scholarly journals Bioprospecting of Beneficial Bacteria Traits Associated With Tomato Root in Greenhouse Environment Reveals That Sampling Sites Impact More Than the Root Compartment

2021 ◽  
Vol 12 ◽  
Author(s):  
Alice Anzalone ◽  
Mario Di Guardo ◽  
Patrizia Bella ◽  
Farideh Ghadamgahi ◽  
Giulio Dimaria ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Sampling Site ◽  
Root Surface ◽  
Antagonistic Activity ◽  
Rrna Gene ◽  
In Planta ◽  
Bacterial Strains ◽  
Tomato Root ◽  
Rdna Sequencing

Tomato is subject to several diseases that affect both field- and greenhouse-grown crops. To select cost-effective potential biocontrol agents, we used laboratory throughput screening to identify bacterial strains with versatile characteristics suitable for multipurpose uses. The natural diversity of tomato root–associated bacterial communities was bioprospected under a real-world environment represented by an intensive tomato cultivation area characterized by extraseasonal productions in the greenhouse. Approximately 400 tomato root–associated bacterial isolates, in majority Gram-negative bacteria, were isolated from three compartments: the soil close to the root surface (rhizosphere, R), the root surface (rhizoplane, RP), and the root interior (endorhizosphere, E). A total of 33% of the isolates produced siderophores and were able to solubilize phosphates and grow on NA with 8% NaCl. A total of 30% of the root-associated bacteria showed antagonistic activity against all the tomato pathogens tested, i.e., Clavibacter michiganesis pv. michiganensis, Pseudomonas syringae pv. tomato, Pseudomonas corrugata and Xanthomonas euvesicatoria pv. perforans, and Fusarium oxysporum f. sp. lycopersici. We found that the sampling site rather than the root compartment of isolation influenced bacterial composition in terms of analyzed phenotype. This was demonstrated through a diversity analysis including general characteristics and PGPR traits, as well as biocontrol activity in vitro. Analysis of 16S rRNA gene (rDNA) sequencing of 77 culturable endophytic bacteria that shared multiple beneficial activity revealed a predominance of bacteria in Bacillales, Enterobacteriales, and Pseudomonadales. Their in vitro antagonistic activity showed that Bacillus species were significantly more active than the isolates in the other taxonomic group. In planta activity against phytopathogenic bacteria of a subset of Bacillus and Pseudomonas isolates was also assessed.

scholarly journals Endophytic Bacterial Isolates From Halophytes Demonstrate Phytopathogen Biocontrol and Plant Growth Promotion Under High Salinity

2021 ◽  
Vol 12 ◽  
Author(s):  
Christos A. Christakis ◽  
Georgia Daskalogiannis ◽  
Anastasia Chatzaki ◽  
Emmanouil A. Markakis ◽  
Glykeria Mermigka ◽  
...  
Keyword(s):  
Plant Growth ◽  
Plant Growth Promotion ◽  
High Salinity ◽  
Growth Promotion ◽  
Antagonistic Activity ◽  
Rrna Gene ◽  
In Planta ◽  
Promoting Effect ◽  
Cakile Maritima

Halophytic endophytes potentially contribute to the host’s adaptation to adverse environments, improving its tolerance against various biotic and abiotic stresses. Here, we identified the culturable endophytic bacteria of three crop wild relative (CWR) halophytes: Cakile maritima, Matthiola tricuspidata, and Crithmum maritimum. In the present study, the potential of these isolates to improve crop adaptations to various stresses was investigated, using both in vitro and in-planta approaches. Endophytic isolates were identified by their 16S rRNA gene sequence and evaluated for their ability to: grow in vitro in high levels of NaCl; inhibit the growth of the economically important phytopathogens Verticillium dahliae, Ralstonia solanacearum, and Clavibacter michiganensis and the human pathogen Aspergillus fumigatus; provide salt tolerance in-planta; and provide growth promoting effect in-planta. Genomes of selected isolates were sequenced. In total, 115 endophytic isolates were identified. At least 16 isolates demonstrated growth under increased salinity, plant growth promotion and phytopathogen antagonistic activity. Three showed in-planta suppression of Verticillium growth. Furthermore, representatives of three novel species were identified: two Pseudomonas species and one Arthrobacter. This study provides proof-of-concept that the endophytes from CWR halophytes can be used as “bio-inoculants,” for the enhancement of growth and stress tolerance in crops, including the high-salinity stress.

