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为了开发番茄对番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)抗病性鉴定的新技术,设计了在番茄试管苗中接种TYLCV的试验。将对TYLCV感病的品种‘Moneymaker’、‘中蔬6号’、‘丽春’和抗病品系‘AVRDC CLN2123 A’在试管中进行组织培养,21～25d后切取长2.0～2.5cm的嫩枝,将嫩枝基部在含TYLCV-[CN:SH2]侵染性克隆质粒的农杆菌EHA105菌株的悬浮液(OD 600 0.5)中浸润1min接种。14d后,感病品种试管苗开始出现TYLCV症状并且愈来愈重,28d后TYLCV症状十分典型,56d后症状极其严重:叶片窄小、黄化、极度卷曲,植株严重矮化,停止生长或死亡;而同样接种的抗病品系试管苗却无一发病。接种的番茄试管苗经PCR检测显示,从感病品种试管苗中均能扩增到356bp的特异片段,而在抗病品系中无此条带。这些结果显示了在试管里鉴定番茄品种或材料对TYLCV抗性的可能性。
In order to develop a new technology for tomato disease resistance identification against Tomato yellow leaf curl virus (TYLCV), a test was designed to inoculate TYLCV into tomato plantlets. TYLCV susceptible cultivars ’Moneymaker’, ’Zhongshuang 6’, ’Laichun’ and disease resistant line ’AVRDC CLN2123 A’ were cultured in vitro in test tubes. After 21 ~ 25 days, cut tender 2.0-2.5cm long Shoots, the shoot base was infiltrated into a suspension of Agrobacterium strain EHA105 containing TYLCV- [CN: SH2] infectious cloning plasmid (OD600 0.5) for 1 min. The symptom of TYLCV began to appear more and more severe on the 14th day after inoculation. The symptoms of TYLCV were very typical after 28 days and the symptoms were extremely serious after 56 days: the leaves were narrow and yellowing, extremely curly, the plants were dwarfed, the growth or death was stopped ; While the same vaccination resistant strains of test tube seedlings but no one disease. Inoculated tomato in vitro plantlets by PCR test showed that, from the susceptible varieties of plantlets can be amplified to 356bp specific fragments, but no such strains in the resistant strains. These results show the possibility of identifying TYLCV resistance of tomato cultivars or materials in vitro.