摘要
【目的】明确农杆菌介导的黄梁木遗传转化体系对筛选物的敏感浓度,为黄梁木遗传转化研究奠定基础.【方法】以黄梁木子叶、胚轴和无菌苗为材料,探讨筛选物卡那霉素、潮霉素、头孢霉素对试验材料生长的影响,以及对农杆菌LBA4404的抑菌效果.[结果]在不定芽分化、生根阶段,卡那霉素的临界浓度分别为20、10 mg/L,潮霉素的临界浓度均为10 mg/L;当头孢霉素的浓度为100 mg/L时,完全抑制农杆菌生长,而且该浓度对黄梁木不定芽分化、生根影响较小.【结论】在黄梁木子叶和胚轴为试验材料的遗传转化体系中,20和10 mg/L的卡那霉素可作为不定芽分化、生根阶段的筛选选择压,10 mg/L的潮霉素可作为两个阶段的筛选选择压;而100~200 mg/L的头孢霉素可作为农杆菌LBA4404的抑菌浓度.
【Objective】To determine the sensitive concentration of agrobacterium-mediated genetic transformation system to screening substances for Neolamarckia cadamba,and to lay a foundation for the research on genetic transformation of N.cadamba.【Method】The effectsof screening substances(kanamycin,hygromycin and cefotaxime)on the differentiation,growth of cotyledon,hypocotyl and aseptic seedling,and the bacteriostatic effect of cefotaxime on agrobacterium LBA4404 were investigated.【Result】At the stage of adventitious bud differentiation and rooting,the critical concentration of kanamycin was 20 and 10 mg/L,and the critical concentration of hygromycin was 10 mg/L,respectively.The reproduction of agrobacterium was completely inhibited when the concentration of cefotaxime was 100 mg/L,and the concentration had little effect on the adventitious buds differentiation and rooting of N.cadamba.【Conclusion】In the genetic transformation system with cotyledon and hypocotyl as test materials,20 and 10 mg/L kanamycin could be used as the selection pressure for the stage of advection buds differentiation and rooting,and 10 mg/L hygromycin could be used as the selection pressure for the both of two stages.The 100-200 mg/Lcefotaxime can be used asbacteriostatic concentration of agrobacteriumLBA4404.
作者
黄浩
韦莹
翟勇进
白隆华
陈晓阳
HUANG Hao;WEI Ying;ZHAI Yongjin;BAI Longhua;CHEN Xiaoyang(Guangxi Botanical Garden ofMedicinal Plants,Nanning 530023,China;College of Forestry and Landscape Architecture,South China Agricultural University,Guangzhou 510642,China)
出处
《四川农业大学学报》
CSCD
北大核心
2020年第1期58-64,共7页
Journal of Sichuan Agricultural University
基金
广西自然科学基金（2018GXNSFAA138094）
广东省科技计划项目（2017B020201008）
国家林业局林业公益性行业科研专项（201004020）
关键词
黄梁木
遗传转化
筛选物
农杆菌
抗生素
Neolamarckia cadamba
genetic transformation
screening substances
agrobacterium
antibiotics
