《植物生理学报》 2015, 51(6): 853-859

参薯DaANS基因克隆及表达差异分析

陈跃华, 许云, 吴文嫱, 刘林娅, 黄小龙, 黄东益*

海南大学农学院, 海口570228

通信作者：黄东益；E-mail: dioscorea@163.com；Tel: 0898-66279037

摘　要：

通过研究参薯块茎花青素合成途径中DaANS基因的功能以为参薯分子育种打下基础。利用RT-PCR与RACE技术从参薯块茎中扩增得到1 320 bp的花青素合成酶cDNA序列, 其编码356个氨基酸, 命名为DaANS (登录号为KP729182)。基因组序列全长1 534 bp, 具有一个内含子。使用荧光定量PCR技术对参薯不同组织及其块茎不同发育时期的DaANS基因相对表达量进行测定。结果表明: DaANS基因具有明显的组织特异性和时空性, 不同组织中块茎表达量最高, 且在块茎生长的前期具有很高的表达量, 在8月份到顶峰, 随后急剧降低。

关键词：花青素; 参薯; 花青素合成酶; 基因克隆; 实时荧光定量PCR

收稿：2015-03-20   修定：2015-06-03

资助：2013年海南省重大科技专项(ZDZX2013023)和海南大学地方服务项目(HDSF201304)。

Cloning and Analysis of Differential Expression of DaANS Gene in Dioscorea alata

CHEN Yue-Hua, XU Yun, WU Wen-Qiang, LIU Lin-Ya, HUANG Xiao-Long, HUANG Dong-Yi*

College of Agronomy, Hainan University, Haikou 570228, China

Corresponding author: HUANG Dong-Yi; E-mail: dioscorea@163.com; Tel: 0898-66279037

Abstract:

The function of DaANS gene in anthocyanin biosynthesis pathway was studied to lay the foundation for molecular breeding of Dioscorea alata. In this paper, the cDNA sequence of DaANS gene (1 320 bp) was cloned by RT-PCR and RACE techniques from the tuber of D. alata. The gene was named as DaANS (accession number: KP729182) encoding a protein of 356 amino acids. The full-length of DNA sequence was 1 534 bp, containing one intron. The relative expression of DaANS was determined by real-time quantitative PCR in six different organizations and different developmental stages of tubers. The results showed the expression of DaANS gene had obvious characters of space and time. The expression of tuber was the highest in different tissues. And the early stage of tuber growth had a high amount of expression, in August to the peak, and then decreased sharply.

Key words: anthocyanidin; Dioscorea alata; anthocyanidin synthase; gene cloning; real-time quantitative PCR