甘蔗生长素结合蛋白ScABP4基因的克隆与表达

发布时间：2019-08-17 12:07

【摘要】：生长素结合蛋白(auxin binding protein,ABP)在生长素信号转导、植物生长发育调控等方面发挥重要作用。该研究利用RT-PCR方法从甘蔗品种Badila中克隆得到了1个ABP基因,序列分析显示该基因的开放读码框(ORF)长度为615bp,编码204个氨基酸。多序列比对和系统进化树分析表明它与高粱(Sorghum bicolor)和玉米(Zea mays)的ABP4蛋白的亲缘关系最近,因此将该基因命名为ScABP4。将其构建到原核表达载体pGEX-6P-1中,成功表达出一个分子量约为22.5kD的蛋白。亚细胞定位结果显示ScABP4主要定位于细胞质网状结构和细胞膜;生物信息学分析显示ScABP4是一个带有信号肽的分泌蛋白,说明ScABP4蛋白可能储存在内质网中并在质膜上执行其生物学功能。荧光定量PCR分析结果表明,ScABP4基因在甘蔗的芽、根、茎、叶中均有表达,其中在芽中相对表达量最高。生长素和黑暗处理下ScABP4基因的表达上调,说明ScABP4基因可能参与甘蔗对生长素的响应及与光信号通路的互作。此外,ABA、JA、SA、CuCl2胁迫能够诱导ScABP4基因的表达,CdCl_2胁迫可抑制ScABP4的表达。由此推测,ScABP4基因可能参与甘蔗对病害、干旱、渗透、重金属等胁迫的应答过程。该研究结果为探讨甘蔗生长素结合蛋白基因的功能提供了实验证据。
[Abstract]:Auxin binding protein (auxin binding protein,ABP) plays an important role in auxin signal transduction and plant growth and development regulation. In this study, a ABP gene was cloned from sugarcane variety Badila by RT-PCR. Sequence analysis showed that the open reading frame (ORF) of the gene was 615bp and encoded 204amino acids. Multiple sequence alignment and phylogenetic tree analysis showed that it was closely related to ABP4 protein of sorghum (Sorghum bicolor) and corn (Zea mays), so the gene was named ScABP4.. A protein with molecular weight of about 22.5kD was successfully expressed in prokaryotic expression vector pGEX-6P-1. The results of subcellular localization showed that ScABP4 was mainly located in cytoplasmic reticular structure and cell membrane, and bioinformatics analysis showed that ScABP4 was a secretory protein with signal peptide, indicating that ScABP4 protein might be stored in endoplasmic reticulum and performed its biological function on plasma membrane. The results of fluorescence quantitative PCR analysis showed that ScABP4 gene was expressed in bud, root, stem and leaf of sugarcane, and the relative expression was the highest in bud. The expression of ScABP4 gene was up-regulated under auxin and dark treatment, suggesting that ScABP4 gene may be involved in the response of sugarcane to auxin and the interaction with light signal pathway. In addition, ABA,JA,SA,CuCl2 stress could induce the expression of ScABP4 gene, and CdCl_2 stress could inhibit the expression of ScABP4. It is speculated that ScABP4 gene may be involved in the response of sugar cane to disease, drought, infiltration, heavy metal and other stresses. The results provided experimental evidence for exploring the function of auxin binding protein gene in sugar cane.
【作者单位】：福建农林大学/农业部福建甘蔗生物学与遗传改良重点实验室;
【基金】：国家高技术研究发展计划(863计划)项目(2013AA102604) 国家自然科学基金(31171605;31371688) 国家现代农业产业技术体系项目(CARS-20-1-1)
【分类号】：S566.1

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