污染严重的海带配子体分离纯化  

Separation and purification of heavily contaminated Saccharina japonica gametophytes

作　　者：武瑞娜 罗世菊 赛珊 赵聚萍 李晓捷 WU Rui-na;LUO Shi-ju;SAI Shan;ZHAO Ju-ping;LI Xiao-jie(National Engineering and Technique Research and Development Center of Algae and Sea Cucumber of China, Key Laboratory of Genetic Improvement and Efficient Culture of Marine Algae of Shandong Province, Shandong Oriental Ocean Sci-tech Co., Ltd., Yantai, 264003, China)

机构地区：[1]山东东方海洋科技股份有限公司、国家海藻与海参工程技术研究中心、山东省海藻遗传育种与栽培技术重点实验室,山东烟台264003

出　　处：《河北渔业》2020年第6期36-37,51,共3页

基　　金：国家重点研发计划“蓝色粮仓科技创新”重点专项(2018YFD0901500);现代农业产业技术体系专项资金(CARS-50);烟台市科技计划项目(2018LJRC067);山东省农业良种工程项目(2016LZGC024);山东省重点实验室专项建设计划(SDKL2017009)。

摘　　要：针对海带(Saccharina japonica)配子体保存过程中严重污染材料的分离纯化进行研究。(1)液相培养分离纯化法:将克隆材料经超声波细胞粉碎仪打碎后重新附着到90 mm培养皿中,利用微毛细吸管在显微镜下分离出状态好且无菌丝附着的细胞段,继续保存。(2)固相培养分离纯化法:将克隆材料4000 r/min离心5~10 min,弃上清,加灭菌PESI培养液重新悬浮藻液,利用5 mL无菌注射器吸取2 mL粉碎后的配子体细胞,逐点注射入固相平板上,接种后利用Parafilm封口膜密封培养皿,平板培养约60~120 d后观察,将未长出菌落且藻斑直径达2 mm的克隆团挑出,然后在PESI液体培养基中复苏继续保存。This study focused on the separation and purification of heavily contaminated Saccharina japonica gametophytes during the preservation.(1)Liquid phase separation and purification.The clone material was treated by ultrasonic,reattached to the 90 mm dish,and then isolated the clear cells segments by micro-capillary pipette to continue preservation.2)Solid phase separation and purification.The contaminated material was centrifuged by 4000 rpm for 5~10 min,and the precipitation was resuspended by PESI media and infected point by point to the solid phase plate by 5 mL sterile syringe.The plates were inoculated for 60~120 days with sealing by parafilm.The sterile clones of gametophyte with 2 mm diameter were picked out and recovered in PESI media.

关 键 词：海带(Saccharina japonica) 配子体 液相培养 固相培养 分离纯化 

分 类 号：TS2[轻工技术与工程—食品科学与工程]