小麦甾醇14α-去甲基酶Ta CYP51H6d基因克隆及其对丙硫菌唑处理的应答  

Identification of obtusifoliol 14α-demethylase gene TaCYP51H6d from wheat and its response to propionazole treatment

作　　者：刘旭 杜红旭 王放放 左宁 尹钧[1] 牛洪斌[1] LIU Xu;DU Hongxu;WANG Fangfang;ZUO Ning;YIN Jun;NIU Hongbin(College of Agronomy,Henan Agricultural University,Zhengzhou 450046,China)

机构地区：[1]河南农业大学农学院,郑州450046

出　　处：《植物生理学报》2021年第10期2033-2042,共10页Plant Physiology Journal

基　　金：国家重点研发计划项目(2017YFD0301101)。

摘　　要：甾醇14α-去甲基酶属于CYP51亚族,在植物甾醇和油菜素内酯合成过程中发挥重要作用。为明确小麦CYP51基因的功能,本研究从小麦中克隆获得TaCYP51H6d基因(序列号为MT912028),全长2 994 bp,包含2个内含子, cDNA序列长1 849 bp,编码491个氨基酸,蛋白序列包括P450家族结构域和跨膜结构域序列。氨基酸相似性比较显示, TaCYP51H6d与水稻OsCYP51H6相似性最高,达59%;与小麦来源的TaW51和TaW516相似性只有31%和41%。烟草瞬时表达Ta CYP51H6d::GFP融合蛋白显示TaCYP51H6d分布于细胞膜上,属于膜蛋白。荧光定量分析表明,丙硫菌唑拌种处理对幼苗期小麦根系和叶片中TaCYP51H6d的表达具有上调作用。研究同时发现,丙硫菌唑拌种处理下幼苗期小麦叶片的可溶性糖含量显著提高,纤维素含量显著降低。推测TaCYP51H6d可能在小麦甾醇合成中发挥重要作用,丙硫菌唑可能对TaCYP51H6d表达以及纤维素合成具有调控作用。Obtusifoliol 14α-demethylases belong to CYP51 subfamily and play an important role in the synthesis of phytosterol and brassinolide. To investigate the function of CYP51 gene in wheat, Ta CYP51 H6 d gene(GenBank No. MT912028) was cloned. Sequence analysis showed that the total length of Ta CYP51 H6 d is 2 994 bp and contains two introns, the cDNA sequence is 1 849 bp and encodes 491 amino acids. The protein sequence includes P450 family domain and transmembrane domain. The resultes showed that TaCYP51 H6 d had the highest similarity with rice OsCYP51 H6(59%), and lower similarity with TaW51(31%) and TaW516(41%) from wheat. Subcellular localization showed that TaCYP51 H6 d was distributed on cell membrane and belonged to membrane protein. qRT-PCR results showed that propionazole seed-dressing could induce the expression of TaCYP51 H6 d in leaves at seedling stage. Simultaneously, it was found that the soluble sugar content of wheat leaves increased significantly, and the cellulose content decreased significantly under the treatment of seed dressing with propionazole. Collectively, we proposed that TaCYP51 H6 d may play an important role in sterol synthesis in wheat, and that propionazole regulates the expression of Ta CYP51 H6 d and cellulose synthesis.

关 键 词：TaCYP51H6d 丙硫菌唑 表达分析 可溶性糖 纤维素 

分 类 号：S512.1[农业科学—作物学]