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1.
Curr Microbiol ; 77(8): 1945-1954, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32350604

RESUMO

The genus Metakosakonia, as the closest phylogenetic neighbor of the genus Kosakonia within the family Enterobacteriaceae, when proposed in 2017, consisted of M. massiliensis JC163T and Metakosakonia spp. strains CAV1151 and GT-16. The strain CAV1151 was later classified into a novel species Phytobacter ursingii. Here, we show that the strain GT-16 shares a digital DNA-DNA hybridization (DDH) similarity of 91.0% with P. diazotrophicus DSM 17806 T and thus also belongs to P. diazotrophicus. M. massiliensis and the strains within the genus Phytobacter formed a monophyletic cluster on a phylogenomic tree based on the core proteins of the family Enterobacteriaceae and on a phylogenetic tree based on the 16S rRNA genes. M. massiliensis and the genus Phytobacter share average amino acid identities of 86.80‒87.41% above the threshold (86%) for genus delimitation within the family Enterobacteriaceae. Moreover, they share conserved signature indels in the intracellular growth protein IgaA and the outer membrane assembly protein AsmA. Therefore, we propose to unite the genus Metakosakonia and the genus Phytobacter to a single genus. Because the genus Phytobacter was validly published earlier in 2017 than the genus Metakosakonia in 2017, the genus name Phytobacter has priority over Metakosakonia. We propose to unite the two genera under the name Phytobacter with the type species P. diazotrophicus and reclassify M. massiliensis as P. massiliensis comb. nov. In addition, the analyses of genome relatedness and phylogenomic relationship identified one potential novel species within the genus Phytobacter and three potential novel species within the genus Kosakonia.


Assuntos
Enterobacteriaceae/classificação , Gammaproteobacteria/classificação , Filogenia , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Tipagem de Sequências Multilocus , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
2.
Int J Syst Evol Microbiol ; 69(7): 1986-1992, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31046894

RESUMO

A Gram-stain-negative, aerobic, nitrogen-fixing bacterium, designated strain L461T, was isolated from leaves of Bryophyllum pinnatum growing at the South China Agricultural University. Phylogenetic analysis of the 16S rRNA gene sequence indicated it as a member of the genus Azotobacter closely related to Azotobacter beijerinckii JCM 20725T (97.82 % similarity) and Azotobacter chroococcum ATCC 9043T (97.34 %). Its major fatty acid components were C16 : 1 ω9c and C16 : 0. Its predominant isoprenoid quinone was Q-9. Its major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, aminophospholipid, phospholipid and one unknown lipid. Its DNA G+C content was 64.9 mol% (Tm). DNA-DNA relatedness values between strain L461T and the reference strains of A. beijerinckii and A. chroococcum were 46.43 and 28.23 %, respectively. Biological and biochemical tests, protein patterns, genomic DNA fingerprinting, and comparison of cellular fatty acids distinguished strain L461T from the closely related Azotobacter species. Based on these data, the novel species Azotobacter bryophylli sp. nov. is proposed, with the type strain L461T (=KCTC 62195T=GDMCC 1.1250T).


Assuntos
Azotobacter/classificação , Kalanchoe/microbiologia , Filogenia , Azotobacter/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , China , Impressões Digitais de DNA , DNA Bacteriano/genética , Ácidos Graxos/química , Hibridização de Ácido Nucleico , Fosfolipídeos/química , Folhas de Planta/microbiologia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ubiquinona/química
3.
Molecules ; 24(12)2019 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-31234369

RESUMO

Bacterial stem and root rot disease of sweet potato caused by Dickeya dadantii recently broke out in major sweet potato planting areas in China and calls for effective approaches to control the pathogen and disease. Here, we developed a simple method for green synthesis of silver nanoparticles (AgNPs) using bacterial culture supernatants. AgNPs synthesized with the cell-free culture supernatant of a bacterium Pseudomonas rhodesiae displayed the characteristic surface plasmon resonance peak at 420-430 nm and as nanocrystallites in diameters of 20-100 nm determined by transmission electron microscopy, scanning electron microscopy, and X-ray diffraction spectroscopy. Functional groups associated with proteins in the culture supernatant may reduce silver ions and stabilize AgNPs. The AgNPs showed antibacterial activities against D. dadantii growth, swimming motility, biofilm formation, and maceration of sweet potato tubers whereas the culture supernatant of P. rhodesiae did not. AgNPs (12 µg∙ml-1) and AgNO3 (50 µg∙ml-1) showed close antibacterial activities. The antibacterial activities increased with the increase of AgNP concentrations. The green-synthesized AgNPs can be used to control the soft rot disease by control of pathogen contamination of sweet potato seed tubers.


