油菜查尔酮异构酶基因的克隆及SNP分析

doi:10.7668/hbnxb.2018.01.016

摘要/Abstract

摘要： 类黄酮是一种重要的植物次生代谢产物，查耳酮异构酶（CHI）是类黄酮生物合成早期阶段的一个关键酶，在种皮发育和颜色形成过程中具有重要的调控作用。为深入研究CHI基因在种皮发育和颜色形成中的作用及生物学功能。以16份三大类型黄、褐籽油菜为试验材料，采用同源克隆法克隆得到CHI基因的序列，并进行分子进化分析。将克隆得到的序列利用NCBI在线软件预测ORF Finder分析该基因的开发阅读框（ORF），结果发现，CHI基因ORF长度为756 bp或759 bp，编码251个或252个氨基酸。利用DNAMAN （v5.0）软件进行序列同源比对分析结果表明，CHI基因在三大类型油菜中的同源率为96.41%；利用NCBI在线软件CDD预测其保守结构域，发现它们都具有查尔酮超家族保守结构域。利用MEGA 5.2软件进行系统进化分析，结果表明，CHI基因在白菜型油菜与甘蓝型油菜中亲缘关系较近，与芥菜型油菜亲缘关系较远，并发现油菜与萝卜、拟南芥的CHI亲缘关系较近。比较三大类型油菜中黄籽油菜与褐籽油菜中CHI基因序列，结果发现，该基因在第202（C/A）位核苷酸处存在差异，可导致第68位氨基酸（P/T）的差异，这可能与油菜种皮的颜色的变化有关。该研究揭示了CHI基因的特征，为阐明CHI基因在油菜种皮颜色形成过程中的作用机制及其功能特征奠定基础。

关键词: 油菜, 查尔酮异构酶, CHI, 克隆, SNP

Abstract: Flavonoids are an important secondary metabolite of plants. Chalcone isomerase(CHI) is a key enzyme in the early stage of flavonoid biosynthesis, which plays an crucial regulatory role in the development and color formation of seed coat.To further study the features and biological function of CHI gene in seed coat development and in seed colour formation. 16 yellow and brown rapeseed of three types as experimental materials. The CHI gene sequence was obtained by homology-based cloning strategy, the open reading frame of CHI was 756 or 759 bp, encoding 251 or 252 amino acids. Furthermore, DNAMAN (v5.0) was used to align the homology sequence.The results showed that the homology of nucleotide sequence was 96.41%; the chalcone superfamily conserved domain was found in these cloned genes using NCBI on-line software CDD; the phylogenetic tree of CHI in many species was constructed by MEGA 5.2. Results showed that compared to Brassica juncea, CHI gene had more closer relationship between Brassica napus and Brassica rapes in three types of rapeseed,besides in Rapes, Radish and Arabidopsis thaliana showed close relationship.By comparing the characteristics of CHI gene sequence between yellow seeded rapeseed and brown seeded rapeseed in three types of rapeseed,there was a difference in nucleotide at position of 202 (C/A),which led to a difference in amino acid (P/T) respectively,as a result,the color of the rape seed coat might change.This study will lay the foundation for clarifying the further study on its action mechanism and function characteristics.

Key words: Rapeseed, Chalcone isomerase, CHI, Cloning, SNP

中图分类号:

Q78
S634.03

王阳, 卢虹, 黄镇, 刘璐, 刘霞, 刘亚萍, 田正书, 徐爱遐. 油菜查尔酮异构酶基因的克隆及SNP分析[J]. 华北农学报, 2018, 33(1): 102-108. doi: 10.7668/hbnxb.2018.01.016.

WANG Yang, LU Hong, HUANG Zhen, LIU Lu, LIU Xia, LIU Yaping, TIAN Zhengshu, XU Aixia. Cloning and SNP Analysis of CHI in Rapeseed[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(1): 102-108. doi: 10.7668/hbnxb.2018.01.016.

http://www.hbnxb.net/CN/Y2018/V33/I1/102

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