厉雪;苏立遥;陈燕;张舒婷;徐小萍;陈晓慧;陈荣珠;赖钟雄;林玉玲;

• 1:福建农林大学园艺植物生物工程研究所

摘要(Abstract)：

【目的】探究LSm14对龙眼体胚发生的影响,克隆RNA结合蛋白DlLSm14e基因并观察其亚细胞定位,分析其表达特性。【方法】以龙眼胚性愈伤组织为材料,依据转录组数据和基因组数据,采用RT-PCR克隆了一个DlLSm14基因,将其命名为DlLSm14e。对该基因进行生物信息学分析、亚细胞定位观察、龙眼非胚性愈伤组织和不同胚性培养物以及不同激素处理下的FPKM值分析和不同浓度细胞分裂素(KT)处理下的qRT-PCR检测。【结果】该基因CDS全长1 707 bp,编码568个氨基酸,该蛋白不含信号肽和跨膜结构,不属于分泌蛋白和跨膜蛋白;此外,该蛋白含有N端保守结构域以及延长的C端,并且含有74个磷酸化修饰位点;系统发育分析表明,DlLSm14e蛋白与漾濞槭(Acer yangbiense)相似度较高,与单子叶植物亲缘关系最远;顺式作用元件分析表明,该基因含有大量光响应元件和多种激素响应元件;亚细胞定位结果表明,该基因为核定位基因;表达谱与实时荧光定量分析显示,DlLSm14e的FPKM值在非胚性愈伤组织(NEC)阶段最低,在球形胚阶段达到最高,推测DlLSm14e大量累积可能有利于龙眼体胚早期形态建成;不同激素处理下,2,4-D抑制该基因表达,KT促进其表达,而KT的浓度能影响该基因的表达。【结论】龙眼LSm14e基因定位于细胞核,该基因FPKM值随龙眼体胚胚性增加而升高,并且响应多种与龙眼体胚发生相关的激素,推测该基因可能参与龙眼体胚早期的形态建成。

关键词(KeyWords)： 龙眼;DlLSm14基因;克隆;亚细胞定位;表达

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(31672127);; 福建省高原学科建设经费(102/71201801101);; 福建农林大学创新基金(CXZX2017187,CXZX2018078)

作者(Author): 厉雪;苏立遥;陈燕;张舒婷;徐小萍;陈晓慧;陈荣珠;赖钟雄;林玉玲;

Email:

DOI: 10.13925/j.cnki.gsxb.20190648

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