摘要
以马铃薯品种‘并薯6号’无菌试管苗茎段作为外植体,通过对遗传转化过程中预培养时间、植物激素的种类及配比、筛选剂浓度和抗生素的种类及浓度进行研究,初步建立了马铃薯品种‘并薯6号’的愈伤再生转化体系。结果表明,预培养时间为2 d时,愈伤组织诱导率、出苗率及转化频率最高。最适外植体愈伤组织诱导的培养基配方为MS+2.0 mg/L 6-苄氨基嘌呤(6-BA)+0.5 mg/L 2,4-二氯苯酚代乙酚(2,4-D)+0.4 mg/L激动素(KT),愈伤组织的诱导率为86.7%,愈伤组织形态较紧密,为深绿色,茎段两端膨大呈哑铃状,表面的白色须状物和毛状根较少。最适愈伤组织分化的培养基配方为MS+1 mg/L 6-苄氨基嘌呤(6-BA)+1 mg/L赤霉素(GA_(3))+1 mg/L玉米素(ZT),幼苗比较粗壮,长势良好。最适的植株再生及生根筛选剂为10 mg/L的潮霉素(Hygromycin B,Hyg),转化频率较高,为20.3%。农杆菌抑制剂的最适浓度为200 mg/L的特美汀(Timentin,TMT),且在愈伤组织诱导、分化及生根阶段都需要加入TMT。综上所述,通过对农杆菌介导的愈伤再生转化过程中各个因素的研究,初步建立了‘并薯6号’的遗传转化体系,为马铃薯分子育种发展提供一定的基础。
The genetic transformation system of Solanum tuberosum cv.'Bingshu 6',using stem segments of plantlets in vitro as explants,was preliminarily established through studying the pre-culture time,plant hormones and their ratios,selection concentrations,and antibiotics and their concentrations.The ratio of callus induction,differentiation and the transformation frequency was the highest when pre-culture time was two days.The most suitable callus induction medium was MS+2.0 mg/L 6-benzylaminopurine(6-BA)+0.5 mg/L 2,4-dichlorophnoxyacetic acid(2,4-D)+0.4 mg/L kinetin(KT),on which the ratio of callus induction was 86.7%,and the shape of callus was dumbbell-shaped and color was dark green,meanwhile,white hairs and hairy roots were less on the surface.The most suitable callus differentiation medium was MS+1 mg/L 6-benzylaminopurine(6-BA)+1 mg/L gibberellin(GA_(3))+1 mg/L zeatin(ZT),on which the shoots were strong and growing well.The optimum screening pressure for plants regeneration and rooting was 10 mg/L hygromycin B,and the transformation frequency was 20.3%.The optimum concentration of agent for Agrobacterium bacteriostatic inhibition was 200 mg/L timentin,and the timentin was needed in callus induction,differentiation and rooting.In conclusion,the genetic transformation system of'Bingshu 6'was preliminarily established through studying the factors at each stage of Agrobacterium-mediated genetic transformation,which would provide the certain basis for the development of potato molecular breeding.
作者
宋倩娜
梅超
霍利光
王慧杰
冯瑞云
SONG Qianna;MEI Chao;HUO Liguang;WANG Huijie;FENG Ruiyun(College of Agriculture,Shanxi Agricultural University/Key Laboratory of Crop Genetics and Molecular Improvement of Shanxi Province,Taiyuan,Shanxi 030031,China)
出处
《中国马铃薯》
2021年第5期385-396,共12页
Chinese Potato Journal
基金
山西省农业科学院作物科学研究所博士基金(ZB1102,ZB1901)
山西省农业科学院应用基础研究计划(YGC2019FZ1)
山西农业大学博士基金(2021BQ44)
山西省博士毕业生来晋工作奖励科研项目(SXBYKY2021084)
山西省重点研发计划项目(201903D221076)
关键词
马铃薯
农杆菌介导的转化
植物激素
遗传转化体系
Solanum tuberosum
Agrobacterium-mediated transformation
plant hormone
genetic transformation system
