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中国农学通报 ›› 2018, Vol. 34 ›› Issue (30): 63-70.doi: 10.11924/j.issn.1000-6850.casb17070067

所属专题: 生物技术

• 生物技术科学 • 上一篇    下一篇

羊踯躅psy基因的克隆及表达分析

肖政,苏家乐,孙晓波,刘晓青,李畅,何丽斯,陈尚平   

  1. 江苏省农业科学院休闲农业研究所/江苏省高效园艺作物遗传改良重点实验室,江苏省农业科学院休闲农业研究所/江苏省高效园艺作物遗传改良重点实验室,江苏省农业科学院休闲农业研究所/江苏省高效园艺作物遗传改良重点实验室,江苏省农业科学院休闲农业研究所/江苏省高效园艺作物遗传改良重点实验室,江苏省农业科学院休闲农业研究所/江苏省高效园艺作物遗传改良重点实验室,江苏省农业科学院休闲农业研究所/江苏省高效园艺作物遗传改良重点实验室,江苏省农业科学院休闲农业研究所/江苏省高效园艺作物遗传改良重点实验室
  • 收稿日期:2017-07-13 修回日期:2018-09-30 接受日期:2017-11-20 出版日期:2018-10-31 发布日期:2018-10-31
  • 通讯作者: 苏家乐
  • 基金资助:
    国家科技支撑计划“重要花卉种质资源发掘与创新利用”(2013BAD01B070403);国家自然科学基金项目“基于转录组学挖掘调控羊踯躅花瓣黄色性 状形成的关键基因”(31600570);江苏省自然科学基金“调控羊踯躅花色突变体花瓣呈色的关键基因挖掘”(BK20150548);江苏省林业科技创新与推广项目“杜 鹃花新品种‘蝶舞’、‘蝶恋’工厂化育苗技术及应用推广”(lykj[2017]48);江苏省农业科技自主创新资金“优新树种选育与高效栽培技术方案”(CX(16)1005)。

Cloning and Expression of psy Gene from Rhododendron molle

  • Received:2017-07-13 Revised:2018-09-30 Accepted:2017-11-20 Online:2018-10-31 Published:2018-10-31

摘要: [目的]为了研究羊踯躅psy基因的功能及其在花瓣着色中的分子机理,[方法]以羊踯躅花瓣的cDNA为模板,采用RT-PCR和RACE方法克隆得到1 638 bp的psy基因全长cDNA序列,命名为RmPSY (GenKank登录号为KX230461)。[结果]序列分析表明,RmPSY开放阅读框为1 296bp,编码432个氨基酸。羊踯躅PSY为亲水性的不稳定蛋白,无信号肽,属于非分泌蛋白;预测的蛋白质分子量48.91 kD,等电点为8.78。RmPSY与GenBank中收录的其它植物PSY蛋白氨基酸的相似性达到81%。构建系统进化树,结果显示羊踯躅PSY与葡萄PSY蛋白的亲缘关系最近。实时荧光定量PCR结果显示,该基因在羊踯躅花发育的不同阶段均有表达,在花蕾期表达量较低,在初开期表达量最高,盛开期表达量有所下降。[结论]试验结果为进一步揭示RmPSY基因参与调控羊踯躅花色形成的分子机制奠定基础。

Abstract: We aim to study the function of psy gene and its molecular mechanism in petal coloring. A psy gene was cloned from the Rhododendron molle cDNA by using RT-PCR and RACE methods. The full length of the psy gene was 1638 bp, named RmPSY (GenBank accession No. KX230461), which contained a 1296 bp open reading frame encoding 432 amino acid residues. RmPSY, a hydrophilic unstable protein with no signal peptide, was a non-secreted protein. The putative protein molecular weight was 48.91 kD and its theoretical isoelectric point was 8.78. Homologous alignment showed that it shared 81% amino acid identities with PSY from other plants in GenBank. The phylogenetic tree constructed on the basis of amino acid sequences of PSY suggested that the relationship between R. molle and Vitis vinifera was the most intimate. The results showed that the RmPSY gene was expressed in the petal during the flower development. The RmPSY transcripts were the most abundant in the initial opening period of flower, low in the bud stage, and decreased during the blooming period of flower. This study may provide references for further research on the molecular mechanism of RmPSY involved in the formation of R. molle flower color.

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