The Research of Antagonistic Endophytic Bacterium Bacillus velezensis CSUFT-BV4 for Growth Promotion and Induction of Resistance to Anthracnose in Camellia oleifera.

Camellia oleifera (C. oleifera) is one of the four main, woody, edible oil tree species in the world, while C. oleifera anthracnose is mainly caused by the fungus Colletotrichum fructicola (C. fructicola), which severely affects the yield of C. oleifera and the quality of tea oil. Bacillus velezensis (B. velezensis) CSUFT-BV4 is an antagonistic endophytic bacterium isolated from healthy C. oleifera leaves. This study aimed to investigate the biocontrol potential of strain CSUFT-BV4 against C. oleifera anthracnose and its possible functional mechanism, and to determine its growth-promoting characteristics in host plants. In vitro, CSUFT-BV4 was shown to have efficient biofilm formation ability, as well as significant functions in the synthesis of metabolic substances and the secretion of probiotic substances. In addition, the CSUFT-BV4 fermentation broth also presented efficient antagonistic activities against five major C. oleifera anthracnose pathogens, including C. fructicola, C. gloeosporioides, C. siamense, C. camelliae, and C. kahawae, and the inhibition rate was up to 73.2%. In vivo, it demonstrated that the growth of C. oleifera treated with CSUFT-BV4 fermentation broth was increased in terms of stem width, plant height, and maximum leaf area, while the activities of various defense enzymes, e.g., superoxide dismutase (SOD), phenylalanine aminotransferase (PAL), and polyphenol oxidase (PPO), were effectively increased. The remarkable antagonistic activities against C. oleifera anthracnose, the growth-promoting characteristics, and the induction of host defense responses indicate that endophytic bacterium CSUFT-BV4 can be effectively used in the biological control of C. oleifera anthracnose in the future, which will have a positive impact on the development of the C. oleifera industry.

Endophytic bacteria for Cd remediation in rice: Unraveling the Cd tolerance mechanisms of Cupriavidus metallidurans CML2.

The utility of endophytic bacteria in Cadmium (Cd) remediation has gained significant attention due to their ability to alleviate metal-induced stress and enhance plant growth. Here, we investigate C. metallidurans CML2, an endophytic bacterial strain prevalent in rice, showing resilience against 2400 mg/L of Cd(II). We conducted an in-depth integrated morphological and transcriptomic analysis illustrating the multifarious mechanisms CML2 employs to combat Cd, including the formation of biofilm and CdO nanoparticles, upregulation of genes involved in periplasmic immobilization, and the utilization of RND efflux pumps to extract excess Cd ions. Beyond Cd, CML2 exhibited robust tolerance to an array of heavy metals, including Mn2+, Se4+, Ni2+, Cu2+, and Hg2+, demonstrating effective Cd(II) removal capacity. Furthermore, CML2 has exhibited plant growth-promoting properties through the production of indole-3-acetic acid (IAA) at 0.93 mg/L, soluble phosphorus compounds at 1.11 mg/L, and siderophores at 22.67%. Supportively, pot experiments indicated an increase in root lengths and a decrease in Cd bioaccumulation in rice seedlings inoculated with CML2, consequently reducing Cd translocation rates from 43% to 31%. These findings not only contribute to the understanding of Cd resistance mechanisms in C. metallidurans, but also underscore CML2's promising application in Cd remediation within rice farming ecosystems.

Pseudoxanthomonas sp. JBR18, a halotolerant endophytic bacterium, improves the salt tolerance of Arabidopsis seedlings.

Salinization of land is globally increasing due to climate change, and salinity stress is an important abiotic stressor that adversely affects agricultural productivity. In this study, we assessed a halotolerant endophytic bacterium, Pseudoxanthomonas sp. JBR18, for its potential as a plant growth-promoting agent with multiple beneficial properties. The strain exhibited tolerance to sodium chloride concentration of up to 7.5 % in the R2A medium. In vitro evaluation revealed that strain JBR18 possessed proteolytic, protease (EC 3.4), and cellulase (EC 3.2.1.4) activities, as well as the ability to produce indole-acetic acid, proline, and exopolysaccharides. Compared with the controls, co-cultivation of Arabidopsis seedlings with the strain JBR18 improved plant growth, rosette size, shoot and root fresh weight, and chlorophyll content under salinity stress. Moreover, JBR18-inoculated seedlings showed lower levels of malondialdehyde, reactive oxygen species, and Na+ uptake into plant cells under salt stress but higher levels of K+. Additionally, seedlings inoculated with JBR18 exhibited a delayed response time and quantity of salt-responsive genes RD29A, RD29B, RD20, RD22, and KIN1 under salt stress. These multiple effects suggest that Pseudoxanthomonas sp. JBR18 is a promising candidate for mitigating the negative impacts of salinity stress on plant growth. Our findings may assist in future efforts to develop eco-friendly strategies for managing abiotic stress and enhancing plant tolerance to salt stress.

