白叶枯病菌效应子XopN在拥有OsSWEET11同源基因的水稻品种上发挥毒性作用

【目的】黄单胞菌细胞外泌蛋白质(Xanthomonas outer proteins, Xops)是植物病原黄单胞菌高度保守的毒性效应子或毒性辅助组分,通过细菌Ⅲ型分泌系统分泌到细菌细胞外部,然后转入植物细胞而发挥病理作用。水稻黄单胞菌水稻致病变种即水稻白叶枯病菌的标准菌株PXO99^A分泌的XopN是一种毒性效应子,通过影响寄主免疫反应而使水稻发病。但是,XopN对病菌毒性的影响是否因水稻品种的不同而异,还有待研究。【方法】利用同源双交换技术,先敲除了PXO99^A的XopN基因,获得了∆XopN突变体,又经过遗传互补,得到了回补菌株∆XopN/XopN。通过营养肉汤液体培养,测定了XopN对病菌繁殖能力的影响;根据文献选用14个水稻品种,通过接种实验,测定了PXO99^A对这些品种的毒性与XopN敲除或回补的影响;在XopN发挥毒性作用的水稻品种上,测定了隐性抗病基因OsSWEET11/xa13与显性感病基因OsSWEET11/Xa13受病菌侵染而表达的情况,分析了XopN敲除或回补的影响。【结果】在营养肉汤液体培养过程中,病菌突变体菌株∆XopN的繁殖速度明显低于野生型PXO99^A。PXO99^A、∆XopN和∆XopN/XopN接种水稻后,根据其在水稻叶片组织内的繁殖量及随后产生的白叶枯病症状的严重程度,将供试的14个水稻品种分为两种情况。一是感病程度与病菌XopN是否敲除或回补无关,这有10个水稻品种（IRBB1、IRBB3、IRBB8、IRBB10、IRBB14、IR24、IRBB203、IRBB204、IRBB205和IRBB211）,PXO99^A、∆XopN和∆XopN/XopN对它们的毒性无明显差别。二是XopN对病菌毒性发挥作用的水稻品种,包括高度感病的3个品种（IRBB208、Asominori和日本晴）和低感品种IRBB13。与PXO99^A或∆XopN/XopN相比,∆XopN对这4个水稻品种的毒性大为降低。在IRBB13上,病菌侵染对隐性抗病基因OsSWEET11/xa13在叶片内的表达发生抑制作用,这一效应与XopN的毒性功能相关。相反,日本晴显性感病基因OsSWEET11/Xa13却受病菌侵染的诱导,在叶片内的表达水平大幅度提高。IRBB208和Asominori携带OsSWEET11/Xa13同源基因,该同源基因在叶片内的表达水平也因病菌侵染而大幅提高。在这4个水稻品种上,PXO99^A和∆XopN/XopN能够诱导OsSWEET11/Xa13或其同源基因表达,但∆XopN无此作用。另外,XopN对病菌在非寄主植物烟草上诱发过敏反应有量变贡献,相比PXO99^A或∆XopN/XopN,∆XopN引起过敏反应的程度有所降低。【结论】XopN是一个有限广谱性效应子,在拥有OsSWEET11同源基因的水稻品种上发挥毒性作用。XopN也是病菌繁殖所需要的,对病菌在非寄主植物上诱导过敏反应有一定贡献。     

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.     

蔗糖水解酶注释基因XC0805与野油菜黄单胞菌8004菌株的蔗糖利用及致病相关

 野油菜黄单胞菌(Xanthomonas campestris pv.campestris,Xcc)是造成十字花科植物减产的重要病原菌,其通过水孔或伤口进入植物体内并在维管束中快速繁殖。光合产物(蔗糖)的运输主要依靠植物维管组织,同时蔗糖也是维管组织重要的能量来源。本研究利用生物信息学和遗传学等方法对Xcc中参与蔗糖利用的基因进行了分析,并研究了这些基因与其致病力的关系。Xcc 8004菌株中注释的参与蔗糖分解蛋白的编码基因有XC1002、XC1642、 XC1645和XC0805。 XC1002和XC1642的编码产物属于糖苷水解酶GH97蛋白超家族,而XC1645和XC0805的编码产物则属于GH13家族。突变分析发现除XC0805外,其他3个都不影响Xcc的蔗糖利用能力。接种实验表明,XC0805参与Xcc的侵染或在植物体内的生长,其他3个不参与致病过程。另外,Xcc 8004中预测参与蔗糖转运的基因(XC0806和XC0807)突变并不影响细菌的蔗糖利用和致病力,表明其可能存在其他的蔗糖转运通路。上述结果说明XC0805可能是Xcc 8004主要的蔗糖水解酶基因,且在致病中起重要作用。     

