浙江宁波雪里蕻花叶病病原鉴定及雪里蕻品种的抗病性测定

Identification of pathogen responsible for the mosaic disease of Brassica juncea var.multiceps in Ningbo,Zhejiang and evaluation of resistance to Turnip mosaic virus infection

从浙江宁波雪里蕻上获得97株呈花叶症状的病毒样品,利用三抗体夹心酶联免疫吸附测定(TAS-ELISA)在97株雪里蕻花叶样品中均检测到芜菁花叶病毒(Turnip m osaic virus,TuMV),所有的样品都没有检测到黄瓜花叶病毒(Cu-cum berm osaic virus,CMV)和烟草花叶病毒(Tobacco m osaic virus,TMV);利用免疫捕捉反转录PCR(IC-RT-PCR)对部分TuMV样品中的CP和HC-Pro基因进行了扩增,所有样品都得到约0.8kb和1.4kb的2条特异条带,因此宁波雪里蕻花叶病的主要病原是TuMV。对53个雪里蕻栽培品种在温室和大田进行了人工接种,共鉴定出抗病品种7个,耐病品种22个,感病品种24个,未发现高抗品种。总的来说,细叶型品种比花叶型和板叶型品种抗病。利用TAS-ELISA方法对接种的53个雪里蕻品种中的TuMV浓度进行测定,在接种后10、15、20和30d,TuMV检出率分别为45.28%、90.57%、100%和100%,大多数雪里蕻OD405值随接种后天数的增加而呈上升趋势,说明TuMV可以在雪里蕻抗性品种内繁殖,抗性品种的抗性主要表现为耐病。

97 samples were collected from Brassica juncea var. multiceps showing mosaic symptoms in Ningbo, Zhejiang Province. Detection with triple antibody sandwich （TAS）-ELISA indicated that all the 97 sampies were infected by Turnip mosaic virus （ TuMV）, while Cucumber mosaic virus （CMV） and Tobacco mosaic virus （TMV） were not detected in any of them. Immunocapture reverse transcription PCR （IC-RT-PCR） was used to amplify the CP and HC-Pro genes of TuMV in selected samples. PCR products with expected length of 0.8 kb and 1.4 kb for the CP and HC-Pro genes were consistently amplified in these samples. The results indicated that TuMV was the pathogen responsible for the mosaic disease in B. juncea var. multiceps in Ningbo. 53 B. juncea var. multiceps cultivars were inoculated with TuMV in greenhouse and field for evaluation of disease resistance. Out of these cultivars, 7 were resistant, 22 were tolerant,24 were susceptible, but none of them was highly resistant to TuMV. In general, cultivars with thin leaves were more resistant than those with wide leaves. Detection of TuMV by TAS-ELISA was positive for all the cultivars 30 days post inoculation, indicating that TuMV can multiply in resistant cultivars of B. juncea var. multiceps, although these cultivars are tolerant to TuMV infection.