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孟加拉国首次报道由[具体病菌名称缺失]引起的水稻细菌性穗枯病(BPB)。

First report of bacterial panicle blight (BPB) in Rice caused by in Bangladesh.

作者信息

Islam Md Rashidul, Jannat Rowzatun, Protic Ismam Ahmed, Happy Mst Nusrat Arobi, Samin Samia Islam, Mita Mamuna Mahjabin, Bashar Samrin, Masud Md Mostafa, Islam Huzzatul, Uddin Md Nasir, Akter Mst Arjina, Alam Md Zahangir

机构信息

Bangladesh Agricultural University, 54492, Plant Pathology, Department of Plant Pathology, Faculty of Agriculture Building., Bangladesh Agricultural University, Mymensingh, Mymensingh, Mymensingh, Bangladesh, 2202;

Bangladesh Agricultural University, 54492, Mymensingh, Bangladesh;

出版信息

Plant Dis. 2023 May 23. doi: 10.1094/PDIS-02-23-0229-PDN.

Abstract

Bacterial panicle blight (BPB) has become one of the most destructive diseases of rice worldwide and and are two major pathogens causing BPB (1). This disease causes several types of damage, most importantly grain spotting, rot, and panicle blight, which can result in yield losses of 75% or more (1,3). In recent years, symptoms including sheath rot, grain spotting, grain rot, and panicle blight have been observed in both inbred and hybrid rice varieties. These symptoms resemble those of BPB and cause cultivar-dependent yield losses. (3) also reported the same symptoms for BPB. To confirm the cause of the disease, 21 rice panicles (Haridhan, a local variety) with typical BPB symptoms were collected from a farmer's field in the region of Mymensingh, Bangladesh during the rainy season in mid-October, 2021. Due to the severity of the outbreak, the panicles became dark brown and produced chaffy grains; nearly 100% of the rice panicles in that field were severely infected. To identify the causal pathogen(s), 1g of rice grains from 20 plants with typical BPB symptoms were surface-sterilized by immersing them in 70% ethanol for a few seconds followed by sodium hypochlorite solution (3%) for 1min. The grains were then rinsed with sterilized distilled water three times. Surface-sterilized grains were then ground with a mortar and pestle; 5mL of sterile distilled water was added during grinding. The extracted suspension (20µL) was then either streaked or spread onto the selective medium (S-PG) (2). Bacterial colonies showing purple color on the S-PG medium were selected and purified as candidate pathogens. For molecular characterization, species specific primers targeting gene were used to perform PCR and resulted in 479bp as reported by (4). To verify further, the PCR products of 16SF & 16SR were amplified and sequenced partially producing around 1400bp (1) and five 16SF partial sequences were deposited into NCBI GenBank (OP108276 to OP108280). 16S rDNA and revealed almost 99% homology with (KU851248.1, MZ425424.1) and (AB220893, CP033430) respectively using BLAST analysis. These purified bacterial isolates produced a diffusible light-yellow pigment on King's B medium indicating toxoflavin production (3). The candidate five bacterial isolates were then confirmed by inoculating 10ml suspension 10CFU/mL into the panicles and sheaths of BRRIdhan28 in net house condition as described previously (1). All of the bacterial isolates obtained from the spotted rice grains produced light brown lesions on the inoculated leaf sheath as well as spotting on the grain. To fulfill Koch's postulates, the bacteria were re-isolated from the symptomatic panicles and were confirmed as by analyzing the sequences of and 16s rDNA genes. Taken together, these results confirmed that is responsible for causing BPB in the rice grain samples that we collected. To our knowledge, this is the first report of BPB caused by in Bangladesh and further research is necessary to develop an effective disease management technique, or else the production of rice will be severely hampered.

摘要

细菌性穗枯病(BPB)已成为全球水稻最具毁灭性的病害之一,[病原菌名称1]和[病原菌名称2]是引发BPB的两种主要病原菌(1)。这种病害会造成多种损害,最重要的是谷粒出现斑点、腐烂以及穗枯,可能导致产量损失达75%甚至更多(1,3)。近年来,在常规稻和杂交稻品种中均观察到包括叶鞘腐烂、谷粒斑点、谷粒腐烂和穗枯在内的症状。这些症状与BPB的症状相似,并导致了因品种而异的产量损失。(3)也报道了BPB的相同症状。为了确定病害的病因,2021年10月中旬雨季期间,从孟加拉国迈门辛地区一位农民的田地里采集了21个具有典型BPB症状的水稻穗(当地品种Haridhan)。由于病害爆发严重,这些稻穗变成深褐色并产生秕谷;该田地中近100%的水稻穗受到严重感染。为了鉴定致病病原菌,从20株具有典型BPB症状的植株上取1克稻粒,将其浸入70%乙醇中几秒钟,然后再浸入次氯酸钠溶液(3%)中1分钟进行表面消毒。然后用无菌蒸馏水冲洗这些谷粒三次。对表面消毒后的谷粒用研钵和研杵研磨;研磨过程中加入5毫升无菌蒸馏水。然后将提取的悬浮液(20微升)划线接种或涂布在选择性培养基(S - PG)(2)上。选择在S - PG培养基上呈现紫色的细菌菌落并进行纯化,作为候选病原菌。为了进行分子鉴定,使用针对[基因名称]基因的种特异性引物进行PCR,如(4)报道的那样得到了479碱基对的产物。为了进一步验证,对16SF和16SR的PCR产物进行扩增并部分测序,产生了约1400碱基对(1),并将五个16SF部分序列存入NCBI基因库(OP108276至OP108280)。使用BLAST分析表明,16S rDNA和[相关基因名称]分别与[参考菌株1](KU851248.1,MZ425424.1)和[参考菌株2](AB220893,CP033430)具有几乎99%的同源性。这些纯化的细菌分离物在King's B培养基上产生了一种可扩散的浅黄色色素,表明产生了毒黄素(3)。然后按照之前描述的方法(1),在网室条件下将10毫升浓度为10CFU/mL的悬浮液接种到BRRIdhan28的稻穗和叶鞘中,对这五种候选细菌分离物进行验证。从有斑点的稻粒中获得的所有细菌分离物在接种的叶鞘上产生了浅褐色病斑,并且谷粒上也出现了斑点。为了满足柯赫氏法则,从有症状的稻穗中重新分离出细菌,并通过分析[相关基因名称]和16s rDNA基因的序列,确认其为[病原菌名称]。综上所述,这些结果证实了[病原菌名称]是我们采集的水稻谷粒样本中导致BPB的病原菌。据我们所知,这是孟加拉国关于由[病原菌名称]引起BPB的首次报道,有必要进一步开展研究以开发有效的病害管理技术,否则水稻生产将受到严重阻碍。

相似文献

1
First report of bacterial panicle blight (BPB) in Rice caused by in Bangladesh.
Plant Dis. 2023 May 23. doi: 10.1094/PDIS-02-23-0229-PDN.

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