甘薯‘福薯604’遗传转化体系的建立

许泳清; 李国良; 张鸿; 李华伟; 林赵淼; 邱永祥; 邱思鑫

甘薯‘福薯604’遗传转化体系的建立

A Genetic Transformation System of Sweet Potato ‘Fushu 604’

摘要

摘要:
目的遗传转化是进行基因功能验证的重要手段，构建较为完善、高效的甘薯遗传转化体系，为基因功能验证和甘薯种质改良提供技术支撑。
方法以‘福薯604’为材料，用携带表达载体pCAMBIA1301-proFLS-GUS的根癌农杆菌转化‘福薯604’胚性细胞，通过探究影响甘薯遗传转化过程中的重要因子，建立以‘福薯604’为基础的甘薯遗传转化体系。
结果结果表明，本试验中最适宜诱导‘福薯604’胚性愈伤组织的培养基配方为：MS+2 mg·L⁻¹ 2,4-D+30 g·L⁻¹蔗糖+3 g·L⁻¹植物凝胶；选择预培养4～5 d的‘福薯604’悬浮细胞团进行转化，侵染效果较佳；潮霉素浓度为5 mg·L⁻¹时较适宜筛选抗性植株。利用以上方法，经共培养、筛选培养和体胚诱导可成功获得拟转基因再生植株，共获得15个株系生根苗，通过PCR检测确定7株为阳性苗，阳性率达46.7%。
结论成功建立了甘薯‘福薯604’的甘薯遗传转化体系，为甘薯关键基因功能验证和种质精准改良提供技术支持。

Abstract:
Objective A comprehensive genetic transformation system was established to facilitate function identification and validation of a sweet potato germplasm.
Methods Non-embryogenic cells carrying the expression vector, pCAMBIA1301-proFLS-GUS, in ‘Fushu 604’ sweet potato were transformed with Agrobacterium tumefaciens. By studying the key factors affecting the sweet potato genetic transformation, a system for genetic improvements was established.
Results For inducing embryogenic callus, MS medium supplemented with 2mg·L⁻¹ of 2,4-D, 30g·L⁻¹ of sucrose, and 3g·L⁻¹ of a plant gel was chosen. The suspension of cell aggregates was pre-cultured for 4-5 days for the transformation. Hygromycin (Hyg) at the concentration of 5mg·L⁻¹ was used to screen for resistant plants. Somatic embryos were then induced to regenerate onto whole plants after co-cultivation and selection to obtain 15 rooted seedlings. By PCR detection, 7 positive seedlings were confirmed positive, at a rate of 46.7%.
Conclusion A genetic transformation system was successfully established for ‘Fushu 604’ to facilitate the function identification and validation on key genes so that improvement on the germplasm could be precisely achieved.

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