中文摘要：

基于体细胞胚胎发生技术平台，利用携带pSuperl300＋质粒，以潮霉素为筛选标记基因的农杆菌GV3101介导日本落叶松遗传转化，对植物受体材料生理状态、农杆菌浓度和浸染时间以及共培养时间等影响因素进行了研究、分析和讨论。结果表明：综合优化各影响因素，生长旺盛的日本落叶松胚性细胞，经浓度为0．4（OD600）的农杆菌浸染10min，共培养2d，再用含400mg／L的头孢霉素的液体培养基清洗脱菌，然后在含400mg／L的头孢霉素固体培养基上恢复培养，并置于含5mg／L潮霉素的固体培养基上多次筛选，最终共获得54个抗性细胞系，转化率平均为0．94个／g。PCR检测鉴定，所有抗性细胞系均为阳性转化体，并排除了农杆菌污染导致的假阳性。研究建立并优化了农杆菌介导的日本落叶松遗传转化技术，为进行遗传改良和基因功能鉴定提供有利平台。

英文摘要：

Based on the technique of L. leptolepis somatic embryogenesis, embryogenic tissues were transformed by A. tumefaciens strain GV3101 carrying a binary plasmid pSuperl300 ＋ with the hpt gene as a selectable marker. Factors that influence transformation were investigated and discussed, including the physiological status of plant calli, the concentration of bacteria, the duration of bacterial infection and co-culture. As a resuh, after infected by 0.4 （ OD6oo ） bacterial solution for lOmin, co-culture for 2d , then washed three times by liquid medium supplemented with 400mg/L cefotaxime, followed by recovery culture for 7d and selection by 5mg/L hygromycin for several times, a total of 54 hygromycin-resistant cell lines had been obtained. The transformation efficiency is 0.94 transgenic cell lines obtained from per gram of plant calli on average. All the resistant ceils were positively transgenic confirmed by polymerase chain reaction （PCR）. The development of such a robust transformation method not only provides a useful approach for genetic improvement but also allows us to conduct a functional identification of genes in L. leptolepis.