Xue Chen, Zheng Jiaxin, Wang Guangli, Feng Liang, Li Feng
College of Life Sciences, Huaibei Normal University, Huaibei, China.
Front Microbiol. 2022 Mar 10;13:842736. doi: 10.3389/fmicb.2022.842736. eCollection 2022.
sp. CTN-16 (GenBank FJ598326) can degrade chlorothalonil (CTN) but not carbendazim (MBC), and sp. MBC-3 (GenBank OK667229) can degrade MBC but not CTN. A functional strain BD2 was obtained by protoplast fusion of CTN-16 and MBC-3 to generate a fusant with improved degradation efficiency of CTN and MBC. Fusant-BD2 with eighth transfer on a medium containing CTN and two antibiotics was obtained. To identify and confirm the genetic relationship between parental strains and fusion strain BD2, scanning electron microscopy (SEM), random amplified polymorphic DNA (RAPD), and 16S ribosomal RNA (rRNA) gene sequences analysis were carried out. SEM analysis illustrated BD2 and its parents had some slight differences in the cell morphology. Fusant-BD2 not only possessed the same bands as parental strains but also had its specific bands analyzed through RAPD. The genetic similarity indices for BD2 and its parental strains CTN-16 and MBC-3 are 0.571 and 0.428, respectively. The degradation rates of CTN and MBC were 79.8% and 65.2% in the inorganic salt solution containing 50 mg·L CTN and 50 mg·L MBC, respectively, and the degradation efficiencies were better than the parental strains CTN-16 and MBC-3. This study provides a prospect for the application of fusion strain BD2 in bioremediation of CTN and MBC contaminated sites.
菌株CTN - 16(GenBank登录号FJ598326)能够降解百菌清(CTN),但不能降解多菌灵(MBC);而菌株MBC - 3(GenBank登录号OK667229)能够降解多菌灵,但不能降解百菌清。通过CTN - 16和MBC - 3的原生质体融合获得了功能菌株BD2,以产生对百菌清和多菌灵降解效率更高的融合体。在含有百菌清和两种抗生素的培养基上转接八次后获得了融合体BD2。为了鉴定并确认亲本菌株与融合菌株BD2之间的遗传关系,进行了扫描电子显微镜(SEM)、随机扩增多态性DNA(RAPD)以及16S核糖体RNA(rRNA)基因序列分析。SEM分析表明BD2与其亲本在细胞形态上存在一些细微差异。通过RAPD分析发现,融合体BD2不仅具有与亲本菌株相同的条带,还具有其特异性条带。BD2与其亲本菌株CTN - 16和MBC - 3的遗传相似性指数分别为0.571和0.428。在含有50 mg·L百菌清和50 mg·L多菌灵的无机盐溶液中,百菌清和多菌灵的降解率分别为79.8%和65.2%,其降解效率优于亲本菌株CTN - 16和MBC - 3。本研究为融合菌株BD2在百菌清和多菌灵污染场地的生物修复中的应用提供了前景。