| 农杆菌介导生防棘孢木霉菌转化体系的优化 |
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| 引用本文： | 刘露,李雅华,咸洪泉. 农杆菌介导生防棘孢木霉菌转化体系的优化[J]. 生物技术, 2012, 22(2): 49-53 |
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| 作者姓名： | 刘露 李雅华 咸洪泉 |
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| 作者单位： | 青岛农业大学生命科学学院,山东青岛,266109 |
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| 摘 要： | 目的:实现棘孢木霉菌T4的遗传转化并优化其转化体系.方法:以潮霉素抗性为选择标记,利用农杆菌转化法介导转化棘孢木霉菌.结果:潮霉素基因成功整合到受体菌基因组中,转化子抗性基因可稳定遗传.结论:最优的转化体系和条件为:IM和CM培养基中AS浓度为200 μg/mL,棘孢木霉T4孢子浓度为106/mL,农杆菌浓度为200 μL( OD600约0.8),共培养时间为48 h,转化效率约为50个转化子/106个孢子.
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| 关 键 词： | 棘孢木霉 农杆菌 遗传转化 潮霉素抗性 体系优化 |
Optimization of Trichoderma asperellum Transformation System with Agrobacterium tumefaciens |
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| LIU Lu , LI Ya-hua , XIAN Hong-quan. Optimization of Trichoderma asperellum Transformation System with Agrobacterium tumefaciens[J]. Biotechnology, 2012, 22(2): 49-53 |
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| Authors: | LIU Lu LI Ya-hua XIAN Hong-quan |
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| Affiliation: | (College of Life Science,Qingdao Agriculture University,Qingdao 266109,China) |
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| Abstract: | Objective:The filamentous fungi Trichoderma asperellum was transformed and the conditions were optimized.Method:Agrobacterium genetic transformation was applied in Trichoderma asperellum T4 and with the selection markers for hygromycin B resistance.Result:The hygromycin B gene had been inserted into the genome of Trichoderma asperellum T4,indicating that the resistance gene is stable genetically.Conclusion:Under the optimized conditions,the concentration of AS in IM and CM medium was 200μg/mL,the spores of T.asperellum was 106/mL,and the concentration of A.tumefaciens was 200μL(OD600=0.8),48h in co-culture medium,which resulted in a reasonable ATMT rates.About 50 transformants /106. |
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| Keywords: | Trichoderma asperellum Agrobacterium tumefaciens genetic transformation hygromycin B-resistant gene transformation optimization |
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