夜来香LIS基因的克隆与表达分析  

Cloning and Expression Analysis of LIS Gene from Cestrum nocturnum

作　　者：吕恃衡[1] 杨华丽[1] 吕科良 蔡韡韡[1] 刘涛[1] 朱梦珠 潘东明[1] 陈桂信[1] Lv Shiheng Yang Huali Lv Keliang Cai Weiwei Liu Tao Zhu Mengzhu Pan Dongming Chen Guixin(College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou, 35000)

机构地区：[1]福建农林大学园艺学院,福州350002

出　　处：《分子植物育种》2017年第7期2525-2531,共7页Molecular Plant Breeding

基　　金：福建省自然科学基金项目(2016J01110)资助

摘　　要：为了研究夜来香生物钟和花香代谢机制,本研究以夜来香花瓣为材料,利用PCR技术,从中获得了夜来香LIS基因的全长cDNA,并命名为CnLIS。生物信息学分析结果表明:该cDNA序列全长为1 800 bp,编码560个氨基酸,其中ORF为1 683 bp,5'UTR为56 bp,3'UTR为61 bp;该序列编码的氨基酸序列与软枣猕猴桃、温州蜜柑LIS基因的同源性分别为55%、50%,属于类异戊二烯合成C1超级家族。实时荧光定量PCR分析结果表明,0~12 h CnLIS表达水平较低,16 h后表达量明显上调,20 h表达量达最大,随后表达量急剧下降,其表达量随花瓣开放与闭合呈节律性变化。推测CnLIS可能参与夜来香香气形成,调控花香形成过程,同时,该基因的表达可能受生物钟基因所调控。A full-length c DNA encoding LIS named CnLIS whose length was 1 800 bp, encoding 560 amino acids, including 1 683 bp ORF, 56 bp 5＇UTR, 61 bp 3＇UTR, was cloned by combination of PCR from Cestrum nocturnum petals, to explore mechanism of biological clock and formation of fragrance in Cestrum nocturnum.Bioinformatics analysis was carried out on the CnLIS amino acid sequence. The results showed that the amino acid sequences were 55% and 50% homology with Actinidia argute and Citrus unshiu, belonging to Isoprenoid_Biosyn_C1 superfamily. Results of analysis of real-time quantitative PCR showed that CnLIS from night to noon,its low expression level, the expression of CnLIS was significantly upregulated at 16 h, the expression maximum was at 20 h, and then the expression amount decreased sharply. CnLIS may play an important role in formation of fragrance in Cestrum nocturnum, which regulate the synthesis and release of terpenoid. The expression level showed a rhythm with petals open and closed, the expression of the gene may be regulated by the circadian clock genes.

关 键 词：夜来香 LIS基因 基因克隆 生物信息学 实时荧光定量PCR 

分 类 号：Q943.2[生物学—植物学] S685.99[农业科学—观赏园艺]