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。
【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.
出处
《中国农业科学》
CAS
CSCD
北大核心
2013年第4期678-685,共8页
Scientia Agricultura Sinica
基金
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)
