摘要
Dof转录因子参与植物对非生物胁迫应答。本研究对大豆GmDof2.2进行克隆及耐盐性功能鉴定。实时荧光定量PCR结果显示GmDof2.2在大豆幼苗中可以响应非生物胁迫,GmDof2.2基因开放阅读框全长867 bp,编码288个氨基酸的蛋白,其分子量31.4 kDa,等电点10.23,蛋白质序列中含有一个保守的Dof结构域。GmDof2.2启动子区含有3种胁迫响应相关的顺式作用元件(ARE、MBS和WUN-motif)和3种激素响应相关顺式作用元件(ABRE、P-box和TCA-element)。将GmDof2.2构建植物表达载体并转化烟草,共获得4株GmDof2.2异源表达的转基因烟草株系(OE1-OE4),OE1中GmDof2.2的表达量最高,其次为OE2。盐胁迫处理后,转基因烟草萎蔫程度高于野生型烟草,转基因烟草叶片的叶绿素、可溶性糖及脯氨酸含量均显著低于野生型烟草,而丙二醛含量则高于野生型烟草。表明GmDof2.2的异源表达提高了转基因烟草株系对盐胁迫的敏感性。本研究为大豆Dof转录因子抗逆机制研究及应用提供理论依据。
Dof transcription factors are involved in plant response to abiotic stresses. In this research, GmDof2.2 was cloned for salt tolerance identification. Real-time quantitative PCR results showed that GmDof2.2 could respond to abiotic stresse in soybean seedlings. The open reading frame of GmDof2.2 was 867 bp, which encodes a protein of 288 amino acids. The molecular weight of GmDof2.2 protein was 31.4 kDa, and isoelectric point was10.23. The GmDof2.2 protein sequence contains a conserved Dof domain. The promoter region of GmDof2.2 contains three types of stress-related cis-elements(ARE, MBS and WUN-motif)and three types of hormone-related cis-elements(ABRE, P-box and TCA-element). GmDof2.2 was constructed into a plant expression vector and transformed into tobacco. Four GmDof2.2 transgenic tobacco lines(OE1-OE4)were obtained. The expression level of GmDof2.2 in OE1 was the highest, followed by OE2. After salt stress treatment, the wilting degree of transgenic tobacco was higher than that of wild tobacco, the contents of chlorophyll, soluble sugar and proline in transgenic tobacco leaves were significantly lower than that of wild tobacco, while the content of malondialdehyde was higher than that of wild tobacco. These results showed that GmDof2.2 heterologous expression increased the sensitivity to salt stress in transgenic tobacco. This study provided theoretical basis for the research and application of soybean Dof transcription factor resistance mechanism.
作者
翟莹
马婷婷
何佳琦
张军
李铭杨
陈炯辛
于海伟
李珊珊
孙天国
ZHAI Ying;MA Ting-ting;HE Jia-qi;ZHANG Jun;LI Ming-yang;CHEN Jiong-xin;YU Hai-wei;LI Shan-shan;SUN Tian-guo(Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Ar-eas,College of Life Science and Agro-Forestry,Qiqihar University,Qiqihar 161006;Branch of Animal Husbandry and Veterinary of Heilongjiang Academy of Agricultural Sciences,Qiqihar 161005)
出处
《中国油料作物学报》
CAS
CSCD
北大核心
2022年第6期1259-1266,共8页
Chinese Journal of Oil Crop Sciences
基金
黑龙江省普通本科高等学校青年创新人才培养计划(UNPYSCT-2017153)
齐齐哈尔大学研究生创新科研项目(YJSCX2020041)。
关键词
大豆
转录因子
转基因烟草
耐盐性
soybean
transcription factor
transgenic tobacco
salt tolerance
