摘要
苯丙氨酸解氨酶(phenylalanine ammonia-lyase,PAL)为茉莉花(Jasminum sambac)花香物质苯丙烷类合成的限速酶。为了解茉莉花花香形成的分子机理,以双瓣茉莉花瓣为材料,采用RT-PCR和RACE技术相结合的方法,克隆获得茉莉花苯丙氨酸解氨酶基因的全长cDNA(GenBank:KM406501.1),命名为JsPAL,其全长cDNA为2220 bp,开放阅读框(ORF)为2140 bp,编码712个氨基酸,含有lyase_I_like超家族蛋白保守域、活性位点及多肽结合位点。采用染色体步移技术,获得该基因的启动子序列。JsPAL基因上游调控序列为1201 bp,其含有开花相关元件(CCAATBOX1)、PAL相关元件(BOXLCOREDCPAL、PALBOXPPC、PALBOXLPC、TATABOXOSPAL)以及花特异苯丙烷类相关元件(MYBPLANT)等与花香形成相关的重要顺式作用元件。实时荧光定量PCR检测结果表明,在不同组织和花瓣发育过程中,JsPAL基因在茉莉花的花蕾、花瓣中表达量较高,并且在花瓣开放过程中的22:00前表达量较高,而后呈下调趋势。
Phenylalanine ammonia-lyase was rate-limiting enzyme of phenylpropanoids components formation of Jasminum sambac.To understand the floral scent formation mechanism,we cloned the full-length cDNA sequence of phenylalanine ammonia-lyase gene from J.sambac by RT-PCR(reverse transcriptase-polymerase chain reaction)and RACE(rapid amplification of cDNA ends)methods(GenBank:KM406501.1).A 2220 bp full-length cDNA sequence was obtained and named JsPAL.The ORF(open reading frame)of JsPAL was 2140 bp and encodes 712 amino acids,containing the conserved domain of lyase_I_like superfamily,active sites and tetramer interface(polypeptide binding site).The promoter of JsPAL was isolated by Genome Walking Method.A 1201 bp sequence was found to be the promoter,which contains many important cis-acting elements involved in flowering and floral scent formation,such as CCAATBOX1(regulating flowering),elements of BOXLCOREDCPAL,PALBOXPPC,PALBOXLPC and TATABOXOSPAL(related to PAL gene),and MYBPLANT(involved in flower-specific phenylpropanoid biosynthetic).The gene expression patterns in different stages of flower opening and different tissues were detected by the real-time PCR.The real-time PCR analysis revealed that the expression of JsPAL gene was high in flower buds and petals.Its expression was high before 22:00 and down-regulated with the development of flower.
作者
孙君
陈雪津
陈笛
王鹏杰
陈桂信
叶乃兴
SUN Jun;CHEN Xuejin;CHEN Di;WANG Pengjie;CHEN Guixin;YE Naixing(College of Horticulture, Fujian Agriculture and Forestry University, Key Laboratory of Tea Science at Universities in Fujian, Fuzhou 350002, China;Institute of Tea, Fujian Academy of Agricultural Sciences, Fuzhou 355015, China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2020年第6期949-956,共8页
Acta Botanica Boreali-Occidentalia Sinica
基金
福州市科技计划项目(2019-N-15)
福建农林大学茶产业链科技创新与服务体系建设项目。
