摘要
【目的】对新疆推广种植的水稻品种进行组织培养,探索适合新疆水稻品种的遗传转化条件,为新疆水稻的遗传改良工作奠定基础。【方法】以新疆3个水稻品种成熟胚诱导的良好胚性愈伤组织为受体,以SiPEBP为目的基因,应用农杆菌介导法对水稻进行遗传转化,同时以抗性愈伤率为依据,对影响转化的愈伤诱导率、农杆菌侵染浓度、时间和共培时间进行优化研究。【结果】在NB培养基中添加2 mg/L的2,4-D的愈伤诱导率高;相比25℃、暗培养,28℃、持续光照培养可使诱导率提高到100%,且缩短了诱导周期;在OD6 0 0值为0.3的农杆菌菌液浓度,侵染愈伤30 min,共培养2 d,同时添加一张无菌滤纸,有利于提高转化效率;通过培养条件的优化,3个水稻品种的抗性愈伤率均达到40%以上。【结论】新疆水稻农杆菌转化的几个主要因素,建立了农杆菌转化体系。
【Objective】The tissue culture of seeds of rice varieties of Xinjiang was conducted for exploring genetic transformation for rice cultivars and laying the basis for genetic improvement.【Method】Callus induced from mature embryos of 3 rice varieties were transformed by Agrobacterium tumefaciens. Transformation efficiency was affected by factors of pre-culture time,density of bacteria,duration for inoculation and co-cultivation. 【Result】The findings showed that the induction rate of callus could be increased to 100% and the period of induction could be shortened under the condition of continuous light and 28℃ compared with the condition of dark and 25℃,in NB medium supplemented with 2 mg/L 2,4-D. Additionally,the higher transformation efficiency was obtained with the infected concentration of A. tumefaciens to an OD600 of 0. 3,30 minutes of infection and 2 days of cocultivation by placing a filter moistened. By the optimization of the conditions,the acquisition rate of resistant callus was up to 40%.【Conclusion】The major factors were analyzed by Agrobacterium including medium,and a high transformation frequency of Xinjiang rice was set up.
出处
《新疆农业科学》
CAS
CSCD
北大核心
2014年第8期1457-1462,共6页
Xinjiang Agricultural Sciences
基金
新疆维吾尔自治区自然科学基金项目(2011211B39)
关键词
水稻
根癌农杆菌
遗传转化
胚性愈伤组织
rice
Agrobacterium tumefacien
genetic transformation
embryonic callus
