茉莉花JsPAL基因及其启动子克隆与表达分析
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福州市科技计划项目(2019N15);


Cloning and Expression Analysis of Phenylalanine Ammonialyase Gene from Jasminum sambac and Isolation of Its Promoter
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    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2 220 bp,开放阅读框(ORF)为2 140 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1 201 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。

    Abstract:

    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 2 220 bp fulllength cDNA sequence was obtained and named JsPAL. The ORF (open reading frame) of JsPAL was 2 140 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 1 201 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.

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孙 君,陈雪津,陈 笛,等.茉莉花JsPAL基因及其启动子克隆与表达分析[J].西北植物学报,2020,40(6):949-956

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  • 在线发布日期: 2020-07-30
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