马铃薯‘华颂66号’遗传转化体系的建立  

Establishment of genetic transformation system for‘Huasong 66’

作　　者：武小娟 吴娟 王沛捷 聂虎帅 刘杰 魏巍 马艳红[1] WU Xiaojuan;WU Juan;WANG Peijie;NIE Hushuai;LIU Jie;WEI Wei;MA Yanhong(Agricultrual College,Inner Mongolia Agricultural University,Hohhot 010019,China;Inner Mongolia Huasong Agricultural Technology Co.,Ltd.,Chifeng 025150,China;Inner Mongolia Tiansong Modern Agriculture Co.,Ltd.,Hohhot 010010,China)

机构地区：[1]内蒙古农业大学农学院,呼和浩特010019 [2]内蒙古华颂农业科技有限公司,内蒙古赤峰025150 [3]内蒙古田颂现代农业有限公司,呼和浩特010010

出　　处：《中国农业大学学报》2024年第1期21-30,共10页Journal of China Agricultural University

基　　金：内蒙古自然科学基金(2022JQ06);内蒙古农业大学基本科研业务费专项(BR22-11-16);呼和浩特市科技计划项目(2022-农-10)。

摘　　要：为探究马铃薯的最佳遗传转化体系,以紫色马铃薯‘华颂66号’无菌组培苗茎段为外植体,用农杆菌介导法将含有植物沉默载体质粒pBWA(V)KS-miR8019转化马铃薯外植体,测定不同预培养时间、不同激素组合及抗生素对马铃薯愈伤再生转化体系的影响。结果表明:1)茎段预培养时间为2 d时,愈伤组织诱导率最高;2)最适的外植体愈伤组织诱导的培养基配方为:MS干粉式培养基+2.0 mg/L 6-BA+1.0 mg/L 2,4-D,愈伤组织的诱导率为96.7%。最适愈伤组织分化的培养基为:MS干粉式培养基+4.0 mg/L 6-BA+0.1 mg/L NAA+1.0 mg/L GA3;3)植株再生及生根的抗生素最适筛选浓度为75 mg/L的卡那霉素;4)农杆菌抑制剂的最适浓度为300 mg/L的特美汀,且在愈伤组织诱导、分化及生根阶段都需要加入。综上,通过对农杆菌介导的愈伤再生转化过程中各个影响因素进行试验,初步建立了马铃薯‘华颂66号’的遗传转化体系。In order to explore the optimal genetic transformation system for potatoes,the genetic transformation system of purple potato‘Huasong 66’,using stem segments of plantlets in vitro as explants.Plant silencing vector plasmid pBWA(V)KS-miR8019 was used to investigate the effects of different preculture times,plant hormones and their ratios,selection concentrations,and antibiotics using Agrobacterium tumefaciens transformation method.The results indicate that:1)The pre-culture time of stem segments was 2 d,and the callus induction ratewas the highest.2)The optimum callus induction medium was MS+2.0 mg/L 6-BA+1.0 mg/L 2,4-D and the ratio of callus induction reached 96.7%.The optimum callus differentiation medium was MS+4.0 mg/L 6-BA+0.1 mg/L NAA+1.0 mg/L GA3.3)The optimum screening pressure for plants regeneration and rooting was 75 mg/L Kan.4)The optimum concentration of agent for Agrobacterium bacteriostatic inhibition was 300 mg/L timentin,and the timentin was needed in callus induction,differentiation and rooting.In conclusion,the genetic transformation system of‘Huasong 66’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.

关 键 词：紫色马铃薯 华颂66号 农杆菌介导 遗传转化体系 

分 类 号：S532[农业科学—作物学]