天麻抗真菌蛋白GAFP基因转化油葵的研究

以培养6 h的油葵自交系6B6的子叶为外植体,利用农杆菌介导法将GAFP基因导入该自交系。通过对其遗传转化主要因素的研究,发现将外植体在OD600=0.7的农杆菌中浸泡10~15 min后,在含有AS 100μmol/L、pH 5.4的预培养培养基上22±1℃暗培养2 d,遗传转化率高。研究建立了油葵遗传转化体系,获得了4株经PCR检测为阳性的转基因植株。研究为通过植物抗病基因工程方法防治油葵菌核病提供了1条新的途径。

Study on Transferring Gastrodia Antifungal Protein (GAFP) Gene into Oil Sunflower (Helianthus annuus L.)

The GAFP genes were transferred into the cotyledons of oil sunflower,which were cultured on the medium for 6 hours,mediated by Agrobacterium tumefaciens.The main factors influencing transformation rate were discussed and the best conduction of transformation were determined.The cotyledons of oil sunflower were immersed in Agrobacterium tumefaciens for 10-15 minutes,co-cultivated in the medium including 100 μmol/L AS at pH 5.4 for 2 days in the dark.The temperatures was maintained at 22±1℃,and thus the higher performance of regeneration was obtained.4 of positive regenerated plants were obtained and confirmed from polymerase chain reaction.The dissertation provided a new approach to the anti-fungal disease of oil sunflower method by use plant genetic engineering technology.