Ren Y Z, Yue Y L, Jin G X, Du Q
Key University Laboratory for Oasis Agricultural Pest Management and Plant Protection Resource Utilization in the Xinjiang Uygur Autonomous Region, College of Agriculture, Shihezi University, Shihezi City, Xinjiang 832003, China.
Plant Dis. 2014 Jun;98(6):840. doi: 10.1094/PDIS-09-13-0983-PDN.
Bacterial blight was observed on field-grown guar (Cyamopsis tetragonoloba L.) for the first time in China. The disease outbreak occurred in the Xinjiang Uyghur Autonomous Region after several weeks of unusually heavy rains during late summer 2013. The disease incidence was generally 40 to 50%, although values as high as 80% were observed in several fields. Initial field symptoms included water-soaked spots on leaves, pods, petioles, and stems. During later stages of infection, the color of the spots became dark. We also observed large, angular, necrotic lesions at leaf tips, black streaks on petioles and stems, split stems, defoliation, wilting or top withering, vascular necrosis, and dieback. Samples of diseased leaves, stems, petioles, pods, and seeds were surface sterilized, ground, and then plated onto King's B medium. Plates were incubated at 28°C for 72 h. Fifteen bacterial strains with yellow-pigmented, opaque, and round colonies were isolated. These strains were aerobic, gram-negative rods with a single, polar flagellum. They were positive for HS, esculin, oxidase, tobacco hypersensitivity, indole production from tryptophan, nitrate reduction to nitrite, and the utilization of glucose, mannose, trehalose, galactose, and starch. The maximum salt tolerance of the strains was 2 to 3%. Pathogenicity tests using eight strains were conducted in July 2013. A bacterial culture was suspended in sterile water with a final concentration of 10 CFU/ml. Eight 4-week-old guar plants were inoculated by (i) spraying the bacterial suspension on the leaves until runoff, or (ii) puncturing the stems with a needle that had been dipped into the bacterial suspension. Sterile water was used as a negative control. Plants were kept in a mist room with 100% relative humidity for 24 h. Stem and leaf symptoms similar to those of the original plants were observed on the inoculated guar plants within 10 days of inoculation. No symptoms developed on the negative control plants. Yellow bacterial colonies re-isolated from inoculated plant tissues were morphologically identical to the original. 16S rDNA was amplified using universal primers (Pa 5'-AGTTTGATCCTGGCTCAG-3' and Ph 5'-TACCTTGTTACGACTTCGTCCCA-3') and sequenced. A BLAST search of the NCBI GenBank database indicated that the 16S rDNA sequences of three strains (accession nos. KF563926, KF563927, and KF563928) had 99.9% identity to Xanthomonas axonopodis strain XV938 (AF123091). Under greenhouse conditions, bacterial strains wilted asparagus bean and pea but rarely infected bean, kidney bean, faba bean, mung bean, soybean, red bean, pea, garbanzo bean, and peanut. Based on morphology, pathogenicity tests, 16S rDNA sequencing, and host plant specificity, the pathogen was confirmed as X. axonopodis pv. cyamopsidis (synonym: X. campestris pv. cyamopsidis [Patel et al., 1953]). To our knowledge, this is the first report of bacterial blight of guar caused by X. axonopodis pv. cyamopsidis in China. Guar has recently been introduced in Xinjiang Province. Our findings indicate that bacterial blight may pose a threat to the economic sustainability of guar production in the region. References: (1) I. A. Milyutina et a1. FEMS Microbiol. Lett. 239:17, 2004. (2) I. M. G. Almeida et al. Summa Phytopathol. 18:255, 1992. (3) J. D. Mihail et al. Plant Dis. 69:811, 1985.
在中国,首次在田间种植的瓜尔豆(Cyamopsis tetragonoloba L.)上观察到细菌性疫病。2013年夏末,新疆维吾尔自治区在经历了数周异常的暴雨后,该病爆发。尽管在几个田块中观察到发病率高达80%,但该病的发病率一般为40%至50%。最初的田间症状包括叶片、豆荚、叶柄和茎上出现水渍状斑点。在感染后期,斑点颜色变深。我们还观察到叶尖出现大的、角状的坏死斑,叶柄和茎上有黑色条纹,茎干开裂,叶片脱落,枯萎或顶部枯萎,维管束坏死以及枝条枯死。将患病叶片、茎、叶柄、豆荚和种子的样本进行表面消毒、研磨,然后接种到金氏B培养基上。平板在28°C下培养72小时。分离出15个具有黄色色素、不透明且圆形菌落的细菌菌株。这些菌株为需氧革兰氏阴性杆菌,具单根极鞭毛。它们对硫化氢、七叶苷、氧化酶、烟草过敏反应、从色氨酸产生吲哚、硝酸盐还原为亚硝酸盐以及利用葡萄糖、甘露糖、海藻糖、半乳糖和淀粉呈阳性反应。这些菌株的最大耐盐性为2%至3%。2013年7月使用8个菌株进行了致病性测试。将细菌培养物悬浮于无菌水中,终浓度为10 CFU/ml。八株4周龄的瓜尔豆植株通过以下方式接种:(i)将细菌悬浮液喷洒在叶片上直至径流,或(ii)用蘸有细菌悬浮液的针刺茎。无菌水用作阴性对照。植株在相对湿度100%的雾室中放置24小时。接种后10天内,在接种的瓜尔豆植株上观察到与原始植株相似的茎和叶症状。阴性对照植株未出现症状。从接种植物组织中重新分离出的黄色细菌菌落形态与原始菌落相同。使用通用引物(Pa 5'-AGTTTGATCCTGGCTCAG-3'和Ph 5'-TACCTTGTTACGACTTCGTCCCA-3')扩增16S rDNA并进行测序。对NCBI GenBank数据库的BLAST搜索表明,三个菌株(登录号KF563926、KF563927和KF563928)的16S rDNA序列与野油菜黄单胞菌菌株XV938(AF123091)的序列一致性为99.9%。在温室条件下,细菌菌株使长豇豆和豌豆枯萎,但很少感染菜豆、芸豆、蚕豆、绿豆、大豆、红豆、豌豆、鹰嘴豆和花生。基于形态学、致病性测试、16S rDNA测序和寄主植物特异性,确认病原菌为野油菜黄单胞菌瓜尔豆致病变种(同义词:野油菜黄单胞菌瓜尔豆致病变种[帕特尔等人,1953年])。据我们所知,这是中国首次关于野油菜黄单胞菌瓜尔豆致病变种引起瓜尔豆细菌性疫病的报道。瓜尔豆最近被引入新疆地区。我们的研究结果表明,细菌性疫病可能对该地区瓜尔豆生产的经济可持续性构成威胁。参考文献:(1)I. A. 米柳蒂娜等人,《FEMS微生物学快报》239:17,2004年。(2)I. M. G. 阿尔梅达等人,《植物病理学综述》18:255,1992年。(3)J. D. 米哈伊尔等人,《植物病害》69:811,1985年。