| 利用正交设计优化金钗石斛的转化体系 |
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| 引用本文： | 杨翠芹,秦耀国,罗玉容,刘运权,黎扬辉,刘伟. 利用正交设计优化金钗石斛的转化体系[J]. 西南大学学报, 2009, 31(6) |
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| 作者姓名： | 杨翠芹 秦耀国 罗玉容 刘运权 黎扬辉 刘伟 |
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| 作者单位： | 杨翠芹,YANG Cui-qin(华南农业大学,生命科学学院,广州,510642;四川农业大学,农学院,四川,雅安,625014);秦耀国,QIN Yao-guo(四川农业大学林学园艺学院,四川,雅安,625014);罗玉容,刘运权,刘伟,LUO Yu-rong,LIU Yun-quan,LIU Wei(华南农业大学,生命科学学院,广州,510642);黎扬辉,LI Yang-hui(广州花卉研究中心,广州,510360) |
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| 基金项目： | 国家自然科学基金，广东省自然科学基金，广州市农业科技招标资助项目 |
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| 摘 要： | 为优化金钗石斛的转化体系,以其类原球茎(PLBs)为材料,利用正交设计研究了影响农杆菌介导转化的因素.结果表明:PLBs预培养1 d、侵染的菌液OD600值1.2、乙酰丁香酮(AS)100μmol/L、共培养5 d时,每100mg鲜物质量材料上的GUS染色蓝点数最多;单因子试验表明AS适宜的浓度为50μmol/L;PCR结果显示得到了转化呈阳性的PLBs;初步表明此体系可用于金钗石斛的遗传转化研究.Abstract：In order to optimize the Agrobacterium-mediated transformation system for Dendrobium nobile Lindl, effects of preculture time, bacterial concentration, acetosyringon (AS) concentration and co-cultivation time on Agrobacterium-mediated transformation of protocorm like bodies (PLBs) of D. nobile Lindl were studied in an experiment with orthogonal design. The results showed that number of GUS staining blue spots per 100 mg flesh weight of PLBs was the greatest in the treatment of the combination of preculture for one day, 1.2 of bacterial concentration (OD600), 100 μmol/L of concentration of AS and co-culture for five days. Single factor experiment showed that the appropriate concentration of AS was 50 μmol/LPCR confirmed that positive transformed PLBs were obtained. It is tentatively concluded that this system can be used in the study of transformation of D. nobile Lindl.
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| 关 键 词： | 金钗石斛 农杆菌 转化 正交设计 |
Optimization of Agrobacterium-Mediated Transformation System for Dendrobium nobile Lindl by Orthogonal Design |
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| YANG Cui-qin,QIN Yao-guo,LUO Yu-rong,LIU Yun-quan,LI Yang-hui,LIU Wei. Optimization of Agrobacterium-Mediated Transformation System for Dendrobium nobile Lindl by Orthogonal Design[J]. Journal of Southwest Agricultural University, 2009, 31(6) |
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| Authors: | YANG Cui-qin QIN Yao-guo LUO Yu-rong LIU Yun-quan LI Yang-hui LIU Wei |
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