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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (16): 219-224.doi: 10.11924/j.issn.1000-6850.2012-0003

所属专题： 生物技术

• 林学 园艺 园林 • 上一篇    下一篇

牡丹类psDHN-YSK2基因全长cDNA的克隆与表达分析

张扬 盖树鹏 刘春英 郑国生 华芳霞 张玉喜   

  • 收稿日期:2011-12-30 修回日期:2012-04-16 出版日期:2012-06-05 发布日期:2012-06-05
  • 基金资助:

    牡丹内休眠中PsMADS-box基因对开花基因和花芽后发育的调控;牡丹花芽内休眠解除的分子机制

Cloning and Expression Analysis of psDHN-YSK2-like Dehydrin Gene in Tree Peony

  • Received:2011-12-30 Revised:2012-04-16 Online:2012-06-05 Published:2012-06-05

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摘要：

为了获得牡丹类脱水素基因的全长cDNA序列，推测其在休眠解除进程中的生物学功能，以不同低温处理时间的牡丹花芽为供试材料，末端快速扩增方法克隆全长cDNA序列，实时定量PCR分析其表达模式。结果表明牡丹类脱水素基因全长cDNA序列为1187 bp，包括831 bp的开放阅读框，114 bp的5’非编码区和242 bp的3’非编码区。其编码的蛋白具有两个植物脱水素蛋白特征性K片段，根据Close的分类方法，属于YSK2类脱水素蛋白。系统发生分析表明psDHN-YSK2基因与葡萄亲缘关系最近。在花芽内休眠解除前期随着低温处理时间的延长psDHN-YSK2表达呈上调趋势。本研究克隆了牡丹psDHN-YSK2的全长cDNA序列，分析了其在花芽内休眠解除过程的表达趋势，暗示了其参与了牡丹花芽的内休眠过程。

关键词: 株高, 株高

Abstract:

The objective of this study was to obtain the full length cDNA of dehydrin gene, and to conclude its role during endo-dormancy release in tree peony (Paeonia suffruticosa). The flower buds endured different time of chilling were used as materials, the full length cDNA of dehydrin gene was obtained using the method of Rapid Amplification of cDNA Ends (RACE) PCR, and its expression pattern were analyzed using real-time PCR. Full length cDNA of dehydrin gene was 1187 bp including a 831 bp open reading frame (ORF), 114 bp 5’ terminal untranslated region (UTR) and 242 bp 3’ terminal UTR followed by 26 bp polyA tail. The deduced amino acid sequence had two conservative K segments, which was the character of dehydrin gene family. According to the classification method of Close, it should be YSK2 type. Phylogenetic analysis indicated that the deduced psDHN-YSK2 was firstly clustered with that of grape. The expression of psDHN-YSK2 was steady up-regulated early in chilling requirement fulfillment. The full length cDNA of dehydrin gene was obtain and it expression trend was analyzed. All results suggested that it played an important role in the regulation of dormancy release in tree peony.