MA Hui,LIN Fuyong,LI Fan,et al.Cloning and Expression of F3′H Gene in Impatiens uliginosa[J].Northern Horticulture,2025,(3):1-8.[doi:10.11937/bfyy.20242796]
滇水金凤F3′H基因的克隆及表达分析
- Title:
- Cloning and Expression of F3′H Gene in Impatiens uliginosa
- 文章编号:
- 1001-0009(2025)03-0001-08
- Keywords:
- Impatiens uliginosa; F3′H gene; flower color; gene cloning; expression analysis
- 分类号:
- S 681.1
- 文献标志码:
- A
- 摘要:
- 以滇水金凤（Impatiens uliginosa）为试材，采用RT-PCR技术，成功克隆出3个F3′H基因，并分别命名为IuF3′H1、IuF3′H2和IuF3′H3，利用DNAMAN和MEGA软件对F3′H基因所编码的蛋白序列进行同源性分析和系统进化分析，并利用qRT-PCR分析IuF3′H基因的表达模式，研究了滇水金凤F3′H基因对花色的调控机理，以期为后续的基因功能验证及对植物的花色育种提供参考依据。结果表明：IuF3′H1、IuF3′H2和IuF3′H3的cDNA全长分别为1 560、1 545 bp和1 521 bp，编码519、514、506个氨基酸；IuF3′H1、IuF3′H2和IuF3′H3基因均不具有内含子，其蛋白均属于CYP450家族。系统进化分析表明，IuF3′H1、IuF3′H2和IuF3′H3基因均与喜马拉雅凤仙花聚为一支，亲缘关系最近，但处于2个不同分支。qRT-PCR分析表明，3个拷贝在滇水金凤2种不同花色（白色、红色）和花苞期、始花期、盛花期、谢花期中均有表达，其中IuF3′H1基因的表达在红色花的谢花期达到最强；IuF3′H2基因2个花色的表达量随着花不断发育总体呈现升高趋势；IuF3′H3基因在红色花的盛花期表达显著高于其他时期。因此，推测IuF3′H基因均在滇水金凤花色苷合成中发挥着重要的作用且在花发育的中后期作用更为显著。
- Abstract:
- Taking Impatiens uliginosa as the test material,three F3′H gene，named IuF3′H1，IuF3′H2 and IuF3′H3，were successfully cloned by RT-PCR technique.Using DNAMAN and MEGA software，homology and phylogenetic analyses of the protein sequences encoded by the F3′H gene，and the expression patterns of the IuF3′H gene by qRT-PCR was analyzed.The mechanism of the F3′H gene of Impatiens uliginosa in the regulation of the flower colours in the plant was researched,in order to provide reference for the subsequent validation of the functions of the genes and the breeding of the flower colours in the plant.The results showed that the full lengths of the cDNAs of IuF3′H1，IuF3′H2 and IuF3′H3 were 1 560 bp，1 545 bp and 1 521 bp，respectively，and encoded 519，514 and 506 amino acids；none of the IuF3′H1，IuF3′H2 and IuF3′H3 genes had introns，and all their proteins belonged to the CYP450 family.Phylogenetic analysis showed that the IuF3′H1，IuF3′H2 and IuF3′H3 genes were all clustered with Impatiens glandulifera，which were closest in affinity but in two different branches.qRT-PCR analysis showed that the three copies were expressed in two different flower colors (white and red) and bud stage，primordial stage，blooming stage，flowering shedding stage in Impatiens uliginosa，among which the expression of IuF3′H1 gene reached the strongest in the flower shedding stage of red flowers；the expression of IuF3′H2 gene in two flower colors showed an increasing trend with the continuous development of flowers;the expression of IuF3′H3 gene was significantly higher in the blooming period of red flowers than other periods.Therefore，it was hypothesized that IuF3′H gene play important roles in the synthesis of the flower color glycosides of Impatiens uliginosa，and their roles were more significant in the middle and late stages of flower development.
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备注/Memo
第一作者简介：马慧（1999-），女，硕士研究生，研究方向为园林植物栽培和遗传育种。E-mail:mahui04179@swfu.edu.cn.责任作者：黄美娟（1972-），女，博士，教授，博士生导师，现主要从事园林植物栽培和遗传育种与分子生物学等研究工作。E-mail:meijuanhuang@swfu.edu.cn.基金项目：国家自然科学基金资助项目（32060364，32060366，31860230）；云南省农业联合专项重点资助项目（202101BD070001-018）；云南省重大科技专项资助项目（202102AE090052）；云南省园林植物遗传改良与高效繁育博士生导师团队资助项目。收稿日期：2024-07-04