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高良姜1-脱氧-D-木酮糖5-磷酸还原异构酶cDNA克隆与表达调控

Keywords: 高良姜,单萜生物合成,1-脱氧-D-木酮糖5-磷酸还原异构酶,茉莉酸甲酯

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Abstract:

目的:克隆高良姜1-脱氧-D-木酮糖5-磷酸还原异构酶(DXR)的全长cDNA,分析其组织表达模式及茉莉酸甲酯(MeJA)的调控模式,为高良姜有效成分的基因调控及基因工程育种奠定基础。方法:应用简并引物RT-PCR和RACE技术从高良姜根茎中克隆DXR全长cDNA,运用生物信息学解析其编码的蛋白质结构,实时荧光定量PCR法分析其组织表达模式和MeJA的调控模式。结果:克隆了高良姜DXR全长cDNA序列(AoDXR),开放读码框长1419bp,编码的蛋白质含472个氨基酸残基、相对分子质量约51.48kDa。推导的AoDXR氨基酸序列与其他高等植物的DXR具有高度的序列一致性(73%~99%)。AoDXR在高良姜叶片中表达量最强,而在根茎中表达量较弱。外源茉莉酸甲酯(MeJA)处理提高了根茎AoDXR的转录水平和1,8-桉油精含量。结论:AoDXR在高良姜根茎中的表达水平与1,8-桉油精的积累不一致,反应了AoDXR催化的终产物的多样性和表达调控的复杂性。外源MeJA可促进根茎AoDXR的表达和1,8-桉油精的积累,对提高药材品质有应用价值。

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