Assuntos
Meios de Cultura/farmacologia , Enterobacteriaceae/efeitos dos fármacos , Nanopartículas Metálicas/química , Pseudomonas/química , Antibacterianos , China , Meios de Cultura/química , Enterobacteriaceae/patogenicidade , Enterobacteriaceae/ultraestrutura , Química Verde , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Prata/química , Espectroscopia de Infravermelho com Transformada de Fourier , Ressonância de Plasmônio de Superfície
4.
J Basic Microbiol ; 56(1): 78-84, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26471769

RESUMO

It was difficult to differentiate Klebsiella pneumoniae, K. quasipneumoniae and K. variicola by biochemical and phenotypic tests. Genomics increase the resolution and credibility of taxonomy for closely-related species. Here, we obtained the complete genome sequence of the K. variicola type strain DSM 15968(T) (=F2R9(T)). The genome of the type strain is a circular chromosome of 5,521,203 bp with 57.56% GC content. From 540 Klebsiella strains whose genomes had been publicly available as at 3 March 2015, we identified 21 strains belonging to K. variicola and 8 strains belonging to K. quasipneumoniae based on the genome average nucleotide identities (ANI). All the K. variicola strains, one K. pneumoniae strain and five K. quasipneumoniae strains contained nitrogen-fixing genes. A phylogenomic analysis showed clear species demarcations for these nitrogen-fixing bacteria. In accordance with the key biochemical characteristics of K. variicola, the idnO gene encoding 5-keto-D-gluconate 5-reductase for utilization of 5-keto-D-gluconate and the sorCDFBAME operon for catabolism of L-sorbose were present whereas the rbtRDKT operon for catabolism of adonitol was absent in the genomes of K. variicola strains. Therefore, the genomic analyses supported the ANI-based species delineation; the genome sequence of the K. variicola type strain provides the reference genome for genomic identification of K. variicola, which is a nitrogen-fixing species.


Assuntos
Genoma Bacteriano , Klebsiella/genética , Klebsiella/metabolismo , Fixação de Nitrogênio , Proteínas de Bactérias/genética , Sequência de Bases , Mapeamento Cromossômico , DNA Bacteriano/genética , Gluconatos/metabolismo , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/metabolismo , Nitrogênio/metabolismo , Nitrogenase/genética , Nitrogenase/metabolismo , Óperon , Oxirredutases/genética , Filogenia , Reação em Cadeia da Polimerase , Análise de Sequência de DNA
5.
J Basic Microbiol ; 56(8): 934-40, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27059698

RESUMO

N2 -fixing bacteria belonging to the genus Raoultella of the family Enterobacteriaceae are widely associated with plants. Raoultella sp. strain L03 was isolated from surface-sterilized sugarcane roots. In this study, we inoculated the strain L03 to microbe-free micropropagated plantlets of the main sugarcane cultivar ROC22 grown in Guangxi, China and determined N2 -fixation and association between strain L03 and sugarcane plants. Inoculation of strain L03 increased plant biomass, total N, N concentration and chlorophyll, and relieved N-deficiency symptoms of plants under an N-limiting condition. An (15) N isotope dilution assay revealed (15) N isotope dilution in the inoculated sugarcane plants and incorporation of the fixed (14) N from air into chlorophyll. Moreover, a gfp-tagged and antibiotic-resistant L03 strain was reisolated from surface-sterilized sugarcane plants and was detected in plant tissues by fluorescent microscopy. This study for the first time demonstrates that a Raoultella bacterium is able to fix N2 in association with the plant host.


