基于抗生素和除草剂敏感性筛选适宜裂叶地黄遗传转化系统的抗性标记  

Screening of Selectable Markers for Genetic Transformation System of Rehmannia piasezkii Based on Antibiotic and Herbicide Sensitivity

作　　者：叶峥秀 罗湘胤 游磊 张泽志 石宪铭 李琳 郑兰兰 张勇洪 YE Zheng-xiu;LUO Xiang-yin;YOU Lei;ZHANG Ze-zhi;SHI Xian-ming;LI Lin;ZHEN Lan-lan;ZHANG Yong-hong(Laboratory of Medicinal Plants and Evolutionary Genetics,School of Basic Medical Science,Hubei University of Medicine;Hubei Key Laboratory of Wudang Local Chinese Medicine Research;Shiyan Academy of Agricultural Sciences,Shiyan,Hubei 442000,China)

机构地区：[1]湖北医药学院基础医学院药用植物实验室 [2]武当特色中药研究湖北省重点实验室 [3]十堰市农业科学院,湖北十堰442000

出　　处：《湖北医药学院学报》2024年第4期358-363,共6页Journal of Hubei University of Medicine

基　　金：国家自然科学基金项目(32000609,32200680);湖北省教育厅项目(B2021163,Q20222106)。

摘　　要：目的:筛选适宜裂叶地黄遗传转化系统的抗性标记。方法:以裂叶地黄的种子为材料,采用不同浓度抗生素和除草剂对裂叶地黄的种子外植体进行处理,对比不同浓度的卡那霉素、潮霉素和除草剂草铵膦(Basta)的筛选效果,为裂叶地黄遗传转化体系的构建提供基础。结果:裂叶地黄种苗对Basta和潮霉素敏感性的强度强于卡那霉素。Basta浓度为1 mg/L和2 mg/L时,地黄种苗培养30 d存活率为38.03%和5.07%,对裂叶地黄种苗生长有明显的抑制作用;浓度为5 mg/L时,完全抑制裂叶地黄种苗的生长,可作为遗传转化抗性苗筛选的选择压。潮霉素浓度为10 mg/L时,地黄种苗存活率为34.17%,对裂叶地黄种苗生长有明显的抑制作用;浓度为20 mg/L时,将裂叶地黄种苗全部致死,可作为抗性苗筛选的选择压。卡那霉素浓度为50 mg/L时,种苗白化率达到89.17%,种苗生长受到明显抑制。结论:Basta适宜筛选浓度为5 mg/L,潮霉素适宜筛选浓度为20 mg/L,卡那霉素适宜筛选浓度为不低于50 mg/L。Objective To screen selectable markers for genetic transformation system of Rehmannia piasezkii.Methods With the seeds of R.piasezkii as materials,different concentrations of antibiotics and herbicides were used to treat the seed ex⁃plants.The screening effects of kanamycin,hygromycin,and Basta of different concentrations were compared to provide a theoretical basis for the construction of the genetic transformation system of R.piasezkii.Results The seedlings of R.piasez⁃kii were more sensitive to Basta and hygromycin than kanamycin.With 1 mg/L and 2 mg/L Basta,the survival rates of the seedlings after 30 days of treatment were 38.03%and 5.07%,respectively,showing significant inhibitory effects on the growth of seedlings.At 5 mg/L,Basta completely inhibited the growth of the seedlings,making it a suitable selection pres⁃sure for screening resistant seedlings in genetic transformation.At a concentration of 10 mg/L hygromycin,the survival rate of the seedlings was 34.17%,indicating a significant inhibitory effect on the growth of the seedlings.At 20 mg/L,hygromy⁃cin killed all the seedlings,which made it a suitable selection pressure for screening resistant seedlings.With 50 mg/L ka⁃namycin,the seedlings had an albinism rate of 89.17%,and their growth was markedly inhibited.Conclusion The suitable screening concentrations for Basta,hygromycin,and kanamycin are 5 mg/L,20 mg/L,and above 50 mg/L,respectively.

关 键 词：裂叶地黄 遗传转化 抗生素 除草剂 

分 类 号：S567.239[农业科学—中草药栽培]