and siderophore production, indicating that they are not controlled by the GacS/GacA system. The sequences reported in this article have been deposited in the DDBJ database under accession numbers AB219364 and AB219365.     

托拉斯假单胞杆菌弱毒菌株诱导平菇抗细菌性褐斑病的研究

为了明确平菇细菌性褐斑病病原菌托拉斯假单胞杆菌（Pseudomonas tolaasii）弱毒菌株JPG250303的诱导抗病作用、弱致病性及诱导抗病作用的遗传稳定性,通过对其诱导抗病性表达、诱导接种浓度及诱导间隔期的研究,明确该弱毒菌株具有较好的诱导抗病效果。利用形态学及分子生物学技术,鉴定出该弱毒菌株为托拉斯假单胞杆菌（P. tolaasii）,将弱毒菌株经平菇子实体连续5代培养后均具有弱致病性,说明其弱致病性可以稳定遗传;同时对5代菌株分别进行了诱导抗病活性的验证,5代菌株对平菇细菌性褐斑病诱导抗病效果分别为67.2%、66.3%、69.1%、68.6%和65.0%,说明该弱毒菌株诱导抗病活性也具有稳定性,这一研究为该弱毒菌株作为生防菌株防治平菇细菌性褐斑病的应用提供了理论基础。     

黄瓜细菌性角斑病菌交叉引物恒温扩增检测技术

黄瓜细菌性角斑病是我国黄瓜生产上的重要病害之一,其病原菌为Pseudomonas syringae pv.lachrymans。根据该病原菌甘油醛-3-磷酸脱氢基因保守序列设计引物和探针,建立了交叉引物恒温扩增和核酸试纸条检测技术。菌体DNA检测灵敏度可达0.55 ng,纯菌直接检测灵敏度基本可达到单个细菌。所测试的5株黄瓜细菌性角斑病菌和染病黄瓜叶片均为阳性,其他13株对照菌株均为阴性。该方法灵敏度高,且操作简单,对设备要求低等,适合基层实验室应用。     

荧光假单胞菌2P24调控基因突变体定殖能力和生防效果分析

在温室盆栽条件下,研究了分离自小麦全蚀病自然衰退土壤、可防治多种土传病害的荧光假单胞杆菌2P24菌株的2,4-二乙酰基间苯三酚合成基因突变体、GacS/GacA双因子调控系统突变体,以及PcoR-PcoI群体感应系统的突变体菌株在植物根围定殖能力和防病效果。结果表明,2,4-二乙酰基间苯三酚生物合成突变体不影响菌株在植物根部的定殖;GacS/GacA双因子调控系统中任一因子的突变菌株和pcoI^-突变菌株显著降低了菌株在植物根部的定殖能力。温室病害防治试验结果表明,与野生菌2P24相比,phlD^-,gacS^-,gacA^-,pcoI^-突变体对小麦全蚀病和番茄青枯病生防效果均显著降低;而相应的互补菌株生防效果恢复到野生型的水平。结合前期工作结果,我们推测GacS/GacA双因子调控系统和QS系统在2P24菌株定殖和防病过程的信号传递上形成级联途径。     

自毒物质对羟基苯甲酸降解细菌ZH2的分离与应用

自毒物质是造成植物连作障碍的主要因子,研究旨在筛选能够降解土壤中自毒物质的细菌.采用选择性分离方法从土壤中筛选自毒物质对羟基苯甲酸降解菌;结合形态特征、生理生化特征和16S rRNA测序鉴定菌种;采用紫外分光光度法测定菌株降解对羟基苯甲酸能力,并通过盆栽实验验证解毒效果.结果表明,分离到1株有降解对羟基苯甲酸能力的菌株...