Assuntos
Endófitos/metabolismo , Enterobacteriaceae/metabolismo , Fixação de Nitrogênio/fisiologia , Saccharum/crescimento & desenvolvimento , Saccharum/metabolismo , Biomassa , Endófitos/isolamento & purificação , Enterobacteriaceae/isolamento & purificação , Raízes de Plantas/microbiologia , Saccharum/microbiologia , Simbiose
6.
Chemosphere ; 356: 141904, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38582174

RESUMO

Rice blast, an extremely destructive disease caused by the filamentous fungal pathogen Magnaporthe oryzae, poses a global threat to the production of rice (Oryza sativa L.). The emerging trend of reducing dependence on chemical fungicides for crop protection has increased interest in exploring bioformulated nanomaterials as a sustainable alternative antimicrobial strategy for effectively managing plant diseases. Herein, we used physiomorphological, transcriptomic, and metabolomic methods to investigate the toxicity and molecular action mechanisms of moringa-chitosan nanoparticles (M-CNPs) against M. oryzae. Our results demonstrate that M-CNPs exhibit direct antifungal properties by impeding the growth and conidia formation of M. oryzae in a concentration-dependent manner. Propidium iodide staining indicated concentration-dependent significant apoptosis (91.33%) in the fungus. Ultrastructural observations revealed complete structural damage in fungal cells treated with 200 mg/L M-CNPs, including disruption of the cell wall and destruction of internal organelles. Transcriptomic and metabolomic analyses revealed the intricate mechanism underlying the toxicity of M-CNPs against M. oryzae. The transcriptomics data indicated that exposure to M-CNPs disrupted various processes integral to cell membrane biosynthesis, aflatoxin biosynthesis, transcriptional regulation, and nuclear integrity in M. oryzae., emphasizing the interaction between M-CNPs and fungal cells. Similarly, metabolomic profiling demonstrated that exposure to M-CNPs significantly altered the levels of several key metabolites involved in the integral components of metabolic pathways, microbial metabolism, histidine metabolism, citrate cycle, and lipid and protein metabolism in M. oryzae. Overall, these findings demonstrated the potent antifungal action of M-CNPs, with a remarkable impact at the physiological and molecular level, culminating in substantial apoptotic-like fungal cell death. This research provides a novel perspective on investigating bioformulated nanomaterials as antifungal agents for plant disease control.


Assuntos
Quitosana , Nanopartículas , Oryza , Doenças das Plantas , Transcriptoma , Quitosana/química , Nanopartículas/toxicidade , Nanopartículas/química , Transcriptoma/efeitos dos fármacos , Oryza/microbiologia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Metabolômica , Antifúngicos/toxicidade , Antifúngicos/farmacologia , Ascomicetos/efeitos dos fármacos , Ascomicetos/genética
7.
J Hazard Mater ; 465: 133266, 2024 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-38118201

RESUMO

The negative impacts of soil heavy metals composite pollution on agricultural production and human health are becoming increasingly prevalent. The applications of green chelating agents and microorganisms have emerged as promising alternate methods for enhancing phytoremediation. The regulatory effects of root secretion composition, microbial carbon source utilization, key gene expression, and soil microbial community structure were comprehensively analyzed through a combination of HPLC, Biolog EcoPlates, qPCR, and high-throughput screening techniques. The application of EDDS resulted in a favorable rhizosphere ecological environment for the king grass Piriformospora indica, characterized by a decrease in soil pH by 0.41 units, stimulation of succinic acid and fumaric acid secretion, and an increase in carbon source metabolic activity of amino acids and carbohydrates. Consequently, this improvement enhanced the bioavailability of Cd/Cr and increased the biomass of king grass by 25.7%. The expression of dissimilatory iron-reducing bacteria was significantly upregulated by 99.2%, while there was no significant difference in Clostridium abundance. Furthermore, the richness of the soil rhizosphere fungal community (Ascomycota: 45.8%, Rozellomycota: 16.7%) significantly increased to regulate the proportion of tolerant microbial dominant groups, promoting the improvement of Cd/Cr removal efficiency (Cd: 23.4%, Cr: 18.7%). These findings provide a theoretical basis for the sustainable development of chelating agent-assisted plants-microorganisms combined remediation of heavy metals in soil.