Biocontrol potential of endophytic bacterium Bacillus altitudinis GS-16 against tea anthracnose caused by Colletotrichum gloeosporioides.

Background: As one of the main pathogens causing tea anthracnose disease, Colletotrichum gloeosporioides has brought immeasurable impact on the sustainable development of agriculture. Given the adverse effects of chemical pesticides to the environment and human health, biological control has been a focus of the research on this pathogen. Bacillus altitudinis GS-16, which was isolated from healthy tea leaves, had exhibited strong antagonistic activity against tea anthracnose disease. Methods: The antifungal mechanism of the endophytic bacterium GS-16 against C. gloeosporioides 1-F was determined by dual-culture assays, pot experiments, cell membrane permeability, cellular contents, cell metabolism, and the activities of the key defense enzymes. Results: We investigated the possible mechanism of strain GS-16 inhibiting 1-F. In vitro, the dual-culture assays revealed that strain GS-16 had significant antagonistic activity (92.03%) against 1-F and broad-spectrum antifungal activity in all tested plant pathogens. In pot experiments, the disease index decreased to 6.12 after treatment with GS-16, indicating that strain GS-16 had a good biocontrol effect against tea anthracnose disease (89.06%). When the PE extract of GS-16 treated mycelial of 1-F, the mycelial appeared deformities, distortions, and swelling by SEM observations. Besides that, compared with the negative control, the contents of nucleic acids, protein, and total soluble sugar of GS-16 group were increased significantly, indicating that the PE extract of GS-16 could cause damage to integrity of 1-F. We also found that GS-16 obviously destroyed cellular metabolism and the normal synthesis of cellular contents. Additionally, treatment with GS-16 induced plant resistance by increasing the activities of the key defense enzymes PPO, SOD, CAT, PAL, and POD. Conclusions: We concluded that GS-16 could damage cell permeability and integrity, destroy the normal synthesis of cellular contents, and induce plant resistance, which contributed to its antagonistic activity. These findings indicated that strain GS-16 could be used as an efficient microorganism for tea anthracnose disease caused by C. gloeosporioides.

Isolation of cadmium-resistance and siderophore-producing endophytic bacteria and their potential use for soil cadmium remediation.

Endophyte-assisted phytoremediation is an emerging technique for soil heavy metals (HMs) remediation and has become a research focus in the world because of the benefits of endophytes on plant growth and uptake of HMs. In this study, multifunctional endophytic bacteria strains were isolated and screened, and the feasibility of these strains for soil cadmium (Cd) remediation was investigated by soil incubation experiments and pot experiments. All endophytic bacteria were isolated from the roots of woody plants grown on Cd-contaminated soil. Seven endophytic bacteria strains had capacities to tolerate Cd toxicity and produce siderophores, and sequence analysis of the 16S rRNA gene classified these strains as belonging to the genera Burkholderia, Pseudomonas, Pantoea, and Herbaspirillum. All strains were able to produce hydroxamate siderophores (32.40%-91.49%) and had three or more plant growth promoting properties such as phosphorus solubilization, nitrogen fixation, indole acetic acid and 1-aminocyclopropane-1-carboxylate deaminase production. They were all strongly resistant to Cd2+ toxicity, with the minimum inhibitory concentration in LB medium ranging from 1.5 mM to 9.0 mM. Except for strain Burkholderia contaminans JLS17, other strains showed decreasing removal rates within continuously elevated Cd2+ concentration of 10-100 mg L-1. Compared with the uninoculated treatment, the inoculation of strains B.contaminans JLS17, Pseudomonas lurida JLS32, and Pantoea endophytica JLS50 effectively increased the concentration of acid-soluble Cd and decreased the concentration of reducible, oxidizable, and residual Cd in the soils of different Cd contamination levels. In pot experiments, inoculation of strains JLS17 and YTG72 significantly (p < 0.05) promoted the growth of above-ground parts and root system of slash pine (Pinus elliottii) under Cd stress. This study provides a valuable biological resource for endophyte-assisted phytoremediation and a theoretical basis for the application of endophytic bacteria for remediation of Cd-contaminated soil.

Sphingomonas abietis sp. nov., an Endophytic Bacterium Isolated from Korean Fir.

