Current Microbiology

Gao, Tantan; Wang, Xudong; Qin, Yanqiu; Ren, Zhengguang; Zhao, Xiaoyan

doi: 10.1007/s00284-023-03206-2pmid: 36802037

Bacillus amyloliquefaciens TR2, one of plant growth-promoting rhizobacteria (PGPR), is capable of colonizing plant roots in a large population size. However, the interaction of watermelon root exudates and colonization of the strain TR2 has not yet been clearly elucidated. In this investigation, we demonstrated that B. amyloliquefaciens TR2 promoted watermelon plants growth and exhibited biocontrol efficacy against watermelon Fusarium wilt under greenhouse conditions. Collected watermelon root exudates significantly induced chemotaxis, swarming motility, and biofilm formation of the strain TR2. We also tested the components of root exudates (organic acids: malic acid, citric acid, succinic acid, and fumaric acid; amino acids: methionine, glutamic acid, alanine, and aspartic acid; phenolic acid: benzoic acid) and the results showed that a majority of these compounds could promote chemotactic response, swarming motility, and biofilm formation in a different degree. Benzoic acid induced the strongest chemotactic response; however, the swarming motility and biofilm formation of the strain TR2 were maximumly enhanced by supplement of fumaric acid and glutamic acid, respectively. In addition, the root colonization examination indicated that the population of B. amyloliquefaciens TR2 colonized on watermelon root surfaces was dramatically increased by adding concentrated watermelon root exudates. In summary, our studies provide evidence suggesting that root exudates are important for colonization of B. amyloliquefaciens TR2 on plant roots and help us to understand the interaction between plants and beneficial bacteria.

Han, Dong Min; Baek, Ju Hye; Choi, Dae Gyu; Jin, Myeong Seo; Jeon, Che Ok

doi: 10.1007/s00284-023-03222-2pmid: 36841905

A strictly aerobic Gram-negative bacterium, designated R8T, isolated from garden soil in South Korea was subjected to a taxonomic study. The cells were non-spore-forming, oxidase-positive and catalase-negative, and non-motile rods (without flagella). Growth was observed between 10 °C and 40 °C (optimum, 30 °C) and between pH 6.0 and 9.0 (optimum, pH 7.0) and in the presence of 0%–1.5% (w/v) NaCl (optimum, 0%). The G + C content of the genomic DNA was 49.9% and the major isoprenoid quinone was found to be menaquinone-7. The major fatty acids of strain R8T were iso-C15:0, C16:1ω5c, and summed feature 3 (comprising iso-C15:0 2-OH and/or C16:1 ω7c/ω6c). Phosphatidylethanolamine was identified as a major polar lipid. Comparative 16S rRNA gene sequence analysis showed that strain R8T had the highest 16S rRNA gene sequence similarity of 98.3% with Chitinophaga sedimenti TFL-3 T. Phylogenetic analyses using 16S rRNA gene sequences and concatenated 92 marker protein sequences revealed that strain R8T formed a robust phylogenetic lineage with C. sedimenti within the genus Chitinophaga. Average nucleotide identity and digital DNA–DNA hybridization values of strain R8T to Chitinophaga species were less than 77.9% and 21.1%, respectively. The phenotypic, phylogenetic, and chemotaxonomic properties support that strain R8T represents a novel species of the genus Chitinophaga, for which the name Chitinophaga horti sp. nov. is proposed. The type strain is R8T (= KACC 19895 T = JCM 33215 T).

Shimoyama, Yu; Sasaki, Daisuke; Ohara-Nemoto, Yuko; Nemoto, Takayuki K.; Nakasato, Manami; Sasaki, Minoru; Ishikawa, Taichi

doi: 10.1007/s00284-023-03212-4pmid: 36797528

Porphyromonas gingivalis is an asaccharolytic, Gram-negative, anaerobic bacterium representing a keystone pathogen in chronic periodontitis. The bacterium’s energy production depends on the metabolism of amino acids, which are predominantly incorporated as dipeptides via the proton-dependent oligopeptide transporter (Pot). In this study, the localization of dipeptidyl-peptidases (DPPs) and Pot was investigated for the first time in P. gingivalis using immunoelectron microscopy with specific antibodies for the bacterial molecules and gold-conjugated secondary antibodies on ultrathin sections. High-temperature protein G and hemin-binding protein 35 were used as controls, and the cytoplasmic localization of the former and outer membrane localization of the latter were confirmed. P. gingivalis DPP4, DPP5, DPP7, and DPP11, which are considered sufficient for complete dipeptide production, were detected in the periplasmic space. In contrast, DPP3 was localized in the cytoplasmic space in accord with the absence of a signal sequence. The inner membrane localization of Pot was confirmed. Thus, spatial integration of the nutrient acquisition system exists in P. gingivalis, in which where dipeptides are produced in the periplasmic space by DPPs and readily transported across the inner membrane via Pot.

