| 刘桃李刘清童建华彭琼.籼型稻V20B 高效遗传转化体系研究初报[J].,2012,12(18):3460-3463 |
| 籼型稻V20B 高效遗传转化体系研究初报 |
| Preliminary Study on High Efficient System of Genetic Transformationof Indica Rice V20B |
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| DOI： |
| 中文关键词: 水稻 成熟胚 愈伤组织 分化 |
| 英文关键词: Rice Mature embryo Callus Differentiation |
| 基金项目:湖南省科技重大专项（2009FJ1004-2）；湖南省高校创新平台开放基金项目（09k052） |
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| 中文摘要: |
| 目的：建立籼型杂交稻亲本V20B 成熟胚愈伤组织培养的高效遗传转化体系。方法：以V20B 的成熟胚作为外植体，研究比
较不同培养基、不同生长物质类型及浓度、不同培养条件对愈伤组织诱导、继代、分化的影响。结果：诱导培养基以N6 为基本培养
基，添加2.5 mg·L-1 2,4-D 和0.2 mg·L-1 6-BA，光照条件下诱导，成熟胚愈伤组织诱导率达到93.44 %；继代培养基以MS 为基本培
养基，添加2.0 mg·L-1 2,4-D，黑暗条件下培养；分化培养基以DL 为基本培养基，激素配比为2.0 mg·L-1KT、2.0 mg·L-16-BA、0.2
mg·L-1 NAA、0.2 mg·L-1 IAA；采用农杆菌介导法对该体系获得的愈伤组织侵染后能获得的抗性愈伤组织，经PCR 检测潮霉素基
因转化率为53.89 %。结论：建立了适宜于籼型稻V20B 的高效组织培养体系。 |
| 英文摘要: |
| Objective: To construct a highly efficient system of genetic transformation of hybrids rice of parent V20B using culturing
mature embryo callus. Methods: Mature embryo of indica rice V20B is used as explants to investigate the influence of different basic
mediums (MS, N6, CC, NMB, NB), phytohormones and culture conditions on callus' induction, subculturation and differentiation. Results:
When N6 as basic induction medium supplemented with 2.5 mg·L-1 2,4-D and 0.2 mg·L-1 6-BA, the induction rate of mature callus tissue
was as high as 93.44 % under light condition. In subculture process, MS were used as basic medium, then supplemented with 2.0 mg·L-1
2,4-D, under dark condition. DL was used in differentiation, as basic medium, adding different hormones as 2.0 mg·L-1 KT, 2.0 mg·L-1
6-BA, 0.2 mg·L-1 NAA, 0.2 mg·L-1 IAA. Callus was immerged and infected by Agrobacterium tumefactiens, and the transformation efficiency
was 53.89 % by PCR analysis. Conclusion: A highly efficient plant regeneration system was established. It was suitable for indica
rice V20B. |
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