苦荞FtHCT的基因克隆与生物信息学分析  

Cloning and Bioinformatics Analysis of Gene FtHCT from Tatary Buck- wheat

作　　者：杨晓琳 段迎 蔡苏云 贺润丽 尹桂芳[2] 王艳青[2] 卢文洁[2] 孙道旺[2] 王莉花[2] Yang Xiaolin;Duan Ying;Cai Suyun;He Runli;Yin Guifang;Wang Yanqing;Lu Wenjie;Sun Daowang;Wang Lihua(College of Traditional Chinese Medicine and Food Engineering,Shanxi University of Traditional Chinese Medicine,Taiyuan030619;Key Labora tory of Southwestern Crop Gene Resources and Germplasm Innovation(Ministry of Agriculture and Rural Affairs),Yunnan Provincial Key Laboratory of Agricultural Biotechnology,Biotechnology and Genetic Germplasm Resources Research Institutes,Yunnan Academy of Agricultural Sciences,Kunming650201)

机构地区：[1]山西中医药大学中药与食品工程学院,太原030619 [2]云南省农业科学院生物技术与种质资源研究所,云南省农业生物技术重点实验室,农业农村部西南作物基因资源与种质创制重点实验室,昆明650201

出　　处：《分子植物育种》2023年第1期102-109,共8页Molecular Plant Breeding

基　　金：国家自然科学基金地区科学基金项目(31460379);国家燕麦荞麦产业技术体系项目(CARS-07-C-2);山西省重点研发计划项目(201803D221012-6);2019年中医药公共卫生服务补助专项“全国中药资源普查项目(财社[2019]68号)”共同资助

摘　　要：本研究克隆木质素生物合成途径关键酶莽草酸/奎宁酸羟基肉桂酰转移酶(HCT),探究苦荞HCT基因的表达调控以及对苦荞果壳发育形成的影响。运用RT-PCR技术克隆获得两条苦荞HCT基因,命名为Ft HCT-1、FtHCT-2,对两基因编码蛋白进行生物信息学分析,对不同组织器官及果壳不同时期混合材料进行RT-qPCR表达分析。结果表明:FtHCT-1、FtHCT-2开放阅读框序列长度分别为1 347、1 278 bp,各编码448和425个氨基酸,蛋白分子量分别为49.66和46.57 kD,理论等电点为5.72和6.64,均为亲水性蛋白,处在细胞质内,均属于PLNO2481和Trasferase超家族,具有高度同源。两基因表达存在明显差异。本研究克隆所得两条HCT基因在苦荞果壳生长发育中特定表达,推断其表达影响苦荞果壳木质素合成,为薄果壳形成的关键基因。To clone the key enzyme shikimate/quinic hydroxycinnamyl transferase(HCT) in the biosynthetic pathway of lignin, and explore the expression regulation of the HCT gene of tartary buckwheat and its influence on the development of tartary buckwheat husk. Two HCT genes were cloned from Tartary buckwheat by RT-PCR technology, named FtHCT-1 and FtHCT-2. Bioinformatics analysis of the two genes encoding proteins, The expression analysis of mixed materials of different tissues and organs and husks at different periods through RT-qPCR.The results showed that the open reading frame sequence length of FtHCT-1 and FtHCT-2 were 1 347 and 1 278 bp,respectively, encoding 448 and 425 amino acids. Protein molecular weight were 49.66 and 46.57 kD, respectively,isoelectric point were 5.72 and 6.64. They are all hydrophilic proteins, located in the cytoplasm. Both belong to the superfamily of PLNO2481 and Trasferase. The two genes are highly homologous. There are obvious differences in the expression of the two genes. The two HCT genes cloned in this study are specifically expressed during the growth and development of tartary buckwheat husks. It is inferred that their expression affects the lignin synthesis of tartary buckwheat husks and are key genes for the formation of thin husks.

关 键 词：苦荞 HCT 基因克隆 生物信息学 表达分析 

分 类 号：S517[农业科学—作物学]