PAMB 00755T, a bacterial strain, was isolated from Korean fir leaves. The strain exhibits yellow colonies and consists of Gram-negative, non-motile, short rods or ovoid-shaped cells. It displays optimal growth conditions at 20°C, 0% NaCl, and pH 6.0. Results of 16S rRNA gene-based phylogenetic analyses showed that strain PAMB 00755T was most closely related to Sphingomonas chungangi MAH-6T (97.7%) and Sphingomonas polyaromaticivorans B2-7T (97.4%), and ≤96.5% sequence similarity to other members of the genus Sphingomonas. The values of average nucleotide identity (79.9-81.3%), average amino acid identity (73.3-75.9%), and digital DNA-DNA hybridization (73.3-75.9%) were significantly lower than the threshold values for species boundaries; these overall genome-related indexes (OGRI) analyses indicated that the strain represents a novel species. Genomic analysis revealed that the strain has a 4.4-Mbp genome encoding 4,083 functional genes, while the DNA G+C content of the whole genome is 66.1%. The genome of strain PAMB 00755T showed a putative carotenoid biosynthetic cluster responsible for its antioxidant activity. The respiratory quinone was identified as ubiquinone 10 (Q-10), while the major fatty acids in the profile were identified as C18:1ω7c and/or C18:1ω6c (summed feature 8). The major polar lipids of strain PAMB 00755T were diphosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid, and phosphatidylcholine. Based on a comprehensive analysis of genomic, phenotypic, and chemotaxonomic characteristics, we proposed the name Sphingomonas abietis sp. nov. for this novel species, with PAMB 00755T as the type strain (= KCTC 92781T = GDMCC 1.3779T).

The Zoige pioneer plant Leymus secalinus has different endophytic bacterial community structures to adapt to environmental conditions.

Background: Leymus secalinus is a pioneer plant grown in the Zoige desertified alpine grassland and it is also one of the dominant plant species used for environmental remediation. L. secalinus plays a large role in vegetation reconstruction in sandy land, but the abundance and diversity of its endophytes have not yet been investigated. Objectives: This study was performed to investigate the changes in the endophytic bacterial community structure of L. secalinus under different ecological environments and to analyze the effects of environmental changes and different plant tissues on the L. secalinus endophytic bacteria. Methods: Leaf, stem, and root tissue samples of L. secalinus were collected from Zoige Glassland (Alpine sandy land) and an open field nursery (Control). DNA was extracted and the 16S ribosomal DNA was amplified. The sequence library was sequenced on an Illumina MiSeq platform and clustered by operational taxonomic units (OTUs). α-diversity and ß-diversity analyses, species diversity analyses, functional prediction, and redundancy (RDA) analyses for the soil physicochemical properties were conducted. Results: α-diversity and ß-diversity analyses showed that the endophytic bacteria in L. secalinus varied in different areas and tissues. The abundance of Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, which is related to nitrogen fixation, increased significantly in the L. secalinus found in the Zoige Grassland.Moreover, the abundance of nutrition metabolism and anti-stress abilities increased in functional prediction in the desert samples. The soil physicochemical properties had an insignificant influence on bacterial diversity. Conclusion: The changes in the endophytic bacterial community structure in L. secalinus were significant and were caused by environmental alterations and plant choice. The endophytic bacteria in L. secalinus grown in alpine sandy land may have greater anti-stress properties and the ability to fix nitrogen, which has potential value in environmental remediation and agricultural production.

Ancylobacter mangrovi sp. nov., a novel endophytic bacterium isolated from mangrove plant.

Two novel strains GSK1Z-4-2T and MQZ15Z-1 were isolated from branches of mangrove plants collected from Guangxi Zhuang Autonomous Region, China. Both strains were Gram-negative, aerobic, non-flagellated and non-spore-forming bacteria. The comparison of 16S rRNA gene sequences initially indicated that the two strains were assigned to the genus Ancylobacter with sharing the highest similarity to Ancylobacter pratisalsi DSM 102029T (97.3%). The 16S rRNA gene sequence similarity, average nucleotide identity (ANI) and in silico DNA-DNA hybridization (isDDH) values between strains GSK1Z-4-2T and MQZ15Z-1 were 99.9%, 97.4% and 77.4%, respectively, which revealed that the two strains belonged to the same species. Phylogenetic analyses based on 16S rRNA gene sequences and the core proteome showed that the two strains formed a well-supported cluster with A. pratisalsi DSM 102029T. Moreover, the ANI and isDDH values between strain GSK1Z-4-2T and A. pratisalsi DSM 102029T were 83.0% and 25.8%, respectively, demonstrating that strain GSK1Z-4-2T was a previously undescribed species. Meanwhile, strains GSK1Z-4-2T and MQZ15Z-1 exhibited most of chemotaxonomic and phenotypic features consistent with the description of the genus Ancylobacter. Based on the polyphasic data, strains GSK1Z-4-2T and MQZ15Z-1 should represent a novel species of the genus Ancylobacter, for which the name Ancylobacter mangrovi sp. nov. is proposed. The type strain is GSK1Z-4-2T (=MCCC 1K07181T = JCM 34924T).

