超声辅助提高长春花毛状根诱导效率及转基因毛状根构建
摘要： 在不同菌种、不同诱导部位、不同培养基以及不同的诱导方法对长春花进行毛状根诱导基础上，尝试改变侵染用试剂以及超声辅助的新方法，通过比较毛状根诱导效率以及毛状根的形态差异，筛选最优的长春花毛状根诱导方法与条件，并建立转基因毛状根体系。最终以C58C1为侵染用菌株，以无菌苗的叶片为外植体材料，1/2 MS培养基为诱导培养基，预培养时间为2天，使用共培养缓冲液侵染，5 s超声辅助为诱导最优的诱导方法，成功获得乳白色、较为浓密的、无向地性并且多分枝的毛状根，并应用此条件成功构建转GUS基因的毛状根，为以后代谢调控途径相关基因的功能研究以及长春花中次生代谢产物合成的调控奠定基础。
关键词: 长春花, 毛状根诱导, 发根农杆菌, 超声辅助, GUS基因
Abstract: Based on the induction of hairy roots of Catharanthus roseus by different strains, different inducing sites, different media and different induction methods, we tried to change the infective reagents and the new ultrasound-assisted method. By comparing the hairy root induction efficiency and hair Morphological differences in root shape, we screened the optimal method and conditions for the induction of hairy roots of C.roseus, and established transgenic hairy roots. Finally, we used C58C1 as the inoculative strain. The leaves of the aseptic seedlings were used as explant materials, 1/2 MS medium was used as the induction medium for 2-day preculture time. The co-culture buffer was used to infect for 5 s. Ultrasound-assisted induction of optimal induction method succeeded in obtaining milky white, dense, non-directional and multi-branched hairy roots, and successfully constructing the hairy roots of GUS gene, which are related to metabolic regulation pathways later. This study laid the foundation on gene function and the synthesis of secondary metabolites of the C.roseus.
Key words: Catharanthus roseus, hairy root-inducing, Agrobacterium rhizogenes, ultrasound assisted, GUS gene
中图分类号:
Q949.776.5
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