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。     

水稻白叶枯病菌转录调控因子OxyRxoo对细菌抗氧化系统的调控作用

病原细菌对活性氧的降解是抗氧化和侵染中的重要毒性机制之一。阐述了水稻白叶枯病菌（Xanthomonas oryzaepv.oryzae）的转录调控因子OxyRxoo在病菌抗氧化系统中的调控作用,以及目前对OxyRxoo的研究进展。     

芒果细菌性黑斑病研究现状及展望

细菌性黑斑病严重影响芒果产量和商品质量, 是芒果生产上的重要生物灾害之一。该病由野油菜黄单胞菌芒果致病变种（Xanthomonas campestris pv. mangiferaeindicae）引起，几乎遍布热带亚热带芒果产区，常造成严重经济损失。本文主要综述了芒果细菌性黑斑病的症状、分布与危害、发生规律、防治方法、病原及其遗传多样性和检测技术等方面的研究进展，以期为该病的进一步研究及有效控制提供参考。     

柑橘溃疡病菌LAMP实时浊度检测方法的建立及应用

以柑橘溃疡病菌特异性高的假定蛋白基因（登录号：AE008923.1）为靶标，设计并筛选4条引物来特异性识别靶标序列中的6个独立区域，建立柑橘溃疡病菌LAMP实时浊度检测方法。该方法利用实时浊度仪检测反应过程中产生的焦磷酸镁白色沉淀，实现对LAMP整个反应过程的实时监控。从镁离子浓度、甜菜碱浓度、dNTPs浓度、引物浓度和反应温度5个变量参数对 LAMP检测体系进行优化，并对方法的特异性、灵敏度、稳定性及实际样品检测效果进行评价。结果显示，该体系经优化后稳定性良好，可在66 ℃ 恒温条件下，60 min内完成检测；DNA检测下限可达0.02 ng/μL，灵敏度与荧光定量PCR方法相同，比常规PCR方法高1个数量级；能快速、准确地对田间疑似样品进行检测，与荧光定量PCR方法的检出情况一致，没有漏检。结果说明柑橘溃疡病菌实时浊度LAMP检测方法操作简便、所需时间短，特异性与灵敏度高，为柑橘溃疡病菌的快速筛查提供较好的途径。     

水稻细菌性条斑病及其抗性研究进展

本文简要综述了近年来国内外对水稻抗细菌性条斑病在病原菌鉴定、水稻抗源筛选、抗性生理遗传以及抗性的分子生物学等方面的研究进展，以期为水稻细条病的深入研究提供参考。     

Inheritance of resistance to Xanthomonas campestris pv phaseoli in tetary bean (Phaseolus acutifolius)

Summary The F1, F2 and F3 from two crosses within Phaseolus acutifolius were exposed to Xanthomonas campestris pv phaseoli to analyse the inheritance of resistance. The resistant parent, PI 319.443, gave a hypersensitive reaction in leaves and pods with small necrotic lesions. Based on the resistance of F1, the segregation in F2 and the reaction of F3 plants and lines, it is concluded that resistance in leaves and pods is governed by one dominant gene. Comparisons are made with the resistance to X. campetris in P. vulgaris.     

Residual effects of the Xa-4 resistance gene in three rice cultivars when exposed to a virulent isolate of Xanthomonas campestris pv. oryzae

Summary F₂ plants of five, and F₃ plants of three, crosses between genotypes carrying the race-specific resistance gene Xa-4 and genotypes not carrying this gene were inoculated with two isolates of Xanthomonas campestris pv. oryzae. Half the tillers of each plant received isolate PX061, avirulent on the Xa-4 gene, the other half of the tillers received isolate PX099, virulent for the Xa-4 gene. The F₂ and F₃ populations segregated for a single dominant resistance gene, Xa-4.The parental, F₂ and F₃ genotypes not carrying Xa-4 had mean lesion lengths between 28 and 29 cm for both isolates. The Xa-4 carrying parents showed a mean lesion length of 2.7 cm with the avirulent isolate and of 12.4 cm with the virulent isolate. The Xa-4 carrying F₂ and F₃ genotypes had mean lesion lengths of 5.2 and 20.1 cm for the two isolates, respectively. These observations strongly indicate that the Xa-4 gene, carried by the rice genotypes studied (IR28, Cisadane and BR51-282-8), had a considerable residual effect when exposed to virulent isolate PXO99.