文冠果XsSCL6基因的克隆及表达分析
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1.北京林业大学 省部共建森林培育与保护教育部重点实验室;2.辽宁文冠实业开发有限公司

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国家自然科学基金(32071738);国家重点研发计划(2022YFD2200400)


Cloning and Expression Analysis of XsSCL6 in Xanthoceras sorbifolium Bunge
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The National Natural Science Foundation of China (32071738); The National Key Technologies R&D Program of China (2022YFD2200400)

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    摘要:

    GRAS家族HAM亚家族(Scarecrow-like 6,SCL6)是一类植物特有的转录因子,在植物生长发育中具有关键作用。文冠果(Xanthoceras sorbifolium Bunge)是我国特有的重要林木能源植物,雄花的雌蕊败育是造成产量低的主要原因。为解析XsSCL6基因在文冠果(Xanthoceras sorbifolium Bunge)雌蕊发育中的调控作用,采用RT-PCR方法克隆文冠果4个XsSCL6同源基因,并对关键XsSCL6a蛋白进行亚细胞定位;对XsSCL6基因进行生物信息学分析,利用qRT-PCR分析XsSCL6基因在文冠果雌蕊败育过程中的表达模式。结果表明:(1)获得文冠果XsSCL6a、XsSCL6b、XsSCL6c和XsSCL6d基因ORF全长,分别编码751、669、717和717个氨基酸,均属于亲水不稳定蛋白。(2)文冠果XsSCL6蛋白主要由无规则卷曲和α-螺旋构成,定位于细胞核。(3)XsSCL6a、XsSCL6c和XsSCL6d均包含GRAS保守结构域, 文冠果XsSCL6蛋白和克莱门柚及甜橙的亲缘关系最近。(4)文冠果XsSCL6基因启动子中包含多种激素响应元件、植物发育相关元件。(5)qRT-PCR结果显示XsSCL6基因各个时期在雄花中的表达量显著高于雌花,表达量的趋势和转录组FPKM值一致。(6)xso-miR171和XsSCL6基因均存在裂解方式的靶向关系,TPM值和FPKM值为负相关关系。XsSCL6基因可能受到xso-miR171调控,共同参与文冠果雌蕊发育过程,对文冠果花发育具有重要作用。本研究为今后研究进一步文冠果中XsSCL6基因的功能和雌蕊败育机制提供了参考,为采用分子育种方法提高种子产量提供了理论依据。

    Abstract:

    The GRAS family HAM subfamily (Scarecrow-like 6, SCL6) is a class of plant-specific transcription factors with key roles in plant growth and development. Xanthoceras sorbifolium is an important woodyenergy plant species endemic to China, and pistil abortion in its male flowers is a key cause of low yield. To resolve the regulatory role of the XsSCL6 genes in the pistil development of yellow-horn (Xanthoceras sorbifolium Bunge), four XsSCL6 homologous genes were cloned by RT-PCR and subcellular localization of key XsSCL6a protein was performed. We used bioinformatics to analyze XsSCL6 genes. In addition, qRT-PCR was used to analyze the expression pattern of XsSCL6 genes during pistil abortion in yellow-horn. The results showed that: (1) The full length of the ORFs of XsSCL6a, XsSCL6b, XsSCL6c and XsSCL6d gene, encoding 751, 669, 717 and 717 amino acids, respectively, were obtained for the hydrophilic unstable proteins. (2) XsSCL6 protein mainly consists of irregular curls and α- Helical structure, located in the nucleus. (3) XsSCL6a, XsSCL6c and XsSCL6d all contained GRAS conserved structural domains. XsSCL6 proteins were most closely related to Clemenegro and sweet orange. (4) The promoter of XsSCL6 genes contained various phytohormone response elements and plant development-related elements. (5) The qRT-PCR results showed that the expression of XsSCL6 genes was significantly higher in male flowers than in female flowers at all times, and the trend of expression was consistent with the transcriptome FPKM values. (6) Both xso-miR171 and XsSCL6 genes were targeted by cleavage mode and their TPM and FPKM values were negatively correlated. XsSCL6 genes might be regulated by xso-miR171. And they jointly participated in the pistil development of yellow-horn, which had an important role in yellow-horn flower development. This study provides a reference for future studies focusing on XsSCL6 genes functions and the mechanism of pistil abortion in X. sorbifolium and a theoretical basis for adopting molecular breeding methods to improve seed yield.

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赵雅欣,陈雨欣,卢涵冰,等.文冠果XsSCL6基因的克隆及表达分析[J].西北植物学报,2023,43(10):1640-1652

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  • 收稿日期:2023-03-24
  • 最后修改日期:2023-07-24
  • 录用日期:2023-08-17
  • 在线发布日期: 2023-10-20
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