The genetic characterization of Xanthomonas arboricola pv. juglandis,the causal agent of walnut blight in Poland

Leaves and fruits of walnut trees exhibiting symptoms of bacterial blight were collected from six locations in Poland. Isolations on agar media resulted in 18 bacterial isolates with colony morphology resembling that of the Xanthomonas genus. PCR using X1 and X2 primers specific for Xanthomonas confirmed that all isolates belonged to this genus. In pathogenicity tests on unripe walnut fruits, all isolates caused typical black necrotic lesions covering almost the entire pericarp. Results of selected phenotypic tests indicated that characteristics of all isolates were the same as described for the type strain of Xanthomonas arboricola pv. juglandis. Genetic analyses (PCR MP, ERIC‐, BOX‐PCR and MLSA) showed similarities between the studied isolates and the reference strain of X. arboricola pv. juglandis CFBP 7179 originating from France. However, reference strains I‐391 from Portugal and LMG 746 from the UK were different. MLSA analysis of partial sequences of the fyuA, gyrB and rpoD genes of studied isolates and respective sequences from GenBank of pathotype strains of other pathovars of X. arboricola showed that the X. arboricola pv. juglandis isolates consisted of different phylogenetic lineages. An incongruence among MLSA gene phylogenies and traces of intergenic recombination events were proved. These data suggest that the sequence analysis of several housekeeping genes is necessary for proper identification of X. arboricola pathovars.     

Thermal tolerance of propagative anthurium stem cuttings to disinfestation by heat treatment for burrowing nematodes and bacterial blight

Hot water and hot air treatments were evaluated for disinfesting anthurium, Anthurium andraeanum Lind., stem cuttings of the bacterial blight pathogen, Xanthomonas axonopodis pathovar dieffenbachiae (Xa pv. dieffenbachiae), and burrowing nematodes, Radopholus similis, and their effect on viability of the cuttings. Xa pv. dieffenbachiae suspended in distilled water in 1.5 ml microcentrifuge tubes, lost at least 6 logs of viability when exposed to hot water at 50 °C for 12 min or hot air at 50 °C, 60% RH for 35 min administered in commercial-sized heat treatment facilities. Stem cuttings exposed to hot air at 50 °C, 60% RH were disinfested of R. similis when their core temperatures attained 50 °C. Plant response to heat treatments varied among cultivars; however, all evaluated cultivars exhibited high tolerance to hot water at 50 °C for up to 24 min with equal or enhanced sprouting rates as compared to untreated checks. Sprouting rates of three of the four cultivars treated with hot air at 50 °C, 60% RH for up to 125 min were equal to or higher than untreated checks, while cuttings from the less tolerant cultivar ‘Tropic Fire’ registered lower sprouting rates for all hot air treatment durations as compared to untreated checks, Flower quality parameters, including average spathe size, stem diameter and number of flowers harvested from plants heat-treated as cuttings, were comparable to or higher than untreated checks for all treatments and cultivars. Disinfestation of anthurium stem cuttings for bacterial blight and the burrowing nematode can be achieved in hot water at 50 °C for 24 min without loss of sprouting rate or flower quality.     

7种检疫性植物病原黄单胞菌III型分泌系统和效应蛋白编码基因的差异性分析

 γ-变形菌纲(γ-Proteobacteria)的黄单胞菌属(Xanthomonas)的大多数种类可引起植物病害,多数是我国检疫对象。与其他革兰氏阴性植物病原细菌一样,植物病原黄单胞菌可通过高度保守的III型分泌系统(type-III secretion system, T3SS)分泌效应蛋白(T3SS-secreted effectors, T3SEs)进入植物细胞,在非寄主植物和抗病寄主植物上产生过敏反应(hypersensitive response, HR)以及在感病寄主植物上具有致病性。尚不清楚哪些种类的黄单胞菌具有T3SS和缺少哪些T3SE是否可作为检疫的依据。搜集7种检疫性植物病原黄单胞菌,通过PCR和Southern杂交试验结果发现:香蕉细菌性青枯病菌(X. campestris pv. musacearum)的ICMP287和ATCC49084菌株、甘蔗流胶病菌(X. axonopodis pv. vasculorum)ATCC13901菌株、洋葱细菌性叶枯病菌(X. axonopodis pv. allii)的LMG576和LMG578菌株中不含有tale基因,并且ATCC13901菌株既不含有T3SS基因也不含有hpa1和xopQ基因;菜豆细菌性疫病菌(X. campestris pv. phaseoli)ATCC49119菌株不含有hpa1基因。相应地,推测含有2~12个tale基因的黄单胞菌有:大豆斑疹病菌(X. axonopodis pv. glycines)ICMP5732和ATCC43911菌株、豌豆细菌性疫病菌(X. axonopodis pv. vignicola)ATCC11648菌株、棉花细菌性角斑病菌(X. campestris pv. malvacearum)ATCC12131和(X. campestris pv. phaseoli)ATCC49119菌株。大豆细菌性斑疹病菌ATCC43911菌株尽管含有hpa1、xopQ和hrcC基因,但在非寄主烟草上不能激发HR反应;而甘蔗流胶病菌ATCC13901菌株不含有hpa1、xopQ和hrcC基因,却激发烟草产生HR反应。这些结果对于分析比较不同植物病原黄单胞菌的致病性因子和设计特定的植物检疫靶点提供了科学线索。     

