植物组织培养中抗污染培养基新配方的探索  

A New Anti-Contamination Medium Formula in Plant Tissue Culture

作　　者：白一苇 张世壮 王雁楠[2] 高晓茹 牛金彪 何绍贞 Bai Yiwei;Zhang Shizhuang;Wang Yannan;Gao Xiaoru;Niu Jinbiao;He Shaozhen(Key Laboratory of Sweet Potato Biology and Biotechnology,Ministry of Agriculture and Rural Affairs,College of Agronomy,China Agricultural University,Beijing 100193;Cereal Institute,Henan Academy of Agricultural Sciences,Zhengzhou 450002)

机构地区：[1]中国农业大学农学院,农业农村部甘薯生物学与生物技术重点实验室,北京100193 [2]河南省农业科学院粮食作物研究所,郑州450002

出　　处：《中国农学通报》2021年第6期89-96,共8页Chinese Agricultural Science Bulletin

基　　金：现代农业产业技术体系北京市创新团队项目(BAIC09-2019)。

摘　　要：组织培养中培养基的污染问题一直是该领域的技术盲区,本实验旨在研发更高效抑制杂菌污染的植物组织培养系统。通过在MS培养基中添加不同浓度及组合的头孢霉素、利福平、青霉素等抗生素以及化学药物进行抗菌筛选,确定了以头孢霉素、百菌清、代森锰锌为主的抗菌培养基配方。之后评估甘薯(Ipomoea batatas)、烟草(Nicotiana tabacum)与拟南芥(Arabidopsis thaliana)三种组培材料在该培养基上的生长与污染情况,进而探索出一种高效培养基抗污染新方法,即药液包衣。利用含有25 mg/L头孢霉素+30 mg/L的代森锰锌与百菌清1:1混合液的培养基,并在外植体插入培养基前利用100 mg/L多菌灵药液或100 mg/L代森锰锌与百菌清1:1混合液进行浸润包衣,可显著降低组培材料被污染的风险,且对除愈伤组织以外的外植体的生长发育影响较弱。该方法的使用在对外植体伤害较小的情况下,对于已污染组培材料的挽救以及大田种质材料的实验室保存有重要借鉴意义。The contamination of culture medium in tissue culture has always been a technical blind area in the field.The purpose of this experiment is to find a more efficient plant tissue culture system to inhibit the pollution of miscellaneous bacteria.By adding different concentrations and combinations of cefomycin,rifampicin,penicillin and other antibiotics and chemical drugs to MS medium for antibacterial screening,the antibacterial medium formula with cefomycin,chlorothalonil and mancozeb was determined.After that,the growth and contamination of Ipomoea batatas,Nicotiana tabacum and Arabidopsis thaliana on the medium were evaluated,and a new anti-contamination method of highly effective medium was obtained,namely liquid coating.Before explants were transplanted onto the medium with 25 mg/L cephalosporin and 30 mg/L 1:1 mixture of mancozeb and chlorothalonil,infiltrated and coated them with 100 mg/L carbendazim solution or the mixture of mancozeb and chlorothalonil at 1:1.By doing so,the risk of contamination risk of tissue culture materials would be significantly reduced with the growth of explants except callus being slightly affected.In general,the use of this method has less damage on the explant,and reference significance for rescuing contaminated tissue culture materials and the laboratory preservation of field germplasm materials.

关 键 词：植物组织培养 高效抗污染配方 药液包衣 代森锰锌 百菌清 头孢霉素 

分 类 号：S481.9[农业科学—农药学]