邓高毅1,彭 宇2,王远亮1.降烟碱解淀粉芽孢杆菌和边缘假单胞菌
原生质体融合选育高效降烟碱菌株[J].广东农业科学,2016,43(6):143-149 |
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降烟碱解淀粉芽孢杆菌和边缘假单胞菌
原生质体融合选育高效降烟碱菌株 |
Protoplast fusion between nicotine-degrading Bacillusamyloliquefaciens and Pseudomonas marginalis tobreed efficient nornicotine strain |
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DOI:10.16768/j.issn.1004-874X.2016.06.025 |
中文关键词: 解淀粉芽孢杆菌T11 边缘假单胞菌 原生质体 融合子 烟碱 |
英文关键词: Bacillus amyloliquefaciens T11 Pseudomonas marginalis protoplast fusant nicotine |
基金项目:湖南中烟工业有限责任公司(KY2012Y
C0001) |
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中文摘要: |
获取1 株高效降解烟碱的融合子,以解淀粉芽孢杆菌T11 和边缘假单胞菌作为亲本菌株,使用溶
菌酶破除亲株细胞壁后以40%PEG-6000 诱导解淀粉芽孢杆菌T11 原生质体和已热力灭活的边缘假单胞菌原
生质体进行融合,并检测了融合子的降烟碱能力。对融合子利用青霉素抗性初筛获得了176 株再生融合子,利
用烟碱降解率为指标进行降烟碱复筛实验后获得5 株烟碱降解能力较高的融合子。根据融合子遗传稳定性分
析及与亲本菌株烟碱降解效率的对比,获得了1 株烟碱降解率比亲本菌株更强的融合子A2。获得的高效降解
融合子A2 在第5 天后其烟碱降解率基本稳定在93%。 |
英文摘要: |
In order to acquire fusant with high nicotine-degrading ability,Bacillus amyloliquefaciens T11 and
Pseudomonas marginalis were used for parent strains. After using lysozyme to break the cell walls of parent strains,
the Bacillus amyloliquefaciens T11 protoplast and heat-inactivated Pseudomonas marginalis protoplast were induced by
40% PEG-6000 for protoplast fusion experiment,then the nicotine-degrading ability of fusant was measured. Through
preliminary screening,176 fusion strains were obtained. After rescreening by nicotine selective culture medium,5
fusion strains with higher nicotine-degrading ability were acquired. Genetic stability analysis of the 5 fusion strains and
comparision with parent strains’ nicotine-degrading rate were further measured. A better fusant A2 with higher nicotinededrading
ability than parent strains was obtained finally,93% nicotine in culture medium was degraded in five days. |
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