CAJ | 학술논문

通过建立遗传转化体系，培育具有特定功能的转基因作物品种对改善作物的生存能力和生态环境适应力具有重要意义。参照农杆菌划胚介导植物萌发种子基因转化方法，以农杆菌菌株 LBA4404（含质粒pCAMBAR．CH I．11）侵染转化燕麦种子，对转化体系中的菌液浓度、超声波功率、种子处理方式和抗除草剂筛选等不同转化条件分别进行了研究，以探索农杆菌介导的燕麦最适宜的遗传转化体系。结果表明：燕麦种子浸泡4 h后采用穿刺法划伤种子，1000 W超声波处理划伤种子10 min，在OD600＝0．4的农杆菌溶液中共培养2 d，最有利于种子转化；用0．8 mg／L草丁膦溶液与种子共培养能够有效抑制种子萌发进行初筛，用1 mg／L草丁膦溶液喷施幼苗进行除草剂抗性筛选，能够明显抑制假阳性苗的生长；将来自链霉菌（ Streptomyces hygroscopicus）的Bar基因导入燕麦，经除草剂筛选以及Bar基因的PCR检测和Southern杂交分析证明获得转基因植株。
통과건립유전전화체계，배육구유특정공능적전기인작물품충대개선작물적생존능력화생태배경괄응력구유중요의의。삼조농간균화배개도식물맹발충자기인전화방법，이농간균균주 LBA4404（함질립pCAMBAR．CH I．11）침염전화연맥충자，대전화체계중적균액농도、초성파공솔、충자처리방식화항제초제사선등불동전화조건분별진행료연구，이탐색농간균개도적연맥최괄의적유전전화체계。결과표명：연맥충자침포4 h후채용천자법화상충자，1000 W초성파처리화상충자10 min，재OD600＝0．4적농간균용액중공배양2 d，최유리우충자전화；용0．8 mg／L초정련용액여충자공배양능구유효억제충자맹발진행초사，용1 mg／L초정련용액분시유묘진행제초제항성사선，능구명현억제가양성묘적생장；장래자련매균（ Streptomyces hygroscopicus）적Bar기인도입연맥，경제초제사선이급Bar기인적PCR검측화Southern잡교분석증명획득전기인식주。
Cultivating transgenic crop varieties with specific function through the establishment of genetic transformation system is significant for the improvement of crop survival ability and ecological environment adaptability.Referring to the gene conversion method of seed germination with agrobacterium mediated , the oat seeds was infected and transformed with agrobacterium strain LBA4404 ( contain plasmid pCAMBAR.CH I.11 ) , the transformation conditions including concentration of bacteria, ultrasonic power, processing method of seeds, selection of herbicide resistance and so on in the transformation system were studied to explore the most appropriate genetic transformation system of oats mediated by agrobacterium.The results showed that after immersed for four hours, oat seeds were scratched by puncture method, 1 000 W ultrasonic treatment by 10 min was done for the scratched seeds, then they were cultured in agrobacterium liquid concentration OD600 =0.4 for 2 days, it was the best condition for genetic transformation.Seeds cultured with 0.8 mg/L glufosinate concentration could effectively inhibit germination of oat seeds.Seedlings were sprayed by 1 mg/L glufosinate concentration to select herbicide , it could obviously inhibit the growth of false positive seedlings .The Bar gene was introduced into the oat , and transgenic plants were first screened by Basta then detected by PCR and Southern blot approaches.