Department of Microbiology, Daegu Catholic University School of Medicine, Daegu, Republic of Korea.
Department of Chemistry, Chonnam National University, Gwangju, Republic of Korea.
Commun Biol. 2021 May 14;4(1):582. doi: 10.1038/s42003-021-02107-z.
The conjugative plasmid (pBV71) possibly confers a selective advantage to Bacillus velezensis strain GH1-13, although a selective marker gene is yet to be identified. Here we show that few non-mucoid wild-type GH1-13 cells are spontaneously converted to mucoid variants with or without the loss of pBV71. Mucoid phenotypes, which contain or lack the plasmid, become sensitive to bacitracin, gramicidin, selenite, and tellurite. Using the differences in antibiotic resistance and phenotype, we isolated a reverse complement (COM) and a transconjugant of strain FZB42 with the native pBV71. Transformed COM and FZB42p cells were similar to the wild-type strain GH1-13 with high antibiotic resistance and slow growth rates on lactose compared to those of mucoid phenotypes. RT-PCR analysis revealed that the expression of plasmid-encoded orphan aspartate phosphatase (pRapD) was coordinated with a new quorum-sensing (QS) cassette of RapF2-PhrF2 present in the chromosome of strain GH1-13, but not in strain FZB42. Multi-omics analysis on wild-type and plasmid-cured cells of strain GH1-13 suggested that the conjugative plasmid expression has a crucial role in induction of early envelope stress response that promotes cell morphogenesis, biofilm formation, catabolite repression, and biosynthesis of extracellular-matrix components and antibiotics for protection of host cell during exponential phase.
尽管尚未鉴定出选择标记基因,但共轭质粒 (pBV71) 可能使 Bacillus velezensis 菌株 GH1-13 具有选择优势。在这里,我们表明,在没有或没有失去 pBV71 的情况下,很少有自发转化为粘液表型的非粘液野生型 GH1-13 细胞。含有或不含质粒的粘液表型对杆菌肽、短杆菌肽、亚硒酸盐和碲酸盐敏感。利用抗生素抗性和表型的差异,我们分离出了一个反向互补(COM)和一个带有天然 pBV71 的菌株 FZB42 的转导子。转化的 COM 和 FZB42p 细胞与野生型 GH1-13 菌株相似,与粘液表型相比,它们对乳糖的抗生素抗性高,生长速度慢。RT-PCR 分析表明,质粒编码的孤儿天冬氨酸磷酸酶(pRapD)的表达与染色体中存在的 RapF2-PhrF2 新的群体感应(QS)盒协调一致,但在菌株 FZB42 中不存在。对野生型和质粒去除的 GH1-13 细胞的多组学分析表明,共轭质粒的表达在诱导早期包膜应激反应中起着关键作用,促进细胞形态发生、生物膜形成、分解代谢物抑制和细胞外基质成分的生物合成以及抗生素的合成,以保护宿主细胞在指数生长期。