不同番茄品种对番茄黄化曲叶病毒的抗病性分析

doi:10.3864/j.issn.0578-1752.2016.14.020

摘要/Abstract

摘要： 【目的】番茄黄化曲叶病毒病 (Tomato yellow leaf curl virus，TYLCV) 是一种影响全世界番茄生产的病害，培育抗病品种是最有效、经济和持久的防控手段。目前生产上应用的抗病品种繁多，但存在对TYLVC不同抗性基因的抗性水平研究甚少、了解不够全面的问题。为了解带有不同抗性基因的番茄品种对番茄黄化曲叶病毒病的抗性，对北京市农林科学院蔬菜中心培育的番茄品种抗病性进行检测，并利用半定量RT-PCR方法分析抗病基因和抗病蛋白的表达水平。【方法】选取携带有Ty3a/Ty3a的番茄材料秋光285（Ty3a 纯合）、棚137×246（Ty3a 杂合），携带Ty1+Ty3a纯合的秋光120、携带Ty1+Ty3a杂合的秋光81和感病对照M82为试验材料，观察番茄植株自然条件下发病情况。通过调查发病率和病情指数，考查这些番茄品种的抗病性，并利用RT-PCR方法分析抗病基因Ty1和Ty3a以及病程相关蛋白葡聚糖酶和几丁质酶基因的表达水平。【结果】携带抗性基因的杂合体、纯合体抗病性差异不明显，含有两个抗性基因和含有单一抗病基因品种的抗病性稍有差异，但差异不显著。半定量RT-PCR结果显示不同番茄品种在感染TYLCV后，体内抗性基因表达量随着发病时间的增加，Ty-3a和Ty-1的表达量逐渐增强。Ty-3a在4个番茄品种中表达水平由弱到强依次为秋光285（Ty-3a纯合）、棚137×246（Ty-3a杂合）、秋光120（Ty-1+Ty-3a纯合）和秋光81（Ty-1+Ty-3a杂合），Ty-1在秋光81中的表达显著高于在秋光120中的表达。同时，植株体内葡聚糖酶和几丁质酶基因表达量均比对照显著增加，并随着发病时间的增加逐渐增强。在发病20 d和30 d后，携带Ty-1+Ty-3a杂合的秋光81的葡聚糖酶基因表达显著强于其他几个品种。【结论】几个番茄材料对TYLCV的抗性由弱到强依次为秋光285（Ty3a 纯合）、棚137×246（Ty3a杂合）、秋光120（Ty1+Ty3a纯合）和秋光81（Ty1+Ty3a杂合）。不同番茄抗病品种感染TYLCV后，体内抗性基因表达量随发病时间增加而增强，葡聚糖酶和几丁质酶基因表达量均比感病对照显著增加，表明番茄抗病品种在感染TYLCV后的抗病性除了与抗性基因的表达有关外，同时也与抗病相关蛋白表达增强有关。

关键词: 番茄黄化曲叶病毒, 抗病基因, 病程相关蛋白, 半定量RT-PCR

Abstract: 【Objective】Tomato yellow leaf curl virus (TYLCV) disease is a serious threat to tomato production worldwide, breeding resistant varieties is the most economical, effective and friendly way to control this disease. Most of the tomato varieties resistant to TYLCV used in production of China were introduced from abroad. Many new tomato varieties resistant to TYLCV have been obtained by breeding, but the resistance is unknown. In this paper, the resistance levels of the new tomato varieties resistant to TYLCV bred by Vegetable Research Center, Beijing Academy of Agriculture and Forestry Sciences were assessed and the expression of resistance genes Ty-1/Ty-3a and the pathogenesis-related proteins glucanase/chitinase were analyzed. The purpose is to provide a reference for rational utilization of germplasm resources and reduce the harm of TYLCV.【Method】Four tomato varieties Qiuguang 285, Peng 137×246, Qiuguang 120 and Qiuguang 81 were used as the experimental materials. TYLCV disease incidence was observed in the field. The disease resistance of these tomato varieties was examined by investigating the disease incidence and index. The resistance levels of the new tomato varieties resistant to TYLCV were assessed and the expression of resistance genes and the pathogenesis-related proteins were analyzed by RT-PCR. 【Result】 The resistance of the hybrid was no obvious differences with that of the pure and the resistance of the two genes was a little higher than that of single resistance genes, but yet no significant differences. Semi-quantitative RT-PCR results showed that the expression of genes Ty-3a and Ty-1 increased with the increase of the incidence of TYLCCV in different tomato varieties. The expression level of Ty-3a in four tomato varieties from weak to strong is Qiuguang 285, Peng 137×246, Qiuguang 120 and Qiuguang 81. The expressionof Ty-1 in Qiuguang 81 was obviously higher than that of in Qiuguang 120. The gene expression of glucanase and chitinase in vivo was significantly higher than that in the control. The expression of the two genes increased with the increase of the time. In the 20 and 30 days after appearing of disease, the expression of glucanase gene in Qiuguang 81 with the Ty-1+Ty-3a was significantly higher than that in other varieties.【Conclusion】The resistance of four tomato varieties to TYLCV from weak to strong is Qiuguang 285, Peng 137×246, Qiuguang 120 and Qiuguang 81. After the infection of TYLCV, the expression levels of resistance genes increased with the time of disease in different tomato varieties. The expression levels of glucanase and chitinase genes in vivo were significantly higher than those in the control. The results suggested that the high resistance of variety to TYLCV is related to the high expression of resistance gene and pathogenesis-related proteins.

Key words: Tomato yellow leaf curl virus, resistance gene, pathogenesis-related proteins, semi-quantitative RT-PCR

田兆丰，刘伟成，厚凌宇，谢华，罗晨，柴敏. 不同番茄品种对番茄黄化曲叶病毒的抗病性分析[J]. 中国农业科学, 2016, 49(14): 2850-2856.

TIAN Zhao-feng, LIU Wei-cheng, HOU Ling-yu, XIE Hua, LUO Chen, CHAI Min. Analysis of Resistance of Different Tomato Varieties to Tomato yellow leaf curl virus in Tomato[J]. Scientia Agricultura Sinica, 2016, 49(14): 2850-2856.

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