银杏干细胞培养体系的建立及细胞特性初步考察  

A Preliminary Study on Establishment of Ginkgo Stem Cell Culture System and its Cell Characteristics

作　　者：张文锦 卢华卫 姜海晶 ZHANG Wenjin;LU Huawei;JIANG Haijing(Guizhou University of Traditional Chinese Medicine,Guiyang 550025,Guizhou,China)

机构地区：[1]贵州中医药大学,贵州贵阳550025

出　　处：《贵州中医药大学学报》2025年第1期35-41,共7页Journal of Guizhou University of Traditional Chinese Medicine

基　　金：2019年贵州中医药大学博士启动基金(贵中医博士启动[2019]114号)。

摘　　要：目的通过植物干细胞诱导技术获得银杏离体干细胞,由于干细胞保留了原植物最完整的遗传信息,结合生物反应器大规模发酵细胞,可有目的的生产银杏萜内酯类化合物。方法以银杏干细胞和愈伤组织悬浮细胞作为材料,考察细胞特性以及对博来霉素的敏感性。结果银杏干细胞多为圆形,单个细胞状态,具有小而多的液泡。将银杏干细胞液体培养在100 mL的摇瓶中,第15 d的生长率达到最大值为478%。HPLC-ELSD法检测银杏干细胞培养体系样品,发现其中含银杏内酯A、B、C和白果内酯。银杏干细胞和愈伤组织的死亡率随博来霉素的浓度和孵育时间的增加而增加,但银杏干细胞的死亡率显著高于愈伤组织。结论本实验所建立的银杏干细胞培养体系,具有遗传信息稳定、生长迅速、细胞性状稳定等特点,可以表达特征性化合物银杏萜内酯,能够稳定继代培养,为后续研究银杏萜内酯生物合成途径提供了基础。Objective Ginkgo biloba vitro stem cells were obtained by plant stem cell induction technology,which retained the most complete genetic information of original plant.Combined with bioreactors and largescale fermentation of cells,purposeful production of Ginkgo biloba terpenoids can be achieved.Methods Ginkgo biloba stem cells and callus suspension cells were used as materials to investigate the cell characteristics and sensitivity to bleomycin.Resultss Ginkgo biloba stem cells are mostly in a circular,single cell state,with small and numerous vacuoles.Ginkgo biloba stem cells liquid culture reached its maximum growth rate of 478% on the 15th day in a 100mL shake flask.HPLC-ELSD detection of Ginkgo biloba stem cell culture system samples revealed the presence of Ginkgolide A,B,C,and Ginkgo biloba lactone.The mortality rate of Ginkgo biloba stem cells and callus tissue increased with the concentration of bleomycin and incubation time,but the mortality rate of Ginkgo biloba stem cells was significantly higher than that of callus tissue.Conclusion Ginkgo biloba stem cells culture system established in this experiment has the characteristics of stable genetic information,rapid growth and stable cell traits,and can express the characteristic compound of ginkgo terpenoid lactone,which can stabilize the subculture,and provides a basis for the subsequent study of ginkgo terpenoid lactone biosynthesis pathway.

关 键 词：银杏 植物组织培养 植物干细胞 萜内酯 HPLC-ELSD 

分 类 号：S792.95[农业科学—林木遗传育种] R284.2[农业科学—林学]