文献类型：期刊文献

中文题名：孝顺竹种胚愈伤组织悬浮培养条件优化

英文题名：Suspension Culture Optimization of Seed Embryo Callus from Bambusa multiplex

作者：袁金玲[1] 张朵[1,2] 顾小平[1] 岳晋军[1] 陈益泰[1]

第一作者：袁金玲

机构：[1]中国林业科学研究院亚热带林业研究所;[2]河南农业大学林学院

年份：2009

卷号：7

期号：4

起止页码：839-844

中文期刊名：分子植物育种

外文期刊名：Molecular Plant Breeding

收录：CSTPCD;;CSCD:【CSCD_E2011_2012】；

基金：浙江省重大科技专项重点项目;中国林科院亚热带林业研究所基本科研业务费(RISF6908)共同资助

语种：中文

中文关键词：孝顺竹;愈伤组织;悬浮培养;转速;2;4-D

外文关键词：Bambusa multiplex, Callus, Suspension culture, Rotation speed, 2,4-dichlorophenoxyacetic acid

分类号：S513;X172

摘要：本研究利用孝顺竹种胚诱导出的淡黄色胚性愈伤组织为材料,对影响细胞悬浮培养的摇床转速、培养液2,4-D浓度、继代天数、接种细胞浓度(W/V)等因素进行了研究。结果表明,悬浮培养时的摇床转速以120r/min时培养效果较好,转速低于100r/min时,愈伤组织容易结团,分散性差;适宜的2,4-D浓度为4～6mg/L,低于4mg/L时愈伤组织易褐变;接种后培养7d是最佳的转接时期,至多不能超过10d;接种细胞密度以4.0%最为合适,可保持较高的增殖速度,同时培养产物较多。在50mL培养液(NB+500mg/L Pro-line+500mg/L Glutamine+300mg/L Hydrolyzed casein+30mg/L Sucrose+4～6mg/L2,4-D)中接种4.0%的愈伤组织,于120r/min的转速下经过7d培养,可增殖一倍以上,以后每7d继代转接一次,培养大约一个月,便可以建立生长迅速、分散性好、均一的悬浮细胞系。
Yellowish embryogenic callus induced from mature seed embryo of Bambusa multiplex was used as material. Rotation speed, concentration of 2,4-dichlorophenoxyacetic acid （2,4-D）, days of subculture, and inoculation cell density were optimized in this research. The results showed that a rotation speed of 120 r/min was good, callus blocks cling at a rotation speed of less than 100 r/rain. The optimal 2,4-D concentration was 4-6 mg/L for the suspension culture system, otherwise browning happened at a concentration of no more than 4 mg/L. Subculture every 7 days was best for suspension culture, at the longest of 10 days. An inoculation density of 4.0% was effective for callus increase. Adding 4.0% callus into 50 mL culture fluid （Wu＇s NB medium supplemented with 500 mg/L Proline, 500 mg/L Glutamine, 300 mg/L Hydrolyzed casein, 30 g/L Sucrose and 4-6 mg/L 2,4-D）, suspension cultured at a rotation speed of 120 r/min, the callus doubled at 7 days, then subculture every 7 days, a good suspension culture system could be established during one month.