芋花叶病毒的RT-PCR检测及外壳蛋白基因序列分析  

The Detection of Dasheen mosaic virus by RT-PCR and Sequence Analysis of Its Coat Protein Gene

作　　者：施世明[1] 王国平[1,2] 徐文兴[1] 王利平[1] 洪霓[1,2] 

机构地区：[1]华中农业大学植物科技学院,湖北省作物病害监测与安全控制重点实验室,武汉430070 [2]华中农业大学,农业微生物学国家重点实验室,武汉430070

出　　处：《园艺学报》2012年第3期509-515,共7页Acta Horticulturae Sinica

基　　金：国家农业行业专项计划项目(nyhyzx200903017-08)

摘　　要：采用RT-PCR技术对采自中国湖北、浙江和山东的91份芋[Colocasia esculenta（L.）Schott]样品的芋花叶病毒（Dasheen mosaicvirus,DsMV）进行了检测,检出率为26.4%。对其中14个DsMV分离物的317bp扩增产物（为外壳蛋白基因的一部分）序列分析的结果显示,各分离物内的核苷酸变异相对较低,而分离物间存在较大的分子变异,相似性为68.3%~97.8%。对来自湖北和浙江的2个DsMV分离物DsMV-SCS和DsMV-JH的外壳蛋白基因（coatproteingene,cp）进行了测序,全长分别为951bp和987bp,二者cp核苷酸和氨基酸序列相似性分别为79.0%和82.3%,与已报道DsMV的cp核苷酸和氨基酸序列相似性分别为73.0%~92.1%和74.8%~98.2%,在构建的系统发育树上聚在两个不同的簇,各DsMV分离物的系统进化关系与其寄主和地理来源无显著相关性。Reverse transcription PCR（RT-PCR）was used for the detection of Dasheen mosaic virus （DsMV）in taro[Colocasia esculenta（L.）Schott] from Hubei,Zhejiang and Shandong Provinces. Results revealed that the average viral infection frequency in 91 collected taro samples was 26.4%. RT-PCR products of 317 bp（covering partial coat protein gene）from 14 isolates were sequenced. Results showed that the obtained sequences had high intra-isolate nucleotide similarities,but inter-isolate nucleotide similarities were varied from 68.3% to 97.8%. The cp genes of two DsMV isolates named DsMV-SCS and DsMV-JH were sequenced,and their sizes were 951 bp and 987 bp,respectively. Their cp genes shared similarities of 79.0% at nucleotide（nt）level and 82.3% at amino acid（aa）level to each other and similarities of 73.0%–92.1% at nt level and 74.8%–98.2% at aa level to reported cp sequences of DsMV,respectively. The phylogenetic trees constructed based on nucleotide and deduced amino acid sequences of the cp of DsMV showed that isolates DsMV-SCS and DsMV-JH clustered into two different groups. There was no obvious correlation between phylogenetic positions and host or geographical origins of different DsMV isolates.

关 键 词：芋 芋花叶病毒 RT-PCR 外壳蛋白基因 序列分析 

分 类 号：S632.3[农业科学—蔬菜学]