摘要
为了研究精氨酰-tRNA合成酶(arginyl-tRNA synthetase,RS)在植物响应镉胁迫中的作用,本研究扩增本氏烟草NbRS基因,进行生物信息学分析,并通过实时荧光定量PCR和BSP技术对镉胁迫下基因的表达和甲基化状态进行分析;利用病毒诱导的基因沉默(VIGS)技术下调该基因表达,分析该基因下调对植物发育、对镉的耐受能力的影响。结果表明,NbRS基因大小为1947 bp,编码649个氨基酸,与茄科植物精氨酰-tRNA合成酶亲缘关系最近;不同浓度Cd^(2+)处理会影响该基因表达,且Cd^(2+)处理下基因甲基化状态发生变化;NbRS基因下调导致植物发育异常,且植物对Cd^(2+)胁迫的敏感性增加。研究表明,NbRS基因在植物耐受Cd^(2+)胁迫中起重要作用,可以作为潜在的培育耐受或低积累Cd^(2+)工程植物的靶基因做进一步研究。
To explore the role of arginyl-tRNA synthetase(RS)in response to cadmium stress,the full length of Nicotiana benthamiana RS gene was amplified and bioinformatics analysis was performed.Meanwhile,the expression and methylation status of RS gene under cadmium stress were analyzed by real-time PCR and BSP.The expression of RS gene was down-regulated by virus-induced gene silencing(VIGS),and the effects of this down-regulated gene on plant development and cadmium tolerance were also analyzed.The results showed that the length of NbRS gene was 1947 bp and encoded 649 amino acids,which had the closest relationship with arginyl-tRNA synthetase of Solanaceae family.Expression of NbRS was affected by different concentrations of Cd^(2+),and Cd^(2+) treatment led to alteration of DNA methylation status.And,the down-regulation of NbRS resulted in abnormal development and the sensitivity to Cd^(2+) stress.The above results indicated that NbRS played an important role in response to Cd^(2+) stress,and could be used as a potential target gene for developing genetic modified plants that were tolerant or low-accumulating Cd^(2+).
作者
娄冰冰
赵一博
辛翠花
卞晓燕
郭江波
LOU Bingbing;ZHAO Yibo;XIN Cuihua;BIAN Xiaoyan;GUO Jiangbo(School of Life Science and Technology,Inner Mongolia University of Science and Technology,Baotou Inner Mongolia 014010,China;Snow Valley Agricultural Development Co.,LTD,Zhangjiakou Hebei 076481,China)
出处
《种子》
北大核心
2022年第3期6-11,共6页
Seed
基金
国家自然科学基金项目(32060498,31660064)
内蒙古自然科学基金项目(2018 MS 03063)。
