异源表达大豆转录因子 GmNF-YA19 提高转基因烟草抗 旱性. (Chinese)

Nuclear factor（NF-Y）is a kind of transcription factor widely existing in eukaryotes, and can play an important role in the regulation of abiotic stress response in plants. Exploring the drought resistance and mechanism of soybean GmNF-YA19 will lay a foundation for the application of GmNF-YA19 in drought resistant breeding. Real-time fluorescence quantitative PCR（RT-qPCR）was used to detect the expression of GmNF-YA19 under abiotic stress. GmNF-YA19 was cloned. Its plant expression vector was constructed and transformed into tobacco, and the drought resistance of transgenic tobacco was identified. RT-qPCR results showed that GmNF-YA19 responded to drought, salt, cold and exogenous ABA in soybean, and the expression of GmNF-YA19 increased most significantly under drought stress. GmNF-YA19 was cloned by PCR using soybean leaf cDNA as template. Sequence analysis revealed that GmNF-YA19 encoded a protein containing 213 amino acids, with predicted molecular weight of 22.99 kD and predicted isoelectric point of 9.40. GmNF-YA19 protein sequence contained a conserved CBF domain.Phylogenetic analysis demonstrated that GmNF-YA19 protein was closely related to OsNF-YA7, AtNF-YA4 and AtNF YA7 proteins. Four transgenic tobacco plants were obtained. The heterologous expression of GmNF-YA19 enhanced the drought resistance of transgenic tobacco plant under drought stress. Compared with wild-type tobacco, the expressions of three stress-related genes（NtABA2, NtNCED3-2 and NtP5CS）in GmNF-YA19 transgenic tobacco significantly increased. [ABSTRACT FROM AUTHOR]

核因子（NF-Y）广泛存在于真核生物中, 是一类能够在植物非生物胁迫反应中发挥重要调控作用的转录因子。探 究大豆 GmNF-YA19 的抗旱性及作用机制, 为 GmNF-YA19 在抗旱植物育种中的应用奠定基础。运用实时荧光定量 PCR（RT-qPCR） 对 GmNF-YA19 在非生物胁迫下的表达量进行检测, 克隆 GmNF-YA19, 构建 GmNF-YA19 植物表达载体并转化烟草, 对转基因烟草 的抗旱性进行鉴定。RT-qPCR 结果显示 GmNF-YA19 在大豆中能够响应干旱、高盐、低温及外源 ABA, 且干旱胁迫下 GmNF-YA19 的表达量升高最显著。以大豆叶片 cDNA 为模板, 通过 PCR 克隆 GmNF-YA19。序列分析结果显示, GmNF-YA19 编码一个含有 213 个氨基酸的蛋白质, 预测分子量 22.99 kD, 预测等电点 9.40。GmNF-YA19 蛋白序列包含一个保守的 CBF 结构域。蛋白系统进化分 析表明, GmNF-YA19 蛋白与 OsNF-YA7 蛋白、AtNF-YA4 蛋白和 AtNF-YA7 蛋白亲缘关系较近。共获得 4 棵转基因烟草植株。在 干旱胁迫下, GmNF-YA19 的异源表达增强了转基因烟草的抗旱性。与野生型烟草相比, GmNF-YA19 转基因烟草中 3 个抗逆相关基 因（NtABA2、NtNCED3-2 和 NtP5CS）的表达量显著升高。GmNF-YA19 的异源表达增强了转基因烟草的抗旱性 [ABSTRACT FROM AUTHOR]

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