Paenibacillus mangrovi sp. nov., a novel endophytic bacterium isolated from bark of Kandelia candel.

A endospore-forming bacterium, designated strain KQZ6P-2T, was isolated from surface-sterilized bark of the mangrove plant Kandelia candel, collected from Maowei Sea Mangrove Nature Reserve in Guangxi Zhuang Autonomous Region, China. Strain KQZ6P-2T was able to grow at NaCl concentrations in the range of 0-3 % (w/v) with optimum growth at 0-1 % (w/v) NaCl. Growth occurred at 20-42 °C (optimal growth at 30-37 °C) and pH 5.5-6.5 (optimal growth at pH 6.5). The 16S rRNA gene sequence similarity between strain KQZ6P-2T and its closest phylogenetic neighbour Paenibacillus chibensis JCM 9905T was 98.2 %. Phylogenetic analyses using 16S rRNA gene sequences showed that strain KQZ6P-2T formed a distinct lineage with Paenibacillus chibensis JCM 9905T. The draft genome of strain KQZ6P-2T was 5 937 633 bp in size and its DNA G+C content was 47.2mol%. Comparative genome analysis revealed that the average nucleotide identity, digital DNA-DNA hybridization and average amino acid identity values among strain KQZ6P-2T and its related species were below the cut-off levels of 95, 70 and 95.5%, respec-tively. The cell-wall peptidoglycan of strain KQZ6P-2T contained meso-diaminopimelic acid as the diagnostic diamino acid. Major cellular fatty acids were anteiso-C15:0 and C16:0. The polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two unidentified aminophospholipids, four unidentified phospholipids, an unidentified aminolipid and five unidentified lipids. Based on phylogenetic, phenotypic and chemotaxonomic data, strain KQZ6P-2T represents a novel species of the genus Paenibacillus, for which the name Paenibacillus mangrovi sp. nov. is proposed. The type strain is KQZ6P-2T (=MCCC 1K07172T =JCM 34931T).

Allorhizobium sonneratiae sp. nov., an endophytic bacterium isolated from the root of Sonneratia apetala.

A novel endophytic bacterium, designated strain BGMRC 0089T, was isolated from a surface-sterilized root of Sonneratia apetala. Cells were observed to be Gram-negative, rod-shaped and motile with polar flagella. Strain BGMRC 0089T was found to grow optimally at 28-30 °C, pH 7.0-8.0 and in the presence of 1 % (w/v) NaCl. Strain BGMRC 0089T contained ubiquinone Q-10 and the predominant fatty acid was summed feature 8. The polar lipid profile of strain BGMRC 0089T was found to contain diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmonomethylethanolamine and phosphatidylethanolamine. Based on the results of 16S rRNA gene analysis, this isolate has the closest phylogenetic relationships with Rhizobium lemnae L6-16T (96.5 %) and Allorhizobium oryziradicis N19T (96.4 %). Average nucleotide identity, amino acid identity and digital DNA-DNA hybridization values of the isolate with the type strains of the genera Rhizobium and Allorhizobium were below 84.6, 73.9 and 22.1  %, respectively. Analysis the 4.55 Mb draft genome of strain BGMRC 0089T revealed several plant-associated genes, which may play important roles for the plant in the adaptation to the mangrove habitat. Based on its distinct phylogenetic, phenotypic and chemotaxonomic characteristics, strain BGMRC 0089T is proposed to represent a novel Allorhizobium species, for which the name Allorhizobium sonneratiae sp. nov. is proposed (type strain BGMRC 0089T=DSM 100171T=MCCC 1K04805T).

Endophytic Bacterium Lysobacter firmicutimachus Strain 5-7 Is a Promising Biocontrol Agent Against Rice Seedling Disease Caused by Pythium arrhenomanes in Nursery Trays.

