桑树皱叶病毒vⅡ变种侵染性克隆的构建及其致病性鉴定  

Establishment of Infectious Clone of Mulberry Crinkle Leaf Virus vll Variant and Identification of Its Pathogenicity

作　　者：韩涛涛 张朋 唐佳璇 马宇 钟馗 卢全有 Han Taotao;Zhang Peng;Tang Jiaxuan;Ma Yu;Zhong Kui;Lu Quanyou(School of Biotechnology,Jiangsu University of Science and Technology,Zhenjiang Jiangsu 212100,China;Key Laboratory of Genetic Improvement of Silkworm and Mulberry,Ministry of Agriculture and Rural Affairs,Sericultural Research Institute,Chinese Academy of Agricultural Sciences,Zhenjiang Jiangsu 212100,China)

机构地区：[1]江苏科技大学生物技术学院,江苏镇江212100 [2]中国农业科学院蚕业研究所农业农村部蚕桑遗传改良重点实验室,江苏镇江212100

出　　处：《蚕业科学》2023年第1期8-14,共7页ACTA SERICOLOGICA SINICA

基　　金：江苏省高校自然科学研究重大项目(18KJA210001)。

摘　　要：桑树皱叶病毒vⅡ变种是对桑树皱叶病毒(mulberry crinkle leaf virus,MCLV)进行遗传进化分析发现的,其对桑树的致病性尚不清楚。利用PCR和同源重组的方法构建了农杆菌介导的含有2倍大基因间隔区(large intergenic region,LIR)的MCLV vⅡ侵染性克隆,分别接种到其天然宿主桑树和实验植物番茄植株上,以期明确MCLV vⅡ变种对桑树的致病性及对实验植物的侵染性和致病性。结果显示,桑树和番茄分别在接种45 d和25 d检测到全部侵染成功的植株,说明构建的含有2倍LIR的MCLV vⅡ侵染性克隆具有侵染性;MCLV vⅡ的侵染性克隆可以侵染番茄,且草本植物番茄接种成功率和接种响应速度都要高于和快于木本植物桑树,说明番茄可以作为MCLV研究的实验宿主;接种后检测为阳性的桑树在90 d的观察期内,其叶部没有表现出病毒感染样症状,接种后检测为阳性的番茄在45 d的观察期内未表现出任何病毒感染样症状。这些发现为双生病毒侵染性克隆的构建提供了一种新的思路,并为进一步研究MCLV与寄主互作和MCLV的应用奠定基础。Mulberry crinkle leaf virus variant ll(MCLV vll)was discovered by genetic evolution analysis of mulberry crinkle leaf virus(MCLV),and its pathogenicity to mulberry remained unknown.Agrobacterium-mediated MCLV vll infectious clones containing 2-repeats large intergenic region(LIR)were constructed by PCR and homologous recombination and inoculated into their natural host mulberry and experimental plant tomato,respectively,in order to clarify the pathogenicity to mulberry and the infectivity and pathogenicity to experimental plants.The results showed that all successfully infected plants were detected in mulberry and tomato at 45 d and 25 d post inoculation,respectively,indicating that the constructed MCLV vll infectious clone containing 2-repeats LIR was infectious.The infectious clones of MCLV vll could infect tomato,and the inoculation success rate and inoculation response rate of herbaceous tomato were higher and faster than those of the woody plant mulberry,indicating that tomato could be used as an experimental host for MCLV study.Mulberry plants that were tested positive after inoculation did not show virus infection-like symptoms in their leaves during the 90-days observation period,and tomatoes that were tested positive after inoculation did not show any virus infection-like symptoms during the 45 d observation period.These findings provide a new idea for the construction of geminivirus infectious clones,and lay the foundation for further study of interaction between MCLV and host and application of MCLV.

关 键 词：桑树皱叶病毒 侵染性克隆 致病性 桑树 番茄 

分 类 号：S888.712[农业科学—特种经济动物饲养]