Assuntos
Basidiomycota , Metais Pesados , Poluentes do Solo , Humanos , Cádmio/metabolismo , Solo/química , Poaceae/metabolismo , Rizosfera , Poluentes do Solo/metabolismo , Metais Pesados/análise , Microbiologia do Solo , Quelantes , Carbono , Biodegradação Ambiental
8.
Carbohydr Polym ; 334: 122023, 2024 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-38553222

RESUMO

Rice blast disease (RBD) caused by Magnaporthe oryzae, threaten food security by cutting agricultural output. Nano agrochemicals are now perceived as sustainable, cost-effective alternatives to traditional pesticides. This study investigated bioformulation of moringa chitosan nanoparticles (M-CsNPs) and their mechanisms for suppressing RBD while minimizing toxic effects on the microenvironment. M-CsNPs, sized 46 nm with semi-spherical morphology, significantly suppressed pathogen growth, integrity, and colonization at 200 mg L-1in vitro. Greenhouse tests with foliar exposure to the same concentration resulted in a substantial 77.7 % reduction in RBD, enhancing antioxidant enzyme activity and plant health. Furthermore, M-CsNPs improved photosynthesis, gas exchange, and the nutritional profile of diseased rice plants. RNA-seq analysis highlighted upregulated defense-related genes in treated rice plants. Metagenomic study showcased reshaping of the rice microbiome, reducing Magnaporthe abundance by 93.5 %. Both healthy and diseased rice plants showed increased microbial diversity, particularly favoring specific beneficial species Thiobacillus, Nitrospira, Nocardioides, and Sphingomicrobium in the rhizosphere and Azonexus, Agarivorans, and Bradyrhizobium in the phyllosphere. This comprehensive study unravels the diverse mechanisms by which M-CsNPs interact with plants and pathogens, curbing M. oryzae damage, promoting plant growth, and modulating the rice microbiome. It underscores the significant potential for effective plant disease management.


Assuntos
Quitosana , Microbiota , Oryza , Resistência à Doença , Oryza/genética , Quitosana/farmacologia , Bactérias , Doenças das Plantas/prevenção & controle
9.
Int J Syst Evol Microbiol ; 63(Pt 7): 2577-2582, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23291881

RESUMO

Five nitrogen-fixing bacterial strains (SP1(T), NN143, NN144, NN208 and HX148) were isolated from stem, root or rhizosphere soil of sugar cane (Saccharum officinarum L.) plants. Cells were Gram-negative, motile, rods with peritrichous flagella. DNA G+C content was 55.0 ± 0.5 mol%. Sequence determinations and phylogenetic analysis of 16S rRNA gene and rpoB indicated that the strains were affiliated with the genus Enterobacter and most closely related to E. radicincitans DSM 16656(T) and E. oryzae LMG 24251(T). Fluorimetric determination of thermal denaturation temperatures after DNA-DNA hybridization, enterobacterial repetitive intergenic consensus PCR and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry differentiated the whole-genome, genotype and protein profiles from those of E. radicincitans and E. oryzae. The strains' cell fatty acid composition differentiated them from E. radicincitans and E. oryzae by containing a higher level of summed feature 2 (C16 : 1ω7c and/or C16 : 1ω6c) and a lower level of C17 : 0 cyclo. Their physiological and biochemical profiles differentiated them from E. radicincitans by being positive for methyl red test, ornithine decarboxylase and utilization of putrescine, D-arabitol, L-fucose and methyl α-D-glucoside and being negative for arginine dihydrolase, and differentiated them from E. oryzae by being positive for aesculin hydrolysis and utilization of putrescine, D-arabitol and L-rhamnose and being negative for arginine dihydrolase, lysine decarboxylase and utilization of mucate. The five strains therefore represent a novel species, for which the name Enterobacter sacchari sp. nov. is proposed, with the type strain SP1(T) ( = CGMCC 1.12102(T) = LMG 26783(T)).


