摘要
根据长春花叶绿体全序列中的CratpA基因编码序列,设计引物并扩增获得特异沉默片段sCratpA,构建基因沉默(VIGS)载体pTRV2-sCratpA,农杆菌瞬时转化长春花幼苗顶端分生组织。结果表明处理后的长春花小苗与空白对照(EV)相比叶片失绿叶色变浅。q-RT-PCR结果表明,CrD4H、Cr7DLS、CrLAMT、CrDL7H相对表达量分别较对照(EV)升高13.85、4.42、18.91和4.31倍。HPLC结果表明,文多灵和长春质碱两种生物碱分别升高了1.05和5.83倍。由此说明,CratpA的基因沉默对长春质碱与文多灵的合成和积累有较大的影响,且呈负相关,即CratpA表达受阻时,长春质碱与文多灵积累量升高。
According to the coding sequence of CratpA gene in the whole chloroplast sequence of Catharanthus roseus,primers were designed and specific silencing fragment sCratpA was amplified to construct gene silencing vector pTRV2-sCratpA.Agrobacterium tumefaciens transformed the apical meristem of Catharanthus roseus seedling instantaneously.The results showed that the color of the leaves of the treated seedlings was lighter than that of the control(EV).The results of q-RT-PCR showed that the relative expression of CrD4H,Cr7DLS,CrLAMT and CrDL7H were 13.85,4.42,18.91 and 4.31 times higher than that of EV,respectively.HPLC results showed that the alkaloids of vindoline and catharanthine increased by 1.05 and 5.83 times,respectively.It’s indicated the gene silencing of CratpA has a significant effect on the synthesis and accumulation of catharanthine and Ventolin,which has a negative correlation,that is,when the expression of CratpA is blocked,the accumulation of catharanthine and vindoline increased.
作者
李娅
周海龙
李琳
陈秋骏
吴琼
刘志文
LI Ya;ZHOU Hailong;LI Lin;CHEN Qiujun;WU Qiong;LIU Zhiwen(School of Biological Engineering, Dalian Polytechnic University, Dalian 116034, China;China Biotech Fermentation Industry Association, Beijing 100833, China)
出处
《大连工业大学学报》
CAS
北大核心
2020年第3期163-168,共6页
Journal of Dalian Polytechnic University
基金
国家自然科学基金项目(31570303)。
