摘要
利用已构建的番木瓜环斑病毒(Papaya ringspot virus,PRSV)基因,通过农杆菌介导法,建立美中红番木瓜(Carica papaya L.)体胚的转化体系。结果表明:共培养时,当农杆菌EHA105携带pCAMBIA2300质粒时,农杆菌的浓度(OD600 nm)应小于或等于0.1,其共培养后抑制农杆菌的羧苄青霉素浓度应以750 mg/L为宜;当农杆菌EHA105携带pBI121质粒时,则农杆菌的浓度(OD600 nm)应小于或等于0.8,其共培养后抑制农杆菌的羧苄青霉素的浓度则为500 mg/L;加上滤纸处理,抗生素洗涤以及洗涤后的干燥处理,可完全抑制体胚表面的农杆菌生长。对再生的番木瓜体胚进行PCR检测,结果表明PRSV基因已转入体胚中,但还需要成苗后的进一步检测。
The genetic transformation system of somatic embryos of papaya （Carica papaya L. ） cultivar Meizhonghong mediated by Agrobacteriurn turnefaciens using genes from Papaya ringspot virus （PRSV） was established. The results showed that the concentration of Agrobacteriurn（OD600nm） harboring plant expression vector of pCAMBIA2300 was less than or equal to 0. 1 in co-culture with the somatic embryos of papaya and the concentration of Carbenicillin for inhibiting Agrobacterium was 750 mg/L after co-cultivation, while the concentrations of agrobacterium and carbenicillin were less than or equal to 0. 8 and 500 mg/L, respectively when Agrobacteriurn harboring plant expression vector of pBI121 was used. Three steps could control the overgrowth of Agrobacterium which were a piece of filter paper on co-culture media, washing with antibiotics after co-cultivation and desiccation after washing with antibiotics. Results of PCR indicated that the genes of PRSV were transformed into regenerative somatic embryos of papaya. It should be further confirmed.
出处
《华中农业大学学报》
CAS
CSCD
北大核心
2007年第3期293-296,共4页
Journal of Huazhong Agricultural University
基金
广东省重大科技专项(2002A208010202)
华南农业大学资源环境学院院长基金资助
关键词
番木瓜
番木瓜环斑病毒
体胚
转化
papaya（Carica papaya L. ）
Papaya ringspot virus （PRSV）
somatic embryo
transformation
作者简介
饶雪琴，女，1969年生，博士，副教授．工作单位：华南农业大学资源环境学院，广州510642.
通讯作者．E-mail：huaping@scau．edu．cn.
