摘要
[目的]建立以农杆菌介导的吴屯杨遗传转化体系。[方法]以吴屯杨无菌苗叶片为遗传转化受体材料,采用农杆菌介导的转化方法,结合草甘膦(PPT)筛选,探讨影响吴屯杨叶片遗传转化效率的重要因素。[结果]吴屯杨叶片分化不定芽和生根的草甘膦临界浓度分别为0.8、0.6 mg/L。对预培养2～3 d的叶片,用OD600=0.4～0.5的菌液侵染10～15 min、共培养培养基中添加乙酰丁香酮(AS)200μmol/L、共培养3 d可有效提高吴屯杨遗传转化效率。对成活的转基因植株进行PCR检测,结果表明目的基因已转入吴屯杨中。[结论]该研究为进一步加强吴屯杨分子育种研究奠定了坚实的基础。
[ Objeqtive ] The study aimed to establish Agrobacterium mediated transformation system of Populus wutunensis. [ Method ] With the Populus wutunensis leaves as explants, combining with glyphosate （PPT） screening, the key factors which affected transformation frequency of Populus wutunensis leaves were discussed by using Agrobacterium mediated transformation methods. [ Result] The Populus wutunensis leaves bud differentiation and rooting critical concentration of glyphosate were 0.8,0.6 mg/L. With OD600 = 0.4 - 0.5 of the pre-cuhure broth 2 - 3 d infes- ted leaves 10- 15min, adding acetosyringone （AS） 200 p, mol/L of medium, co-culture 3 d, can effectively improve the efficiency of genetic transformation of Populus wutunensis. Through the survival of transgenic plants by PCR, the results showed that the exogenous gene has been transferred to Populus wutunensis. [ Conclusion] The study will lay a foundation to the further research of Populus wutunensis molecular breeding.
出处
《安徽农业科学》
CAS
2013年第14期6134-6136,共3页
Journal of Anhui Agricultural Sciences
基金
大连民族学院大学生创新创业训练计划
国家自然科学基金(31100452)
关键词
吴屯杨
农杆菌
外源基因
遗传转化
Populus wutunensis
Agrobacterium
Exogenous gene
Genetic transformation
