农杆菌介导法将ACC合成酶和氧化酶反义基因转入日本甜柿的研究

Transformation of ACC Synthase and ACC Oxidase Antisense Gene to Jirou Japanese Persimmon

以日本甜柿品种次郎(Diospyros kaki Thunb.cv.Jirou)的叶片为外植体,通过农杆菌介导法,建立了次郎柿遗传转化体系。结果表明,菌液OD600值为0.6,乙酰丁香酮浓度为200mg·L-1,筛选培养3d,头孢霉素为300mg·L-1的转化率最高。在此条件下,将ACC合成酶和氧化酶的反义联合基因导入到次郎柿的组培苗中,经PCR检测,得到了8株转基因植株。

Using young leaf of Diospyros kaki Thunb. cv. Jirou as explant, the transformation conditions were studied systemically in this paper. A high efficient and stable genetic transformation system of Jirou in vitro was established with the OD600 of engineering bacterial suspension of 0.6, the concentration of AS of 200 mg·L^-1, the filt-cuhured time of 3 days and the concentration of Cef of 300 mg·L^-1. 8 strains of Jirou transgenetic plants carrying ACC synthase and ACC oxidase antisense gene were identified by PCR.