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苹果MdCPS基因的克隆、定位及其在柱型/普通型间的表达差异分析

DOI: 10.7668/hbnxb.2014.06.005, PP. 21-27

Keywords: 苹果,赤霉素,柯巴焦磷酸合酶基因(CPS),柱型性状

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Abstract:

为研究赤霉素合成途径关键酶基因,了解果树的矮化机理。以苹果品种富士当年生新梢的茎尖为试材,以苹果基因组数据库为依据,克隆了编码苹果CPS的基因MdCPS(GenBanK登录号:KC433942.1)。该基因gDNA序列含有15个外显子和14个内含子,其编码序列(Codingsequence,CDS)长度为2400bp,共编码799个氨基酸。在已发表的金冠苹果基因组中,与该基因相对应的转录本是MDP0000147908,它的定位区间为chr11:32433834~32439214。同源性分析表明,MdCPS与其他植物的CPS间有较高的相似性(49%~67%)。以杂交组合富士(普通型)×舞姿(柱型)的亲本品种及F1后代当年生新梢的茎尖组织为试材,实时荧光定量PCR(qRT-PCR)分析表明,尽管该基因在柱型亲本中的表达水平显著低于普通型亲本,但在其后代的柱型与普通型群体间差异并不明显。同时,在柱型杂种及其普通型突变体间的分析结果也表明,MdCPS的表达水平与柱型性状没有明显的相关性。可见,柱型苹果茎尖组织中活性赤霉素含量偏低受其合成早期步骤关键酶基因MdCPS的影响不大。

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