小麦F-box/Kelch类基因TaFKOR23的抗逆相关表达模式及分子互作蛋白鉴定  

Stress-related Expression Profile of F-box/Kelch Gene TaFKOR23 in Wheat and Molecular Characterization of the Interacting Target Protein

作　　者：魏春茹 孟钰玉 范润侨 赵梦伊 于秀梅 赵伟全[3] 康振生 刘大群[3] WEI Chun-ru;MENG Yu-yu;FAN Run-qiao;ZHAO Meng-yi;YU Xiu-mei;ZHAO Wei-quan;KANG Zhen-sheng;LIU Da-qun(College of Life Sciences,Hebei Agricultural University/Hebei Key Laboratory of Plant Physiology and Molecular Pathology,Baoding 071001;College of Plant Protection,Northwest A&F University/State Key Laboratory of Crop Stress Biology for Arid Areas,Shaanxi Yangling 712100;Technological Innovation Center for Biological Control of Crop Diseases and Insect Pests of Hebei Province,Baoding 071001)

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

出　　处：《植物遗传资源学报》2020年第3期695-705,共11页Journal of Plant Genetic Resources

基　　金：旱区作物逆境生物学国家重点实验室开放课题基金(CSBAAKF2018008);河北省高等学校科学技术研究项目(ZD2019086);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：为了解F-box成员在小麦中响应生物和非生物逆境的表达情况及作用机制,本研究自小麦抗叶锈病近等基因系TcLr15中克隆了F-box基因TaFKOR23,该基因编码一个由421个氨基酸残基组成的蛋白,N端具有F-box结构域,中间带有2个明显的Kelch结构域,属于F-box/Kelch类型基因。系统进化分析表明,TaFKOR23与粗山羊草(Aegilops tauschii Coss.)和二穗短柄草(Brachypodium distachyon(L.)P.Beauv.)中的F-box/Kelch-repeat protein OR23同源性均较高。利用qRTPCR对接种亲和及非亲和叶锈菌、激素处理、非生物逆境胁迫后TcLr15植株中该基因的表达模式进行分析。研究结果表明,TaFKOR23基因表达受叶锈菌侵染而略升高,但在亲和与非亲和组合间的表达量无明显差异;受脱落酸(ABA)、水杨酸(SA)和茉莉酸甲酯(MeJA)3种激素处理后,该基因均呈先升高后降低的表达趋势,且表达量于处理后12 h达到最高,MeJA对该基因的诱导表达程度略高于SA和ABA;盐胁迫处理后,除了12 h外,TaFKOR23整体呈现上升的表达趋势,最高表达峰出现在处理后48 h;TaFKOR23受聚乙二醇(PEG)处理的影响较小;该基因在旗叶中的表达量远高于其他部位。利用酵母双杂交文库筛选并验证与该基因编码蛋白互作的上游靶蛋白。经文库筛选得到11类可能与TaFKOR23互作的靶蛋白,进一步回转验证及β-半乳糖苷酶检测结果表明TaFKOR23与小麦S-期激酶相关蛋白(TaSkp1)、SEC1家族运输蛋白SLY1(TaSLY1)和几丁质酶2(TaChitinase 2)均存在相互作用。研究结果为深入解析小麦中Kelch类F-box基因的功能及代谢网络奠定了基础,并拓宽了对植物中Kelch类F-box基因的功能认识。To understand the expression profile and molecular mechanism of F-box members in response to biotic/abiotic stresses in wheat,TcLr15-a near isogenic line against leaf rust pathogen,was used as the material to obtain a wheat F-box gene TaFKOR23.The complete coding region encodes a polypeptide of 421 aa with 1 F-box domain at N-terminal and 2 typical Kelch domains in the middle,which means that the wheat TaFKOR23 belongs to F-box/Kelch subfamily.Phylogenetic tree showed that wheat TaFKOR23 had extremely high homology with F-box/Kelch-repeat protein OR23 from Aegilops tauschii Coss.and Brachypodium distachyon(L.)P.Beauv.Expression profiles of the gene in TcLr15 plants after treatment with leaf rust pathogens,3 hormones and abiotic stresses were analyzed by qRT-PCR in the present study.TaFKOR23 was slightly up-regulated by infection of leaf rust pathogen,but no significant difference was observed for TaFKOR23 transcripts accumulation between compatible and incompatible combinations.TaFKOR23 showed a trend of rise first,then fall,and the expression peak reached at 12 h after treatment with 3 homones,and MeJA caused slightly expression of TaFKOR23 than that of SA and ABA.TaFKOR23 transcript increased gradually after treatment with NaCl at all time course except 12 h,and the maximum expression occurred at 48 h.PEG had less effection on TaFKOR23.TaFKOR23 prominently expressed in flag leaves than in other parts of wheat.Target proteins interacting with TaFKOR23 were screened and identified by yeast two-hybrid,11 kinds of target proteins were obtained by library screening,and further results showed that TaFKOR23 can interact with TaSkp1,TaSLY 1 and TaChitinase 2.The research results laid a solid foundation for further understanding the function and metabolic network of F-box/Kelch genes in wheat,and broadened the functional knowledge of Kelch repeat-containing F-box genes in plant.

关 键 词：小麦 F-box/Kelch TaFKOR23 表达模式 蛋白互作 

分 类 号：S512.1[农业科学—作物学]