大豆转录因子基因GmNF-YCa可提高转基因拟南芥渗透胁迫的耐性

发布时间：2018-07-02 11:19

本文选题：大豆 + 核因子Y　；参考：《作物学报》2017年08期

【摘要】：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。
[Abstract]:Plant nuclear factor Y (NF-Y) is composed of three subunits, Agna B and C, and plays an important role in response to abiotic stress. In this study, a soybean c DNA library was established with soybean Tiefeng 8 as material, and pGBKT7-Gm Di19-5 was used as bait to screen soybean c DNA library by yeast two-hybrid technique. A member of soybean NF-Y transcription factor family subunit C was obtained and named as GmNF-YCa. The full length of the gene is 864 BP, encoding 287 amino acids, belonging to the NF-YC subfamily. The molecular weight of GmNF-YCa is 31.6 KD, the isoelectric point is 5.07 hydrophilic protein, it has a transmembrane domain and no signal peptide. Sequence analysis showed that the NF-YC subfamily was highly conserved. The promoter of GmNF-YCa contained stress and photoresponse elements such as ACERE and CAT-Box. Tissue specific analysis showed that GmNF-YCa gene was the highest in seed germination. The real time quantitative results showed that GmNF-YCa was up-regulated by sucrose and mannitol. The GmNF-YCa gene of soybean was introduced into Arabidopsis thaliana by Agrobacterium tumefaciens and its function was analyzed. The germination rate analysis showed that GmNF-YCa transgenic enhanced the tolerance of transgenic Arabidopsis thaliana to osmotic stress during germination, and improved the root growth and lateral root development of transgenic Arabidopsis thaliana under sucrose and mannitol treatments.
【作者单位】：西北农林科技大学农学院/旱区作物逆境生物学国家重点实验室;中国农业科学院作物科学研究所/国家农作物基因资源与基因改良重大科学工程/农业部麦类生物学与遗传育种重点实验室;宁夏农林科学院农作物研究所;
【基金】：国家转基因生物新品种培育重大专项(2014ZX0800916B) 国家自然科学基金项目(31371620)资助~~
【分类号】：S565.1

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