肯尼亚南裂谷地区玉米致命坏死病病毒的分子特征

R. Leitich, J. Korir, J. Muoma, A. Wangai, K. Bong, G. Johal, S. Loesch-Fries
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摘要

非洲首次报告玉米致命坏死(MLN)病是9年前在肯尼亚的Bomet。此后,它在东非和中非(ECA)国家蔓延,造成了巨大的产量损失。目前,肯尼亚农民种植的首选商业玉米种质中有90%易患MLN病。因此,这种疾病继续对非洲经委会区域的粮食安全构成严重挑战。本研究试图表征从南裂谷地区收集的玉米叶片样本中存在的引起病毒的MLN。利用从Bomet、Kericho和Kisumu县采集的60份叶片样品中提取的总RNA,进行逆转录聚合酶链反应(RT-PCR)。对条带最强的PCR产物进行纯化,并采用Sanger测序技术进行测序。结果表明,3个县的玉米绿斑病毒(MCMV) (MH645622和MH645621)和甘蔗花叶病毒(SCMV) (MH645623、MH645624和MH645625)检测呈阳性,小麦条纹花叶病毒(WSMV)检测呈阴性。MCMV分离株的外壳蛋白(CP)序列与先前报道的东部和撒哈拉以南非洲MCMV分离株的序列密切相关。在SCMV分离株的CP序列中,KCO59株只有一个序列与肯尼亚分离株(JX286708)的序列相似。分离株KCO5和KCO24的序列与肯尼亚分离株JX286708的序列不相同。基于这些结果,在所调查的县中,SCMV分离株具有遗传多样性,而MCMV分离株则没有。存在一种SCMV的新变种,它似乎是与MCMV协同引起肯尼亚MLN病的主要痘病毒。关键词:玉米致死性坏死(MLN),玉米褪绿斑驳病毒(MCMV),甘蔗花叶病毒(SCMV),小麦条纹花叶病毒(WSMV)
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Molecular characterization of viruses causing maize lethal necrosis disease in South-Rift region, Kenya
The first report of Maize lethal necrosis (MLN) disease in Africa was in Bomet, Kenya, nine years ago. It has since spread in East and Central African (ECA) countries, causing massive yield losses. Currently, 90% of the preferred commercial maize germplasm grown by farmers in Kenya is susceptible to MLN disease. As such, the disease has continued to pose a serious challenge to food security in the ECA region. This study sought to characterize the MLN, causing viruses present in the maize leaf samples collected from the South-Rift region. Using total RNA extracted from 60 leaf samples collected from Bomet, Kericho, and Kisumu Counties, reverse transcription-polymerase chain reaction (RT-PCR) was carried out. The PCR products with the strongest bands were purified and sequenced using the Sanger sequencing technique. The results showed that samples from the three counties were positive for maize chlorotic mottle virus (MCMV) (MH645622 and MH645621) and sugarcane mosaic virus (SCMV) (MH645623, MH645624 and MH645625) and negative to wheat streak mosaic virus (WSMV). The coat protein (CP) sequences of MCMV isolates were closely related to the sequences of MCMV isolates, which had been previously reported from Eastern and Sub-Saharan Africa.  For the CP sequences of SCMV isolates, only one sequence of the isolate KCO59 was similar to the sequence of a Kenyan isolate (JX286708). Sequences of isolates KCO5 and KCO24 were not identical to those of the Kenyan isolate (JX286708). Based on these results, in the surveyed counties, isolates of SCMV are genetically diverse, while those of MCMV are not.  There exists a new variant of SCMV, which appears to be the main potyvirus in synergism with MCMV in causing MLN disease in Kenya.   Key words: Maize lethal necrosis (MLN), maize chlorotic mottle virus (MCMV), sugarcane mosaic virus (SCMV), wheat streak mosaic virus (WSMV).
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