番茄愈伤组织的诱导与遗传转化体系的建立  

Tomato callus induction and establishment of genetic transformation system

作　　者：徐青青 张懿丹 李静源 姜立波 王娜[1] XU Qingqing;ZHANG Yidan;LI Jingyuan;JIANG Libo;WANG Na(School of Life Sciences and Medicine,Shandong University of Technology,Zibo 255049,China)

机构地区：[1]山东理工大学生命与医药学院,山东淄博255049

出　　处：《山东理工大学学报（自然科学版）》2025年第1期73-78,共6页Journal of Shandong University of Technology:Natural Science Edition

基　　金：国家自然科学基金项目(32202518)。

摘　　要：番茄是植物研究领域常用的模式植物之一,番茄高效稳定的遗传转化体系是研究番茄基因生物学功能的基础,但常规番茄遗传转化耗时过长。以番茄愈伤组织为侵染对象,最终获得转基因番茄愈伤组织,可极大地缩短获得番茄转基因材料的时间。本文以番茄Micro-Tom(Solanum lycopersicum cv. Micro-Tom)为实验材料,探索番茄愈伤诱导条件,建立标准的番茄愈伤组织遗传转化体系。结果表明,胚根为外植体、激素配比2,4-二氯苯氧乙酸(2,4-Dichlorophenoxyacetic acid, 2,4-D)1 mg/L+6-苄氨基腺嘌呤(6-Benzylaminopurine, 6-BA)1 mg/L、蔗糖浓度30 g/L、光照强度3 000 lx为最佳诱导条件。标准番茄愈伤遗传转化体系包括愈伤组织培养、悬浮细胞建立、农杆菌侵染、共培养、抗性愈伤筛选、核酸鉴定等。该实验结果能够为以番茄为研究对象的分子机理研究及其所涉及的免疫沉淀、高通量测序、亚细胞定位等实验提供科研实验材料和方法参考。Tomato is one of the commonly used model plants in the field of plant research.The efficient and stable genetic transformation system of tomato serves as the foundation for studying the functional biology of tomato genes.However,conventional genetic transformation of tomato is time-consuming.Using tomato callus tissue as the infestation target and finally obtaining the transgenic tomato callus tissue can greatly shorten the time required to obtain transgenic tomato materials.In this study,the tomato Micro-Tom(Solanum lycopersicum cv.Micro-Tom)was used as the experimental material to explore the induction conditions for tomato callus and to establish a standardized genetic transformation system for tomato callus tissue.The results show that the primary root as the explant,a hormone ratio of 2,4-D 1 mg/L+6-BA 1 mg/L,a sucrose concentration of 30 g/L,and a light intensity of 3000 lx are the optimal induction conditions.The standard genetic transformation system for tomato callus includes callus culture,establishment of suspension cells,agrobacterium infection,co-cultivation,selection of resistant callus,and nucleic acid identification.The above experimental results can serve as method references and provide scientific research materials for molecular mechanistic studies on tomatoes,as well as related experiments involving immunoprecipitation,high-throughput sequencing,and subcellular localization.

关 键 词：番茄 愈伤组织 遗传转化 农杆菌介导法 亚细胞定位 

分 类 号：TB532.1[理学—物理] TB553[理学—声学]