园艺学报 ›› 2011, Vol. 38 ›› Issue (6): 1071-1080.doi:

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徐红霞1,陈俊伟1,*,杨 勇2,孙骏威3,严成其2   

  1. (1浙江省农业科学院园艺研究所,杭州 310021;2浙江省农业科学院病毒学与生物技术研究所,杭州 310021;3中国计量学院生命科学学院,杭州 310018)   
  • 收稿日期:2010-11-26 修回日期:2011-04-26 出版日期:2011-06-25 发布日期:2011-06-25
  • 通讯作者: 陈俊伟

Isolation and Expression Analysis of DHN Gene in Loquat Fruit Under Cold Stress

XU Hong-xia1,CHEN Jun-wei1,*,YANG Yong2,SUN Jun-wei3,and YAN Cheng-qi2   

  1. (1Institute of Horticulture,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China;2Institute of Virology and Biotechnology,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China;3College of Life Sciences;China Jiliang University,Hangzhou 310018,China)   
  • Received:2010-11-26 Revised:2011-04-26 Online:2011-06-25 Published:2011-06-25
  • Contact: CHEN Jun-wei

摘要: 以白沙枇杷(Eriobotrya japonica Lindl.)‘宁海白’幼果为试材,通过RT-PCR与RACE扩增,获得两个具有脱水素基因典型结构域特征的cDNA序列。其中DHN1(GenBank登录号:FJ472835)全长639 bp,编码188个氨基酸,属于Y2SK3类型;DHN2(GenBank登录号:FJ472836)全长858 bp,编码273个氨基酸,属于SK3类型。用低温胁迫处理枇杷幼果,–3 ℃处理24 h后电导率明显升高,表明细胞膜开始受到破坏,–6 ℃处理24 h后电导率显著高于–3 ℃处理。半定量RT-PCR和Western-blotting分析结果表明这两个DHN基因在低温胁迫下的枇杷幼果中都上调表达,表明其与枇杷抗低温胁迫有一定关系。   

关键词: 枇杷, DHN, 基因克隆, 低温胁迫, 表达分析

Abstract: The full length of two dehydrin(DHN)genes from the young fruit of loquat(Eriobotrya japonica Lindl.‘Ninghaibai’)were obtained using RT-PCR and RACE-PCR approach. The deduced amino acid sequences of the DHN1 and DHN2 possessed the typical characters of DHN family. However,they differ from each other significantly in the structure characteristics. DHN1 cDNA sequence length is 639 bp,which contains an open reading frame that would encode 188 amino acid residues,and it is Y2SK3 type dehydrin;DHN2 cDNA sequence length is 858 bp,which contains an open reading frame that would encode 273 amino acid residues,and it is SK3 type dehydrin. The relative electric conduction was significantly increased in the loquat fruit treated with–3 ℃ for 24 h,which suggested that the cell membrane was injured,and it was more serious in the loquat fruit treated with–6 ℃ for 24 h. Semi-quantitative RT-PCR and Western-blotting analysis showed that the expression of both DHN1 and DHN2 were induced by cold stress. The results suggested that both DHN1 and DHN2 involved in cold resistant in loquat young fruit   

Key words: loquat, DHN, gene clone, cold stress, expression