摘要
研究建立了根癌农杆菌介导的矮牵牛遗传转化体系。结果表明最佳的不定芽诱导分化和继代培养基为:MS+6-BA1.5mg/L+NAA0.2mg/L;选择培养基中30mg/L卡那霉素和300mg/L的头孢霉素是适宜的抗生素使用浓度;侵染过程中0.05MPa负压处理5min和共培养过程中添加20mg/L的乙酰丁香酮均可提高转化效率。卡那霉素抗性植株经2次选择继代培养后,PCR检测全部为阳性.对部分PCR阳性植株进行PCR-Southern杂交,证实外源基因已整合进入矮牵牛基因组中。
An Agrobacterium tumeficens-mediated transformation system has been developed for Petunia hybrida Vilm.The results showed the MS medium with 6-BA 1.5 mg/L and NAA 0.2 mg/L was the most suitable for the shoots induction and sub-cultivation.The optimun antibiotics concentration in seletive medium was 30 mg/L Kan and 300 mg/L Cef.And transformation efficiency was improved either by subjecting explants to-0.05 MPa pressure for 5 min while they were being infected or by adding AS at 20 mg/L into co-culture medium.After a 2-round selection of Kan-resistance,selected Kan-resistant plants were further identified by PCR and PCR based Southern blot analysis.The results demonstrated that all the Kan-resistant plants were transgenic ones and that the exogenous target gene,the GmNHX1 gene,has been integrated into the genomic DNA of all transformants obtained.
出处
《北方园艺》
CAS
北大核心
2007年第8期177-179,共3页
Northern Horticulture
基金
廊坊师范学院博士专项基金资助项目(LSZB200603)
关键词
矮牵牛
遗传转化
再生体系
Petunia hybrida Vilm
Genetic transformation system
Regeneration system
