摘要
NF-YA是组成NF-Y转录因子的3个亚基之一,在植物中参与生长发育、胁迫响应以及与微生物互作等重要生物过程。通过克隆BnNF-YA1,并对BnNF-YA1进行生物信息学分析和表达模式分析,分离克隆其编码序列,构建过表达载体,转化甘蓝型油菜,通过阳性鉴定获得BnNF-YA1的转基因阳性植株,并对过表达BnNF-YA1的转基因植株进行了逆境表型鉴定。结果表明,BnNF-YA1的开放读码框为858 bp,编码285个氨基酸;烟草瞬时表达分析发现BnNF-YA1蛋白定位于细胞核内;酵母生化分析表明BnNF-YA1全长蛋白没有转录激活活性;表达分析结果显示,BnNF-YA1受PEG处理诱导表达,且在幼叶、花苞和受精后20 d的角果中表达水平较高;过表达BnNF-YA1的甘蓝型油菜对PEG和MV处理的耐受性提高,表明BnNF-YA1可能参与调控甘蓝型油菜对渗透胁迫和氧化胁迫的响应。
NF-YA is one of the three subunits of NF-Y(nuclear factor Y)transcription factors,which is involved in plant growth and development,stress response,and interaction with microorganisms in plants.The bioinformatics and expression pattern analysis of cloned BnNF-YA1 were carried out.The coding sequence of the BnNF-YA1 was isolated and cloned.The overexpression vector of the BnNF-YA1 was constructed and transformed into rapeseed.The positive transgenic plants were identified,and the performances of the transgenic plants overexpressing BnNF-YA1 under stress treatments were evaluated.The results showed that the length of open reading frame of BnNF-YA1 was 858 bp,encoding 285 amino acids.Tobacco-transient expression analysis uncovered that BnNF-YA1 protein was localized in the nucleus.Biochemical analysis using yeast system showed that there was no transactivation activity in BnNF-YA1 full-length protein.Expression analysis revealed that the transcript abundance of BnNF-YA1 was induced by PEG treatment,and BnNF-YA1 showed a relatively high expression in the seedling leaf,bud,and 20 DAP silique.The overexpression of BnNF-YA1 increased the tolerance of transgenic rapeseed plants to PEG and methyl viologen(MV)treatments,suggesting that BnNF-YA1 may be involved in regulating the responses of rapeseed to osmotic and oxidative stress.
作者
林科运
段钰晶
王高升
孙念礼
方玉洁
王幼平
LIN Ke-yun;DUAN Yu-jing;WANG Gao-sheng;SUN Nian-li;FANG Yu-jie;WANG You-ping(College of Bioscience and Biotechnology,Yangzhou University,Yangzhou 225009)
出处
《生物技术通报》
CAS
CSCD
北大核心
2022年第4期106-116,共11页
Biotechnology Bulletin
基金
国家自然科学基金项目(31872874),扬州大学“拔尖人才成长计划”。