| 摘要: |
| 【目的】克隆大豆GmCYP85A2基因并进行表达特性分析，为进一步研究GmCYP85A2基因的功能及其对大豆叶柄角的调控机制奠定基础。【方法】以郑豆196作为试验材料，利用同源克隆技术克隆GmCYP85A2基因，对其编码蛋白进行生物信息学分析，采用实时荧光定量PCR(qRT-PCR)方法检测GmCYP85A2基因在大豆植株不同组织（根、茎、叶片、叶柄、花、花芽和豆荚）及不同光照处理时间（0，4，8，12，16，20，24 h）下叶柄中的相对表达量，分析该基因的表达特性。【结果】成功克隆了大豆GmCYP85A2基因，其长1 524 bp，开放阅读框长1 401 bp，编码466个氨基酸；编码蛋白分子质量为53.4 ku，理论等电点（pI）为9.00，具有典型的P450超家族保守结构和蛋白三级结构，亚细胞定位结果表明该蛋白为分泌型蛋白。启动子分析结果表明，该基因转录起始位置ATG上游2 kb的基因组序列含有多个响应光照以及与生长发育、逆境胁迫相关的顺式作用元件。基因表达特性分析结果表明，GmCYP85A2基因主要在花、花芽、豆荚及叶柄中表达，且以花中的表达量最大；在营养生长时期，叶柄中GmCYP85A2的表达量受光照影响而上调，并伴随着叶柄角的变大。【结论】克隆了大豆GmCYP85A2基因，并研究了该基因的表达特性，分析了其在叶柄角调控中的可能机制。 |
| 关键词: 大豆 叶柄角 细胞色素P450 GmCYP85A2 油菜素内酯 |
| DOI： |
| 分类号: |
| 基金项目:河南省科技攻关计划（农业领域）项目（192102110125） |
|
| Cloning and expression characteristics of GmCYP85A2 gene in soybean |
|
LIU Panpan,MENG Xiaobing,FU Rao,et al
|
| Abstract: |
| 【Objective】The soybean GmCYP85A2 gene was cloned and its expression characteristics were analyzed to provide basis for further study on its function and mechanism in regulation of petiole angle in soybean.【Method】GmCYP85A2 gene of Zhengdou 196 was cloned by homologous cloning technique and the physical properties and structure of its encoded protein were analyzed.The qRT-PCR was conducted to study its expression characteristics in different tissues (root,stem,leaf,petiole,flower,flower buds and pod) of soybean and in the petiole under different lengths of illumination time (0,4,8,12,16,20 and 24 h).【Result】The GmCYP85A2 gene was successfully cloned,which contained 1 524 bp and 1 401 bp open reading frame and encoded a protein of 466 amino acids.The molecular weight of its encoded protein was 53.4 ku and the theoretical isoelectric point (pI) was 9.00 with a typical conservative domain and tertiary structure of P450 superfamily.The subcellular localization indicated that the protein was a secretory protein.According to promoter analysis,there were several cis acting elements responding to light and relating to the development and stress in the transcriptional initiation site of 2 kb upstream of ATG.GmCYP85A2 gene was mainly expressed in flowers,unopened flowers,pods and petioles with the highest expression in flowers.During the growth period,the expression of GmCYP85A2 in petiole was up-regulated by light,accompanied by the increase of petiole angle.【Conclusion】The soybean GmCYP85A2 gene was cloned,its expression characteristics were studied,and its possible mechanism in regulation of petiole was analyzed. |
| Key words: soybean petiole angle cytochrome P450 GmCYP85A2 brassinolide |
