萱藻(Scytosiphon lomentaria)无菌种质获取技术研究  

Studies on the Method of Obtaining the Axenic Germplasm of Scytosiphon lomentaria

作　　者：沈世军 宫相忠[1] 张红霞 杨儒谦 王吉[1] 张静[1] 高伟[1] 庄英瑞 SHEN Shijun;GONG Xiangzhong;ZHANG Hongxia;YANG Ruqian;WANG Ji;ZHANG Jing;GAO Wei;ZHUANG Yingrui(College of Marine Life Sciences,Ocean University of China,Qingdao 266003,China)

机构地区：[1]中国海洋大学海洋生命学院

出　　处：《海洋湖沼通报》2019年第5期149-155,共7页Transactions of Oceanology and Limnology

基　　金：山东省重点研发计划(2016GSF115042);国家“八六三”高技术研究发展计划(2012AA10A413)资助

摘　　要：本研究将抗生素处理法与固体平板培养法相结合,对获取无菌萱藻种质的技术进行探索研究。萱藻种质首先经过匀浆机处理(50Hz,59s)、无菌海水冲洗过滤,然后在黑暗条件下,以抗生素混合液(头孢噻肟钠800μg/L、红霉素200μg/L、亚胺培南200μg/L、乙酰磺胺1600μg/L和制霉菌素16μg/L)处理7d,之后转移至固体平板培养后光照条件下培养形成藻落。经过固体培养平板、液体培养基及DAPI染色三种方式确认20个样品中有19个达到了无菌状态,成功率达95%。结果表明这种获取无菌萱藻种质的方法是成熟可靠的。In order to develop an efficient and reliable procedure of obtaining axenic germplasm of Scytosiphon lomentaria, antibiotic treatment and agar plate culture were combined in this study. Germplasm of S. lomentaria was chopped into 0.1-0.2 mm segments with tissuelyser(50 Hz, 59 s), and then repeatedly washed and antibiotic treated with a mixture of cefotaxime sodium(800 μg/mL), erythrocin(200 μg/mL), imipenem(200 μg/mL), sulfacetamide(1600 μg/mL) and nystatin(16 μg/mL) in dark for seven days. The antibiotic mixture which contains(50±10) segmented filaments were spread onto Zobell 2216 e agar plate 0.2 mL each, and then cultured under light. According to the results of agar plates, F1 medium enriched with 15 mg/L peptone and 20 mg/L glucose and epifluorescence microscopy with DAPI staining, nineteen of the twenty samples were demonstrated to be axenic. This method is efficient and reliable in the acquisition of axenic S. lomentaria germplasm.

关 键 词：萱藻 丝状体 无菌化 抗生素 固体培养 DAPI染色 

分 类 号：Q331[生物学—遗传学]