摘要
应用DAS-ELISA和RT-PCR方法从褪绿和银色斑驳的西瓜叶片中检测到病毒分离物(WSMoV-YN),感病样品能与WSMoV/GBNV复合抗血清(Agdia)呈阳性反应。获得WSMoV N蛋白的多克隆抗体,抗体能与WSMoV血清组成员CaCV和TZSV反应,但不能与INSV、TSWV、HCRV和GYSV反应。为明确引起该病害的病毒种类,采用Tospovirus通用引物对样品的总RNA进行RT-PCR扩增,获得长度为3 554 nt的S RNA全序列,经Blastn比对分析与WSMoV中国台湾分离物同源性最高,为95.8%,其N和NSs蛋白氨基酸序列同源性分别为99%和97.6%。构建系统进化树发现,西瓜银灰斑驳病毒云南分离物(WSMoV-YN)与其他WSMoV聚为一支。确定引起云南西瓜病害的病毒为WSMoV。
Using DAS-ELISA and RT-PCR, a viral isolate （WSMoV-YN） was detected in watermelon show- ing chlorotic and silver mottle. Diseased samples were positive for antibodies to Watermelon silver mottle virus （WSMoV） and Groundnut bud necrosis virus （GBNV） （Agdia, USA）. The polyclonal antiserum of N protein were obtained and could react with Capsicum chlorosis virus （CaCV） and Tomato zonate spot virus （TZSV）, the members of Watermelon silver mottle virus （ WSMoV ） serogroup, but not react with Impatiens necrotic spot virus （INSV）, Tomato spotted wilt virus （TSWV）, Hippeastrum chlorotic ringspot virus （HCRV） and Ground- nut yellow spot virus （GYSV）. To further confirm the presence of WSMoV, the samples were examined by RT- PCR using tospovirus universal primer pairs. Full-length sequence of WSMoV-YN S RNA with a length of 3 554 nt was obtained and sequence alignment indicated that WSMoV-YN shares 95.8% identity with WSMoV isolates from Taiwan, China （NC_003843） at the nucleotide level. The nucleocapsid protein （N） and non-structual （NSs） shared 99% and 97.6% amion acid identity with the Taiwan（China） isolates, respectively. Taken together, our results suggested that the WSMoV-YN formed a distinct cluster with the other WSMoV isolates.
出处
《植物病理学报》
CAS
CSCD
北大核心
2016年第4期461-468,共8页
Acta Phytopathologica Sinica
基金
农业部公益性行业科研专项(201303028)
关键词
西瓜
西瓜银灰斑驳病毒
S
RNA
血清学特性
watermelon
Watermelon silver mottle virus
S RNA
serological characterization
