| 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。 |
| 关键词: 紫色马铃薯 华颂66号 农杆菌介导 遗传转化体系 |
| DOI：10.11841/j.issn.1007-4333.2024.01.02 |
| 投稿时间：2023-07-03 |
| 基金项目: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） |
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| Establishment of genetic transformation system for ‘Huasong 66’ |
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WU Xiaojuan1, WU Juan1, WANG Peijie1, NIE Hushuai1, LIU Jie2, WEI Wei3, MA Yanhong1*
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| (1.Agricultrual College, Inner Mongolia Agricultural University, Hohhot 010019, China;2.Inner Mongolia Huasong Agricultural Technology Co., Ltd., Chifeng 025150,China;3.Inner Mongolia Tiansong Modern Agriculture Co., Ltd., Hohhot 010010,China) |
| Abstract: |
| 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 pre-culture 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. |
| Key words: purple potato Huasong 66 Agrobacterium-mediated transformation genetic transformation system |
