园艺学报 ›› 2019, Vol. 46 ›› Issue (3): 444-452.doi: 10.16420/j.issn.0513-353x.2018-0651

• 研究论文 • 上一篇    下一篇


王孝娣,冀晓昊,郑晓翠,王莹莹,宋 杨*,刘凤之*   

  1. (中国农业科学院果树研究所,农业部园艺作物种质资源利用重点实验室,辽宁省落叶果树矿质营养与肥料高效利用重点实验室,辽宁兴城 125100)
  • 出版日期:2019-03-25 发布日期:2019-03-25

Cloning and Functional Analysis of a Cold Treatment Response Factor Gene PdCIbHLH in Peach

WANG Xiaodi,JI Xiaohao,ZHENG Xiaocui,WANG Yingying,SONG Yang*,and LIU Fengzhi*   

  1. (Research Institute of Pomology,Chinese Academy of Agricultural Sciences/Key Laboratory of Fruit Germplasm Resources Utilization,Ministry of Agriculture,Laboratory of Mineral Nutrition and Efficient Fertilization for Deciduous Fruits,Liaoning Province,Xingcheng,Liaoning 125100,China)
  • Online:2019-03-25 Published:2019-03-25

摘要: 以山桃(Prunus davidiana Franch)为试材,克隆获得冷处理响应基因PdCIbHLH(cold-induced bHLH),其ORF为1 617 bp,编码含539个氨基酸的蛋白质,含有典型的bHLH结构域。系统发生分析表明,PdCIbHLH与苹果MdICE1的同源性最高。PdCIbHLH在山桃不同器官中均可表达,在叶片中表达最高,在枝条中表达较弱。叶片中PdCIbHLH受4 ℃处理诱导表达。在拟南芥中超量表达PdCIbHLH,其低温抗性较野生型显著升高。酵母杂交试验结果显示,PdCIbHLH可与AtCBF3的启动子相互作用。烟草瞬时表达试验结果表明,PdCIbHLH可激活AtCBF3的表达。推测PdCIbHLH通过直接激活CBF的表达正调控植株对低温处理的响应。

关键词: 山桃, 低温胁迫, PdCIbHLH, 表达分析, 功能鉴定

Abstract: This study is aiming at cloning PdICbHLH from Prunus davidiana Franch,to analyze its expression pattern and reveal biological functions involved in cold stress. PdCIbHLH gene was obtained,and the open reading frame(ORF)contained 1 617 bp,encoding a protein of 539 amino acid residues. Sequence analysis indicated that PdCIbHLH exhibited the structures of bHLH domain. Phylogenetic relationship analysis showed that the homology of PdCIbHLH was close to the MdICE1. The qRT-PCR analysis indicated that PdCIbHLH was expressed in different organs. The expression showed high level in leaves,and the lowest in stems,and then expression of PdCIbHLH in leaves was induced by 4 ℃ treatment. The PdCIbHLH-overexpressing Arabidopsis exhibited increased tolerance to cold stress. Yeast one-hybrid assay showed that PdCIbHLH interacted with promoter of AtCBF3. The transient expression assays showed that PdCIbHLH could induce the expression of AtCBF3. It is speculated that PdCIbHLH was an important regulatory in response to cold treatment by directly activating the transcript of CBF.

Key words: Prunus davidiana, cold stress, PdCIbHLH, expression analysis, functional identification