提高埃及糖橙遗传转化效率的研究  

Further study on improving genetic transformation efficiency of succari orange

作　　者：胡威[1,2] 杨莉[1,2] 谢玉明[1,2] 周潇[1,2] 葛红娟[1,2] 李大志[1,2] 邓子牛[1,2] 

机构地区：[1]湖南农业大学园艺园林学院,湖南长沙410128 [2]国家柑橘改良中心长沙分中心,湖南长沙410128

出　　处：《湖南农业大学学报（自然科学版）》2013年第4期371-376,F0003,共7页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：现代农业(柑橘)产业技术体系岗位科学家项目(CAR–27);湖南省重点项目(2011WK2007);国家自然科学青年基金项目(31201596)

摘　　要：为提高农杆菌介导的甜橙遗传转化效率，以埃及糖橙实生苗上胚轴节间茎段为外植体，对实生苗生长过程中的光照条件和农杆菌侵染时的共培养条件进行优化，并将无核基因转入糖橙中。结果表明，暗培养20d后光照10d实生苗外植体的再生率和转化率均较高；乙酰丁香酮（As）浓度是影响转化效率的主要因素，共培养基中加入质量浓度20mg／L的AS、共培养温度控制在19℃、共培养基pH为5．4～5．7时最有利于转化，经GUS染色分析，最高GUS阳性率可达29．4％；经PCR进一步分析，可以初步确证外源基因已经整合到糖橙基因组中。To increase the genetic transformation efficiency of sweet orange infected by Agrobacterium, an expiant derived from internode stem segments of sucarri orange seedling was adopted to optimize light regimes and co-cultivation condition for the growth. Meantime, seedless gene was transferred to succari orange. The results revealed that explant from seedling had higher regeneration and transformation rate under the condition of cultured 20 d in darkness first and then exposed 10 d in lighting environment. Acetosyringone （AS） concentration was a key factor impacting transformation rate, and the optimal performance of transformation was under 20 mg/L concentration of co-cultivated medium with a temperature 19 ℃ and pH 5.4-5.7. The positive rate could reach 29.4% through the analysis of GUS staining, which were further proved by PCR analysis that the foreign gene was indeed integrated into the host genome.

关 键 词：埃及糖橙 农杆菌介导 遗传转化 光照 共培养 

分 类 号：S571.1[农业科学—茶叶生产加工]