摘要
为了揭示白菜(Brassica campestris L.ssp.chinensis Makino)开花调控转录因子(MADS AFFECTING FLOWERING 2)MAF2在开花过程中的作用,该研究通过同源克隆的方法获得BcMAF2基因的全长序列。结果表明:(1)BcMAF2基因含有1个长度为588bp开放阅读框,编码196个氨基酸;将BcMAF2蛋白氨基酸序列与其他物种MAF2氨基酸比较表明,BcMAF2基因与其他物种中该基因具有高度保守的结构域。(2)将BcMAF2与YFP和HA标签融合,构建亚细胞定位载体pEarleyGate101-BcMAF2-YFP-HA,采用农杆菌介导法将其瞬时表达于本氏烟草(Nicotiana tabacum)叶片中,激光共聚焦显微镜观察发现BcMAF2蛋白定位于细胞核中,表明BcMAF2符合作为转录因子的功能。(3)将BcMAF2基因遗传转化拟南芥中进行功能验证,通过蛋白印迹试验获得5个过表达株系,且在选取的蛋白表达量较高的第8、10株拟南芥中均表现出明显延迟其抽薹开花表型。研究推测BcMAF2基因可能参与植物开花的春化途径。
The NHCC004 of Brassica campestris was used as the experimental material.A pair of primers were designed according to the BcMAF2 gene sequence of NHCC004.The results showed that：（1）BcMAF2 contained an open reading frame（ORF）of 588 bp which encodes 196 amino acids.The comparison of amino acid sequence of BcMAF2 protein with other species showed that the BcMAF2 gene had a highly conserved domain with other species.（2）The subcellular localization vector pEarleyGate101-BcMAF2-YFP-HA was expressed transiently in the leaves of Nicotiana tabacumvia Agrobacterium tumefaciens-mediated method.The results showed that BcMAF2 protein was located in the nucleus,acting as a transcription factor.（3）The plant expression vector of pEarleyGate101-BcMAF2-YFP-HA and then introduced into Arabidopsis by dipping method and 5 strains of overexpression lines were obtained by Western blot.Late bolting was observed in the line 8 and 10 transgenic Arabidopsis thaliana.This research predicted result shows that BcMAF2 participates in vernalization pathway of flowering in plants.
作者
吴小婷
邵帅旭
任海波
李林
侯喜林
刘同坤
WU Xiaoting;SHAO Shuaixu;REN Haibo;LI Lin;HOU Xilin;LIU Tongkun(State Key Laboratory of Crop Genetics and Germplasm Enhancement,Key Laboratory of Biology and Germplasm Enhancement of IIorticultural Crops in East China,Ministry of Agriculture,Nanjing Agricultural University,Nanjing 210095,China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2018年第9期1571-1577,共7页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家自然科学基金(31301782)
中央高校基本科研业务费自主创新重点项目(Y0201700179)
江苏省自然科学基金(BK20171374)
关键词
白菜
BcMAF2
开花
亚细胞定位
转基因
Brassica campestris L. ssp. chinensis
BcMAF2
flowering
subcellular localization
transgene
