摘要
为了建立农杆菌介导的高效毛白杨遗传转化体系,并大量获得转MdSPDS1基因的转基因毛白杨,对MdSPDS1基因导入毛白杨的遗传转化体系中关键转化因子进行了优化。获得的优化转化体系如下:叶片预培养3d后,用OD600为0.4的菌液侵染7min,再共培养3d。卡那霉素抗性芽的二次筛选中,卡那霉素的选择压分别为20和50mg/L。实验获得毛白杨卡那霉素抗性植株共43株,经PCR检测,其中9株呈阳性,占抗性植株总数的20.9%,优化转化系统的阳性植株转化率达到9.38%。本研究为下一步鉴定基因功能和转基因植株耐盐性的分析奠定了基础。
In order to establish the high efficiency genetic transformation system of Populus tomentosa Carr.by Agrobacterium tumefaciens,and to obtain mass-produced transgenic P.tomentosa with MdSPDS1 gene,several key factors of P.tomentosa transferring system were optimized during MdSPDS1 transformation.The optimized conditions for transformation of P.tomentosa were obtained as follows：the explant was precultivated for 3 days and infected with Agrobacterium at the concentration of OD600 = 0.4 for about 7 min,the foliages were co-cultured with Agrobacterium for 3 days,and the prophase and anaphase selecting methods were applied under the selection pressure of 20 and 50 mg /L Kan,respectively.Consequently,9 out of 43（20.9%） Kan-resistant plantlets were positive by PCR identification,suggesting that MdSPDS1 gene had been integrated into P.tomentosa genome.And the transformation frequency was increased to 9.38%.Our work laid a foundation for appraisal of gene function and the analysis of salttolerant capability of transgenic plants in the future.
出处
《北京林业大学学报》
CAS
CSCD
北大核心
2010年第5期21-26,共6页
Journal of Beijing Forestry University
基金
"863"国家高技术研究发展计划项目(2009AA10Z107)
"十一五"国家科技支撑计划项目(2006BAD01A15-02)
关键词
MdSPDS1
遗传转化
毛白杨
农杆菌
MdSPDS1
transformation
Populus tomentosa
Agrobacterium tumefaciens