Survival of the Anthurium Blight Pathogen, Xanthomonas axonopodis pv. Dieffenbachiae, in Field Crop Residues

Bacterial blight, caused by Xanthomonas axonopodis pv. dieffenbachiae (Xad), is a major threat to the anthurium cut flower industry worldwide. Two field trials in Hawaii evaluated the long-term persistence of Xad in artificially-infested crop residues. Xad survived in leaf, petiole, and root residues for as long as 4 months when tissues were left on the surface or buried 15cm deep. Survival was considerably shorter (approximately 20 days) outside of residues. Xad that was recovered from residues over a period of 4 months retained pathogenicity. Xad was isolated from living roots of naturally-infected plants which further suggests that roots left in the field after culling may be particularly important, but overlooked, inoculum source. This information is key to determining minimum fallow periods before replanting devastated fields.     

脐橙溃疡病综合防控技术规范

根据江西省赣南脐橙溃疡病的发生特点,对新老病区脐橙溃疡病提出了相应的综合治理对策。     

在水稻悬浮细胞系中非亲和白叶枯病菌繁殖受到抑制的现象及其机理的研究

 研究了水稻白叶枯病菌在水稻悬浮细胞系中的繁殖动力学特征以及悬浮细胞胞外液中各类物质对白叶枯病菌繁殖的影响。结果表明,水稻-白叶枯病菌非亲和互作与亲和互作相比病原菌繁殖的数量明显减少。胞外液中粗提蛋白对白叶枯病菌繁殖不具有抑制作用。非亲和互作胞外液中二元酚含量显著增高,并且水稻细胞防卫反应基因的转录和翻译活性明显增强。表明非亲和互作中白叶枯病菌繁殖受到抑制与水稻细胞防卫反应的启动相关。     

东北水稻白叶枯病菌生理小种及水稻品种对9号小种的抗性

对东北地区水稻白叶枯病菌生理小种群体的构成及分布,以及水稻品种对白叶枯病菌9号小种的抗性进行了研究。结果表明,东北地区水稻白叶枯病菌生理小种群体由6个小种,即Race 1、Race 2、Race 3、Race 6、Race 8和Race 9构成。小种Race 1、Race 2和Race 9在东北三省吉林、辽宁和黑龙江都有分布,小种Race 3和Race 8只存在黑龙江省,Race 6只存在辽宁省。在30个水稻品种中,对9号生理小种表现抗病的有13个,出现频率为43.3%;中抗7个,频率23.3%;中感6个,频率20.0%;感病4个,频率13.3%。本研究为在东北粳稻地区挖掘白叶枯病的抗源,以及利用抗病品种控制白叶枯病危害提供依据。     

Multiplex PCR for specific and robust detection of Xanthomonas campestris pv. musacearum in pure culture and infected plant material

The present study developed a pathovar‐specific PCR for the detection of Xanthomonas campestris pv. musacearum (Xcm), the cause of banana xanthomonas wilt, by amplification of a 265‐bp region of the gene encoding the general secretion pathway protein D (GspD). A distinct DNA fragment of the expected size was amplified from genomic DNA from all of 12 Xcm isolates tested and no amplification of DNA was observed from other xanthomonads or plant‐associated bacteria, including the two closely related species Xanthomonas vasicola pv. holcicola and Xanthomonas axonopodis pv. vasculorum. The Xcm‐specific PCR was successfully multiplexed with internal control primers targeting 16S rDNA for application on DNA from bacterial cultures and with primers targeting plant mitochondrial 26S rDNA for application on DNA extracted from plant material. Diagnostic discrimination of healthy and infected plants was subsequently demonstrated in tests on artificially inoculated screenhouse cultivars of banana and field bananas with and without symptoms sampled from different parts of Uganda. This study therefore demonstrated a robust and specific Xcm diagnostic tool with the added advantage of applying internal PCR controls for direct quality assessment of results.     

三套鉴别品种对云南高原粳稻白叶枯病菌致病型的鉴定比较

为了明确高原粳稻白叶枯病菌的致病型及其致病性变异,采用剪叶接种法,用三套鉴别品种对云南高原粳稻白叶枯病菌10个代表性菌株进行致病型鉴定比较.结果表明,云南高原粳稻区菌株的致病性具有特殊性,以毫糯扬、TN1、黄玉、珍珠矮、IR26、南粳33和金南风组成的鉴别品种对其致病型的鉴定效果最佳;菌株Q-1-2在国内公认的两套鉴别品种上的抗性反应模式分别是SRRSR和SRRRSR,不同于已发表的所有菌株;稻种资源毫糯扬、IRBB14、IRBB3、IRBB4和IRBB5等对所用菌株都表现抗病,其所携带的抗性基因在云南高原粳稻育种中具有利用价值.