石榴赤霉素合成相关基因PgKO的克隆及表达分析
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国家自然科学基金(30900971);


Cloning and Expression Analysis of Gibberellin BiosynthesisRelated Gene PgKO in Pomegranate
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    摘要:

    该研究以‘红玉石籽’(Punica granatum cv. Hongyushizi) 石榴为试验材料,采用RTPCR和RACE技术,从石榴花中克隆得到赤霉素合成关键酶——内根贝壳杉烯氧化酶(KO)基因,命名为PgKO(GenBank登录号为MG208017)。PgKO全长cDNA为1 729 bp,包含1 542 bp开放阅读框(ORF),编码513个氨基酸,相对分子量为141.16 kD,理论等电点为4.9。多重序列对比显示,PgKO与苹果MdKO、白梨PcKO、葡萄VvKO的相似性分别为61.28%、64.56% 和73%, 并且具有CYP450结构域(脯氨酸富含域、氧还原位点和血红素结合域)。系统进化树分析表明,PgKO与其他物种的起源相同,并与桉树EgKO的亲缘关系最近。荧光定量PCR发现,在石榴蕾期和盛花期,PgKO基因的表达量均为筒状花高于钟状花,且PgKO基因在两种花型的子房、花药、花萼中均有表达,但筒状花中子房的表达量最高,花萼的表达量最低;钟状花中花药的表达量最高,子房的表达量最低。

    Abstract:

    In this study, PgKO, the gibberellin biosynthesisrelated gene, was cloned from flowers of pomegranate using RTPCR and RACE technology methods, with ‘Punica granatum cv. Hongyushizi’ as the experimental material. Its accession of GenBank is MG208017. The cDNA fulllength sequence of PgKO was 1 729 bp, with 1 542 bp open reading flame encoding 513 amino acids. The protein molecular weight and isoelectric point were 141.16 kD and 4.9 respectively. Multiple alignment showed that PgKO had 61.28%, 64.56% and 73% similarity with Malus × domestica MdKO, Pyrus communis PcKO and Vitis vinifera VvKO respectively, also included CYP450 domain embracing prolinerich domain, redio sites and hemebinding domain. Phylogenetic tree analysis indicated that PgKO shared the same origin with other species, and had the closest relationship with Eucalyptus grandis EgKO. Real time quantitative PCR showed that no matter the bud stage or flowering stage, PgKO expression in cylindricshaped flowers were higher than that in trumpetshaped flowers. Sometimes PgKO expression was detected in ovary, anther and calyx. In cylindricshaped flowers, the expression in ovary is the highest, while lowest in calyx; In trumpetshaped flowers, the expression in anther is the highest, while lowest in ovary.

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黄 蓉,刘 娜,王 琦,等.石榴赤霉素合成相关基因PgKO的克隆及表达分析[J].西北植物学报,2018,38(4):631-636

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  • 在线发布日期: 2018-05-21
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