| 陆荣生，韩美丽，吴耀军，杜晓莉，杨玉霞，马跃峰.农杆菌介导AFT基因转入四季桔的研究[J].广东农业科学,2013,40(11):145-148 |
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农杆菌介导AFT基因转入四季桔的研究 |
| Studies on ATF gene transformation mediated by Agrobacterium in Citrus mitis |
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| DOI： |
| 中文关键词: 四季桔 不定芽 转化 |
| 英文关键词: citrus mitis adventitious bud transformation |
| 基金项目:广西农业科学院基本科技业务专项也200808Z(基)页 |
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| 摘要点击次数: 1126 |
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| 中文摘要: |
| 以四季桔上胚轴为外植体，研究农杆菌介导转化过程中的影响因素。结果显示，转化研究所用抑菌剂抗菌素中，替门的汀杀菌效果较好，且对外植体出芽无明显抑制作用。转化芽选择初期，2~3 d的预培养及选择培养初期3~5d的暗培养有利于卡那霉素抗性芽的发生。PCR扩增检测结果显示，AFT基因已转化至四季桔，转基因植株-4℃处理6d时，SOD酶与脯氨酸含量均高于对照。该试验建立了四季桔上胚轴遗传转化再生系统，其再生条件为院MT基本培养基附加TDZ0.5~1.0mg/L，替门汀300mg/L，卡那霉素100mg/L；预培养时间为2~3d，共培养后3~5d的暗培养有利于转化率的提高。 |
| 英文摘要: |
| Epicotyls from Citrus mitis seedling were used as explants to investigate the factors which had an influence on differentiation and regeneration of anti-Kan adventitious buds in transformation mediated by Agrobacterium. Among antibiotics, Timentin was the best for differentiation of adventitious buds and growth control of agrobacterium. Growing in darkness for 3~5 days after explants co-culture with agrobacterium was help to anti-Kan buds developing. It was better for anti-Kan adventitious buds developing when explants were cultured in differentiation medium for 2~3 days before infected by agrobacterium. Transgenic plants were obtained by PCR assay, and SOD enzyme activity and content of proline were raised in transgenic plant treated by -4益for 4~6 days. The result showed that transformation system was established for epicotyls in Citrus mitis. The transgenic plant could be obtain in medium with TDZ 0.5~1.0 mg/L, Timentin 300 mg/L, Km 100 mg/L. Frequency of transformation would be increased as explants precultured for 2~3 d and dark culture for 3~5 d after co-culture. |
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