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Plant Diversity ›› 2010, Vol. 32 ›› Issue (02): 151-157.DOI: 10.3724/SP.J.1143.2010.09190

• 研究论文 • 上一篇    下一篇

珙桐中一个与低温相关基因的克隆及其表达研究

季红春1,2,苏智先1,2, 杨军1,边春象1,
胡进耀2,齐刚2
  

  1. 1 西华师范大学生命科学学院,四川 南充637002;2 绵阳师范学院生命科学与技术学院,四川 绵阳621000
  • 收稿日期:2009-09-28 出版日期:2010-04-25 发布日期:2010-06-29
  • 通讯作者: 苏智先

Cloning and Expression of a Coldinduced Gene (DiRCI) from Davidia involucrata (Davidiaceae)

JI HongChun1,2, SU ZhiXian1,2, YANG Jun1, BIAN ChunXiang1,
HU JinYao2, QI Gang 2
  

  1. 1 College of Life Science, China West Normal University, Nanchong 637002, China; 2 College of Life Science & Biotechnology, Mianyang Normal University, Mianyang 621000, China
  • Received:2009-09-28 Online:2010-04-25 Published:2010-06-29
  • Contact: SU ZhiXian

摘要: 低温诱导膜蛋白是由低温诱导表达蛋白基因编码的一类疏水性蛋白,在植物抵御寒冷环境时起着一定的保护作用。从珙桐 cDNA文库中获得一个未知基因(DiRCI),该基因长539 bp,其中包括174 bp的开放阅读框,92 bp的5′末端非编码区和273 bp的 3′ 末端非编码区,编码57个氨基酸残基蛋白,在氨基酸水平上同源性较高的是车前草的低温诱导膜蛋白(登入号:ACA662471),其相似性为895%。半定量 RTPCR 分析发现,25℃以上,该基因在珙桐各器官中基本上均未表达,但经8℃低温处理时,在成熟叶片、叶柄和成熟未萌发的种子中均有表达,但是在根中基本没有表达,进一步研究发现该基因在24 h~48 h内表达量增多并达到最高值,48 h后其表达量逐渐减弱直至消失,说明该基因确实与低温诱导相关,从而初步推测该基因为低温诱导膜蛋白基因。该基因的克隆丰富和保存了珙桐基因资源,并为进一步研究冷胁迫的分子机制奠定了基础。

关键词: 珙桐;基因;克隆;低温诱导;半定量 RTPCR

Abstract: A coldinduced plasma membrane protein which plays a protective role in avoiding freezing injury for some plants is one of hydrophobic proteins. A gene, named as DiRCI, was isolated from the cDNA library of Davidia involucrata, containing an open reading frame of 174 bp flanked by a 92 bp 5′untranslated sequence and a long 273 bp 3′noncoding region. Alignment analysis indicated that the deduced amino acid sequence of 57 amino acids was high conserved with the MpRCI from Plantago asiatica, and the identity was 895%. Semiquantitative reverse transcriptionpolymerase chain reaction (RTPCR) analysis showed that DiRCI wasn′t detected in various organs of Dinvolucrata above 25℃. When the plant grew under 8℃, the gene expressed in leaves, leafstalk and seeds, but not the roots. The expression level gradually increased after 4 h of the cold treatment and reached a maximum during 24-48 h, and then decreased. The results indicated the gene did have a relation with cold stress in plant and was predicted to be one DiRCI gene. The present data not only enrich gene resources of Dinvolucrata, but also laid a foundation for the research on molecular mechanism of plant acclimation to cold stress.

Key words: Davidia involucrata

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