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小麦与叶锈菌互作过程中TaGRF1-A基因功能的研究OACSTPCD

Study on the Function of TaGRF1-A in the Interaction Between Wheat(Triticum aestivum)and Puccinia triticina

中文摘要英文摘要

克隆小麦(Triticum aestivum)抗叶锈病相关基因,深入研究其表达特征,对阐释小麦抵抗叶锈菌(Puccinia triticina)的分子机制意义重大.通过对本实验室前期得到的RNA-seq转录组数据库的分析,发掘了1个14-3-3家族基因,与小麦抵抗叶锈菌侵染密切相关,在亲和与不亲和组合中表达差异明显.为深入了解该基因在小麦抵御叶锈菌侵染过程中的功能,本研究从小麦抗叶锈近等基因系TcLr26中克隆了该基因,经序列比对显示其为小麦中的TaGRF1-A基因.制备TaGRF1-A蛋白的多克隆抗体,Western blot检测分析表明,该抗体能够与小麦中的TaGRF1-A蛋白呈特异性结合,并且在不亲和互作过程中受叶锈菌诱导呈现先上调后下降的表达趋势.利用病毒诱导的基因沉默(virus-induced gene silencing,VIGS)技术降低不亲和组合中TaGRF1-A的表达量,接种叶锈菌后结果显示,叶锈菌侵染部位的过敏性反应(hypersentive reaction,HR)面积和吸器母细胞(haustorium mother cell,HMC)数量明显增加,在接种叶锈菌15d后叶片表面出现了叶锈菌夏孢子堆,并且致病相关(pathogenesis-related,PR)基因表达降低.结果表明,TaGRF1-A在小麦抵抗叶锈菌侵染过程中正调控寄主的抗锈性.本研究为进一步探究TaGRF1-A在小麦与叶锈菌互作过程的作用机制提供了参考.

The cloning of genes related to wheat(Triticum aestivum)resistance leaf rust and the in-depth study of their expression characteristics are of great significance for elucidating the molecular mechanisms of wheat resistance Puccinia triticina.By analyzing the RNA-seq transcriptome database obtained in the laboratory in the early stage,a gene belong to 14-3-3 family closely related to resistance to P.triticina infection was discovered,with significant differences in expression between compatible and incompatible combinations.In order to deeper understanding of the mechanism of this gene in wheat resistance to P.triticina infection,this study cloned the gene from wheat near isogenic lines TcLr26,the result of sequence alignment showed that the gene was TaGRF1-A in wheat.Preparation of polyclonal antibodies against TaGRF1-A protein,Western blot analysis showed that the antibody was able to bind specifically to TaGRF1-A protein in wheat,and showed an upregulation and then decrease expression trend induced by Puccinia triticina in the incompatible combinations.The virus induced gene silencing(VIGS)technology was used to reduce the expression of TaGRF1-A in incompatible combinations,inoculate with leaf rust showed a significant increase in the area of hypersensitive reaction(HR)and the number of haustorium mother cells(HMCs)at the site of P.triticina infection.After 15 d of inoculation with leaf rust,a spore of P.triticina appeared on the surface of the leaves,and the expression of pathogenesis-related(PR)genes were reduced.The results indicated that TaGRF1-A was positive regulating the wheat resistance to leaf rust infection.This study provides reference for further exploring the mechanism of TaGRF1-A in the interaction between wheat and leaf rust.

顾佳;牛泽霖;王茜;王冬梅

河北农业大学华北作物改良与调控国家重点实验室/河北省植物生理与分子病理学重点实验室/生命科学学院,保定 071001

农业科学

小麦叶锈菌TaGRF1-A多克隆抗体病毒诱导的基因沉默(VIGS)

WheatPuccinia triticinaTaGRF1-APolyclonal antibodyVirus-induced gene silencing(VIGS)

《农业生物技术学报》 2024 (002)

自噬在小麦受叶锈菌侵染诱发的过敏性反应中的作用

247-258 / 12

国家自然科学基金(31171472;31871548);河北省研究生创新资助项目(CXZZBS2022246);河北省自然科学基金(C2023204054)

10.3969/j.issn.1674-7968.2024.02.001

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