银叶真藓外植体组织培养消毒方法的筛选

Screening of Sterilization Methods for Tissue Culture Using Bryum argenteum as Explants

为给银叶真藓全基因组序列测定提供无污染材料,本研究对银叶真藓外植体组织培养的消毒方法进行筛选。分别使用酒精、次氯酸钠溶液和青霉素溶液作为消毒剂,在不同浓度和不同时间下处理银叶真藓外植体,然后分别将处理后的材料接种在改良的Knop培养基上进行培养。一定时间后,通过形态观察银叶真藓的生长状况。实验结果表明,酒精和次氯酸钠溶液对银叶真藓的消毒效果欠佳,而青霉素溶液对银叶真藓的消毒效果较好,由此筛选出银叶真藓外植体组织培养的最佳消毒方法为:用浓度为300 mg/L青霉素溶液浸泡1.5 h。利用本研究筛选出的消毒方法可以获得形态观察上无杂菌污染的银叶真藓材料,为后续全基因组序列测定前银叶真藓DNA的获取提供物质基础。

The disinfection methods for tissue culture using Bryum argenteum as explants were screened in this study to provide non-polluting materials for the whole genome sequencing of B. argenteum. Using ethanol, sodium hypochlorite solution and penicillin solution as disinfection reagents, B. argenteum explants were treated under different concentrations and time, and then the treated explants were inoculated on the modified Knop medium for culture. After a certain time, the growth status of B. argenteum was observed through morphology. The results showed that the disinfection effect of penicillin solution on B. argenteum was better than that of ethanol and sodium hypochlorite solution. Thus, the best disinfection method for B. argenteum explant tissue culture was that samples were soaked in 300 mg/L penicillin solution for 1.5 h. B. argenteum material without microbial contamination on morphological observation could be obtained by using the disinfection method screened in this experiment, which could provide a material basis for further acquisition of B. argentium DNA before whole genome sequencing.