水稻FELBs胚胎模式的新型遗传转化体系的构建

摘要：本研究以籼稻MH63为试材，通过无菌苗培养，外植体筛选和激素组合配比，最终确立水稻根为外植体，诱导得到MH63的FELB。以此为转化受体材料，通过小瓶真空抽提法对水稻FELB不同胚龄，转化抽提时间、次数和时间间隔、预培养和筛选培养等条件进行优化，将转化后的FELB置于筛选培养基中进行光照诱导抗性芽。结果表明：FELB在相同转化介质的情况下对不同抽提次数与时间的试验结果进行分析，转化率分别为39.63%，65.63%和18.30%。抽提3次，间隔为10min，筛选培养基为MS+0.2mg/L NAA+2.0mg/L 6-BA+200mg/L 特美汀+50.0mg/L K+的催萌效果最佳。通过GUS染色检测和PCR检测，最终成功构建了籼稻MH63FELB高效、稳定新型遗传转化体系。31178
毕业论文关键词：籼稻；FELB；小瓶真空抽提；遗传转化体系
Establishment of Novel Genetic Transformation System of Rice on account of FELB Model
Abstract: The indica rice MH63 was taken as experimental material in the study, the FELB induction system of MH63 were established by aseptic seedlings culturation, different explants screening and different plant growth regulators combination. The plasmid transformation was operated through vacuum extraction in vial after analysis extraction number, time and extraction interval. The experimental results of extraction numbers and times were analyzed under the same transformation solution circumstance. The extracted FELBs were transferred onto MS+0.2mg/L NAA+2.0mg/L 6-BA+200mg/L Timentin+50.0mg/L K+ for resistant buds. The results showed that, the beat extraction transformation method was three times with 10 minutes interval (each extraction until no bubble), and the worst was the one time. And the transformation efficiency were respectively 39.63%，65.63% and 18.30%. The novel transformation system on MH63 on account of FELBs was successfully constructed by analyzing the GUS dyeing and PCR of FELB.
Key words: Indica rice; FELB; Vial vacuum extraction; Genetic transformation system
目    录
摘要   1
Abstract   1
引言   2
1 材料与方法   2
1.1 材料   2
1.2 培养准备   2
1.2.1 无菌苗培养   2
1.2.2 FELB诱导   2
1.2.3 FELB胚萌诱导   3
1.2.4 转化质粒提取   3
1.2.5 FELB转化渗透介质   3
1.3 方法   3
1.3.1 无菌苗获得   3
1.3.2 外植体选择   3
1.3.3 愈伤组织和FELB的诱导培养   3
1.3.4 FELB的质粒转化   4
1.3.5 FELB直接和离体再生诱导   4
1.3.6 FELB的筛选培养   4
1.3.7 FELB转化效率的鉴定   4
1.3.8 统计分析   4
2 结果与分析   4
2.1 诱导结果   4
2.2 生长素类似物对愈伤组织和FELB诱导的影响   5
2.3 FELB的胚性分析   5
2.4 FELB的再生诱导   6
2.5 FELB的抽提转化   6
3 讨论   7
3.1 激素配比对FELB诱导效率的影响   7
3.2 FELB胚龄对转化效率的影响   8
3.3 抽提时间与抽提次数对转化效率的影响   8
参考文献   10
致谢   12
水稻FELBs胚胎模式的新型遗传转化体系的构建
引言
水稻（Oryza sativa）是草本稻属的一种，也是稻属中作为粮食的最主要最悠久的一种，又称为亚洲型栽培稻，简称稻。原产于亚洲热带，在我国广为种植，并逐渐传播到世界各地。水稻原产于中国，是世界主要粮食作物之一，中国水稻播种面积占全国粮食作物的1/4，而产量则占一半以上。根据不同的方法，水稻可以分为粳稻与籼稻、早稻与中晚稻，糯稻与非糯稻。栽培历史已有14000-18000年。作为重要粮食作物，除食用颖果外，可制淀粉、酿酒、制醋，米糠可制糖、榨油、提取糠醛，可供工业及医药用；稻秆为良好饲料及造纸原料和编织材料，谷芽和稻根可供药用。水稻FELBs胚胎模式的新型遗传转化体系的构建:http://www.youerw.com/shengwu/lunwen_27243.html