甘薯无症状病毒1号韩国分离株农染性克隆的构建及其与本菌烟中农染性的比较

IF 1.8 3区 农林科学 Q2 PLANT SCIENCES Plant Pathology Journal Pub Date : 2023-06-01 DOI:10.5423/PPJ.OA.12.2022.0168
Phuong T Ho, Hee-Seong Byun, Thuy T B Vo, Aamir Lal, Sukchan Lee, Eui-Joon Kil
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摘要

甘薯无症状病毒1 (SPSMV-1)是一种单链环状DNA病毒,属于双病毒科Mastrevirus属,于2012年首次在韩国的甘薯植物上发现。虽然SPSMV-1在甘薯植株中不会引起明显的症状,但它与不同甘薯病毒的共同感染非常普遍,从而威胁到韩国的甘薯生产。本研究采用Sanger测序方法,对在水原地区采集的甘薯植株进行PCR扩增,获得了SPSMV-1韩国分离株的全基因组序列。构建SPSMV-1侵染克隆(1.1-mer),将其克隆到植物表达载体pCAMBIA1303中,利用3株农杆菌(GV3101、LBA4404和EHA105)接种到本烟中。虽然在模拟组和感染组之间没有观察到视觉上的差异,但通过PCR检测到SPSMV-1在根、茎和新产生的叶片中积累。在将SPSMV-1基因组转移到benthamiana上,产瘤杆菌菌株LBA4404最有效。我们通过使用病毒粒子感和互补感特异性引物组进行链特异性扩增,证实了病毒在benthamiana样品中的复制。
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Construction of an Agroinfectious Clone of a Korean Isolate of Sweet Potato Symptomless Virus 1 and Comparison of Its Infectivity According to Agrobacterium tumefaciens Strains in Nicotiana benthamiana.

Sweet potato symptomless virus 1 (SPSMV-1) is a single-stranded circular DNA virus, belonging to the genus Mastrevirus (family Geminiviridae) that was first identified on sweet potato plants in South Korea in 2012. Although SPSMV-1 does not induce distinct symptoms in sweet potato plants, its co-infection with different sweet potato viruses is highly prevalent, and thus threatens sweet potato production in South Korea. In this study, the complete genome sequence of a Korean isolate of SPSMV-1 was obtained by Sanger sequencing of polymerase chain reaction (PCR) amplicons from sweet potato plants collected in the field (Suwon). An infectious clone of SPSMV-1 (1.1-mer) was constructed, cloned into the plant expression vector pCAMBIA1303, and agro-inoculated into Nicotiana benthamiana using three Agrobacterium tumefaciens strains (GV3101, LBA4404, and EHA105). Although no visual differences were observed between the mock and infected groups, SPSMV-1 accumulation was detected in the roots, stems, and newly produced leaves through PCR. The A. tumefaciens strain LBA4404 was the most effective at transferring the SPSMV-1 genome to N. benthamiana. We confirmed the viral replication in N. benthamiana samples through strand-specific amplification using virion-sense- and complementary-sense-specific primer sets.

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来源期刊
Plant Pathology Journal
Plant Pathology Journal 生物-植物科学
CiteScore
4.90
自引率
4.30%
发文量
71
审稿时长
12 months
期刊介绍: Information not localized
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