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洋葱(Allium cepa L.)MADS-Box 基因的克隆与表达分析及细胞质雄性不育的调控机制。

Cloning and Expression Analysis of Onion (Allium cepa L.) MADS-Box Genes and Regulation Mechanism of Cytoplasmic Male Sterility.

机构信息

School of Forestry, Northeast Forestry University, 26 Hexing Road, Harbin, 150040, China.

Medical Laboratory College of Youjiang Medical College for Nationalities, Baise, 533000, Guangxi, China.

出版信息

Biochem Genet. 2023 Oct;61(5):2116-2134. doi: 10.1007/s10528-023-10360-6. Epub 2023 Mar 22.

Abstract

Flower organ development is one of the most important processes in plant life. However, onion CMS (cytoplasmic male sterility) shows an abnormal development of floral organs. The regulation of MADS-box transcription factors is important for flower development. To further understand the role of MADS-box transcription factors in the regulation of cytoplasmic male sterility onions. We cloned the full-length cDNA of five MADS-box transcription factors from the flowers of onion using RACE (rapid amplification of cDNA ends) technology. We used bioinformatics methods for sequence analysis and phylogenetic analysis. Real-time quantitative PCR was used to detect the expression patterns of these genes in different onion organs. The relative expression levels of five flower development genes were compared in CMS onions and wild onions. The results showed that the full-length cDNA sequences of the cloned MADS-box genes AcFUL, AcDEF, AcPI, AcAG, and AcSEP3 belonged to A, B, C, and E MADS-box genes, respectively. A phylogenetic tree construction analysis was performed on its sequence. Analysis of MADS-box gene expression in wild onion and CMS onion showed that the formation of CMS onion was caused by down-regulation of AcDEF, AcPI, and AcAG gene expression, up-regulation of AcSEP3 gene expression, and no correlation with AcFUL gene expression. This work laid the foundation for further study of the molecular mechanism of onion flower development and the molecular mechanism of CMS onion male sterility.

摘要

花器官的发育是植物生命过程中最重要的过程之一。然而,洋葱 CMS(细胞质雄性不育)表现出花器官的异常发育。MADS-box 转录因子的调控对于花发育很重要。为了进一步了解 MADS-box 转录因子在调控细胞质雄性不育洋葱中的作用。我们使用 RACE(快速扩增 cDNA 末端)技术从洋葱花中克隆了五个 MADS-box 转录因子的全长 cDNA。我们使用生物信息学方法进行序列分析和系统发育分析。实时定量 PCR 用于检测这些基因在不同洋葱器官中的表达模式。比较了 CMS 洋葱和野生洋葱中五个花发育基因的相对表达水平。结果表明,克隆的 MADS-box 基因 AcFUL、AcDEF、AcPI、AcAG 和 AcSEP3 的全长 cDNA 序列分别属于 A、B、C 和 E MADS-box 基因。对其序列进行了系统发育树构建分析。野生洋葱和 CMS 洋葱中 MADS-box 基因表达分析表明,CMS 洋葱的形成是由于 AcDEF、AcPI 和 AcAG 基因表达下调、AcSEP3 基因表达上调,与 AcFUL 基因表达无关。这项工作为进一步研究洋葱花发育的分子机制和 CMS 洋葱雄性不育的分子机制奠定了基础。

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