Assuntos
Enterobacter/classificação , Fixação de Nitrogênio , Filogenia , Saccharum/microbiologia , Proteínas de Bactérias/análise , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Enterobacter/genética , Enterobacter/isolamento & purificação , Ácidos Graxos/análise , Genes Bacterianos , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Raízes de Plantas/microbiologia , Caules de Planta/microbiologia , RNA Ribossômico 16S/genética , Rizosfera , Análise de Sequência de DNA
10.
Microorganisms ; 11(4)2023 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-37110240

RESUMO

Polymyxin-producing bacteria within the Paenibacillus polymyxa complex have broad-spectrum activities against fungi and bacteria. Their antibacterial activities against soft rot Dickeya and Pectobacterium phytopathogens containing multiple polymyxin-resistant genes were not clear. Here, we selected nine strains within the P. polymyxa complex having broad-spectrum antagonistic activities against phytopathogenic fungi and a polymyxin-resistant D. dadantii strain causing stem and root rot disease of sweet potato and did antagonistic assays on nutrient agar and sweet potato tuber slices. These strains within the P. polymyxa complex showed clear antagonistic activities against D. dadantii in vitro and in vivo. The most effective antagonistic strain P. polymyxa ShX301 showed broad-spectrum antagonistic activities against all the test Dickeya and Pectobacterium strains, completely eliminated D. dadantii from sweet potato seed tubers, and promoted the growth of sweet potato seedlings. Cell-free culture filtrate of P. polymyxa ShX301 inhibited D. dadantii growth, swimming motility, and biofilm formation and disrupted D. dadantii plasma membranes, releasing nucleic acids and proteins. Multiple lipopeptides produced by P. polymyxa ShX301 may play a major role in the bactericidal and bacteriostatic actions. This study clarifies that the antimicrobial spectrum of polymyxin-producing bacteria within the P. polymyxa complex includes the polymyxin-resistant Dickeya and Pectobacterium phytopathogens and strengthens the fact that bacteria within the P. polymyxa complex have high probability of being effective biocontrol agents and plant growth promoters.

11.
Plants (Basel) ; 12(9)2023 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-37176882

RESUMO

Nanomaterials are increasingly being used for crop growth, especially as a new paradigm for plant disease management. Among the other nanomaterials, silver nanoparticles (AgNPs) draw a great deal of attention because of their unique features and multiple usages. Rapid expansion in nanotechnology and utilization of AgNPs in a large range of areas resulted in the substantial release of these nanoparticles into the soil and water environment, causing concern for the safety of ecosystems and phytosanitary. In an attempt to find an effective control measure for sweet potato soft rot disease, the pathogen Dickeya dadantii was exposed to AgNPs, the cell-free culture supernatant (CFCS) of Bacillus amyloliquefaciens alone, and both in combination. AgNPs were synthesized using CFCS of Bacillus amyloliquefaciens strain A3. The green synthesized AgNPs exhibited a characteristic surface plasmon resonance peak at 410-420 nm. Electron microscopy and X-ray diffraction spectroscopy determined the nanocrystalline nature and 20-100 nm diameters of AgNPs. Release of metal Ag+ ion from biosynthesized AgNPs increases with time. AgNPs and CFCS of B. amyloliquefaciens alone exhibited antibacterial activity against the growth, biofilm formation, swimming motility, and virulence of strain A3. The antibacterial activities elevated with the elevation in AgNPs and CFCS concentration. Similar antibacterial activities against D. dadantii were obtained with AgNPs at 50 µg·mL-1, 50% CFCS alone, and the combination of AgNPs at 12 µg·mL-1 and 12% CFCS of B. amyloliquefaciens. In planta experiments indicated that all the treatments reduced D. dadantii infection and increased plant growth. These findings suggest that AgNPs along with CFCS of B. amyloliquefaciens can be applied to minimize this bacterial disease by controlling pathogen-contaminated sweet potato tuber with minimum Ag nano-pollutant in the environment.

12.
J Bacteriol ; 194(24): 6963-4, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23209221

RESUMO

Enterobacter sp. strain SP1 is an endophytic nitrogen-fixing bacterium isolated from a sugarcane stem and can promote plant growth. The draft genome sequence of strain SP1 presented here will promote comparative genomic studies to determine the genetic background of interactions between endophytic enterobacteria and plants.


