小麦转录因子TaMYB5-like转录自激活能力及其参与生理型花粉败育过程表达模式研究  

Transcription Self-Activation Ability of the Wheat Transcription Factor TaMYB5-like and Its Expression Pattern Analysed during Pollen Abortion in the Physiological Male Sterilization

作　　者：王芮 唐鹏 单思聪 牛娜[1] 闵东红[1] 马守才[1] 王军卫[1] 孙联合 张改生[1] 宋瑜龙[1] WANG Rui;TANG Peng;SHAN Sicong;NIU Na;MIN Donghong;MA Shoucai;WANG Junwei;SUN Lianhe;ZHANG Gaisheng;SONG Yulong(College of Agronomy,Northwest A&F University/National Yangling Agricultural Biotechnology&Breeding Center/Yangling Branch of State Wheat Improvement Center/Wheat Breeding Engineering Research Center,Ministry of Education/Key Laboratory of Crop Heterosis of Shaanxi Province,Yangling,Shaanxi 712100,China;Zhumadian Academy of Agricultural Sciences,Zhumadian,Henan 463000,China)

机构地区：[1]西北农林科技大学农学院/国家杨凌农业生物技术育种中心/国家小麦改良中心杨凌分中心/小麦育种教育部工程研究中心/陕西省作物杂种优势研究与利用重点实验室,陕西杨凌712100 [2]驻马店市农业科学院,河南驻马店463000

出　　处：《麦类作物学报》2023年第9期1093-1104,共12页Journal of Triticeae Crops

基　　金：国家自然科学基金项目(31701500);陕西省重点研发计划一般项目(2022NY-176);中央高校基本科研业务费专项(Z1090322148)。

摘　　要：化学杂交剂SQ-1诱导的小麦花粉败育是一个复杂的生理代谢过程,TaMYB5-like被发现参与此过程。为了解TaMYB5-like基因在SQ-1诱导小麦生理型花粉败育过程中的作用,以SQ-1诱导的小麦生理型雄性不育系PHYMS-1376和对照可育系CK-1376四分体、单核早期、单核晚期、二核期和三核期花药为试验材料,克隆TaMYB5-like基因,并对其进行序列结构、生物信息学、表达模式、亚细胞定位及转录自激活效应分析。结果发现,TaMYB5-like基因序列长5 207 bp,其cDNA长为1 133 bp,其中CDS序列长819 bp,编码272个氨基酸,含有2个SANT结构域,分子量为29.36 kDa,无信号肽,无跨膜结构,亚细胞定位在细胞核内;TaMYB5-like基因启动子涉及光响应、逆境胁迫响应、茉莉酸甲酯响应等顺式作用元件。自激活效应发现,TaMYB5-like蛋白的自激活区段位于N端,合适浓度的AbA和3-AT可以抑制BD-TaMYB5-like和BD-TaMYB5-like-N-SANT融合蛋白的自激活性;随着蛋白质氨基酸序列的截断,自激活性呈降低趋势。与CK-1376相比,PHYMS-1376四分体和单核早期花药中TaMYB5-like表达量差异不显著;花药发育到单核晚期、二核期和三核期时,TaMYB5-like表达量显著增加;CK-1376和PHYMS-1376三核期时花药内TaMYB5-like表达量均最高。The chemical hybridization agent SQ-1 induces wheat pollen abortion,which is a complex physiological and metabolic process,and TaMYB5-like has been found to be involved in this process.To understand the role of TaMYB5-like gene in the process of SQ-1 induced physiological pollen abortion in wheat,the TaMYB5-like gene was cloned.The SQ-1 induced physiological male sterile line PHYMS-1376 and the control fertile line CK-1376 treated with equal amounts of water were used as experimental materials,including tetrad,early mononuclear,late mononuclear,binuclear,and trinuclear anthers.The sequence structure,bioinformatics,and expression patterns of the TaMYB5-like gene were analyzed and subcellular localization and transcriptional self-activation effect were also performed.The results showed that the TaMYB5-like gene sequence was 5207 bp long,and its cDNA was 1133 bp long.The CDS sequence was 819 bp long,encoding 272 amino acids,containing two SANT domains,with a molecular weight of 29.36 kDa,but without signal peptide,or transmembrane structure.Subcellular localization showed TaMYB5-like is located in the nucleus.The promoter of TaMYB5-like gene involves cis acting elements such as light response,stress response and methyl jasmonic acid response.The self activation effect found that the self activation region of TaMYB5-like protein is located at the N-terminus,and appropriate concentrations of AbA and 3-AT can inhibit the self activation activity of BD-TaMYB5-like and BD-TaMYB5-like N-SANT fusion proteins.As the amino acid sequence of the protein was truncated,the self activation activity decreased.Compared with CK-1376,there was no significant difference in TaMYB5-like expression levels between the tetrad and early mononuclear anthers of PHYMS-1376 plants.TaMYB5-like expression levels significantly increased during the late mononuclear,binuclear,and trinuclear stages of anther development.The expression levels of TaMYB5-like in the anthers of CK-1376 and PHYMS-1376 were the highest during the trinuclear stage.

关 键 词：小麦 TaMYB5-like 酵母双杂交 转录自激活 

分 类 号：S512.1[农业科学—作物学] S330