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园艺学报 ›› 2015, Vol. 42 ›› Issue (3): 496-504.doi: 10.16420/j.issn.0513-353x.2014-0906

• 蔬菜 • 上一篇    下一篇

莲藕可溶性淀粉合成酶基因LrSSS的克隆与表达特性分析

张 莉,印 荔,杨见秋,程立宝,李良俊   

  1. 扬州大学园艺与植物保护学院,江苏扬州 225009
  • 出版日期:2015-03-25 发布日期:2015-03-25
  • 基金资助:

    江苏省科技支撑计划项目(BE2013388);江苏省普通高校研究生科研创新计划项目(CXLX13-918)

Cloning and Expression Profiling Analysis of Soluble Starch Synthase Gene in Lotus Rhizome

ZHANG Li,YIN Li,YANG Jian-qiu,CHENG Li-bao,and LI Liang-jun   

  1. School of Horticulture and Plant Protection,Yangzhou University,Yangzhou,Jiangsu 225009,China
  • Online:2015-03-25 Published:2015-03-25

摘要: 以莲藕品种‘美人红’叶片为试材,利用RACE结合RT-PCR技术克隆得到全长4 080 bp的莲藕可溶性淀粉合成酶基因(LrSSS)cDNA序列(GenBank登录号:KP201636),其中开放阅读框3 696 bp,编码1 231个氨基酸;该序列与甜瓜、葡萄SSS基因编码氨基酸序列同源性较高,分别达79%、69%。LrSSS 所编码的氨基酸序列包含3个典型的碳水化合物结合结构域(CBM_25)和1个淀粉合成酶催化域(Glyco_transf_5)。LrSSS 表达分析表明,莲藕根状茎膨大具3节段时,在终止叶叶片中的表达量最高,其次为后把叶叶片,终止叶叶柄表达量最少;在根状茎膨大至4节段时,LrSSS在第1、2节段根状茎中表达量较高,在第3、4节段根状茎中表达量较低,表明在第1、2节段根状茎形态基本建成后LrSSS在调控产物转化为淀粉过程中可能起到重要作用。

关键词: 莲藕, 淀粉, LrSSS, 克隆, 表达

Abstract: The soluble starch synthase gene(LrSSS)was cloned from‘Meirenhong’,a species of lotus rhizome based on reverse transcription polymerase chain RT-PCR and RACE methods with leaf as template in the present study. The full-length of LrSSS(GenBank accession number KP201636)was 4 080 bp in nucleotide containing an open reading frame of 3 696 bp which encoding 1 231 amino acid. Phylogenetic analysis showed that LrSSS had 79%,69% homolog with melon,grape respectively. LrSSS contained three typical carbohydrate domains(CBM_25)and one catalyzing domains(Glyco_transf_5)relevant to starch synthesis. The expression in different temporal and spatial of LrSSS was determined by RT-PCR method. The results was that LrSSS showed the highest expression in the last leaf,and then followed by the stalk of the penultimate leaf,the lowest expression was found in the the stalk of the last leaf. In addition,the expression of LrSSS was higher in the first and second internodes than in the third and fourth internodes of rhizome,suggesting that LrSSS might play an important role in starch synthase processes.

Key words: lotus rhizome, starch, LrSSS, clone, expression

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