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植物的核因子Y(NF-Y)是由3个亚基A、B和C组成,在响应非生物胁迫过程中起着重要的作用。本研究以大豆铁丰8号为材料,建立大豆c DNA文库,以p GBKT7-Gm Di19-5为诱饵,通过酵母双杂交技术筛选大豆c DNA文库,获得了大豆NF-Y转录因子家族亚基C的一个成员,命名为GmNF-YCa。该基因全长为864 bp,编码287个氨基酸,属于NF-YC亚家族。GmNF-YCa分子量为31.6 k D,等电点为5.07亲水性蛋白,具有一个跨膜结构域,无信号肽。序列分析表明,NF-YC亚家族具有很高的保守性。GmNF-YCa基因启动子含有ARE、Box4、GATA-motif、Box I、ACE、ABRE和CAT-Box等胁迫和光响应元件。组织特异性分析表明,GmNF-YCa基因在种子萌发期表达量最高。实时定量结果表明,GmNF-YCa受蔗糖和甘露醇的诱导上调表达。使用农杆菌介导法,将大豆GmNF-YCa基因导入拟南芥,并进行了功能分析。发芽率试验分析表明,GmNF-YCa的转基因提高了转基因拟南芥萌发期对渗透胁迫的耐性;改良了在蔗糖和甘露醇处理下转基因拟南芥的根系生长和侧根发育。
Plant nuclear factor Y (NF-Y) is composed of three subunits, A, B and C, play an important role in the response to abiotic stress. In this study, a soybean c DNA library was constructed with soybean Tiefeng 8 as a material. The soybean c DNA library was screened by yeast two-hybrid technique using p GBKT7-Gm Di19-5 as bait, and the soybean NF-Y transcription factor family subunit A member of C, named GmNF-YCa. The full length of this gene is 864 bp, encoding 287 amino acids, belonging to the NF-YC subfamily. GmNF-YCa has a molecular weight of 31.6 kD and an isoelectric point of 5.07 hydrophilic protein with a transmembrane domain and no signal peptide. Sequence analysis showed that the NF-YC subfamily is highly conserved. The GmNF-YCa promoter contains stress and light-responsive elements such as ARE, Box4, GATA-motif, Box I, ACE, ABRE and CAT-Box. Tissue-specific analysis showed that the GmNF-YCa gene was the highest in the seed germination period. Real-time quantitative results showed that GmNF-YCa up-regulated by the induction of sucrose and mannitol. Using Agrobacterium-mediated method, the soybean GmNF-YCa gene was introduced into Arabidopsis thaliana and its function was analyzed. The germination rate analysis indicated that GmNF-YCa transgene enhanced the tolerance to osmotic stress during the germination of transgenic Arabidopsis thaliana and improved the root growth and lateral root development of transgenic Arabidopsis thaliana under sucrose and mannitol treatment.