生物技术通报2024，Vol.40Issue(9)：33-41,9.DOI:10.13560/j.cnki.biotech.bull.1985.2024-0536
CRISPR/Cas9介导的高效四倍体马铃薯试管薯基因编辑体系的建立
Establishment of CRISPR/Cas9-mediated Highly Efficient Gene Editing System in Microtubers of Potatoes
摘要
Abstract
[Objective]The development of gene editing technology has made it possible to achieve precise molecular breeding in potato.Microtubers are ideal materials for genetic transformation,but its induction and genetic transformation are of genotype-dependence.Thus,establishing an efficient and widely used gene editing technology may provide technical support for precise molecular breeding for potatoes in future.[Method]Cultivated tetraploid potato variety Qingshu No.9 and Bingshu No.6 were used to explore microtuber induction and genetic transformation.Concurrently,the CRISPR/Cas9 gene editing vector was transformed for genome editing.In addition,the screening system was tested in another three potato varieties.[Result]The efficient tuberizations of five potato varieties were obtained using stem segment with two leaves on solid media including 10%sucrose and 5 mg/L kinetin under dark condition,while induction effect varied.The optimal ratio of hormones for the regeneration of Qingshu No.9 was 0.5 mg/L 6-benzylaminopurine,0.2 mg/L indole-3-acetic acid,0.2 mg/L gibberellin,and 2 mg/L zeatin,under which the regeneration rate,the transformation frequency,genome editing rate was 41.5%,51.9%,and 82.1%,respectively.Using the above screened the formula of hormones,genetic transformation and regeneration was efficiently achieved in another four potato varieties,and the genome editing rate was 63.2%,33.3%,and 10%for Bingshu No.6,Desiree,and Jingshu No.16,respectively.[Conclusion]We established the highly efficient genetic transformation and regeneration system for five potato varieties,and genome editing can be completed for four potato varieties.关键词
马铃薯/试管薯诱导/遗传转化再生/GFP/基因组编辑Key words
potato/microtubers inducing/genetic transformation and regeneration/GFP/genome editing引用本文复制引用
宋倩娜,段永红,冯瑞云..CRISPR/Cas9介导的高效四倍体马铃薯试管薯基因编辑体系的建立[J].生物技术通报,2024,40(9):33-41,9.基金项目
山西省农业科学院作物科学研究所博士基金项目(ZB1102),山西农业大学博士科研启动项目(2021BQ44),山西省博士毕业生来晋工作奖励资金科研项目(SXBYKY2021084),山西省基础研究计划项目(202203021211272) （ZB1102）