Assuntos
Enterobacter/genética , Genoma Bacteriano , Saccharum/microbiologia , Técnicas de Tipagem Bacteriana , Sequência de Bases , DNA Bacteriano/genética , Endófitos/classificação , Endófitos/genética , Endófitos/isolamento & purificação , Enterobacter/classificação , Enterobacter/isolamento & purificação , Dados de Sequência Molecular , Fixação de Nitrogênio/genética , Análise de Sequência de DNA
13.
J Bacteriol ; 194(24): 6995-6, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23209242

RESUMO

Most Herbaspirillum seropedicae strains are beneficial to plants. In contrast, H. seropedicae strain Os45, isolated from rice roots, is pathogenic. The draft genome sequence of strain Os45 presented here allows an in-depth comparative genome analysis to understand the subtle mechanisms of beneficial and pathogenic Herbaspirillum-plant interactions.


Assuntos
Genoma Bacteriano , Herbaspirillum/genética , Oryza/microbiologia , Raízes de Plantas/microbiologia , Sequência de Bases , DNA Bacteriano/genética , Fímbrias Bacterianas/genética , Herbaspirillum/isolamento & purificação , Interações Hospedeiro-Patógeno , Dados de Sequência Molecular , RNA Bacteriano/genética , Análise de Sequência de DNA
14.
J Bacteriol ; 194(24): 6993-4, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23209241

RESUMO

Most Herbaspirillum seropedicae strains are beneficial endophytes to plants. In contrast, H. seropedicae strain Os34, isolated from rice roots, is pathogenic. The draft genome sequence of strain Os34 presented here allows in-depth comparative genome analyses to understand the specific mechanisms of beneficial and pathogenic Herbaspirillum-plant interactions.


Assuntos
Genoma Bacteriano , Herbaspirillum/genética , Oryza/microbiologia , Raízes de Plantas/microbiologia , Sistemas de Secreção Bacterianos/genética , Sequência de Bases , DNA Bacteriano/genética , Fímbrias Bacterianas/genética , Herbaspirillum/isolamento & purificação , Herbaspirillum/metabolismo , Interações Hospedeiro-Patógeno/genética , Ácidos Indolacéticos/metabolismo , Dados de Sequência Molecular , Fixação de Nitrogênio/genética , Análise de Sequência de DNA , Sideróforos/biossíntese
15.
Appl Microbiol Biotechnol ; 93(3): 1185-95, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22002067

RESUMO

Members of the genus Microbacterium lineage of Gram-positive actinobacteria are increasingly being reported to display significant traits associated with environmental biotechnology and bioengineering. 16SH is a nitrogen-fixing bacterial strain isolated from a surface-sterilized stem of sugarcane grown in Guangxi, China. Analysis of 16S rRNA gene sequences revealed that 16SH belonged to the genus Microbacterium. pPROBE-pTet(r) plasmids were constructed by cloning the promoter region of the Tet(r) gene into the promoterless pPROBE-AT, -OT, and -TT vectors derived from the pBBR1 plasmid that has a broad host range of Gram-negative bacteria and sequence similarities to plasmids from Gram-positive bacteria. The pPROBE-pTet(r) plasmids expressed the gfp reporter gene and were stably maintained in 16SH cells without antibiotic selection in free-living state and in planta. Confocal microscopy on intact roots of micropropagated sugarcane plantlets showed that gfp-tagged 16SH cells formed biofilms on root maturation and elongation zones but not on root meristem zones and root caps, and colonized in intercellular spaces of root cortices. Inoculation of 16SH significantly increased biomass and nitrogen content of micropropagated sugarcane seedlings grown with a nitrogen fertilization of 6.3 mg N/kg soil. ¹5N isotope dilution assays demonstrated that biological nitrogen fixation contributed to this plant growth promotion. This study for the first time demonstrated that the pBBR1-based pPROBE plasmids provided an efficient genetic transfer system for a Gram-positive Microbacterium strain, and that a nitrogen-fixing Microbacterium endophyte colonized in intact host plants and fixed N2 associated with the host plants.


