石榴4香豆酸辅酶A连接酶基因的克隆和表达分析
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国家自然科学基金(30900971)


Cloning and Expression Analysis of Pomegranate 4coumarateCoA Ligase Pg4CL
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    摘要:

    4香豆酸辅酶A连接酶(4coumarateCoA ligase,4CL)是木质素合成途径的关键酶。该研究以‘红玉石籽’ (Punica granatum cv. Hongyushizi)石榴为试验材料,采用RACE和RTPCR方法获得4CL同源基因Pg4CLPg4CL基因cDNA全长2 121 bp,编码544个氨基酸;序列比对和功能域分析发现,Pg4CL包含有2个保守域SSGTTGLPKGV和GEICIRG;系统进化树分析显示,Pg4CL与其他物种起源相同,而与赤桉 Ec4CL亲缘关系最近。荧光定量PCR分析表明,Pg4CL基因在叶、果皮、种皮和茎等组织中均有表达,其中果皮和叶片中表达量较高,种皮中表达量最低;Pg4CL基因在6个石榴品种的种皮中均有表达,其中在‘突尼斯软籽’中表达较高,‘会理软籽’表达量最低;Pg4CL 基因在‘红玉石籽’籽粒的不同时期均有表达,在15~60 d相对表达量逐步上升,并在 60 d时达到最高;75 d以后表达量急剧下降,仅维持较低水平表达。

    Abstract:

    4coumaric acid coenzyme A ligase is a key enzyme in lignin biosynthesis pathway. In this study, Pg4CL,the homologous gene of 4CL, was obtained with RACE and RTPCR method, with ‘Punica granatum cv. Hongyushizi’ as the test material. The cDNA fulllength sequence of Pg4CL was 2 121 bp, with reduced 544 amino acids. Sequence alignment and functional domain analysis revealed that Pg4CL contained two conserved domains: SSGTTGLPKGV and GEICIRG. Phylogenetic tree analysis indicated that Pg4CL shared the same origin with other species, and had the closest relationship with Eucalyptus camaldulensis Ec4CL. The fluorescent quantitative PCR analysis showed that Pg4CL expression was detected in leaves, peel, seed coat and stems. The expression in peel and leaves were higher, while in seed coat was the lowest; Pg4CL expression was detected in six cultivars of pomegranate seeds, and the expression of ‘Tunisiruanzi’ was higher, while ‘Huiliruanzi’ was lower; Pg4CL expression was detected in different development periods of ‘Fenpi’ seeds. The relative expression of Pg4CL gradually increased from 15 d-60 d, and reached a maximum at 60 d. While sharply declined from 75 d, and maintained a lower level of expression.

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董丽丽,阿不都热扎克·依沙克,户 倩,等.石榴4香豆酸辅酶A连接酶基因的克隆和表达分析[J].西北植物学报,2016,36(11):2146-2151

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  • 在线发布日期: 2016-12-12
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