KMITL

Permanent URI for this communityhttps://dspace.kmitl.ac.th/handle/123456789/1

Browse

Settings

Author: search.filters.author.Kittiwongwattana, ChokchaiSubject: search.filters.subject.Bacillus

Search Results

Now showing 1 - 3 of 3
  • Loading...
    Thumbnail Image
    Some of the metrics are blocked by your 
    consent settings
    Item type:Publication,
    Biocontrol Potential, Genome and Nonribosomal Peptide Synthetase Gene Expression of Bacillus velezensis 2211
    (2023-01-01)
    Niemhom, Nantawan
    ;
    Kittiwongwattana, Chokchai
    Members of the genus Bacillus produced a diverse group of antimicrobial compounds. Here, we presented the antifungal activity and genome sequence analysis of Bacillus sp. 2211, a potential plant-growth-promoting bacterium. Bacterial supernatants from strain 2211 cultures in nutrient broth (NB) and potato dextrose broth (PDB) suppressed the mycelial growth of Pyricularia oryzae, Colletotrichum aenigma, Colletotrichum fructicola and Fusarium oxysporum. The supernatants were also able to suppress spore germination of these fungi, except for F. oxysporum. However, the supernatant from PDB displayed a significantly higher inhibition activity than NB. Additionally, the supernatant from PDB significantly reduced the disease severity caused by P. oryzae on rice seedlings. The genome of strain 2211 was sequenced. The highest digital DNA-DNA hybridization (80.1%) and average nucleotide identity (97.57%) levels indicated that strain 2211 was a member of the species Bacillus velezensis. The phylogenomic analysis showed that it clustered with B. velezensis NRRL B-41580<sup>T</sup>, B. velezensis KACC 13105 and B. velezensis subsp. plantarum FZB42<sup>T</sup>. The gene expression analysis showed the up-regulation of nonribosomal peptide synthetase (NRPS) genes bmyA, fenB and dhbE in PDB, compared to NB. This work demonstrated that the culture media affected the antagonistic activity of strain 2211 possibly through the modification of NRPS biosynthesis genes.
  • Loading...
    Thumbnail Image
    Some of the metrics are blocked by your 
    consent settings
    Item type:Publication,
    Validation of reference genes for the normalization of RT-qPCR gene expression in Bacillus siamensis 1021 grown in different culture media
    (2022-01-01)
    Nuwong, Warinya
    ;
    Kittiwongwattana, Chokchai
    Background and Objectives: House-keeping genes are generally selected as reference genes in gene expression analysis. However, some genes may not be stably expressed across all experimental conditions. Thus, this study aimed to validate seven house-keeping genes for gene expression analysis in Bacillus siamensis 1021. Materials and Methods: Strain 1021 was grown in potato dextrose broth, nutrient broth and mineral salt medium. Re-verse-transcription quantitative PCR was used to determine Cq values of seven reference genes including gyrA, gyrB, ssb and dnaB, rpsU, gat_Yqey and udp in these media. Expression stability of these genes was analyzed, using geNorm and Normfinder applications. The target gene ftsZ was used for assessment of the best candidate genes. Results: Based on geNorm and Normfinder, ssb was the most-stably expressed gene, while udp was the least-stably expressed gene. Pairwise variation indicated the combination of ssb, gyrA, gyrB and gatB_Yqey was suitable for the normalization of ftsZ expression. ftsZ expression in potato dextrose broth and mineral salt medium was higher than that in nutrient broth. In contrast, the normalization against udp resulted in an under-and overestimation of ftsZ expression in potato dextrose broth and mineral salt medium, respectively. Conclusion: The combination of ssb, gyrA, gyrB and gatB_Yqey was the best candidate for normalization of target gene expression in B. siamensis 1021 in these media. This study emphasized the significance of reference gene validation for gene expression analysis and provided a guideline for future gene expression studies in B. siamensis.
  • Loading...
    Thumbnail Image
    Some of the metrics are blocked by your 
    consent settings
    Item type:Publication,
    Medium Effect on Antagonistic Activity and Detection of Nonribosomal Peptide Synthetase Genes in Epiphytic Bacillus Strains
    (2021-01-01)
    Apimeteethamrong, Suchitra
    ;
    Kittiwongwattana, Chokchai
    The biosynthesis of non-ribosomal peptides (NRPs) in biocontrol bacteria was one of the major antagonistic mechanisms for their application in agriculture. Bacillus spp. 1021, 2211 and 3210 were previously shown to inhibit mycelial growth of the leaf blast fungus Pyricularia oryzae. Here, we aimed to further study the antagonistic mechanism in those three strains. Cell-free supernatants obtained from bacteria grown in potato dextrose broth (PDB) exhibited a higher degree of inhibition against P. oryzae when compared to those obtained from nutrient broth (NB). This indicated the effect of culture media in the production of extracellular antibiotic compounds by these strains. Phylogenetic analysis of their partial 16S rRNA gene sequences indicated a close relationship between the three strains and Bacillus siamensis KCTC13613<sup>T</sup>, Bacillus amyloliquefaciens DSM7<sup>T</sup> and Bacillus velezensis CR-502<sup>T</sup>. Complete genome sequences of these Bacillus species were analyzed on the antiSMASH server to identify the presence of NRP biosynthesis gene clusters. Non-degenerate primers were designed for the detection of the core biosynthesis genes for surfactin (srfAA), fengycin (fenC) and bacillibactin (dhbF). All three genes were amplified in strains 1021 and 2211, while only srfAA and dhbF were detected in strain 3210. Phylogenetic analysis of the deduced amino acid sequences indicated that the sequences of strain 1021 were distinct from those of strains 2211 and 3210. This result indirectly suggests the possibility of NRP production as the antagonistic mechanism of these three strains.