Rice root rot disease caused by Pythium spp. is a highly destructive disease in rice nurseries. Biocontrol with endophytic bacteria was developed in this study to control rice seedling diseases. An in planta screening assay revealed that two bacterial endophytes, strains 5-7 and 6-4, displayed strong protection of rice seedlings from attack by Pythium arrhenomanes. Phylogenetic analysis indicated that strain 5-7 is Lysobacter firmicutimachus, while strain 6-4 belongs to the Kitasatospora genus. To quickly evaluate the disease severity of the root system damaged by Pythium spp. in nursery trays, a root surface area measurement assay was developed. By using this measurement, the control efficacy in nursery trays was evaluated, and L. firmicutimachus 5-7 showed promising biocontrol activity against Pythium disease. In a field trial, the two endophytes exhibited significant disease control efficacy on rice brown spot disease caused by Bipolaris oryzae naturally occurring in a commercial nursery field. The two endophytes exhibited multiple enzymatic activities and broad-spectrum antagonistic activities against multiple rice pathogens. The two endophytes colonized the root surface and inside of the root. L. firmicutimachus 5-7 primarily colonized the intercellular space and aerenchyma. Antibiosis is the major mechanism used by strain 5-7 to cause Bipolaris hyphal swelling and inhibit Pythium zoospore germination and sporangium formation, while a hyperparasitism-like phenomenon was found in the interaction of strain 6-4 with Pythium and Bipolaris hyphae. In conclusion, we report the promising biocontrol agent L. firmicutimachus 5-7 and the potential biocontrol agent Kitasatospora sp. 6-4 for disease control of rice seedlings in commercial nursery trays and their possible mechanisms of action.

Jiella avicenniae sp. nov., a novel endophytic bacterium isolated from bark of Avicennia marina.

A Gram-stain-negative and short rod-shaped strain CBK1P-4T, isolated from surface-sterilized bark of Avicennia marina was investigated by a polyphasic taxonomic approach to resolve its taxonomic position. Strain CBK1P-4T grew at 10-30 °C (optimum, 25 °C), pH 5.0-9.0 (optimum, pH 5.5) and in the presence of 0-9% (w/v) NaCl (optimum, 1-2%). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain CBK1P-4T belonged to the genus Jiella and was most closely related to species of the genus Jiella (97.4-98.3%). The genome comparisons between strain CBK1P-4T and the closely related species indicated that average nucleotide identity and digital DNA-DNA hybridization values were below the recommended thresholds for assigning strains to the same species (95-96% and 70%, respectively). The cell wall peptidoglycan contained meso-diaminopimelic acid as diagnostic diamino acid. The principal fatty acids were C18:1ω7c and C19:0cycloω8c. The polar lipids were mainly comprised of phosphatidylcholine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, two unidentified aminolipids, one unidentified phospholipid and one unidentified glycolipid. The dominant respiratory quinone was ubiquinone-10. The DNA G + C content of strain CBK1P-4T was 66.7%. Based on the phenotypic features, phylogenetic analysis as well as genome analysis, we conclude that strain CBK1P-4T represents a novel Jiella species, for which the name Jiella avicenniae sp. nov. is proposed. The type strain is CBK1P-4T (= CGMCC 1.18742T = JCM 34330T).

[Colonization performance and pyrene degradation characteristics of Stenotrophomonas maltophilia PX1]. / å—œéº¦èŠ½å¯¡å »å•èƒžèŒPX1的降解芘特性及定殖效能.

We screened and identified an endophytic bacterium that could efficiently degrade PAHs, which would expand the library of polycyclic aromatic hydrocarbons (PAHs) degrading microorganisms and reduce the pollution risk of crops. Its degradation mechanism and colonization performance were preliminarily examined. The results showed that strain PX1 belonged to Stenotrophomonas maltophilia. The strain had broad spectrum ability to remove PAHs. In PAH mineral salt (MS) media, almost 100% naphthalene was degraded by strain PX1 after 7-d incubation. In a cultivation system solely containing phenanthrene of 50.0 mg·L-1, pyrene of 20.0 mg·L-1, fluoranthene of 20.0 mg·L-1 or benzo[a]pyrene of 10.0 mg·L-1, the degradation efficiency of phenanthrene, pyrene, fluoranthene and benzo[a]pyrene by strain PX1 reached 72.6%, 50.7%, 31.9%, and 12.9%, respectively. Pyrene was selected as PAHs model to study the degradation characteristics of strain PX1. Enzyme activity tests showed that the activities of phthalate dioxygenase, catechol-1,2-dioxygenase, and catechol-2,3-dioxygenase in strain PX1 were induced by pyrene. Some metabolic intermediates such as 4,5-epoxypyrene, 4,5-dihydroxypyrene, gentilic acid/protocatechuic acid, salicylic acid, cis-hexadienedioic acid/2-hydroxymyxofuroic acid semialdehyde, cis-2'-carboxyphenylpyruvic acid, 1-hydroxy-2-naphthoic acid, and salicylaldehyde were detected during the degradation of pyrene by strain PX1. Results of the seed soaking experiment showed that strain PX1 could efficiently colonize in Ipomoea aquatic and Triticum aestivum. After inoculated with strain PX1, the growth of I. aquatic and T. aestivum was significantly increased, and the pyrene concentration in I. aquatic, T. aestivum and MS media was reduced by 29.8%-50.7%, 52.4%-67.1% and 8.0%-15.3%, respectively. Our results suggested that strain PX1 degraded pyrene mainly through 'salicylate pathway' and 'phthalate pathway', and could be colonized into plants and promote plant growth.

