Mondal K K, Mani C, Singh J, Kim J-G, Mudgett M B
Division of Plant Pathology, Plant Bacteriology Section, Indian Agricultural Research Institute, New Delhi 110 012, India.
Department of Biology, Stanford University, Stanford, CA 94305-5020.
Plant Dis. 2011 Dec;95(12):1582. doi: 10.1094/PDIS-06-11-0533.
In September 2008, a new blight disease appeared on basmati rice (Oryza sativa L.) in fields in the northern states of India, including Uttar Pradesh, Haryana, and Punjab. First symptoms were water-soaked lesions at the tip of rice leaves. Lesions eventually spread down the leaf blades. Infected leaves turned light brown, exhibiting a blighted appearance. The disease was severe during the post-flowering stage. From 2008 to 2011, yellow-pigmented bacteria were consistently recovered on nutrient agar (beef extract 5 g, peptone 10 g, NaCl 5 g, and agar 20 g) from symptomatic rice leaves. The disease was thought to be caused by Xanthomonas oryzae pv. oryzae, the rice bacterial blight pathogen. However, physiological and molecular analysis of two strains (ITCC B0050 and ITCC B0055) isolated in 2008 revealed that the causal agent was the bacterium Pantoea ananatis. Colonies, raised and translucent with smooth margins, grew well within 24 h at 37°C. The bacteria are gram-negative facultative anaerobes with small rods arranged singly or in a chain of two to five cells. The bacteria are positive for catalase and indole production while negative for oxidase and alkaline reaction in malonate broth. Electron microscopy shows that the bacterial cells were 1.1 to 2.3 × 0.4 to 0.7 μm and have three to six peritrichous flagella. 16S rRNA gene sequence (1,535 nt generated by PCR with primers 5'AGAGTTTGATCATGGCTCAG3' and 5'AAGGAGGTGATCCAACCGCA3') of ITCC B0050 and ITCC B0055 (GenBank Nos. JF756690 and JF756691, respectively) share 99%-nt identity with P. ananatis (GenBank No. DQ512490.1). Biolog microbial identification analysis (version 4.2) of both strains showed similarity indices of 0.842 with P. ananatis (Biolog Inc., Hayward, CA). Pathogenicity was confirmed by employing the leaf tip clipping method to inoculate susceptible basmati rice (cv. Pusa basmati 1). Leaves were inoculated in triplicate with sterile water or a 1 × 10 CFU ml suspension of each isolate in water. The artificially inoculated rice leaves produced water-soaked lesions similar to that observed during natural rice infection in the field. At 10 to 15 days postinoculation, the lesions on the inoculated leaves dried and turned from straw color to light brown. Yellow-pigmented bacteria were reisolated from the infected rice leaves and their identity was confirmed to be identical to the original strain by 16S rRNA sequence analysis and Biolog analysis. Both pathogen isolates elicited hypersensitive reaction in tobacco (Nicotiana tabacum cv. Xanthi) leaves 24 to 48 h postinoculation (1 × 10 CFU ml). These studies indicate that the causal agent of the newly emerged rice leaf blight disease in northern India is P. ananatis. Pantoea spp. are opportunistic pathogens documented to cause different diseases in economically important crop plants including grain discoloration of rice in China (1), leaf blight and bulb decay of onion in the United States (2), and leaf blight of rice in Korea (3). To our knowledge, this is the first report of rice leaf blight caused by P. ananatis in India. The significance of this pathogen to basmati rice production in India was not known until this report. The predominance of the disease in the major basmati-growing belts of northern India would certainly have great impact in reducing the yield potential of basmati rice. References: (1) H. Yan et al. Plant Dis. 94:482, 2010. (2) H. F. Schwartz and K. Otto. Plant Dis. 84:808, 2000. (3) H. B. Lee et al. Plant Dis. 94:1,372, 2010.
2008年9月,印度北部各邦(包括北方邦、哈里亚纳邦和旁遮普邦)的稻田里,一种新的稻瘟病出现在巴斯马蒂水稻(Oryza sativa L.)上。最初症状是水稻叶片尖端出现水渍状病斑。病斑最终蔓延至叶片。受感染叶片变为浅褐色,呈现出枯萎状。该病害在开花后阶段较为严重。2008年至2011年期间,从出现症状的水稻叶片上,在营养琼脂(牛肉浸膏5克、蛋白胨10克、氯化钠5克和琼脂20克)上持续分离出黄色色素细菌。该病害被认为是由水稻白叶枯病菌Xanthomonas oryzae pv. oryzae引起的。然而,对2008年分离的两个菌株(ITCC B0050和ITCC B0055)进行的生理和分子分析表明,致病因子是菠萝泛菌Pantoea ananatis。菌落凸起、半透明,边缘光滑,在37°C下24小时内生长良好。这些细菌是革兰氏阴性兼性厌氧菌,小杆菌单个排列或呈二至五个细胞的链状排列。这些细菌过氧化氢酶和吲哚产生试验呈阳性,而氧化酶和丙二酸肉汤中的碱性反应试验呈阴性。电子显微镜显示,细菌细胞大小为1.1至2.3×0.4至0.7μm,有三至六根周生鞭毛。ITCC B0050和ITCC B0055的16S rRNA基因序列(通过使用引物5'AGAGTTTGATCATGGCTCAG3'和5'AAGGAGGTGATCCAACCGCA3'进行PCR产生1535个核苷酸)(GenBank编号分别为JF756690和JF756691)与菠萝泛菌(GenBank编号DQ512490.1)的序列有99%的核苷酸同一性。对两个菌株进行的Biolog微生物鉴定分析(4.2版)显示,与菠萝泛菌(Biolog公司,加利福尼亚州海沃德)的相似性指数为0.842。通过采用叶尖剪取法接种易感巴斯马蒂水稻(品种Pusa basmati 1)来确认致病性。用无菌水或每种分离物在水中的1×10 CFU/ml悬浮液对叶片进行三次重复接种。人工接种的水稻叶片产生的水渍状病斑与田间自然感染水稻时观察到的相似。接种后10至15天,接种叶片上的病斑干燥,从稻草色变为浅褐色。从受感染的水稻叶片中重新分离出黄色色素细菌,并通过16S rRNA序列分析和Biolog分析确认其身份与原始菌株相同。两种病原菌分离物在接种后24至48小时(1×10 CFU/ml)在烟草(Nicotiana tabacum品种Xanthi)叶片上引发过敏反应。这些研究表明,印度北部新出现的水稻叶瘟病的致病因子是菠萝泛菌。泛菌属是机会性病原菌,据记载可在包括中国水稻籽粒变色(1)、美国洋葱叶枯病和鳞茎腐烂(2)以及韩国水稻叶枯病(3)等经济重要作物上引起不同病害。据我们所知,这是印度首次报道由菠萝泛菌引起的水稻叶瘟病。在本报告之前,这种病原菌对印度巴斯马蒂水稻生产的影响尚不清楚。该病害在印度北部主要巴斯马蒂种植带的盛行肯定会对降低巴斯马蒂水稻的产量潜力产生重大影响。参考文献:(1)H. Yan等人,《植物病害》94:482,2010年。(2)H. F. Schwartz和K. Otto,《植物病害》84:808,2000年。(3)H. B. Lee等人,《植物病害》94:1372,2010年。