Assuntos
Actinomycetales/crescimento & desenvolvimento , Actinomycetales/genética , Fixação de Nitrogênio , Raízes de Plantas/microbiologia , Plasmídeos/genética , Saccharum/microbiologia , Actinomycetales/isolamento & purificação , Actinomycetales/metabolismo , Biotecnologia/métodos , Endófitos , Microbiologia Ambiental , Técnicas de Transferência de Genes , Vetores Genéticos , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Dados de Sequência Molecular , Raízes de Plantas/crescimento & desenvolvimento , RNA Ribossômico 16S/genética , Saccharum/crescimento & desenvolvimento , Análise de Sequência de DNA
16.
Microorganisms ; 10(6)2022 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-35744741

RESUMO

Bacterial panicle blight of rice or bacterial grain rot of rice is a worldwide rice disease. Burkholderia glumae and B. gladioli are the causal agents. The early and accurate detection of seed-borne B. glumae and B. gladioli is critical for domestic and international quarantine and effective control of the disease. Here, genomic analyses revealed that B. gladioli contains five phylogroups and the BG1 primer pair designed to target the 3'-end sequence of a gene encoding a Rhs family protein is specific to B. glumae and two phylogroups within B. gladioli. Using the BG1 primer pair, a 138-bp DNA fragment was amplified only from the tested panicle blight pathogens B. glumae and B. gladioli. An EvaGreen droplet digital PCR (dPCR) assay on detection and quantification of the two pathogens was developed from a SYBR Green real-time quantitative PCR (qPCR). The detection limits of the EvaGreen droplet dPCR on the two pathogens were identical at 2 × 103 colony forming units (CFU)∙mL-1 from bacterial suspensions and 2 × 102 CFU∙seed-1 from rice seeds. The EvaGreen droplet dPCR assay showed 10-fold detection sensitivity of the SYBR Green qPCR and could detect a single copy of the target gene in a 20-µL assay. Together, the SYBR Green qPCR assay allows for routine high-throughput detection of the panicle blight pathogens and the EvaGreen droplet dPCR assay provides a high-sensitive and high-accurate diagnostic method for quarantine of the pathogens.

17.
Viruses ; 14(12)2022 12 02.
Artigo em Inglês | MEDLINE | ID: mdl-36560706

RESUMO

Over the last several decades, kiwifruit production has been severely damaged by the bacterial plant pathogen Pseudomonas syringae pv. actinidiae (Psa), resulting in severe economic losses worldwide. Currently, copper bactericides and antibiotics are the main tools used to control this bacterial disease. However, their use is becoming increasingly ineffective due to the emergence of antibiotic resistance. In addition, environmental issues and the changes in the composition of soil bacterial communities are also concerning when using these substances. Although biocontrol methods have shown promising antibacterial effects on Psa infection under in vitro conditions, the efficiency of antagonistic bacteria and fungi when deployed under field conditions remains unclear. Therefore, it is crucial to develop a phage-based biocontrol strategy for this bacterial pathogen. Due to the specificity of the target bacteria and for the benefit of the environment, bacteriophages (phages) have been widely regarded as promising biological agents to control plant, animal, and human bacterial diseases. An increasing number of studies focus on the use of phages for the control of plant diseases, including the kiwifruit bacterial canker. In this review, we first introduce the characteristics of the Psa-induced kiwifruit canker, followed by a description of the diversity and virulence of Psa strains. The main focus of the review is the description of recent advances in the isolation of Psa phages and their characterization, including morphology, host range, lytic activity, genome characterization, and lysis mechanism, but we also describe the biocontrol strategies together with potential challenges introduced by abiotic factors, such as high temperature, extreme pH, and UV irradiation in kiwifruit orchards. The information presented in this review highlights the potential role of phages in controlling Psa infection to ensure plant protection.


Assuntos
Actinidia , Bacteriófagos , Humanos , Pseudomonas syringae , Especificidade de Hospedeiro , Actinidia/microbiologia , Frutas/microbiologia , Doenças das Plantas/prevenção & controle , Doenças das Plantas/microbiologia
18.
Microorganisms ; 10(2)2022 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-35208879

