CAJ | 학술논문

为了快速验证与梨果实大小、硬度相关基因 fw 2．2、PL 是否能够通过转基因植株顺利表达.本研究利用根癌农杆菌介导 Micro-Tom 番茄技术,以子叶为外植体,分析不同预培养时间、菌液浓度、浸染时间、共培养时间、延迟筛选对抗性芽诱导的影响,得出最优转化体系为：子叶预培养36 h,在 OD 600nm 值为0.4菌液中浸染6 min,共培养60 h,在300 mg/L 头孢霉素(Cef)、25 mg/L 卡那霉素(Kan)延迟筛选14 d,转入300 mg/L Cef、40 mg/L Kan 分化培养.分别得到转基因阳性株为42株、38株,转化率为14.9％,13.1％.初步验证了 fw 2．2、PL 基因已经整合到 Micro-Tom 番茄中.
위료쾌속험증여리과실대소、경도상관기인 fw 2．2、PL 시부능구통과전기인식주순리표체.본연구이용근암농간균개도 Micro-Tom 번가기술,이자협위외식체,분석불동예배양시간、균액농도、침염시간、공배양시간、연지사선대항성아유도적영향,득출최우전화체계위：자협예배양36 h,재 OD 600nm 치위0.4균액중침염6 min,공배양60 h,재300 mg/L 두포매소(Cef)、25 mg/L 잡나매소(Kan)연지사선14 d,전입300 mg/L Cef、40 mg/L Kan 분화배양.분별득도전기인양성주위42주、38주,전화솔위14.9％,13.1％.초보험증료 fw 2．2、PL 기인이경정합도 Micro-Tom 번가중.
This experiment aims at rapidly verifying whether the fw 2.2 and PL genes related to the firm-ness and size of pear fruit and whether they could smoonthly express throuth transgenic plants.In this ex-periment,the tomato Micro-Tom technology mediated by agrobacterium tumefaciens were applied,and cot-yledons were used as explants to analyze the effects of different pre-incubation time,bacterial concentra-tion,dip-dye time,co-culture time and delayed screening on the induced resistance buds and obtain the opti-mal transformation system:the cotyledons were pre-cultured for 36 h,and dipped in bacteria with an OD 600nm of 0.4 for 6 min,totaly co-cultured for 60 h,subsequently delayed screening with Cef of 300 mg/L and Kan of 25 mg/L for 14 d,at last differentiation-cultured with Cef of 300 mg/L and Kan of 40 mg/L. The result showed that the obtained positive transgenic plants were 42 strains and 38 strains respectively, and the transformation rate was 14.9% and 13.1% respectively.Meanwhile,the experiment preliminarily verified that the fw 2.2 and PL genes had been integrated into the Micro-Tom tomato.