红花黄萎病病原菌鉴定

番茄斑萎病毒Tomato spotted wilt virus （TSWV）是我国进境植物检疫性有害生物，近年来相继在国内一些省份发现。利用 TSWV 的通用引物 NF302／NR575对从山东烟台地区收集的15份疑似感染番茄斑萎病毒病的番茄样品进行检测；进一步对特异引物 TSWV-NF2037／TSWV-NR2825扩增的 TSWV 的 N 基因序列克隆、测序，并对 N 基因片段编码氨基酸进行遗传距离及系统发育分析。结果表明，15份疑似样品中有4个样品扩增得到 TSWV 病毒片段；基于 N 基因序列分析发现，山东 TSWV 番茄分离物与云南 TSWV 番茄分离物（AEI70836．1）的遗传距离最近，为0．8％，且与云南 TSWV 番茄分离物聚为一支。这是山东地区首次利用分子标记证实番茄斑萎病毒病的危害。

Identification of the pathogen causing Verticillium wilt on safflower

Tomato spotted wilt virus (TSWV) is an important plant quarantine pest, which has been found in some provinces of China in recent years. In this study, 15 suspected tomato samples collected from Yantai, Shandong were first detected by using PCR with the TSWV-universal primers NF302/NR575. Then, the N gene of TSWV isolates amplified by specific primers TSWV-NF2037/TSWV-NR2825 was sequenced for phylogenetic analysis and genetic distance analysis. The results based on the detection with NF302/NR575 showed that four of the 15 samples were positive for TSWV.Genetic distance analysis based on the N gene sequences showed that the genetic distance between Shandong isolate and Yunnan (AEI70836.1) was the closest (i.e. 0.8%). Phylogenetic analysis showed that the Shandong isolate was closely related to the Yunnan isolate (AEI70836.1). Our results proved the introduction of TSWV into Shandong, which is the first evidence of TSWV disease in Shandong based on molecular markers.