Lieu, Dong My; Tran, Giang Thi Chau; Nguyen, Ngan Thi; Dang, Thuy Thi Kim

doi: 10.1007/s00284-023-03225-zpmid: 36872377

The objective of the study was to determine the efficacy of ultrasound-treatment Saccharomyces cerevisiae and spray drying in preserving the viability of Lactiplantibacillus plantarum. The combination of ultrasound-treated S. cerevisiae and L. plantarum was evaluated. Next, the mixture was blended with maltodextrin and either Stevia rebaudiana-extracted fluid, prior to undergoing spray drying. The L. plantarum viability was assessed after the spray drying process, during storage, and in simulated digestive fluid (SDF) conditions. The results showed that the impact of ultrasound caused the crack and holes in the yeast cell wall. Besides, the moisture content values were not significantly different in all samples after spray drying. Although the amount of powder recovery in stevia-supplemented samples was not higher than that of the control sample, the L. plantarum viability was significantly improved after the spray drying process. The density of L. plantarum tended to be stable during the first 30 days of storage and decreased more rapidly after that. The results reveal that there was no statistically significant difference in the trend of the samples before and after storage. In the SDF test, the L. plantarum viability mixing with ultrasound-treated yeast cells in the spray drying samples was significantly improved. Besides, the presence of Stevia showed positive efficiency on the L. plantarum viability. The L. plantarum viability mixing with ultrasound-treated yeast cells and stevia-extracted fluid by spray drying process showed potential application due to making powder form which helped to improve the L. plantarum stability during the storage time.

Han, Binghua; Yu, Qiaoling; Su, Wanghong; Yang, Jiawei; Zhang, Shiheng; Li, Xiaoshan; Li, Huan

doi: 10.1007/s00284-023-03210-6pmid: 36808560

The deep mechanisms shaping bacterial assembly are a crucial challenge in drinking water ecosystem. However, much less is known about seasonal diversity distributions and assembly mechanisms of abundant and rare bacteria in drinking water. The combination of environmental variables and high-throughput 16S rRNA gene sequencing was conducted to examine the composition, assembly, co-occurrence patterns of abundant and rare bacteria from five drinking water sites across four seasons in one year in China. The results indicated that abundant taxa were mainly composed of Rhizobiales_UG1, Sphingomonadales_UG1, and Comamonadaceae, while rare taxa were Sphingomonadales_UG1, Rhizobiales_UG2, and Rhizobiales_UG1. The richness of rare bacteria was greater than that of abundant ones, and the richness had no differences among seasons. The beta diversity was significantly discrepant in abundant and rare communities and among seasons. Deterministic mechanism accounted for a larger contribution to abundant taxa than rare taxa. Furthermore, water temperature had higher effects on abundant microbiome than rare ones. Co-occurrence network analysis indicated that abundant taxa that occupied frequently in central positions had stronger effect on co-occurrence network. In our study, these results collectively suggested that rare bacteria respond to environmental variables with an analogical pattern to abundant counterparts (similar community assembly), but their ecological diversities, driving forces, and co-occurrence patterns were not equivalent in drinking water.Graphical Abstract[graphic not available: see fulltext]

Yu, Chuan; Dong, Hefan; Li, Qi; Wang, Xiaoli; Mao, Fuchao; Qian, Man; Niu, Junhui; Cheng, Xiangchao; Liao, Chengshui

doi: 10.1007/s00284-023-03229-9pmid: 36853439

TatD is the subunit of the twin-arginine translocation (Tat) pathway. Members of TatD family are multifunctional, conserved and widely presented proteins in most prokaryotes. It has been reported that Tat can affect bacterial motility in some bacteria. This study was conducted to determine the contribution of the TatD protein (herein named LmTatD) to the regulation of flagella in Listeria monocytogenes. We constructed an LmTatD gene mutant in L. monocytogenes strain 10403 s and evaluated its biological characteristics. The results showed no difference in growth or morphology between the wild-type strain and the ΔLmTatD mutant. Intriguingly, the ΔLmTatD mutant showed impaired swimming motility and flagella structure but increased biofilm formation. Comparative proteomic analysis using tandem mass tag (TMT) combined with liquid chromatography–tandem mass spectrometry (LC‒MS/MS) was performed to determine differentially expressed proteins (DEPs). The results revealed that 134 proteins out of 2228 total proteins identified were differentially expressed, among which 18 proteins were upregulated and 116 proteins were downregulated in the ΔLmTatD mutant. Analysis of DEPs indicated that the reduced expression levels of the proteins related to flagellar assembly in the ΔLmTatD mutant correlate with its characteristics. Compared to the wild-type strain, the most downregulated proteins in the ΔLmTatD mutant included FlaA, FliD, FliR, FlgD, FlgL, and FlgG. Collectively, our data suggest that although LmTatD is not required for growth in L. monocytogenes, loss of LmTatD reduces flagellar production and motility by regulating flagellar assembly-related protein expression.

