‘黄花’梨及其芽变‘绿黄花’梨HHT基因克隆与表达分析
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国家自然科学基金(31272140);


Cloning and Expression of HHT Gene in ‘Huanghua’ Pear and Its Bud Mutant ‘Lühuanghua’ Pear (Pyrus pyrifolia Nakai)
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    摘要:

    该试验以砂梨品种‘黄花’梨(果皮褐色)及其芽变‘绿黄花’梨(果皮绿色)盛花后第8周的果皮为试材,利用常规PCR和巢式PCR技术克隆了ω羟基棕榈酸O阿魏酰转移酶(ωhydroxypalmitate Oferuloyl transferase, HHT)基因cDNA的全长,命名为 PpyHHT(登录号为KX131155)。序列分析结果表明,该基因开放阅读框(ORF)为1 335 bp,编码444个氨基酸。生物信息学分析显示,推定的PpyHHT蛋白质相对分子质量为49.91 kD,等电点是4.75,与白梨相似性高达98%,亲缘关系最近。实时荧光定量PCR(qRTPCR)表达分析显示,2种梨果皮中 PpyHHT基因在盛花后6~9周的4个转色关键期表达量不断变化,在‘黄花’梨果皮中的表达量明显高于‘绿黄花’梨。推测 PpyHHT基因可能参与砂梨果实褐色/绿色性状的形成。

    Abstract:

    The study used fruit peel of sand pear (Pyrus pyrifolia Nakai) cultivars ‘Huanghua’ pear (russet fruit) and its bud mutant ‘Lühuanghua’ pear (green fruit) at 8 weeks after full bloom (WAFB) as experiment materials. the cDNA fulllength of HHT gene,which was named PpyHHT (GenBank accession No. KX131155), was cloned by conventional and nest PCR techniques. Sequence analysis showed that the fulllength of the PpyHHT ortholog consisted of a 1 335 bp open reading frame (ORF) encoding a polypeptide containing 444 amino acid residues. The molecular weight of deduced amino acids was 49.91 kD, with an isoelectric point (pI) of 4.75, which had the highest similarity (98%) and closest relationship with that in Pyrus bretschneideri. Realtime fluorescence quantitative PCR (qRTPCR) analysis demonstrated that the expression of PpyHHT gene in ‘Huanghua’ and ‘Lühuanghua’ pear peel were changing constantly during the key period (6-9 weeks), and significantly higher in ‘Huanghua’ pear peel than that in ‘Lühuanghua’. PpyHHT gene involved in the formation of sand pear russet/green traits, its expression level differences may play a role in the formation of sand pear skin color.

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吕照清,任丹丹,周 贺,等.‘黄花’梨及其芽变‘绿黄花’梨HHT基因克隆与表达分析[J].西北植物学报,2016,36(6):1105-1109

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  • 在线发布日期: 2016-07-18
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