RESUMO

Kiwifruit worldwide suffers from the devastating diseases of bacterial canker caused by Pseudomonas syringae pv. actinidiae (Psa) and gray mold caused by Botrytis cinerea. Here, an endophytic bacterium XL17 isolated from a rape crown gall was screened out for its potent antagonistic activities against Psa and B. cinerea. Strain XL17 and its cell-free culture filtrate (CF) inhibited the growth of Psa and B. cinerea, Psa-associated leaf necrosis, and B. cinerea-associated kiwifruit necrosis. Electron microscopy showed that XL17 CF could damage the cell structures of Psa and B. cinerea. Genome-based taxonomy revealed that strain XL17 belongs to Pseudomonas bijieensis within the P. corrugata subgroup of the P. fluorescens species complex. Among the P. corrugata subgroup containing 31 genomospecies, the presence of the phl operon responsible for the biosynthesis of the phenolic polyketide 2,4-diacetylphloroglucinol (DAPG) and the absence of the lipopeptide/quorum sensing island can serve as the genetic marker for the determination of a plant-protection life style. HPLC detected DAPG in extracts from XL17 CF. MALDI-TOF-MS analysis revealed that strain XL17 produced cyclic lipopeptides of the viscosin family and orfamide family. Together, phenotypic, genomic, and metabolic analyses identified that P. bijieensis XL17 producing DAPG and cyclic lipopeptides can be used to control bacterial canker and gray mold pathogens of kiwifruit.

19.
J Hazard Mater ; 422: 126936, 2022 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-34463272

RESUMO

Cadmium (Cd) and dichlorodiphenyltrichloroethane (DDT) are frequently detected in agricultural soils, which poses a threat to public health. This study investigated the effects of inoculation of king grass with Piriformospora indica on the remediation of soils co-contaminated with Cd and DDTs. After treatment for 90 days, the dry shoot and root biomass of king grass inoculated with P. indica markedly increased by 13.0-15.8% and 24.1-46.4%, respectively, compared with those of uninoculated plants. Inoculation with P. indica also increased the uptake of Cd and DDTs by shoots and roots of king grass. The removal efficiency of Cd and DDTs from soils reached 4.88-17.4% and 48.4-51.0%, respectively, in the presence of king grass inoculated with P. indica. Under three Cd-DDTs contamination conditions, root secretion of organic acids, alcohol, and polyamines was distinctively stimulated by P. indica inoculation of king grass compared with planting king grass alone. After phytoremediation, changes in soil bacterial and fungal community composition occurred at different contamination levels. Overall, the results showed that king grass associated with P. indica can be adopted for phytoextraction of Cd and DDTs from moderately contaminated soils by regulating root excretion and reshaping rhizosphere microbial community structure.


Assuntos
Microbiota , Poluentes do Solo , Basidiomycota , Biodegradação Ambiental , Cádmio/análise , Raízes de Plantas/química , Poaceae , Rizosfera , Solo , Poluentes do Solo/análise
20.
Appl Environ Microbiol ; 77(9): 3092-101, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21378048

RESUMO

A novel tomato-infecting begomovirus from Guangxi province, China, was identified and characterized, for which the name Tomato leaf curl China virus (ToLCCNV) was proposed. Phylogenetic and recombination analyses of the virus genomic sequences suggested that ToLCCNV may have arisen by recombination among Tomato leaf curl Vietnam virus (ToLCVV), Tomato leaf curl Gujarat virus (ToLCGV), and an unknown virus. A betasatellite molecule was found to be associated with ToLCCNV (ToLCCNB), and its complete nucleotide sequences were determined. Infectious clones of ToLCCNV and ToLCCNB were constructed and then used for agro-inoculation of plants; ToLCCNV alone infected Nicotiana benthamiana, Nicotiana glutinosa, Petunia hybrida, and Solanum lycopersicum plants, but no symptoms were induced. ToLCCNB was required for induction of leaf curl disease in these hosts. The ßC1 protein of ToLCCNB was identified as a suppressor of RNA silencing and accumulated primarily in the nucleus. Deletion mutagenesis of ßC1 showed that the central part of ßC1 (amino acids 44 to 74) was responsible for both the suppressor activity and nuclear localization.


Assuntos
Begomovirus/classificação , Begomovirus/genética , DNA Satélite/genética , DNA Viral/genética , Doenças das Plantas/virologia , Solanum lycopersicum/virologia , Begomovirus/isolamento & purificação , China , DNA Satélite/química , DNA Viral/química , Evolução Molecular , Dados de Sequência Molecular , Filogenia , Recombinação Genética , Análise de Sequência de DNA , Proteínas Virais/genética , Proteínas Virais/metabolismo
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