Shao, Chenyi; Song, Xiaobo; Wang, Lili; Zhang, Hongying; Liu, Yinhui; Wang, Chunhao; Chen, Shenmin; Ren, Baowei; Wen, Shu; Xiao, Jing; Tang, Li

doi: 10.1007/s00284-023-03224-0pmid: 36877409

Gut microbiota contributes to human health. Plenty of studies demonstrate that antibiotics can disrupt gut ecosystem leading to dysbiosis. Little is known about the microbial variation of appendix and its up/downstream intestine after antibiotic treatment. This study aimed to investigate the microbiome and mucosal morphology of jejunum, appendix, and colon of rats in health and dysbiosis. A rodent model of antibiotic-induced dysbiosis was employed. Microscopy was used to observe mucosal morphological changes. 16S rRNA sequencing was performed for identifying bacterial taxa and microbiome structure. The appendices of dysbiosis were found enlarged and inflated with loose contents. Microscopy revealed the impairment of intestinal epithelial cells. High-throughput sequencing showed the Operational Taxonomic Units changed from 361 ± 33, 634 ± 18, 639 ± 19 in the normal jejunum, appendix, colon to 748 ± 98, 230 ± 11, 253 ± 16 in the disordered segments, respectively. In dysbiosis, Bacteroidetes translocated inversely from the colon and appendix (0.26%, 0.23%) to the jejunum (13.87% ± 0.11%); the relative abundance of all intestinal Enterococcaceae increased, while Lactobacillaceae decreased. Several bacterial clusters were found correlated to the normal appendix, whereas nonspecific clusters correlated to the disordered appendix. In conclusion, species richness and evenness reduced in the disordered appendix and colon; similar microbiome patterns were shared between the appendix and colon regardless of dysbiosis; site-specific bacteria were missing in the disordered appendix. Appendix is likely a transit region involving in upper and lower intestinal microflora modulation. The limitation of this study is all the data were derived from rats. We must be cautious about translating the microbiome results from rats to humans.

Ghare, Uma; Narvekar, Simran; Lodha, Tushar; Mallebhari, Rubiya; Dastager, Syed; Barvkar, Vitthal T.; Dhotre, Dhiraj; Karmalkar, Nitin R.; Pable, Anupama A.

doi: 10.1007/s00284-023-03207-1pmid: 36807001

The Western Ghats is one of India’s mega-diversity hotspots and an ecologically and geologically important area for the diversity of endemic plants and animals. The present study provides insights into the aerobic bacterial diversity and composition of the soils of North Western Ghats located in Maharashtra state (NWGM), India. The samples for the culture-dependent study were collected from 6 different locations namely Malshej Ghat, Bhimashankar, Lonavala, Mulshi, Tail-Baila, and Mahabaleshwar. A total of 173 isolates were obtained from the different samples, which belonged to Proteobacteria (43%), Firmicutes (36%), and Actinobacteria (19%). Sequences of 15 strains shared ≤ 98.7% similarity (a species cut-off) which represent potential novel species. Metagenomic analysis revealed the presence of Actinobacteria and Proteobacteria as the most dominant phyla at both MB and MG. However, both sites showed variation in the composition of rare phyla and other dominant phyla. This difference in bacterial community composition could be due to differences in altitude or other physicochemical properties. The functional prediction from the amplicon sequencing showed the abundance of carbohydrate, protein, and lipid metabolism which was corroborated by screening the isolated bacterial strains for the same. The present study has a unique take on microbial diversity and defines the importance of community assembly processes such as drift, dispersal, and selection. Such processes are relatively important in controlling community diversity, distribution, as well as succession. This study has shown that the microbial community of NWGM is a rich source of polysaccharide degrading bacteria having biotechnological potential.

de Sousa, Leandro Pio; Filho, Oliveiro Guerreiro; Mondego, Jorge Maurício Costa

doi: 10.1007/s00284-023-03236-wpmid: 36890285

The analysis of large-scale sequence data has revealed that plants over time recruit certain microbes that are efficient colonizers of the rhizosphere. This enrichment phenomenon is especially seen in annual crops, but we suggest that there could have been some type of enrichment in perennial crops such as coffee plants. To verify this hypothesis, we performed a metagenomic and chemical analysis in rhizosphere with three different plant ages (young, mature, and old) and cultivated on the same farm. We verified that from mature to old plants, there was a decrease in diversity, particularly Fusarium and Plenodomus, while there was an increase in Aspergillus, Cladosporium, Metarhizium, and Pseudomonas. We also detected that the abundance of anti-microbials and ACC-deaminase grows as plants age, although denitrification and carbon fixation had reduced abundances. In summary, we detected an enrichment in the microbial community, especially in the great increase in the participation of Pseudomonas, passing from 50% of the relative abundance as the plants get older. Such enrichment can occur through the dynamics of nutrients such as magnesium and boron.

Amini, Seyed Mohammad; Hadighi, Ramtin; Najm, Mehdi; Alipour, Maryam; Hasanpour, Hamid; Vosoogh, Mehran; Vosough, Araz; Hajizadeh, Maryam; Badirzadeh, Alireza

doi: 10.1007/s00284-022-03172-1pmid: 36781499