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园艺学报 ›› 2014, Vol. 41 ›› Issue (10): 2125-2131.

• 研究报告 • 上一篇    下一篇

铁炮百合热激转录因子基因HSFA2b的克隆与表达分析

辛海波1,2,连青龙2,3,董爱香1,张华丽1,宋利娜1,义鸣放2,*   

  1. 1北京市园林科学研究院,绿化植物育种北京市重点实验室,北京 100102;2中国农业大学观赏园艺与园林系,北京 100193;3农业部规划设计研究院设施农业研究所,北京 100125
  • 收稿日期:2014-05-29 出版日期:2014-10-25 发布日期:2014-10-25
  • 基金资助:

    国家自然科学基金项目(30972024);农业部公益性行业(农业)科研专项(200903020)

Cloning and Expression Analysis of HSFA2b from Lilium longiflorum

XIN Hai-bo1,2,LIAN Qing-long2,3,DONG Ai-xiang1,ZHANG Hua-li1,SONG Li-na1,and YI Ming-fang2,*   

  1. 1Beijing Institute of Landscape Architecture,Beijing Key Laboratory of Greening Plants Breeding,Beijing100102,China;2Department of Ornamental Horticulture and Landscape Architecture,China Agricultural University,Beijing 100193,China;3Institute of Facility Agriculture,Chinese Academy of Agricultural Engineering,Beijing 100125,China
  • Received:2014-05-29 Online:2014-10-25 Published:2014-10-25

摘要: 以铁炮百合(Lilium longiflorum)‘白天堂’为试材,采用RACE方法克隆到了1个热激转录因子(Heat shock transcription factor,HSF)A家族全长基因。该基因包含1个编码315个氨基酸的开放阅读框。对推定的氨基酸序列进行聚类分析,与已知的水稻5个HSFA2同源性较高,是1个新的HSFA2编码基因,命名为LlHSFA2b。经过与已报道的番茄、拟南芥、水稻和百合的HSFA2序列比对分析,该基因编码的HSF蛋白具有热激转录因子所具备的典型结构域和调控元件,不同的是,它缺少1个转录激活域和核输出信号。半定量RT-PCR分析表明,该基因特异地受高温和H2O2诱导表达。荧光定量PCR表明:该基因在百合根、鳞茎和叶片中均受热激诱导表达,且表达量无明显差异;37 ℃热激处理下,与已报道的LlHSFA2a相比,LlHSFA2b表达时间点和表达量的峰值均提前。这些结果说明,本试验中克隆到了1个新的HSFA2编码基因,与LlHSFA2a相比,该基因可能参与了百合早期热信号传导过程。

关键词: 铁炮百合, 热激转录因子, 热激, 耐热性

Abstract: A full-length Heat shock transcription factor(HSF)gene encoding 315 aa amino acids,belonging to HSFA2,was cloned from Lilium longiflorum‘White Heaven’by RACE(Rapid Amplification of cDNA Ends)technique. Cluster analysis showed that the putative HSFA2b was highly homologous to known OsHSFA2s and a novel member of HSFA2. Furthermore,alignment with HSFA2 from tomato,Arabidopsis,rice and lily indicated there were conserved functional domains except for an activation domain and a nuclear export signal in LlHSFA2b. Semi-quantitative RT-PCR showed that expression of this gene could be induced by heat and H2O2 exclusively. Real-time PCR revealed that LlHSFA2b transcripts were induced by heat in root,bulb and leaf without significant difference and detected earlier than that of LlHSFA2a during heat shock. These data indicate that LlHSFA2b is a novel HSF-encoding genewhich could respond to heat at an early stage.

Key words: Lilium longiflorum, heat shock transcription factor(HSF), heat shock, thermotolerance

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