小蔓长春花组织培养快速繁殖中的遗传稳定性

Analysis of Genetic Stability in Rapid Propagation of Vinca minor L.

以小蔓长春花茎段为外植体,在含1.5mg·L-1NAA的MS固体培养基上直接生根,多次继代培养、移栽,建立小蔓长春花组织培养快速繁殖体系。利用RAPD技术结合高效液相色谱技术对连续三次继代无菌苗和炼苗后小蔓长春花的遗传稳定性、长春胺含量稳定性进行分析。结果显示,实验选取的20条随机引物共扩增出清晰的46个条带,不同植株扩增获得的条带数目和带型一致,在所检测范围内继代培养和炼苗均未影响小蔓长春花DNA序列。同时HPLC测得相应材料中长春胺含量均在0.12%以上,培养过程中长春胺含量稳定。本实验建立的小蔓长春花组织培养快速繁殖体系遗传稳定、长春胺含量稳定,可用于大量繁殖小蔓长春花无菌植株,为长春胺生产的提供资源。

The stem-segmens of Vinca minor were cultured on MS medium supplemented with 1.5 mg.L-1 NAA. A few of roots were induced and then rapid propagation system was established. Random amplified poly-morphic DNA （RAPD） markers were used to assess the genetic stability of regenerated plantlets and HPLC was used to assess the stability ofvincamine contents. The results demonstrated that the 20 screened primers used in this analysis yielded 46 scoreable bands. These 46 bands were monomorphic with identical bands at DNA level among all leaves from the regenerated plantlets, transplants and control plants of Vinca minor. The average vincamine contents of different plants of Vinca minor are all above 0.12% and have no significant difference, This rapid propagation system of Vinca minor might be used for micropropogation ofvincamine.