Stakelama flava sp. nov., a novel endophytic bacterium isolated from a branch of Kandelia candel.

A Gram-stain-negative, aerobic, motile, non-spore-forming, short-rod-shaped strain that did not produce diffusible pigment, designated CBK3Z-3T, was isolated from a branch of Kandelia candel, collected from the Beilun Estuary National Nature Reserve in Guangxi Zhang Autonomous Region, PR China, and investigated by a polyphasic approach to determine its taxonomic position. Strain CBK3Z-3T grew at pH 5.0-10.0 (optimum, pH 8.0), 20-37 °C (optimum, 25-30 °C) and with 0-10 % (w/v) NaCl (optimum, 2-3 %). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain CBK3Z-3T was closely related to species of genus Stakelama and had the highest 16S rRNA gene sequence similarity of 98.7 % to Stakelama pacifica CGMCC 1.7294T. The DNA G+C content value of strain CBK3Z-3T was 62.6 mol%. The diagnostic diamino acid in the cell-wall peptidoglycan was meso-diaminopimelic acid and the polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, sphingoglycolipid, an unidentified aminolipid and an unidentified lipid. The major fatty acids were C18 : 1 ω7c and C16 : 0. The average nucleotide identity, estimated digital DNA-DNA hybridization and average amino acid identity values between strain CBK3Z-3T and the type strain of Stakelama pacifica CGMCC 1.7294T were 80.4, 23.1 and 81.5 %, respectively. Based on the phylogenetic, phenotypic and chemotaxonomic data, strain CBK3Z-3T should be designated as a novel species of the genus Stakelama, for which the name Stakelama flava sp.nov. is proposed. The type strain is CBK3Z-3T (=JCM 34534T=CGMCC 1.18972T).

Pyomelanin-Producing Brevundimonas vitisensis sp. nov., Isolated From Grape (Vitis vinifera L.).

A novel endophytic bacterial strain, designated GR-TSA-9T, was isolated from surface-sterilized grape (Vitis vinifera L.). 16S rRNA gene sequence analyses showed that the isolate was grouped within the genus Brevundimonas, displaying the highest similarity with Brevundimonas lenta DS-18T (97.9%) and Brevundimonas kwangchunensis KSL-102T (97.8%) and less than 97.5% similarity with other members of Brevundimonas. The strain GR-TSA-9T was a gram negative, rod shaped, facultatively anaerobic, catalase and oxidase positive, and motile bacterium. Its growth occurred at 10-37°C (optimally 25-30°C), at pH 7.0-8.0, and in NaCl 0-1% (optimally 0%). It contained ubiquinone-10 as a respiratory quinone, and the major cellular fatty acids (>10% of the total) were C16:0 (14.2%) and summed feature 8 (C18:1ω7c and/or C18:1ω6c, 65.6%). The polar lipids present in the strain were phosphoglycolipids, phosphatidylglycerol, 1,2-di-O-acyl-3-O-[d-glucopyranosyl-(1→4)-α-d-glucopyranuronosyl]glycerol, and unidentified lipids (L1, L2, and L4). The strain had one 2,976,716bp circular chromosome with a G+C content of 66.4%. The digital DNA-DNA hybridization value between strain GR-TSA-9T and B. lenta DS-18T was 20.9%, while the average nucleotide identity value was 76.7%. In addition, the dDDH and ANI values to other members in this genus, whose genome sequences are available, are less than 21.1 and 77.6%. Genome annotation predicted the presence of some gene clusters related to tyrosine degradation and pyomelanin formation. Strain GR-TSA-9T produced a brown melanin-like pigment in the presence of L-tyrosine-containing media. The highest pigment production (0.19g/L) was observed in tryptic soy broth with 1.0mg/ml L-tyrosine at 25°C for 6days of culture. Biophysical characterization by ultraviolet (UV)-visible spectroscopy, Fourier-transform infrared spectroscopy, and electrospray ionization mass spectrometry confirmed that the pigment was pyomelanin. Additionally, melanized GR-TSA-9T cells could protect the cells against UVC exposure. The phylogenetic, genomic, phenotypic, and chemotaxonomic features indicated that strain GR-TSA-9T represents a novel melanin-producing species of Brevundimonas. The type strain was GR-TSA-9T (KCTC 82386T=CGMCC 1.18820T).

