MdSPDS1基因导入毛白杨的遗传转化体系优化研究
A study on the efficient protocol for transforming MdSPDS1 gene into Populus tomentosa Carr.
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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3 d后，用OD600为0.4的菌液侵染7 min，再共培养3 d。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50 mg/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。Abstract: 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 pre-cultivated 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 salt-tolerant capability of transgenic plants in the future.
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Keywords:
- MdSPDS1 /
- transformation /
- Populus tomentosa /
- Agrobacterium tumefaciens
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期刊类型引用(4)
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百度学术
2. 王蓓蓓，张炎，金博文，王泽伟，李亦轩，赵健，张金凤. 农杆菌介导的白杨遗传优化体系研究. 西南林业大学学报(自然科学). 2018(04): 14-21 . 百度学术
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其他类型引用(2)
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