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采用番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)侵染克隆接种技术和实时荧光定量PCR方法,从TYLCV在番茄叶片内复制繁殖的角度,系统研究了不同环境温度下单个和多个抗性基因叠加对病毒复制的影响,以期为合理进行抗病基因整合,选育抗TYLCV的番茄新品种提供理论指导。结果显示,(1)春季温室栽培环境下,含Ty-1/Ty-3的番茄材料能抑制病毒复制,接种后28 d其体内病毒含量仅是感病材料病毒含量的千分之一;秋季温室栽培环境下,这种抑制作用降低,病毒含量与感病材料相当。精确控温种植的含Ty-1/Ty-3的近等基因系番茄材料中病毒的含量变化趋势与此相同。(2)含Ty-2的番茄材料在春秋两季栽培环境下,均表现出对病毒复制的抑制作用,接种后28 d其体内病毒含量仅为感病材料病毒含量的万分之一。(3)同时含有2个基因(Ty-1和Ty-2)和多个基因(Ty-1、Ty-2和Ty-3)的番茄材料在抑制病毒复制方面不存在累加效应。
Tomato yellow leaf curl virus (ToLCV) infection cloning and real-time fluorescence quantitative PCR method from the perspective of TYLCV replication in tomato leaves, the system under different ambient temperature single and multiple In order to provide theoretical guidance for rational integration of resistance genes and selection of new tomato varieties resistant to TYLCV, the effects of superposition of resistance genes on virus replication were studied. The results showed that: (1) The tomato material containing Ty-1 / Ty-3 could inhibit the replication of virus in spring greenhouse environment. The virus content in vivo was only 1/1000 of the susceptible material at 28 days after inoculation; Under greenhouse cultivation, this inhibition is reduced and the virus content is comparable to susceptible materials. The same trend was observed in the content of virus in the tomato material with the near-isogenic line Ty-1 / Ty-3 grown by precise temperature control. (2) The tomato material containing Ty-2 showed inhibitory effect on virus replication in the spring and autumn seasons, and the virus content in vivo was only one ten thousandth of the virus content of the susceptible material 28 days after inoculation. (3) Tomato material containing both genes (Ty-1 and Ty-2) and multiple genes (Ty-1, Ty-2 and Ty-3) had no additive effect in inhibiting viral replication.