小麦F-box/LRR类基因TaFBL14的抗逆相关表达模式及互作蛋白鉴定  

Stress-related expression profiles and identification of the target protein of wheat F-box/LRR gene TaFBL14

作　　者：魏春茹[2] 孟钰玉 范润侨 李虎滢 赵伟全 于秀梅[1,2,3] 康振生 刘大群[3] WEI Chunru;MENG Yuyu;FAN Runqiao;LI Huying;ZHAO Weiquan;YU Xiumei;KANG Zhensheng;LIU Daqun(State Key Laboratory of Crop Stress Biology for Arid Areas,College of Plant Protection,Northwest A&F University,Yangling,Shaanxi 712100,China;Hebei Key Laboratory of Plant Physiology and Molecular Pathology,College of Life Sciences,Hebei Agricultural University,Baoding,Hebei 071001,China;Technological Innovation Center for Biological Control of Crop Diseases and Insect Pests of Hebei Province,Baoding,Hebei 071001,China)

机构地区：[1]旱区作物逆境生物学国家重点实验室,西北农林科技大学植物保护学院,陕西杨凌712100 [2]河北省植物生理与分子病理学重点实验室,河北农业大学生命科学学院,河北保定071001 [3]河北省农作物病虫害生物防治技术创新中心,河北保定071001

出　　处：《植物生理学报》2020年第4期886-894,共9页Plant Physiology Journal

基　　金：河北省高等学校科学技术研究项目(ZD2019086);旱区作物逆境生物学国家重点实验室开放课题基金(CSBAAKF-2018008);国家自然科学基金(31301649);高等学校博士学科点专项科研基金(20121302120010)。

摘　　要：F-box家族是植物中最大的蛋白质家族之一,广泛参与植物生长发育、自交不亲和、衰老、生物/非生物胁迫等多个生物学过程。拟南芥(Arabidopsis thaliana)、水稻(Oryza sativa)等模式植物中F-box家族的研究开展较早,认识比较全面,但对重要的粮食作物小麦(Triticum aestivum)中F-box家族的报道还比较少。为了解F-box家族成员在小麦中响应生物/非生物逆境的情况及具体作用机制,以本实验室前期获得的F-box基因TaFBL14为研究对象,探讨了该基因在3种激素和3种非生物逆境处理下的表达模式。实时定量PCR (qRT-PCR)结果表明,水杨酸(SA)、脱落酸(ABA)、茉莉酸甲酯(MeJA)三种激素以及氯化钠(NaCl)、过氧化氢(H2O2)、聚乙二醇6000 (PEG6000)三种逆境胁迫处理后, Ta FBL14基因均呈现先升高后降低的表达趋势。经三种激素处理后,TaFBL14的表达峰值均为处理后12 h,相对于SA和ABA两种激素处理, MeJA处理可诱导TaFBL14更高的表达水平;在三种逆境胁迫下,相对于H2O2和PEG6000处理, NaCl可诱导TaFBL14更高的表达水平,且峰值出现在处理后6 h。本研究构建了酵母载体pGBKT7-TaFBL14,并以之为诱饵筛选了非亲和叶锈菌(Puccinia triticina) 05-19-43②侵染的小麦TcLr15叶片的酵母cDNA文库;钓取了11类可能与TaFBL14互作的靶蛋白;进一步的回转验证及β-半乳糖苷酶检测结果表明, TaFBL14与TaChitinase 2 (TaChi 2)存在相互作用。研究结果拓宽了对小麦中富含亮氨酸(LRR)类F-box基因功能的认识,拟为深入解析小麦中F-box/LRR基因的功能及代谢网络提供参考。F-box proteins constitute one of the largest protein families in plant, and are widely involved in many biological processes, such as plant growth and development, self-incompatibility, senescence, biotic/abiotic stresses, etc. The F-box family has been characterized earlier and more thoroughly in the model plant, such as Arabidopsis thaliana and rice(Oryza sativa), but less is known in wheat(Triticum aestivum)—an important cereal crop. In order to understand the expression profiles and detailed molecular mechanisms of F-box members in response to biotic/abiotic stresses in wheat, TaFBL14 obtained previously by our group was used as the research target to analyze its expression patterns under treatments with three hormones and three abiotic stresses using quantitative real-time PCR(qRT-PCR). The results show that TaFBL14 expression increased first and then decreased at wheat TcLr15 treated with salicylic acid(SA), abscisic acid(ABA), methyl jasmonate(MeJA), NaCl, H2O2 and polyethylene glycol 6000(PEG6000). The expression reached peak level at 12 h after treatments with three hormones, and MeJA caused higher expression of TaFBL14 compared to treatments with SA and ABA. Among the treatments of three abiotic stresses, NaCl induced relatively stronger expression compared to H2O2 and PEG6000, and the maximal expression occurred at 6 h after treatment. Yeast bait vector pGBKT7-TaFBL14 was constructed, and was used to screen the yeast cDNA library of wheat TcLr15 leaves infected by incompatible leaf rust pathogen(Puccinia triticina) strain 05-19-43② by yeast two-hybridization(Y2 H). Eleven types of target proteins interacting with TaFBL14 were obtained and identified. Y2H and β-galactosidase assays further showed that TaFBL14 could interact with TaChitinase 2(TaChi 2). The research results would broaden knowledge for the function of leucine rich repeat(LRR)-like F-box genes in wheat, and lay a foundation for further understanding the function and metabolic network of wheat F-box/LRR genes.

关 键 词：小麦 F-box/LRR TaFBL14 表达模式 蛋白互作 

分 类 号：S512.1[农业科学—作物学]