Akhter M S, Holkar S K, Akanda A M, Mandal B, Jain R K
Fruit Research Station, Bangladesh Agricultural Research Institute, Rajshahi-6205, Bangladesh.
Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi-110012, India.
Plant Dis. 2012 Jun;96(6):917. doi: 10.1094/PDIS-02-12-0199-PDN.
An unusual disease of tomato characterized by leaf mottling and necrotic streaks on veins, shortened internodes, necrosis of terminal buds, and concentric rings on fruits was observed during 2010 to 2011 surveys in tomato growing regions of Godagari Upzila, Rajshahi district, Bangladesh. Disease incidence in popularly grown F1 hybrid cultivars, which include Sobal, Abhiruchi, Salamat, Bangobir, and BARI hybrid tomato-5 and -6 in about 40 commercial fields, ranged from 40 to 90%. Extracts from the field samples (n = 10) reacted with polyclonal antiserum to Groundnut bud necrosis virus (GBNV) in direct antigen coated ELISA, suggesting the association of a tospovirus antigenically related to serogroup IV topsovirus (1). To identify whether the tospovirus was a distinct virus species, ELISA-positive samples were subjected to total RNA extraction with an RNeasy Plant Mini Kit (Qiagen, Chatsworth, CA) followed by reverse transcription (RT)-PCR with tospovirus-specific primers (5'-ATGGTTGAAAAGAGCAAGAATGATGC-3') and degenerate primer (5'-CTTCTTATGAAGTGTACTCACCATAAGTCATCC-3') derived from the conserved sequences of GBNV, Watermelon bud necrosis virus (WBNV), and Capsicum chlorosis virus (CaCV) (2). The RT-PCR product was cloned into pGEM-T Easy vector (Promega, Madison, WI) and sequenced at Department of Biochemistry, University of Delhi, South Campus, Delhi, India (GenBank Accession No. JQ692083). The sequences of cloned fragments were assembled. Analysis of the 477-bp region of the nucleocapsid protein (N) gene revealed that the tomato tospovirus shared maximum identity both at the nucleotide (96%) and amino acid (97%) levels with the corresponding region of GBNV. In contrast, only 78 to 81% and 85 to 87% identity at nucleotide and amino acid levels, respectively, was observed with the corresponding region of the N genes of CaCV, WBNV, and Watermelon silver mottle virus. These results suggested the association of GBNV with the diseased tomato samples. To our knowledge, this is the first report of GBNV infecting tomato in Bangladesh and regular surveys are necessary to ascertain the prevalence and incidence of GBNV in other crops. References: (1) R. K. Jain et al. J. Virol. Methods 130:162, 2005. (2) M. Tsompana and J. W. Moyer. Tospovirus. Page 157 in: Encyclopedia of Virology. Academic Press, New York, 2009.
2010年至2011年期间,在孟加拉国拉杰沙希县戈达加里乡的番茄种植区进行调查时,发现了一种番茄的罕见病害,其特征为叶片斑驳、叶脉出现坏死条纹、节间缩短、顶芽坏死以及果实上出现同心环。在约40个商业种植田块中,广泛种植的F1杂交品种(包括索巴尔、阿比鲁奇、萨拉马特、班戈比尔以及BARI杂交番茄-5和-6)的发病率在40%至90%之间。在直接抗原包被酶联免疫吸附测定(ELISA)中,田间样本(n = 10)的提取物与花生芽坏死病毒(GBNV)的多克隆抗血清发生反应,这表明存在一种与血清群IV番茄斑萎病毒抗原相关的番茄斑萎病毒(1)。为确定该番茄斑萎病毒是否为一个独特的病毒种,对ELISA呈阳性的样本使用RNeasy植物小提试剂盒(Qiagen公司,加利福尼亚州查茨沃思)进行总RNA提取,随后使用源自GBNV、西瓜芽坏死病毒(WBNV)和辣椒褪绿病毒(CaCV)保守序列的番茄斑萎病毒特异性引物(5'-ATGGTTGAAAAGAGCAAGAATGATGC-3')和简并引物(5'-CTTCTTATGAAGTGTACTCACCATAAGTCATCC-3')进行逆转录(RT)-PCR(2)。RT-PCR产物被克隆到pGEM-T Easy载体(Promega公司,威斯康星州麦迪逊)中,并在印度德里大学南校区生物化学系进行测序(GenBank登录号:JQ692083)。对克隆片段的序列进行了拼接。对核衣壳蛋白(N)基因477 bp区域的分析表明,该番茄斑萎病毒在核苷酸(96%)和氨基酸(97%)水平上与GBNV的相应区域具有最高同源性。相比之下,与CaCV、WBNV和西瓜银斑驳病毒N基因的相应区域在核苷酸和氨基酸水平上的同源性分别仅为78%至81%和85%至87%。这些结果表明GBNV与患病番茄样本有关。据我们所知,这是GBNV在孟加拉国感染番茄的首次报道,有必要进行定期调查以确定GBNV在其他作物中的流行情况和发病率。参考文献:(1)R. K. Jain等人,《病毒学方法杂志》130:162,2005年。(2)M. Tsompana和J. W. Moyer,《番茄斑萎病毒》,载于《病毒学百科全书》,第157页,学术出版社,纽约,2009年。