| 番茄试管苗抗TYLCV 鉴定技术研究 |
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| 引用本文： | 冯兰香,宗园园,白描,龚慧芝,杨宇红,谢丙炎.番茄试管苗抗TYLCV 鉴定技术研究[J].园艺学报,2012,39(12):2515-2522. |
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| 作者姓名： | 冯兰香 宗园园 白描 龚慧芝 杨宇红 谢丙炎 |
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| 作者单位： | （中国农业科学院蔬菜花卉研究所，北京 100081） |
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| 基金项目： | 公益性行业(农业)科研专项，农业部园艺作物遗传改良重点开放实验室项目，国家大宗蔬菜产业技术体系项目 |
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| 摘 要： | 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 CLN 2123 A’在试管中进行组织培养，21 ~ 25 d后切取长2.0 ~ 2.5 cm的嫩枝，将嫩枝基部在含TYLCV-CN：SH2]侵染性克隆质粒的农杆菌EHA 105菌株的悬浮液（OD600 0.5）中浸润1 min接种。14 d后，感病品种试管苗开始出现TYLCV症状并且愈来愈重，28 d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356 bp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。
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| 关 键 词： | 番茄 试管苗 番茄黄化曲叶病毒 侵染性克隆 抗病性鉴定 |
In Vitro Inoculation Technique for the Identification of Resistance to Tomato yellow leaf curl virus in Tomato |
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| FENG Lan-xiang,ZONG Yuan-yuan,BAI Miao,GONG Hui-zhi,YANG Yu-hong,and XIE Bing-yan.In Vitro Inoculation Technique for the Identification of Resistance to Tomato yellow leaf curl virus in Tomato[J].Acta Horticulturae Sinica,2012,39(12):2515-2522. |
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| Authors: | FENG Lan-xiang ZONG Yuan-yuan BAI Miao GONG Hui-zhi YANG Yu-hong and XIE Bing-yan |
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| Institution: | （Institute of Vegetables and Flowers，Chinese Academy of Agricultural Sciences，Beijing 100081，China） |
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| Abstract: | An in vitro-inoculation test was conducted for developing a new technique to identify tomato resistance to Tomato yellow leaf curl virus（TYLCV）. The tomato seeds of the TYLCV-susceptible‘Moneymaker’，‘Zhongshu 6’and‘Lichun’，and the resistant line AVRDC CLN 2123 A were cultured in the media of test tubes. The tomato microshoots（2.0–2.5 cm in length）of each cultivar were excised after 21–25 days cultivated，and their basal parts were dipped in solutions（OD600 of 0.5）containing Agrobacterium tumefaciens EHA 105 transformed with the infectious clone for the TYLCV-CN：SH2]for 1 min. The inoculated in vitro microshoots were monitored for development of TYLCD（Tomato yellow leaf curl disease）symptoms for 8 weeks post-inoculation. The symptoms of TYLCD started to appear on susceptible in vitro-inoculated tomato plants two weeks after inoculation.Four weeks later，the susceptible plants displayed severe symptoms of stunting，upward leaf curling and yellowing. Eight weeks later，the susceptible plants showed extremely serious TYLCD symptoms and resulted in the browning，necrosisor death. But all of the microshoots of resistant tomato line‘AVRDC CLN 2123 A’remained symptomless and continued the growth normally.Agarose gel showed PCR products（356 bp）amplified from DNA extracts of the plants of the susceptible cultivars except the resistant plants of line‘AVRDC CLN 2123 A’. These results indicated the high possibility to identify the resistance against TYLCV in tomato varieties or germplasms in vitro using tomato microshoots. |
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| Keywords: | tomato in vitro-cultured plant Tomato yellow leaf curl virus the infectious clone disease resistance identification |
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