生物技术通报2025，Vol.41Issue(4)：88-97,10.DOI:10.13560/j.cnki.biotech.bull.1985.2024-1178
马铃薯川芋50遗传转化及基因编辑体系的建立
Establishment of Genetic Transformation and Gene Editing System for a Potato Cultivar Chuanyu 50
摘要
Abstract
[Objective]To construct a genetic transformation system and gene editing system of potato Chuanyu 50.[Method]Aseptic histocultured seedling stems and leaves of Chuanyu 50,the main potato(Solanum tuberosum L.)variety promoted in Sichuan province,were used as explants.The potato explants were transformed with Agrobacterium-mediated transformation containing the plant CRISPR/Cas9 vector pJCV55-StU6-200-StUBI10-T#01.Four different media with various phytohormone ratios were analyzed and screened for callus induction and differentiation.CRISRR-Cas9-based editing method was applied for creating mutants.[Result]1)The optimal medium among the tested ones for genetic transformation Chuanyu 50 was B system.The medium formulation for stem and leaf pre-cultivation and callus induction,respectively,were as follows:MS basic salt+20 g/L sucrose+1 mL/L 1 000×N&N vitamins+1.0 mg/L TZR(trans-zeatin-riboside,TZR)+0.027 8 mg/L GA3(gibberellin A3,GA3)+0.02 mg/L NAA(1-naphthaleneacetic acid,NAA)+2.0 g/L phytagel,and MS basic salt+20 g/L sucrose+1 mL/L 1 000×N&N vitamins+0.5 mg/L TZR+2.5 mg/L IAA(indole acetic acid,IAA)+2.0 g/L phytagel,the callus induction rates using stem and leaf were 93%and 88%,respectively.The medium for differentiation was MS basic salt+20 g/L sucrose+1 mL/L 1 000×N&N vitamins+2.0 mg/L TZR+10 mg/L GA3+2.0 g/L phytagel.2)It is demonstrated that the two-round rooting screening method accurately identified transgenic-positive plants with a 100%accuracy.3)An Agrobacterium-mediated CRISPR/Cas9 gene editing system for Chuanyu 50 with 63%editing rate was established.[Conclusion]The genetic transformation and gene editing system of potato Chuanyu 50 are initially established and multiple factors affecting the process of callus induction and regeneration are evaluated.关键词
马铃薯/川芋50/根癌农杆菌/遗传转化/基因编辑/CRISPR/Cas9Key words
potato/Chuanyu 50/Agrobacterium tumefaciens/genetic transformation/gene editing/CRISPR/Cas9引用本文复制引用
文博霖,何卫,曾子贤,万敏,胡建军,王克秀,景晟林,王心悦,朱博,唐铭霞,李兵..马铃薯川芋50遗传转化及基因编辑体系的建立[J].生物技术通报,2025,41(4):88-97,10.基金项目
国家自然科学基金面上项目(32170573),"十四五"四川省科技厅薯类育种攻关项目(2021YFYZ0019) （32170573）
