影响农杆菌介导朝仓花椒遗传转化因素研究

曾晓芳; 赵德刚

影响农杆菌介导朝仓花椒遗传转化因素研究

曾晓芳^1,2,
赵德刚^1,2,3

1 贵州大学贵州省农业生物工程重点实验室2 贵州大学教育部绿色农药与农业生物工程重点实验室3 贵州大学生命科学学院

计量
- 文章访问数: 3644
- HTML全文浏览量: 205
- PDF下载量: 55
出版历程
- 收稿日期: 1899-12-31
- 修回日期: 1899-12-31
- 发布日期: 2011-11-29

Factors affecting transformation of Zanthoxylum piperitum DC. var. inerme Makino via Agrobacterium tumefaciens.

ZENG Xiao-fang^1,2,
ZHAO De-gang^1,2,3

1 Guizhou Key Laboratory of Agrobioengineering，Guizhou University, Guiyang, 550025, P. R. China; 2 Key Laboratory of Green Pesticide and Agricultural Bioengineering of Ministry of Education，Guizhou University, Guiyang, 550025, P. R. China； 3 College of Life Sciences, Guizhou University, Guiyang, 550025, P. R. China.

摘要

摘要: 为建立农杆菌介导的朝仓花椒遗传转化体系，以朝仓花椒叶柄及茎段为转化受体，研究了预培养时间、侵染菌液浓度、侵染时间、共培养时间等因素对转化效率的影响。获得转化体系如下：外植体预培养3 d，在OD600为0.5~0.8菌液侵染10 min，共培养3 d，延迟7 d筛选，以30和50 mg/L卡那霉素(Kan)进行梯度筛选。结果表明：重悬液及共培养基中添加100 μmol/L乙酰丁香酮(AS)，可提高朝仓花椒Kan抗性愈伤率至24.6%；转化植株经GUS组织活性及PCR检测呈阳性，证实外源基因已导入朝仓花椒基因组中，利用本转化体系可为下一步将有重要经济价值外源基因导入花椒打下基础。
- 朝仓花椒 /
- 农杆菌 /
- 遗传转化
Abstract: In order to establish an Agrobacteriummediated genetic transformation system for Zanthoxylum piperitum DC.var.inerme Makino,using petioles and stems as explants, we examined several influencing factors of transformation efficiency, including precultivation time, optimum bacterium concentration and inflection period, and cocultivation time. The efficient transformation system obtained was 3 days precultivation time, 10 minutes inflection using bacterium at the concentration of OD600 0.5~0.8, followed by 3 days cocultivation. Selection of the transgenic plants was carried out after 7 days regeneration using gradient kanamycin at 30 and 50 mg/L, respectively. Adding 100 μmol/L acetosyringone in the resuspending solution and cocultivation medium increased the transformation efficiency to 24.6%. Transgenic plants were confirmed by GUS histochemical assays and polymerase chain reaction. The results suggest that an efficient Agrobacteriummediated transformation system for stable integration of foreign genes into Z. piperitum DC. var. inerme Makino has been developed, which could be applied in future studies on transferring economically important genes into Z. pipertum DC. var. inerme Makino.
- Zanthoxylum piperitum DC. var. inerme Makino /
- Agrobacterium tumefaciens /
- genetic transformation

HTML全文

参考文献(0)

施引文献(8)

期刊类型引用(4)

1.	高巧春. 聚合物基复合材料界面性能的实验设计. 广东化工. 2015(17): 31-32 . 百度学术
2.	朱德钦, 生瑜, 邹寅将, 苏晓芬, 王真. 苯甲酸/硬脂酸复合改性剂对PP/木粉复合材料性能的影响. 福建师范大学学报(自然科学版). 2012(01): 66-72 . 百度学术
3.	生瑜, 朱德钦, 苏晓芬, 邹寅将, 王真, 陈前火. 不同改性剂对PP/木粉复合材料性能的影响. 高分子材料科学与工程. 2012(10): 51-54 . 百度学术
4.	生瑜, 方镇, 朱德钦, 邓爱民, 叶敏, 谢远建, 生亚丰. 竹粉用量对PVC/竹粉复合材料阻燃抑烟性能的影响. 聚氯乙烯. 2011(04): 30-32 . 百度学术

其他类型引用(4)

资源附件(0)

计量

文章访问数: 3644
HTML全文浏览量: 205
PDF下载量: 55
被引次数: 8

影响农杆菌介导朝仓花椒遗传转化因素研究

计量

出版历程

Factors affecting transformation of Zanthoxylum piperitum DC. var. inerme Makino via Agrobacterium tumefaciens.

期刊类型引用(4)

其他类型引用(4)

计量

出版历程

目录