摘要
在生产过程中,杂草会极大影响蓖麻的品质和产量。随着生物技术与基因编辑系统的出现,为实现蓖麻抗除草剂的定向育种带来了可能。为了提高蓖麻抗草铵膦除草剂的能力,以‘通蓖5号’大穗型蓖麻为实验材料,利用根癌农杆菌介导法将bar基因遗传转化蓖麻子叶节,研究了农杆菌菌液侵染浓度、共培养时间和Basta筛选浓度对遗传转化效率的影响。结果表明:当农杆菌菌液浓度OD_(600)=0.7时,侵染15 min,共培养3 d,用1 mg/L的Basta进行抗性筛选时,遗传转化效率最高。生根后移栽的转化子经PCR检测和200 mg/L的Basta除草剂叶片涂抹检测,初步证明抗草铵膦基因bar已成功整合到蓖麻基因组中,得到具有草铵膦抗性的转基因蓖麻植株。
In the production process,weeds can greatly affect the quality and yield of castor.With the advent of biotechnology and gene editing system,it is possible to achieve targeted breeding of castor for herbicide resistance.In order to improve the resistance of castor to glufosinate-ammonium herbicide,the bar gene was introduced into the cotyledon node of castor Tong bi No.5 by Agrobacterium-tumefaciens mediated genetic transformation method.We investigated the effects of bacterial infection concentration,co-culture time and basta screening concentration on the efficiency of genetic transformation.The results showed that:castor cotyledon nodes were infected with bacterium concentration(OD_(600)=0.7),infection for 15 min,co-culture for 3 days,and 1 mg/L basta for resistance screening to improve transformation efficiency.PCR detection and application of basta herbicide(200 mg/L)proved that the target gene bar was successfully transferred into castor genome and transgenic castor plants were obtained.
作者
李思琪
王怡婷
于耸
郑志民
Li Siqi;Wang Yiting;Yu Song;Zheng Zhimin(Key Laboratory of Saline-alkali Vegetation Ecology Restoration(Ministry of Education),College of Life Science,Northeast Forestry University,Harbin,150040;State Key Laboratory of Tree Genetics and Breeding,College of Forestry,Northeast Forestry University,Harbin,150040)
出处
《分子植物育种》
CAS
北大核心
2023年第6期1899-1905,共7页
Molecular Plant Breeding
基金
东北林业大学5211高层次人才引进科研启动基金项目(GCC2016-01)资助。
关键词
蓖麻
草铵膦
BAR基因
转基因
Castor
Glufosinate-ammonium
Bar gene
Transgenic
