小麦类甜蛋白TaTLP1互作蛋白的筛选及其与TaGF14的互作验证  

Screening of Wheat(Triticum aestivum)Thaumatin-like Protein TaTLP1 Interacting Proteins and Verification of Its Interaction with TaGF14

作　　者：申松松 冯燕 孟麟硕 王菲[1] 崔钟池 马秋颖 刘大群[1] 马利松 王海燕[1] SHEN Song-Song;FENG Yan;MENG Lin-Shuo;WANG Fei;CUI Zhong-Chi;MA Qiu-Ying;LIU Da-Qun;MA Li-Song;WANG Hai-Yan(College of Plant Protection/Technological Innovation Center for Biological Control of Crop Diseases and Insect Pests of Hebei Province,Hebei Agricultural University,Baoding 071001,China;College of Horticulture,Hebei Agricultural University,Baoding 071001,China)

机构地区：[1]河北农业大学植物保护学院/河北省农作物病虫害生物防治技术创新中心,保定071001 [2]河北农业大学园艺学院,保定071001

出　　处：《农业生物技术学报》2023年第7期1345-1356,共12页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(32172384,31501623);河北省自然科学基金(C2020204028)。

摘　　要：类甜蛋白(thaumatin-like protein,TLP)是一类重要的病程相关蛋白,在植物抵御生物胁迫和非生物逆境中具有重要作用。课题组前期发现小麦(Triticum aestivum)类甜蛋白基因TaTLP1参与小麦抗叶锈病菌防御反应,为进一步解析其抗病机制,本研究利用GST-pull down和酵母双杂交(yeast two-hybrid,Y2H)技术同时筛选小麦中TaTLP1的互作蛋白,两种技术均筛选到14-3-3类蛋白(TaGF14)为TaTLP1的候选互作蛋白,利用Y2H和共定位技术验证了TaTLP1与TaGF14的互作;qPCR分析发现TaGF14基因与TaTLP1基因受叶锈菌诱导后的表达趋势基本一致,均在叶锈菌诱导前期高表达,且非亲和组合中表达量高于同期亲和组合中表达量;亚细胞定位研究显示TaGF14定位于细胞核。进一步的特征分析表明,TaGF14可能具有潜在的抗病功能。本研究为深入研究TaTLP1参与小麦抗叶锈病防御反应的分子机制以及发掘寄主中更多抗病基因提供了基础,为小麦抗性品种的培育提供更多优秀的抗病资源。Thaumatin-like protein(TLP)belongs to pathogenesis-related proteins(PRPs),which plays an important role in plant resistance to biotic stresses and abiotic stress.Earlier studies have reported that TaTLP1 involved in wheat(Triticum aestivum)defense in response to Puccinia triticina(Pt).To further analyze the resistance mechanism of TaTLP1,in this study,GST-pull down and yeast two hybrid(Y2H)assays were conducted to identify the interacting proteins of TaTLP1 in wheat.14-3-3-like protein(TaGF14)as a candidate interactor of TaTLP1 was identified by both techniques.The interaction between TaTLP1 and TaGF14 was confirmed by Y2H and co-localization.qPCR analysis showed that TaTLP1 and TaGF14 exhibited the similar expression profiles and were highly induced in the early stage of leaf rust infection.Moreover,the expression levels of both genes in the incompatible interaction were higher than that in the compatible interaction.The subcellular localization study showed that TaGF14 was localized in the nucleus.Further characteristic analysis of TaGF14 showed that it might have potential disease resistance function.This study provides a basis for further study on the molecular mechanism of TaTLP1 and participating in the defense response of wheat against leaf rust and exploring more disease resistance genes in the host and provides more excellent disease resistance resources for developing cultivated wheat resistant varieties.

关 键 词：小麦 类甜蛋白 GST-pull DOWN 酵母双杂交(Y2H) TaGF14 基因特征分析 

分 类 号：S432.1[农业科学—植物病理学]