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作物学报 ›› 2014, Vol. 40 ›› Issue (08): 1392-1402.doi: 10.3724/SP.J.1006.2014.01392

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

小麦自噬相关基因ATG10的克隆及白粉菌侵染诱导的ATG10的表达

张微1,孙鸿1,邢莉萍2,卫晓静1,王华忠1,*   

  1. 1 天津师范大学生命科学学院 / 天津市动植物抗性重点实验室, 天津 300387; 2 南京农业大学作物遗传与种质创新国家重点实验室, 江苏南京 210095
  • 收稿日期:2013-11-20 修回日期:2014-04-16 出版日期:2014-08-12 网络出版日期:2014-06-03
  • 通讯作者: 王华忠, E-mail: skywhz@mail.tjnu.edu.cn
  • 基金资助:

    天津市自然科学基金重点项目(12JCZDJC23000), 霍英东教育基金会高等院校青年教师基金(131026)和天津市动植物抗性重点实验室开放基金项目资助。

Cloning of Autophagy-Related Genes, ATG10s, in Wheat and Their Expression Characteristics Induced by Blumeria graminis f. sp. tritici

ZHANG Wei1,SUN Hong1,WEI Xiao-Jing1,XING Li-Ping2,WANG Hua-Zhong1,*   

  1. 1 School of Life Sciences, Tianjin Normal University / Tianjin Key Laboratory of Animals and Plants Resistance, Tianjin 300387, China; 2 National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China
  • Received:2013-11-20 Revised:2014-04-16 Published:2014-08-12 Published online:2014-06-03
  • Contact: 王华忠, E-mail: skywhz@mail.tjnu.edu.cn

摘要:

细胞自噬是一种保守的真核生物细胞内物质分解和循环利用机制, 在植物生长、发育和逆境响应等过程中均扮演了重要角色。自噬相关蛋白ATG10是参与自噬小体形成的关键因子之一。利用同源克隆方法, 从经白粉病菌诱导48 h的小麦材料92R137/扬麦1587中克隆了ATG10基因家族3个成员(TaATG10aTaATG10bTaATG10c)。序列特征分析、物种间的比较和进化分析, 以及酵母功能互补实验结果证实, 3个基因均为酵母ATG10的功能性同源基因。TaATG10aTaATG10b的基因组序列具有相似的6外显子-5内含子基因结构。RT-PCR分析还发现这2个基因都具有2种可变剪接产物。TaATG10aTaATG10bGFP融合蛋白被定位于洋葱表皮细胞的细胞质中。白粉菌侵染能够诱导TaATG10aTaATG10b表达, 因此推测, 小麦针对白粉菌侵染的免疫反应涉及对TaATG10及其参与的自噬过程的调控, 其调控模式因小麦抗、感白粉病反应、不同类型抗病基因介导的免疫反应和不同遗传背景下的感病反应而差异明显, 说明TaATG10及其参与的自噬过程与小麦—白粉菌互作反应关系的复杂性。从外源激素处理诱导的表达情况还发现, 抗、感白粉病的表型差异可能涉及抗、感材料TaATG10基因对同种激素(SA、乙烯或ABA)信号的不同响应模式。

关键词: 小麦, 自噬相关基因, 白粉病

Abstract:

Autophagy, a conserved eukaryotic cellular process functioning in material decomposition and nutrient recycling, is deeply involved in plant growth, development, and response to stresses. ATG10 is one of the key factors required in autophagosome formation. In this study, we identified three members (TaATG10a, TaATG10b, and TaATG10c) of the ATG10 family in common wheat 92R137/Yangmai 1587 induced by Blumeria graminis f. sp. tritici for 48 h, using homologous cloning technique. The three TaAGT10s shared high similarity with their counterparts from other model plants. Expression of TaATG10a or TaATG10b rescued the autophagy process in ATG10-defective yeast mutant, suggesting that both genes are functional homologues of yeast ATG10. TaATG10a and TaATG10b had similar gene models containing six exons and five introns, and both had two alternatively-spliced mRNA isoforms according to RT-PCR assay. TaATG10a- and TaATG10b-GFP fusion structures were located in cytosol of onion epidermal cells. The expression of TaATG10a and TaATG10b was induced by the infection of Blumeria graminis f. sp. tritici (Bgt), implying that TaATG10s and their involved autophagy process are implicated in the wheat immune response to Bgt. This implication is very complex because the regulated expression profiles of TaATG10a and TaATG10b are different between resistant and susceptible reactions, between different types of resistance gene-mediated immune reactions, and between susceptible reactions on different genetic backgrounds. Besides, exogenous phytohormones also modulated TaATG10a and TaATG10b expressions. The difference of Bgt immune reaction between the resistant and susceptible genotypes might result partially from the different response patterns of TaATG10sto exogenous salicylic acid, ethylene, or abscisic acid.

Key words: Triticum aestivum L., Autophagy-related genes, Powdery mildew

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