摘要
芜菁(Brassica rapa var.rapa L.)存在再生困难、转化效率低等问题。以新疆柯坪白皮芜菁为试材,对外植体类型、菌液侵染浓度、侵染与共培养时间及乙酰丁香酮(AS)浓度等进行筛选,建立发根农杆菌介导的芜菁毛状根转化体系。以6 d苗龄的芜菁无根苗为外植体,采用添加200μmol·L^(-1)AS,菌液浓度OD600值为0.4时侵染12min,共培养36h后再转入含有400mg·L^(-1)羧苄青霉素(Carb)的MS抑菌培养基的转化效率最高,并将可视化标记RUBY应用于转基因阳性毛状根高效筛选。基于该体系结合生长发育调节因子(developmental regulators,DRs)成功获得稳定的阳性转基因芜菁植株,阳性转化效率可达10.61%,从种子萌发、芽再生到生根整个过程可以在3个月内完成。
Turnip(Brassica rapa var.rapa L.)has problems such as poor regeneration and low transformation efficiency.The influenced factors including explants type,bacterial infection concentration,infection time,co-cultivation time and acetosyringone(AS)were firstly screened to establish the genetic transformed hairy roots system by Agrobacterium rhizogenes with a white skin turnip variety from Xinjiang Keping.The results showed that 6 d rootless seedlings were selected as explant and the suspension added 200μmol·L^(-1) AS with the bacterial concentration OD 600 of 0.4 was infected for 12 min.After 36 h co-cultivation,hairy roots were exhibited the highest transformation efficiency in MS bacteriostatic medium with 400 mg·L^(-1) carbenicillin,as well as the visual marker RUBY was applied to efficiently screening for transgenic positive hairy roots.Furthermore,based on this system combined with developmental regulators,the stable positive transgenic plants were successfully obtained with a transformation efficiency of 10.61%.The whole process from seed germination,shoot regeneration to rooting could be completed within 3 months.
作者
马曦
张金睿
庄红梅
陈巍
张蜜
袁晓琪
易小芳
王聪聪
王海云
王燕
MA Xi;ZHANG Jinrui;ZHUANG Hongmei;CHEN Wei;ZHANG Mi;YUAN Xiaoqi;YI Xiaofang;WANG Congcong;WANG Haiyun;WANG Yan(State Key Laboratory of Crop Genetics and Germplasm Enhancement and Utilization,Key Laboratory of Horticultural Crop Biology and Genetic Improvement(East China),College of Horticulture,Nanjing Agricultural University,Nanjing 211800,China;Xinjiang Vegetable Engineering Technology Research Center,Institute of Fruit and Vegetable Research,Xinjiang Uyghur Autonomous Region Academy of Agricultural Sciences(Luntai Fruit Tree Resource Garden of the National Fruit Tree Germplasm Resources),Urumqi 830091,China;Jiangsu Engineering and Technology Center for Modern Horticulture,Jiangsu Polytechnic College of Agriculture and Forestry,Zhenjiang,Jiangsu 212400,China)
出处
《园艺学报》
北大核心
2025年第5期1389-1398,共10页
Acta Horticulturae Sinica
基金
国家自然科学基金项目(31960600)
新疆维吾尔自治区重点研发计划项目(2022B02058)
新疆维吾尔自治区自然科学基金面上项目(2022D01A268)
新疆农业科学院青年科技骨干创新能力培养项目(xjnkq-2019003)。
关键词
芜菁
发根农杆菌
毛状根
RUBY
发育调节因子
turnip
Agrobacterium rhizogenes
hairy root
RUBY
developmental regulator
作者简介
共同第一作者:马曦;共同第一作者:张金睿;通信作者:王燕,E-mail:wangyanhs@njau.edu.cn。
