大叶女贞抗寒基因AmEBP1高效转化体系的研究  

STUDY ON THE HIGHLY EFFECTIVE TRANSFORMATION SYSTEM OF COLD-TOLERANT GENE AMEBP1 TRANSFORMED INTO LIGUSTRUM IUCIDUM AIT

作　　者：王长宪[1] 刘静[1] 王斌[2] 孙仲序[3] 刘杰[4] 黄艳艳[1] 赵进红[1] 张虹[1] 

机构地区：[1]泰安市泰山林业科学研究院,山东泰安271000 [2]中国科学院遗传与发育生物学研究所,北京10010 [3]山东农业大学园艺学院,山东泰安271018 [4]青岛科技大学生物系,山东青岛266042

出　　处：《山东农业大学学报（自然科学版）》2009年第4期508-512,共5页Journal of Shandong Agricultural University：Natural Science Edition

摘　　要：本研究以大叶女贞为受体材料,利用农杆菌浸染方法对沙冬青抗寒基因AmEBP1进行转化试验,通过对不同的受体材料浸染部位、菌液浓度和浸染时间对转化材料的死亡率%、分化率%、分化芽数(个)、芽苗长度(cm)以及芽苗颜色影响进行了分析,结果表明:转化材料为茎段,菌液浓度稀释为50%,浸染时间15 min;其中浸染部位是影响基因高效转化的最主要因素,菌液浓度和浸染时间的作用次之;方差分析进一步发现,浸染部位的三个水平对于外源基因在植物体内的转化显著差异,因此选择合适的浸染部位、适宜的菌液浓度和浸染时间能提高基因转化效率;经在50 mg/L浓度的卡那霉素培养基筛选培养,获得一批抗性植株,经PCR检测获得5株转化株系。By means of Agrobacterium contamination, cold - tolerant gene AmEBP1, absorbed form Ammopiptanthus mongolicus, was transformed into Ligustrum lucidum Air. We made an investigation that different contamination position, Agrobacterium concentration and contamination time of the acceptor material affected the transformant death rate, differentiation rate, number of differential buds, buds length and buds color. The result showed that contamination position was the major factor of effects on the gene advanced transformation, the effects of bacterial concentration and contamination time were minor factors when stem part was used as the transformant, Agrobacterium concentration was 50 percent, it was contaminated for 15 minutes. The result of analysis of variance indicated that three levels of contamination position caused obvious difference in transformations of foreign genes into plants. Therefore, selecting suitable position contaminated, Bacterial concentration and contamination time could improve transformation efficiency. A group resistant plants wene obtained through with the oulture medium contains 50 mg/L kanamycin, then five transferplants can he got by PCR examination.

关 键 词：大叶女贞 农杆菌浸染 抗寒基因AmEBP1 高效转化 

分 类 号：S718.46[农业科学—林学]