摘要
本文报告了以甘蓝(bressicaoleracea)子叶为外植体,成功地将抗潮霉素(Hyg~r)基因和抗卡那霉素(Km~r)基因导入甘蓝品种,培育出转基因植物的技术研究成果.以MS为基本培养基,添加IAA2mg/1,BA4mg/1,蔗糖20mg/1和琼脂8.0mg/1,进行高再生率基因型选择,结果19个品种的再生芽发生率在0%~90%之间。对高再生率的品种进一步试验,进行不同激素浓度的适宜培养基的选择。结果表明:以IAA3mg/1和BA4mg/1的MS固体培养基诱导甘蓝子叶,其再生芽发生率高达95%~100%;每个外植体平均可诱导出2.4~3.9个再生芽。抗生素耐性试验结果指出,培养基中含有500mg/1羧苄青霉素几乎时再生芽的发生率和再生芽个数无影响,只是诱导的再生芽相对发育缓慢,体态弱小。用卡那霉素和潮霉素所进行的致死试验结果是:虽然致死率因基因型不同而有所不同,但培养基中含有Km20mg/1和Hyg20mg/1以上基本可以使所有的外植体死亡而无幼芽的再生。选择时期的试验结果显示,在外植体与EHA101农杆菌共培养3~l0d范围内,共培养6d和7d进行选择分别获得3%~5%的转化率。
This paper reported the results of tranfering the genes of resistance to Hygromycinand Karamycin into Cabbage using its cotyledon explomts. The rates of regeneration shootsfrom 19 varieties of cabbage were 0%  ̄ 90% in the basic MS medium added IAA2mg/l,BA4mg/1, sucross 20mg/1 and agrose 8.0mg/1. The selective experiment of hormonal concertratiorsand proportions showed that there were 95%  ̄ 100% shoot regeneration rate and 2.4-3.9regeneration shoots per explant in the MS solid medium containe IAA3mg/1 and BA4mg/1. Wealso found that carbenicill in (500mg/1) in the medium had nearly no influence on shootregeneration rate and shoot reeeneration .number except that the regeneration shoots developpedslowly. The lethal test with Kanamycin and hygromycin in the mrdium could lead to dead of allthe explants. During 3 days-10days of co-culture of the explants with agrobacteriumtumefaciens EHA101 3% ̄ 5% rate of the transgenic plants were obtained on the 6th dayand 7th day
出处
《沈阳农业大学学报》
CAS
CSCD
1995年第4期348-353,共6页
Journal of Shenyang Agricultural University
关键词
甘蓝
子叶
转基因植物
cabbage cotyledon, transgenic plant