大豆GmCWIN3基因的克隆及转基因拟南芥株型鉴定
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海南大学

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国家自然科学基金项目 (32171937);海南省重点研发计划项目(ZDYF2022XDNY142);三亚“崖州湾”菁英人才科技专项(SCKJ-JYRC-2023-17)


Cloning of GmCWIN3 gene and the Identification of Plant Architecture in Transgenic Arabidopsis
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National Natural Science Foundation of China (32171937); the Hainan Province Science and Technology Special Fund (ZDYF2022XDNY142); the Project of Sanya Yazhou Bay Science and Technology City (SCKJ-JYRC-2023-17)

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    摘要:

    植物细胞壁转化酶(CWIN,cell wall invertase)可以催化蔗糖水解为葡萄糖和果糖,在植物生长发育过程中具有重要作用。为探究GmCWINs在植物株高、分枝数等株型发育过程中的功能,本研究克隆到1个该基因家族成员GmCWIN3,序列分析结果显示,GmCWIN3基因的CDS全长为1728bp,其编码575个氨基酸。GmCWIN3蛋白二级结构中无规则卷曲结构占比最大,为48.17%;蛋白质三级结构显示该基因编码了一种细胞壁转化酶。该基因启动子区具有多个光响应元件,同时也具有响应生长素、赤霉素及昼夜节律的顺式作用元件。进一步地分析生长素和赤霉素诱导下GmCWIN3的表达模式,发现GmCWIN3基因能在反应初期快速地响应两种激素的诱导。构建GmCWIN3基因的植物表达载体并完成其在拟南芥中的过表达,对转基因株系及野生型拟南芥的细胞壁转化酶活性和蔗糖含量进行测定,结果表明转基因拟南芥中细胞壁转化酶活性显著提高,且蔗糖含量显著降低。此外,对植株表型进行观察和统计,发现与野生型相比,过表达植株的株高和分枝数均显著增加。综上所述,本研究推测GmCWIN3基因可能通过调节体内蔗糖的水解,从而参与生长素及赤霉素介导的大豆株高及分枝数等性状的形成过程。本研究结果为进一步探究GmCWIN3基因调控大豆株型相关性状的分子机制研究奠定了基础。

    Abstract:

    Cell wall invertase (CWIN) is an enzyme that catalyzes sucrose hydrolysis to glucose and fructose, and plays an important role in plant growth and development. To explore the function of GmCWINs in plant architecture, particular in plant height and branch number, GmCWIN3, a member of GmCWINs family was cloned in our study. The sequence analysis results showed that the total CDS length of GmCWIN3 gene was 1728 bp and it encoded 575 amino acids.The secondary structure of GmCWIN3 protein exhibited that the irregular coil structure accounted for the largest proportion (48.17%), and the tertiary structure of the protein revealed that GmCWIN3 encoded a cell wall invertase. Except for several photo-responsive cis-acting elements, some elements associated with the process of growth and development were also found in the promoter region of GmCWIN3, such as auxin responsive element, gibberellin responsive element and circadian rhythm element, etc. Further, through the assay of exogenous spraying different hormones, it was found that GmCWIN3 gene could rapidly respond to the induction of auxin and gibberellin in the initial reaction period. The GmCWIN3 gene overexpression vector was constructed and the transgenic GmCWIN3 Arabidopsis plants were obtained. The cell wall invertase activity and sucrose content of the transgenic Arabidopsis and wild type plants were measured. The results showed that the cell wall invertase activity of transgenic plants was significantly higher than that in the wild type plants, and the sucrose content was significantly decreased. Additionally, the phenotype identification exhibited that the plant height and branch number of the transgenic plants were significantly increased compared with the wild type plants. In summary, this study speculated that GmCWIN3 gene may be involved in the development of regulating soybean plant height and branch number mediated by hormones signaling process through the regulation of sucrose hydrolysis. Our study laid a foundation for further exploring the molecular mechanism of GmCWIN3 in regulating soybean plant architecture related traits.

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  • 收稿日期:2024-03-08
  • 最后修改日期:2024-04-25
  • 录用日期:2024-08-19
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