Pseudooceanicola endophyticus sp. nov., a novel endophytic bacterium isolated from bark Aegiceras corniculatum.

A Gram-stain-negative, aerobic, short-rod-shaped bacterium, designated strain CBS1P-1T, was isolated from a surface-sterilized bark of Aegiceras corniculatum. Growth of strain CBS1P-1T was observed with between 0 and 12.0 % (w/v) NaCl (optimally with 5.0 %) and at between pH 6.0-9.0. It grew at temperatures between 25-37 °C (optimum, 30 °C). Chemotaxonomic analysis showed that ubiquinone-10 was the respiratory quinone. The lipids comprised diphosphatidylglycerol, phosphatidylglycerol, an unidentified glycolipid, an unidentified phospholipid and an unidentified aminolipid. The major fatty acids of strain CBS1P-1T were C18 : 1 ω7c, C16 : 0 and C19 : 0 cyclo ω8c. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain CBS1P-1T was most related to Pseudooceanicola antarcticus CGMCC 1.12662T with a sequence similarity of 96.5 %. The average nucleotide identity and digital DNA-DNA hybridization values between strain CBS1P-1T and P. antarcticus 1.12662T were 77.5 and 21.1 %, respectively. The G+C content of the genomic DNA was 67.3 mol%. Based on phylogenetic, chemotaxonomic and phenotypic data, strain CBS1P-1T is considered to represent a novel species of the genus Pseudooceanicola, for which the name Pseudooceanicola endophyticus is proposed. The type strain is CBS1P-1T (=KCTC 62836T=CGMCC 1.13743T).

Jiella mangrovi sp. nov., a novel endophytic bacterium isolated from leaf of Rhizophora stylosa.

A Gram-stain negative, non-motile, brilliant yellow and non-spore forming, coccoid- or short rod-shaped bacterium, designated strain KSK16Y-1T, was isolated from surface-sterilised leaf of Rhizophora stylosa collected from Shankou Mangrove Nature Reserve, Guangxi Zhuang Autonomous Region, China. Genome of strain KSK16Y-1T is 4.93 Mb with 68.1% DNA G + C content and encoded 4359 predicted proteins, 4 rRNAs, 45 tRNAs and 4 ncRNA. Comparative 16S rRNA gene sequence analysis showed that the strain KSK16Y-1T has 98.1%, 97.9% and 96.9% 16S rRNA gene similarities with Jiella aquimaris JCM 30119T, J. endophytica CBS5Q-3T and J. pacifica 40Bstr34T, respectively. Whole-genome comparisons between strain KSK16Y-1T and J. aquimaris 22II-16-19i, J. endophytica CBS5Q-3T, J. pacifica 40Bstr34T, using average nucleotide identity (ANI) values (< 82.0%) and digital DNA-DNA hybridization (dDDH) values (< 25.1%), confirmed low genome relatedness. Strain KSK16Y-1T grew at 20-30 °C (optimum, 30 °C), pH 6.0-11.0 (optimum, pH 6.0-7.0) and with 0-10% (w/v) NaCl (optimum, 0-2%). Cell wall contained meso-diaminopimelic acid and the major fatty acid was C18:1ω7c. The polar lipid profile consists of phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylmonomethylethanolamine, one unknown phospholipid, one unknown aminolipid, one unknown aminophospholipid and four unidentified lipids. The predominant respiratory quinone is ubiquinone-10 (Q-10). The polyphasic characterization indicated that strain KSK16Y-1T represents a novel Jiella species. The name Jiella mangrovi sp. nov., type strain KSK16Y-1T (= CGMCC 1.18745T = JCM 34332T) is proposed.

Metabolic pathway analysis of walnut endophytic bacterium Bacillus subtilis HB1310 related to lipid production from fermentation of cotton stalk hydrolysate based on genome sequencing.

