激素和硝酸银对芥菜型油菜下胚轴再生植株的影响  

Effects of hormone and AgNO_3 on regeneration of Brassica juncea hypocotyl explants

作　　者：袁玉辉[1] 刘显军[1] 杨柳[1] 陆赢[1] 官春云[1] 刘忠松[1] 

机构地区：[1]湖南农业大学油料作物研究所,湖南长沙410128

出　　处：《湖南农业大学学报（自然科学版）》2014年第1期1-7,共7页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家自然科学基金项目(31101176;30971799)

摘　　要：为建立芥菜型油菜高效遗传转化体系,以芥菜型油菜品种‘四川黄籽’的下胚轴为外植体,采用农杆菌介导法进行遗传转化,对愈伤诱导、芽分化及生根培养3个阶段的激素、硝酸银及其配比进行优化处理。结果表明:用含1.0 mg/L 2,4?D的预培养基诱导愈伤,愈伤再生率达到100%;用含3.0 mg/L 6-BA+5.0 mg/L AgNO3的分化培养基诱导芽再生,芽再生率达28.33%,平均芽丛数为1.73个;在含2.5 mg/L IBA+0.05mg/L NAA生根培养基中,不定芽生长11 d后开始生根,生根率达到100%;经PCR检测抗卡拉霉素转化苗,阳性率达到23.80%,且GUS染色呈阳性,说明建立的芥菜型油菜遗传转化体系高效可行。Hypocotyls of Sichuan yellow seed of Brassica juncea was used as explants to establish an efficient genetic transformation system by Agrobacterium-mediated transformation. Different combinations of hormone and silver nitrate for callus induction, bud differentiation and root culture were optimized. The results showed that 100% callus regeneration rate was observed with pre-culture medium containing only 1.0 mg/L of 2,4-D; shoot regeneration rate was 28.33% and the average number of bud bundles was 1.73 when using the differentiation medium with 3.0 mg/L 6-BA and 5.0 mg/L AgNO3; the shoot begins to root with 100% rooting rate 11 days after culturing on rooting medium with 2.5 mg/L IBA and 0.05 mg/L NAA. The transformant resistant positive rate was 23.80% as determined by PCR detection and GUS staining, which illustrated the feasibility of this Biassicajuncea genetic transformation system.

关 键 词：芥菜型油菜 硝酸银 农杆菌介导 下胚轴 激素配比 

分 类 号：S634.303.53[农业科学—蔬菜学]