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作物学报 ›› 2014, Vol. 40 ›› Issue (06): 1002-1010.doi: 10.3724/SP.J.1006.2014.01002

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

甘蔗S-腺苷甲硫氨酸合成酶基因(ScSAM)的克隆及表达

宋修鹏1,张保青2,黄杏2,杨丽涛1,2,*,李杨瑞1,2,*   

  1. 1 广西大学农学院 / 亚热带农业生物资源保护与利用国家重点实验室,广西南宁530004;2 广西农业科学院 / 中国农业科学院甘蔗研究中心 / 农业部广西甘蔗生物技术与遗传改良重点实验室 / 广西甘蔗遗传改良重点实验室,广西南宁530007
  • 收稿日期:2013-11-17 修回日期:2014-03-04 出版日期:2014-06-12 网络出版日期:2014-04-09
  • 通讯作者: 杨丽涛, E-mail: liyr@gxu.edu.cn, Tel: 0771-3236407; 李杨瑞, E-mail: liyr@gxaas.net, Tel: 0771-3247689
  • 基金资助:

    本研究由国家高技术研究发展计划(863计划)项目(2013AA102604),国家国际合作项目(2013DFA31600),广西科学研究与技术开发计划项目(桂科产1123008-1、桂科攻1222009-1B、桂科合1347004-2),广西八桂学者、特聘专家专项基金,广西自然科学基金项目(2011GXNSFF018002、2013NXNSFAA019073)和广西农业科学院团队项目(桂农科2011YT01)资助。

Cloning and Expression of Sugarcane S-Adenosylmethionine Synthetase Gene ScSAM

SONG Xiu-Peng1,ZHANG Bao-Qing2,HUANG Xing2,YANG Li-Tao1,2,*,LI Yang-Rui1,2,*   

  1. 1 Agricultural College / State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530004, China; 2 Guangxi Academy of Agricultural Sciences / Sugarcane Research Center, Chinese Academy of Agricultural Sciences / Key Laboratory of Sugarcane Biotechnology and Genetic Improvement (Guangxi), Ministry of Agriculture / Guangxi Key Laboratory of Sugarcane Genetic Improvement, Nanning 530007, China
  • Received:2013-11-17 Revised:2014-03-04 Published:2014-06-12 Published online:2014-04-09
  • Contact: 杨丽涛, E-mail: liyr@gxu.edu.cn, Tel: 0771-3236407; 李杨瑞, E-mail: liyr@gxaas.net, Tel: 0771-3247689

摘要:

利用RT-PCR和RACE技术从甘蔗品种新台糖22号(ROC 22)中克隆获得SAM基因的cDNA序列,命名为ScSAM,GenBank登录号为KC172558。用生物信息学方法预测分析其序列,cDNA全长1466 bp,含有1个1191 bp的完整开放阅读框(ORF),编码396个氨基酸,与高粱和玉米等植物的SAM蛋白有很高的相似性。系统进化树分析显示,甘蔗ScSAM与高粱的SAM蛋白亲缘关系较近。Real-time PCR分析表明ScSAM为组成型表达,在根中的表达量最高,是叶中表达量的3.6倍。其在黑穗病菌胁迫和低温(4℃)、聚乙二醇(PEG)、NaCl非生物胁迫下均被诱导表达,但表达模式不同;在H2O2胁迫下其表达被抑制。推测其可能参与甘蔗抗黑穗病过程,且在甘蔗抗寒、抗旱、抗盐和抗氧化等胁迫过程中也起某种作用。

关键词: 甘蔗, S-腺苷甲硫氨酸合成酶, 克隆, 表达分析

Abstract:

The full-length sequencecDNA of ScSAM (GenBank accession number: KC172558) was cloned from sugarcane variety ROC 22 using RT-PCR combined with RACE techniques. This sequence consists of 1466 bp with an intact open reading frame of 1191 bp, encoding a polypeptide of 396 amino acids. Homology analysis showed that the deduced ScSAM protein was highly homologous to SAM proteins from different species. Phylogenetic analysis indicated that ScSAM was closely related to the SAM of sorghum. Real-time PCR results showed that the ScSAM gene constitutively expressed in plant, with different expression levels in root, stalk and leaf. The transcript of ScSAM in root was the highest among the three organs, which was 3.6 times higher than that in leaf. Furthermore, ScSAM transcription was induced by biotic (smut infection) and abiotic (low temperature, PEG and NaCl) stresses, but the expression patterns were different. Under oxidative stress (H2O2), the expression of ScSAM was inhibited. We suggested that ScSAM might participate in smut-resistant activities in sugarcane, and also play a role in sugarcane resistances to chill, drought, salt and oxidation stresses.

Key words: Sugarcane, S-Adenosylmethionine synthetase (SAM), Clone, Expression analysis

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