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园艺学报 ›› 2020, Vol. 47 ›› Issue (6): 1111-1125.doi: 10.16420/j.issn.0513-353x.2019-0507

• 研究论文 • 上一篇    下一篇

蝴蝶兰PhSVP的克隆及其在花发育过程中的表达分析

张 燕,许申平,梁 芳,蒋素华,袁秀云,牛苏燕,崔 波*   

  1. 郑州师范学院生物工程研究中心,郑州 450044
  • 出版日期:2020-06-25 发布日期:2020-06-25
  • 基金资助:
    河南省高等学校重点科研项目(18A180034);河南省科技攻关计划项目(182102110357);郑州师范学院科技创新团队支持计划项目

Cloning SVP Family Gene PhSVP from Phalaenopsis and Its Expression Analysis During Flower Development

ZHANG Yan,XU Shenping,LIANG Fang,JIANG Suhua,YUAN Xiuyun,NIU Suyan,and CUI Bo*   

  1. Biotechnology Research Center,Zhengzhou Normal University,Zhengzhou 450044,China
  • Online:2020-06-25 Published:2020-06-25

摘要: 从蝴蝶兰杂交品种‘大辣椒’中克隆到SVP同源MADS-box基因PhSVP,使用RACE方法获得PhSVP全长,其开放阅读框为687 bp,编码228个氨基酸。蛋白序列多重比对表明PhSVP蛋白包含一个高度保守的MEF2-like MADS结构域和一个中度保守的K-box结构域,且与兰科的SVP/AGL24同源蛋白相似度高,进化树分析也表明PhSVP蛋白与其他兰科植物的SVP/AGL24同源蛋白亲缘关系最近。PhSVP的空间表达模式表明其在营养器官表达量高,在萼片、侧瓣和唇瓣等生殖器官表达量低。进一步对PhSVP在成花转换和花发育过程中花序顶端的表达进行分析,表明PhSVP的转录水平在营养分生组织向花序分生组织过渡的过程中无显著变化,仅在花序分生组织阶段有少量上升,但在花发育早期花分生组织阶段中的表达水平显著升高,而在花发育后期的萼片原基、花瓣原基和合蕊柱原基阶段持续下降。此外还发现A类基因PhAP1在花发育过程中的花序顶端的表达模式与PhSVP相似,而B类基因PhPI和C类基因PhAG在花瓣原基和合蕊柱原基阶段的表达量最高。检测了PhSVP在抽葶期、花蕾期和盛花期的根、叶和花葶中的转录水平,结果表明PhSVP在花蕾期的根中的表达量稍高于其他时期,在不同阶段的叶中的表达水平无显著差异,而在抽葶期花葶中的表达量显著高于花蕾期和盛花期。试验结果暗示蝴蝶兰PhSVP可能起到调控花分生组织状态的维持、避免花器官原基过早分化的作用。

关键词: 蝴蝶兰, SVP基因, 表达分析, 花发育, 花分生组织决定

Abstract: A SVP-like MADS-box gene PhSVP was isolated from Phalaenopsis hybrida‘Big Chilli’. The full-length cDNA of PhSVP was obtained using the RACE method and the open reading frame was 687 bp which encoded 228 amino acids. Multiple protein sequence alignment revealed that PhSVP protein contained a highly conserved MEF2-like MADS domain and moderately conserved K-box domain and shared high identity with the homologs of SVP/AGL24 from the orchid family. Phylogenetic analysis also showed that PhSVP protein was more closely related to SVP/AGL24 homologues of other orchid species. The spatial expression pattern of PhSVP showed that it was highly expressed in vegetative organs and weakly expressed in reproductive organs such as sepals,lateral petals and lips. Further analysis of PhSVP expression in the inflorescence apex during the floral transition and flower development revealed that there was no significant change in the level of PhSVP transcript during progression from vegetative phase to inflorescence meristem phase and only a slight increase of PhSVP expression level was found in inflorescence meristem phase,whereas the expression of PhSVP was upregulated in the floral meristem phase during the early stage of flower development and gradually decreased in the late stages when sepal,petal and column primordia are sequentially formed. In addition,we also found the expression pattern of A-class MADS-box gene PhAP1 in the inflorescence apex during the flower development was similar to the expression pattern of PhSVP,whereas the expression levels of B-class gene PhPI and C-class gene PhAG were the highest in petal and column primordia stage. Finally,the expression of PhSVP in various tissues such as root,leaf and scape during scape elongation stage,floral bud stage and blooming flower stage was investigated. The result showed that PhSVP expression level in root was slightly higher at floral bud stage and its expression level in leaf at different flowering stages remained unchanged. However,the PhSVP transcript in the scape accumulated to higher levels at scape elongation stage than floral bud stage and blooming flower stage. These results suggest that PhSVP may play a regulatory role in maintaining floral meristem identity and preventing precocious differentiation of floral organ primordia.

Key words: Phalaenopsis, SVP gene, expression analysis, flower development, floral meristem identity

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