Effect of the endophytic plant growth promoting Enterobacter ludwigii EB4B on tomato growth

2020 ◽  
Vol 13 (2) ◽  
pp. 54-65 ◽  
Author(s):  
M.E.A. Bendaha ◽  
H.A. Belaouni
Keyword(s):  
Root Length ◽  
Biocontrol Agent ◽  
Growth Promotion ◽  
Dry Weight ◽  
Antagonistic Activity ◽  
Rrna Gene ◽  
Plant Growth Promoting ◽  
Enterobacter Ludwigii ◽  
Tomato Growth

SummaryThis study aims to develop a biocontrol agent against Fusarium oxysporum f.sp. radicis-lycopersici (FORL) in tomato. For this, a set of 23 bacterial endophytic isolates has been screened for their ability to inhibit in vitro the growth of FORL using the dual plate assay. Three isolates with the most sound antagonistic activity to FORL have been qualitatively screened for siderophore production, phosphates solubilization and indolic acetic acid (IAA) synthesis as growth promotion traits. Antagonistic values of the three candidates against FORL were respectively: 51.51 % (EB4B), 51.18 % (EB22K) and 41.40 % (EB2A). Based on 16S rRNA gene sequence analysis, the isolates EB4B and EB22K were closely related to Enterobacter ludwigii EN-119, while the strain EB2A has been assigned to Leclercia adecarboxylata NBRC 102595. The promotion of tomato growth has been assessed in vitro using the strains EB2A, EB4B and EB22K in presence of the phytopathogen FORL. The treatments with the selected isolates increased significantly the root length and dry weight. Best results were observed in isolate EB4B in terms of growth promotion in the absence of FORL, improving 326.60 % of the root length and 142.70 % of plant dry weight if compared with untreated controls. In the presence of FORL, the strain EB4B improved both root length (180.81 %) and plant dry weight (202.15 %). These results encourage further characterization of the observed beneficial effect of Enterobacter sp. EB4B for a possible use as biofertilizer and biocontrol agent against FORL.

scholarly journals Zinc phosphate protects tomato plants against Pseudomonas syringae pv. tomato

2021 ◽  
Author(s):  
Mara Quaglia ◽  
Marika Bocchini ◽  
Benedetta Orfei ◽  
Roberto D’Amato ◽  
Franco Famiani ◽  
...  
Keyword(s):  
Disease Severity ◽  
Pseudomonas Syringae ◽  
Zinc Phosphate ◽  
Plant Defence ◽  
In Planta ◽  
Tomato Plants ◽  
Pathogenesis Related Protein ◽  
Defence Responses ◽  
Plant Defence Responses

AbstractThe purpose of this study was to determine whether zinc phosphate treatments of tomato plants (Solanum lycopersicum L.) can attenuate bacterial speck disease severity through reduction of Pseudomonas syringae pv. tomato (Pst) growth in planta and induce morphological and biochemical plant defence responses. Tomato plants were treated with 10 ppm (25.90 µM) zinc phosphate and then spray inoculated with strain DAPP-PG 215, race 0 of Pst. Disease symptoms were recorded as chlorosis and/or necrosis per leaf (%) and as numbers of necrotic spots. Soil treatments with zinc phosphate protected susceptible tomato plants against Pst, with reductions in both disease severity and pathogen growth in planta. The reduction of Pst growth in planta combined with significantly higher zinc levels in zinc-phosphate-treated plants indicated direct antimicrobial toxicity of this microelement, as also confirmed by in vitro assays. Morphological (i.e. callose apposition) and biochemical (i.e., expression of salicylic-acid-dependent pathogenesis-related protein PR1b1 gene) defence responses were induced by the zinc phosphate treatment, as demonstrated by histochemical and qPCR analyses, respectively. In conclusion, soil treatments with zinc phosphate can protect tomato plants against Pst attacks through direct antimicrobial activity and induction of morphological and biochemical plant defence responses.

scholarly journals HopA1 Effector from Pseudomonas syringae pv syringae Strain 61 Affects NMD Processes and Elicits Effector-Triggered Immunity

2021 ◽  
Vol 22 (14) ◽  
pp. 7440
Author(s):  
Shraddha K. Dahale ◽  
Daipayan Ghosh ◽  
Kishor D. Ingole ◽  
Anup Chugani ◽  
Sang Hee Kim ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Disease Susceptibility ◽  
Null Mutant ◽  
In Planta ◽  
Host Range Expansion ◽  
Unique System ◽  
Effector Triggered Immunity ◽  
Transcriptomic Changes ◽  
Immune Detection

