Conventional random mutation breeding remains the simplest and most cost-effective approach in enhancing strains in fermentation industry.Nonetheless,extensive research on compound mutagenesis are limited to screening...Conventional random mutation breeding remains the simplest and most cost-effective approach in enhancing strains in fermentation industry.Nonetheless,extensive research on compound mutagenesis are limited to screening target strains,and few studies on the interaction mechanism of compound mutagenesis factors.Herein,transcriptomic was employed to elucidate the molecular mechanism behind helium-neon(He-Ne)laser and ultraviolet(UV)compound mutagenesis in Bacillus subtilis.Transcriptomic sequencing analysis was conducted on five sample groups(control/CK,UV,laser/LA,UV-laser/UL,and laser-UV/LU),facilitating the statistical com-parison of differentially expressed genes(DEGs)across 4 sample relationships(CK-vs-UV,CK-vs-LA,CK-vs-LU,and CK-vs-UL).The DEGs revealed that UV irradiation improved the biosynthesis of pyrimidine nucleotides by upregulating the activity of key enzymes(pyrimidine nucleotides synthetase,pyruvate dehydrogenase,phos-photransferase system(PTS)fructose-specific enzyme),while laser irradiation stimulated metabolism through the regulation of transmembrane transporter activity.Further KEGG enrichment analysis of 1891 DEGs in UV-vs-LU sample relationship unveiled 89 genes significantly enriched in the two-component system,which potentially linked to phytochrome.Moreover,analysis of 784 DEGs in UV-vs-UL sample relationship indicated up-regulation of genes associated with translation,replication,and genetic repair pathways,affirming the laser’s induction of UV damage repair mechanisms.Subsequently,9 pivotal genes(liaS,yvfT,yxdK,cheA,ykoW,uvrC,pcrA,recA,and ruvB)were selected for quantitative reverse transcriptional PCR(qRT-PCR)validation,demonstrating consistent trends with transcriptomic data.Conclusively,our findings unveiled interplay mechanism between UV and laser compound mutagenesis at molecular level,offered valuable insights for mutagenesis mode selection and further elucidation of mutagenesis mechanisms.展开更多
基金supported by the National Key Research and Devel-opment Program of China[Grant No.2021YFC2104200,2022YFF0710101]Jiangsu Provincial Key Research and Development Program[Grant No.BE2018368]Zhenjiang Key Research and Development Plan[Grant No.NY2023001].
文摘Conventional random mutation breeding remains the simplest and most cost-effective approach in enhancing strains in fermentation industry.Nonetheless,extensive research on compound mutagenesis are limited to screening target strains,and few studies on the interaction mechanism of compound mutagenesis factors.Herein,transcriptomic was employed to elucidate the molecular mechanism behind helium-neon(He-Ne)laser and ultraviolet(UV)compound mutagenesis in Bacillus subtilis.Transcriptomic sequencing analysis was conducted on five sample groups(control/CK,UV,laser/LA,UV-laser/UL,and laser-UV/LU),facilitating the statistical com-parison of differentially expressed genes(DEGs)across 4 sample relationships(CK-vs-UV,CK-vs-LA,CK-vs-LU,and CK-vs-UL).The DEGs revealed that UV irradiation improved the biosynthesis of pyrimidine nucleotides by upregulating the activity of key enzymes(pyrimidine nucleotides synthetase,pyruvate dehydrogenase,phos-photransferase system(PTS)fructose-specific enzyme),while laser irradiation stimulated metabolism through the regulation of transmembrane transporter activity.Further KEGG enrichment analysis of 1891 DEGs in UV-vs-LU sample relationship unveiled 89 genes significantly enriched in the two-component system,which potentially linked to phytochrome.Moreover,analysis of 784 DEGs in UV-vs-UL sample relationship indicated up-regulation of genes associated with translation,replication,and genetic repair pathways,affirming the laser’s induction of UV damage repair mechanisms.Subsequently,9 pivotal genes(liaS,yvfT,yxdK,cheA,ykoW,uvrC,pcrA,recA,and ruvB)were selected for quantitative reverse transcriptional PCR(qRT-PCR)validation,demonstrating consistent trends with transcriptomic data.Conclusively,our findings unveiled interplay mechanism between UV and laser compound mutagenesis at molecular level,offered valuable insights for mutagenesis mode selection and further elucidation of mutagenesis mechanisms.
