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园艺学报 ›› 2016, Vol. 43 ›› Issue (6): 1117-1125.doi: 10.16420/j.issn.0513-353x.2016-0301

• 蔬菜 • 上一篇    下一篇

芋淀粉合成酶AGPase基因的克隆及表达分析

王 立,殷剑美*,韩晓勇,张培通,郭文琦,李春宏   

  1. (江苏省农业科学院经济作物研究所,南京 210014)
  • 出版日期:2016-06-25 发布日期:2016-06-25

Gene Cloning and Expression Analysis of ADP-glucose Pyrophosphorylase in Colocasia esculenta

WANG Li,YIN Jian-mei*,HAN Xiao-yong,ZHANG Pei-tong,GUO Wen-qi,and LI Chun-hong   

  1. (Institute of Industrial Crops,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China)
  • Online:2016-06-25 Published:2016-06-25

摘要:

通过对靖江香沙芋转录组数据进行功能注释,克隆获得靖江香沙芋淀粉合成酶AGPase的基因,序列全长为1 596 bp,命名为CeApS1NCBIColocasia KU288757,未公布)。该基因编码532个氨基酸,蛋白含有1NTP_transf_3结构域(Gly104 ~ Pro306)和1PbH1结构域(Gly473 ~ Ser515),编码产生AGPase小亚基,等电点和分子量分别为6.7357.6 kDApS1在单子叶和双子叶植物中广泛存在,CeApS1与紫萍(AIZ00992.1)中相应蛋白的亲缘关系最近,相似性为83.4%,与草莓(AAS00541.1)、苜蓿(XP_003617925.1)、水稻(AAK27313.1)和小麦(CAA46879.1)等植物中相应蛋白的亲缘关系较远。CeApS1靖江香沙芋母芋和子孙芋中表达量较高,在叶片、叶柄和根中低表达。芋球茎中的CeApS1表达水平与球茎发育密切相关,在播种160 d后达到最大值。该基因的表达模式与球茎淀粉含量的积累显著正相关。

关键词: 芋, 淀粉合成酶, 基因克隆, 表达分析

Abstract:

Based on the transcriptome data of Colocasia esculentaL.Schottfull-length cDNA of ApS1ADP-glucose pyrophosphorylase small subunit 1gene was cloned fromJingjiang Xiangsha Taro’,which consisted of 1 596 bp and encoded a protein of 532 amino acidsand tentatively designated as CeApS1NCBIColocasia KU288757unreleased. CeApS1 contained a NTP_transf_3 domainGly104Pro306and a PbH1 domainGly473Ser515. This gene encoded a small subunit of ADP-glucose pyrophosphorylaseand isoelectric point and molecular weight were 6.73 and 57.6 kDrespectively. Results displayed ApS1 existed in monocotyledons and dicotyledons. BlastX showed that ApS1 in Colocasia esculenta shared 83.4% identity with Landoltia punctateand showed farther phylogenetic relationships with Fragaria ananassaAAS00541.1),Medicago truncatulaXP_003617925.1),Oryza sativaAAK27313.1and Triticum aestivumCAA46879.1. The CeApS1 transcript widely presented in the main cormlateral cormelsleavespetioles and roots of Colocasia esculentaand displayed higher expression levels in taro corms than those in leaves. The results indicated that there was a significantly positive correlation between CeApS1 transcript expression and starch contents in taro corms.

Key words: Colocasia esculenta, starch synthase, gene cloning, expression analysis

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