OBJECTIVES: In this study, genome sequencing and metabolic analysis were used to identify and verify the key metabolic pathways for glucose and xylose utilization and fatty acid synthesis in the walnut endophytic bacterium (WEB) Bacillus subtilis HB1310. RESULTS: The genome sequence of WEB HB1310 was generated with a size of 4.1 Mb and GC content of 43.5%. Genome annotation indicated that the Embden-Meyerhof-Parnas, pentose phosphate, and fatty acid synthesis pathways were mainly involved in mixed sugar utilization and lipid production. In particular, diverse and abundant fatty acid synthesis genes were observed in a higher number than in other Bacillus strains. The tricarboxylic acid cycle competitively shared the carbon flux flowing before 48 h, and the acetic acid fermentation competed after 72 h. Moreover, fatty acid synthase activity was highly correlated with lipid titer with a correlation coefficient of 0.9626, and NADPH might be more utilized for the lipid synthesis within 48 h. CONCLUSIONS: This study is the first attempt to explain the metabolic mechanism of mixed sugar utilization and lipid production based on genomic information, which provides a theoretical basis for the metabolic regulation of bacterial lipid production from lignocellulosic hydrolysates.

Characterization, Antimicrobial and Antioxidant Evaluation of Biofabricated Silver Nanoparticles from Endophytic Pantoea anthophila.

Endophyte mediated nanoparticles fabrication were emerging as a new frontier in nanomedicines that produce high biocompatible and functionalized silver nanoparticles. In this study, silver nanoparticles were successfully biosynthesized from the extracellular extract of endophytic bacterium Pantoea anthophila isolated from the stem of Waltheria indica for the first time. The synthesized nanoparticles showed a strong absorption band at 410 nm in the UV-Visible range. The dynamic light scattering and zeta potential analysis indicated that the average particle size was 16 nm at 5.30 mV. FTIR spectrum displayed the presence of various functional groups at 3423.65, 1633.71, 1022.27, 607.58 cm-1 that stabilised the nanoparticle. X-ray diffraction peaks were conferred to 100, 200, 220 and 311 planes of a face centred cubic structure. TEM and SEM micrograph revealed the spherical-shaped, polycrystalline nature with the presence of elemental silver analysed by EDAX. Selected area electron diffraction also confirms the orientation of silver nanoparticles with X-ray diffraction analysis. Antimicrobial activity against 10 different human pathogenic bacteria and fungi showed a broad spectrum inhibition against both Gram-positive and Gram-negative bacteria. Among the bacterial pathogens, B. Subtilis exhibited low activity compared to other pathogens. C. albicans was greatly controlled than other fungal species. A strong free radical scavenging activity of silver nanoparticles with IC50 values 31.29 ± 0.73, 19.83 ± 1.57, 35.64 ± 0.94, 42.07 ± 1.30, 29.70 ± 2.26, 29.10 ± 0.82, 36.80 ± 0.63 µg/ml was obtained in different antioxidant assays that were comparable to the reference. The study suggests that the silver nanoparticles can be biosynthesized from endophytic P. anthophila metabolites with significant therapeutic potential. With proper validation, the biosynthesized silver nanoparticles can be developed as a promising antiviral and anticancer drug candidate.

Herbaspirillum camelliae sp. nov., a novel endophytic bacterium isolated from Camellia sinensis L.

Bacterial strain WT00CT is an endophytic bacterium that was isolated from the tea plant (Camellia sinensis L.). The phylogenetic analysis of 16S rRNA genes demonstrated that strain WT00CT was a member of the genus Herbaspirillum. This strain is microaerobic, gram-negative and non-pigmented, and its cells are rod shaped, with a polar flagellum. It grew optimally at 34-37 °C, pH 5.0-8.0 and 0-1.5% NaCl (w/v). The G + C content of its genomic DNA was 62.36 mol%. C16:0, iso-C15:0, iso-C17:0, anteiso-C15:0 and anteiso-C17:0 were major fatty acids. The strain WT00CT contained six polar lipids, namely DPG (diphosphatidylglycerol), PE (phosphatidylethanolamine), PG (phosphatidylglycerol), PC (phosphatidylcholine), GL (glycolipid) and APL (aminophospholipids), and its respiratory quinone was Q8. The strain WT00CT had a genome size of 6.08 Mb with a total ORF of 5,537, in which one gene cluster (36 genes) encoding a type IV secretion system was absent in other members of the Herbaspirillum genus. ANI values of genomic comparison between the strain WT00CT and other Herbaspirillum species were 75-96%. Based on the phylogenetic, chemotaxonomic and phenotypic data presented here, the strain WT00CT represents a novel species in the Herbaspirillum genus, for which the name Herbaspirillum camelliae sp. nov. is proposed. The type strain of H. camelliae sp. nov. is WT00CT (AB 2018017 T and KCTC 62527 T).