我国不同地区番茄主要病毒病种类的分子检测与分析

Molecular Detection and Analysis of Main Viruses of Tomato in China

为了检测和鉴定我国不同地区番茄上的病毒种类,2015-2018年,在不同蔬菜种植区采集431份番茄疑似感病样品。利用已报道侵染番茄的12种主要病毒的特异性引物对样品进行RT-PCR分子检测与鉴定,结果表明:TYLCV在番茄上普遍存在,检出率为65.20%;而ToCV、TMV、CMV、ToMV、TSWV和PVY侵染番茄相对较少,检出率分别为21.11%,20.19%,17.63%,15.31%,14.15%,11.37%;未检测到TICV、PVX、TYLCVNB和BBWV等病毒。通过对番茄病毒复合侵染现象进行分析发现,病毒复合侵染率达35.73%,其中2种病毒复合侵染现象最多,占复合侵染总比的52.60%;3种病毒复合侵染率占复合侵染总比27.92%;4种及5种病毒复合侵染率略低,分别占复合侵染总比0.65%和18.83%。通过对我国20个地区的番茄病毒进行检测,结果表明,在番茄上检出病毒种类最多的为北京地区,检测到7种病毒,其次是山东地区,检测到5种病毒;河南、甘肃及宁夏地区检测到4种病毒;西藏、河北、浙江、陕西、海南地区检测到3种病毒;四川、天津、黑龙江、山西、内蒙古等检测到2种病毒;其他地区只检测到单一番茄病毒种类。同时根据TYLCV和TSWV部分基因序列构建系统发育树,结果表明,TYLCV北京分离物(MK336782)与中国湖南分离物的亲缘关系最近,TSWV河南分离物(MK318839)与山东分离物的亲缘关系最近。

In order to detect and identify the viral causing agent that infects tomato vegetable plants in China,a total of 431 susceptible tomato plant samples were collected from different vegetable planting areas during 2015-2018.Total RNA were subjected to detect the viruses using reported specific primers of 12 main viruses and demonstrated by reverse transcription-PCR(RT-PCR).The TYLCV had the highest detection rate(65.20%).ToCV,TMV,CMV,ToMV,TSWV and PVY were popular,having the detection rates of 21.11%,20.19%,17.63%,15.31%,14.15%and 11.37%,respectively.No TICV,PVX,TYLCVNB and BBWV was detected.The complex infection phenomenon showed that the compound infection rate with two viruses accounted for 52.60%of the total,the combined infection rate with three viruses accounted for 27.92%,and the combined infection rates with four viruses and five viruses was slightly lower,accounting for 0.65%and 18.83%,respectively.Based on the detection data of tomato viruses from 20 regions of China,the most detected viruses were in Beijing,of which 7 viruses were detected.Five viruses were detected in Shandong Province.Four viruses were detected in Henan,Gansu and Ningxia.Three viruses were detected in Xizang,Hebei,Zhejiang,Shaanxi and Hainan.Two viruses were detected in Sichuan,Tianjin,Heilongjiang,Shanxi and Neimenggu,while only one virus was detected in other areas.Phylogenetic trees were constructed with partial gene sequences of TYLCV and TSWV,respectively.The isolates of TYLCV from Beijing(MK336782)and Hunan had the closest relationship,and the isolates of TSWV from Henan(MK318839)and Shandong shared the closest relationship.