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大连海洋大学学报  2011, Vol. 26 Issue (5): 407-413    DOI: 10.3969/j.issn.1000-9957.2011.05.006
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采用原生质体融合技术选育提高氨氮降解效能的光合细菌
纪敦敦,邱宏端,谢航
福州大学生物科学与工程学院,福建福州350108
The improvement of N-ammonia degradation of photosynthetic bacteria by protoplast fusion
JI Dun-dun,QIU Hong-duan,XIE Hang
College of Biological Science and Technology,Fuzhou University,Fuzhou 350108,China
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摘要 将高效降解氨氮的假丝酵母菌Candida sp.与高效降解亚硝酸盐氮的耐盐红螺菌Rhodospeudomonas capsulate进行原生质体融合,探讨原生质体制备及融合的条件,并对融合子进行了筛选。结果表明,原生质体制备的优化条件如下:耐盐红螺菌,溶菌酶量为1.5 mg/mL,EDTA浓度为0.1 g/L,作用时间为45 min;假丝酵母菌,蜗牛酶量为0.5 mg/mL,巯基乙醇的质量分数为0.1%,EDTA浓度为1 g/L,作用时间为30 min。两种原生质体在聚乙二醇(PEG-6000)和Ca2+的诱导下发生融合,在添加制霉菌素和链霉素的选择培养基上进行初筛,以生长稳定性及对氨氮、亚硝酸盐氮的降解效能等为指标进行复筛,获得了具有较好降解效能的融合子R1菌株。该菌株对亚硝酸盐氮的降解效能与耐盐红螺菌相同,达到90%以上;对氨氮的降解效能为63%,较耐盐红螺菌提高54%。
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纪敦敦
邱宏端
谢航
关键词:  耐盐红螺菌  假丝酵母菌  氨氮降解  原生质体融合    
Abstract: A protoplast fusion was conducted between two efficient photosynthetic bacteria,N-ammonia degrading Candida sp.and N-nitrite degrading Rhodospeudomonas capsulate.The conditions of the protoplast formation and fusion were optimized,and the best fusant was selected.The better protoplast formation for Rhodospeudomonas capsulata was found under the conditions of:lysozyme at 1.5 mg/mL,EDTA 0.1 g/L and 45 min reaction time;for Candida sp.the better conditions of snail enzyme at 0.5 mg/mL,mercaptoethanol 0.1%,EDTA 1 g/L and 30 min reaction time.The fusion was induced by PEG 6000 and Ca2+.Fusants were screened out by selective media containing added nystatin and streptomycin.The fusant R1 with better efficient degradation was selected by growth stability and degradation.The fusant R1 showed N-nitrite degradation at a rate of more than 90%,as good as for Rhodospeudomonas capsulata,but had N-ammonia degradation of only 63%,increased by 54%.
Key words:  Rhodospeudomonas capsulata    Candida sp.    N-ammonia degradation    protoplast fusion
               出版日期:  2011-10-21      发布日期:  2016-12-30      期的出版日期:  2011-10-21
中图分类号:  Q939.99  
引用本文:    
纪敦敦, 邱宏端, 谢航. 采用原生质体融合技术选育提高氨氮降解效能的光合细菌[J]. 大连海洋大学学报, 2011, 26(5): 407-413.
JI Dun-dun, QIU Hong-duan, XIE Hang. The improvement of N-ammonia degradation of photosynthetic bacteria by protoplast fusion. Journal of Dalian Ocean University, 2011, 26(5): 407-413.
链接本文:  
https://xuebao.dlou.edu.cn/CN/10.3969/j.issn.1000-9957.2011.05.006  或          https://xuebao.dlou.edu.cn/CN/Y2011/V26/I5/407
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