中国农业科学 ›› 2013, Vol. 46 ›› Issue (4): 678-685.doi: 10.3864/j.issn.0578-1752.2013.04.003

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

拟南芥T1N6_22在抵抗Pst DC3000侵染过程中的功能分析

 郝丛丛, 郑会欣, 贾娇, 司贺龙, 陈展, 赵斌, 张靖, 邢继红, 董金皋   

  1. 1.河北农业大学生命科学学院真菌毒素与植物分子病理学实验室,河北保定 071001
    2.吉林省农林科学院植物保护研究所,吉林公主岭 136100
    3.河北省农林科学院果树研究所,石家庄 050061
  • 收稿日期:2012-11-05 出版日期:2013-02-15 发布日期:2013-01-14
  • 联系方式: 郝丛丛,E-mail:hcc_622@163.com。郑会欣,E-mail:zhhx329hu@sina.com。郝丛丛和郑会欣为同等贡献作者
  • 基金资助:

    国家自然科学基金(31200203)、河北省自然科学基金(C2012204032)、河北农业大学青年科学基金(QJ201231)

Functional Analysis of T1N6_22 in Arabidopsis thaliana Against the Infection of Pst DC3000

 HAO  Cong-Cong, ZHENG  Hui-Xin, JIA  Jiao, SI  He-Long, CHEN  Zhan, ZHAO  Bin, ZHANG  Jing, XING  Ji-Hong, DONG  Jin-Gao   

  1. 1.College of Life Science, Agricultural University of Hebei/Mycotoxin and Molecular Plant Pathology Laboratory, Baoding 071001, Hebei
    2.Institute of Plant Protection, Jilin Academy of Agricultural Sciences, Gongzhuling 136100, Jilin; 3.Institute of Fruit Tree, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050061
  • Received:2012-11-05 Published:2013-02-15 Online:2013-01-14

摘要: 【目的】明确拟南芥抗灰霉病基因T1N6_22在抗Pst DC3000过程中的功能,分析T1N6_22影响拟南芥对Pst DC3000的抗性原因。【方法】对t1n6_22突变体和转基因回复突变体(t1n6_22/T1N6_22)接种Pst DC3000,检测其症状;采用间苯胺蓝染色法检测接种突变体中胼胝质的积累情况;测定接种叶片中Pst DC3000的生长量,明确T1N6_22在拟南芥抗Pst DC3000过程中的功能。利用RT-PCR技术,检测SA、JA和ET对T1N6_22表达的影响及T1N6_22对抗病防御相关基因表达的影响;利用quantitative real-time PCR技术,检测Pst DC3000对T1N6_22及抗病相关基因表达的影响,分析T1N6_22影响拟南芥对Pst DC3000的抗性原因。【结果】t1n6_22突变体接种Pst DC3000后表现明显的抗病症状,而回复突变体t1n6_22/T1N6_22和拟南芥野生型表现明显的感病症状。SA处理拟南芥野生型,T1N6_22的表达量明显增强,经JA和ACC处理,该基因的表达量无显著变化。t1n6_22突变体中,PAL、PR4、PPO、SOD和CAT的表达水平均明显高于野生型Col-0和转基因回复植株。接种Pst DC3000后,拟南芥野生型中T1N6_22及抗病相关基因PR1、PR3、PR5和PDF1.2的表达量明显增强。【结论】T1N6_22在拟南芥抗Pst DC3000过程中起负调控作用,T1N6_22的表达受SA诱导,可能主要通过调节植物的次生代谢产物的分泌影响拟南芥对Pst DC3000的抗性。

关键词: T1N6_22 , 拟南芥 , Pst DC3000 , 抗病信号途径

Abstract: 【Objective】 The objective of this study is to investigate the function of the T1N6_22, a resistant gene of Arabidopsis against B. cinerea, in Arabidopsis resistance to Pseudomonas syringae pv. tomato DC3000, to further analyze the mechanism of T1N6_22 genes in Arabidopsis resistance to Pst DC3000. 【Method】 The symptoms, increase of the content of callose and bacterial concentration of the t1n6_22 and t1n6_22/T1N6_22 in plants inoculated with Pst DC3000 were investigated to study the function of the T1N6_22 in Arabidopsis resistance to Pst. DC3000. RT-PCR technology was used to analyze the expression of T1N6_22 in Col-0 treatment with SA, JA, ET and the expression of defence-related genes in Col-0, the t1n6_22 and t1n6_22/T1N6_22 plants. Quantitative real-time PCR technology was used to analyze the expression of T1N6_22 and the key genes involved in the SA, JA/ET signal pathway in Col-0 inoculated with Pst DC3000. 【Result】 The t1n6_22 exhibited enhanced resistance to Pst DC3000, the Col-0 and complemental plants showed an obvious susceptibility to Pst DC3000. The expression of T1N6_22 in Col-0 treatment with SA was significantly enhanced, suggesting the expression of T1N6_22 induced by SA. Compared with the Col-0 and t1n6_22/T1N6_22 plants, the expression of PAL, PR4, PPO, SOD and CAT genes were downregulated in the t1n6_22 plants. The expression of T1N6_22 and the key genes involved in the SA, JA/ET signal pathway, such as PR1, PR3, PR5 and PDF1.2 genes, were upregulated in Col-0 after inoculation of Pst DC3000.【Conclusion】T1N6_22 gene is a negative regulatory component of Arabidopsis against Pst DC3000 and the T1N6_22 gene expression is induced by SA. T1N6_22 may be involved in the regulation of plant secondary metabolites in impact of the resistance to Pst DC3000 in Arabidopsis.

Key words: T1N6_22 , Arabidopsis thaliana , Pst DC3000 , disease-resistant pathway