Pseudomonas syringae-secreted HopA1 effectors are important determinants in host range expansion and increased pathogenicity. Their recent acquisitions via horizontal gene transfer in several non-pathogenic Pseudomonas strains worldwide have caused alarming increase in their virulence capabilities. In Arabidopsis thaliana, RESISTANCE TO PSEUDOMONAS SYRINGAE 6 (RPS6) gene confers effector-triggered immunity (ETI) against HopA1pss derived from P. syringae pv. syringae strain 61. Surprisingly, a closely related HopA1pst from the tomato pathovar evades immune detection. These responsive differences in planta between the two HopA1s represents a unique system to study pathogen adaptation skills and host-jumps. However, molecular understanding of HopA1′s contribution to overall virulence remain undeciphered. Here, we show that immune-suppressive functions of HopA1pst are more potent than HopA1pss. In the resistance-compromised ENHANCED DISEASE SUSCEPTIBILITY 1 (EDS1) null-mutant, transcriptomic changes associated with HopA1pss-elicited ETI are still induced and carry resemblance to PAMP-triggered immunity (PTI) signatures. Enrichment of HopA1pss interactome identifies proteins with regulatory roles in post-transcriptional and translational processes. With our demonstration here that both HopA1 suppress reporter-gene translations in vitro imply that the above effector-associations with plant target carry inhibitory consequences. Overall, with our results here we unravel possible virulence role(s) of HopA1 in suppressing PTI and provide newer insights into its detection in resistant plants.

scholarly journals Identification and Genomic Characterization of Pathogenic Bacillus altitudinis from Common Pear Trees in Morocco

Agronomy ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1344
Author(s):  
Naima Lemjiber ◽  
Khalid Naamani ◽  
Annabelle Merieau ◽  
Abdelhi Dihazi ◽  
Nawal Zhar ◽  
...  
Keyword(s):  
16S Rrna Gene Sequencing ◽  
Genomic Islands ◽  
Rrna Gene ◽  
In Planta ◽  
Bacterial Strains ◽  
Bacillus Altitudinis ◽  
Genes Encoding ◽  
Bacillus Spp ◽  
Pear Trees ◽  
Rrna Gene Sequencing

Bacterial burn is one of the major diseases affecting pear trees worldwide, with serious impacts on producers and economy. In Morocco, several pear trees (Pyrus communis) have shown leaf burns since 2015. To characterize the causal agent of this disease, we isolated fourteen bacterial strains from different parts of symptomatic pear trees (leaves, shoots, fruits and flowers) that were tested in planta for their pathogenicity on Louise bonne and Williams cultivars. The results showed necrotic lesions with a significant severity range from 47.63 to 57.77% on leaves of the Louise bonne cultivar inoculated with isolate B10, while the other bacterial isolates did not induce any disease symptom. 16S rRNA gene sequencing did not allow robust taxonomic discrimination of the incriminated isolate. Thus, we conducted whole-genome sequencing (WGS) and phylogenetic analyzes based on gyrA, gyrB and cdaA gene sequences, indicating that this isolate belongs to the Bacillus altitudinis species. This taxonomic classification was further confirmed by the Average Nucleotide Identity (ANI) and the in silico DNA-DNA hybridization (isDDH) analyzes compared to sixty-five Bacillus spp. type strains. The genome was mined for genes encoding carbohydrate-active enzymes (CAZymes) known to play a role in the vegetal tissue degradation. 177 candidates with functions that may support the in planta phytopathogenicity results were identified. To the best of our knowledge, this is the first data reporting B. altitudinis as agent of leaf burn in P. communis in Morocco. Our dataset will improve our knowledge on spread and pathogenicity of B. altitudinis genotypes that appears as emergent phytopathogenic agent, unveiling virulence factors and their genomic location (i.e., within genomic islands or the accessory genome) to induce trees disease.

scholarly journals The algT Gene of Pseudomonas syringae pv. glycinea andNew Insights into the Transcriptional Organization of thealgT-muc Gene Cluster

2006 ◽  
Vol 188 (23) ◽  
pp. 8013-8021 ◽  
Author(s):  
Alexander Schenk ◽  
Michael Berger ◽  
Lisa M. Keith ◽  
Carol L. Bender ◽  
Georgi Muskhelishvili ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Pseudomonas Syringae ◽  
Azotobacter Vinelandii ◽  
Regulatory Gene ◽  
In Planta ◽  
Phytopathogenic Bacterium ◽  
Reverse Transcription Pcr ◽  
Alginate Production ◽  
Alginate Biosynthesis

ABSTRACT The phytopathogenic bacterium Pseudomonas syringae pv. glycinea infects soybean plants and causes bacterial blight. In addition to P. syringae, the human pathogen Pseudomonas aeruginosa and the soil bacterium Azotobacter vinelandii produce the exopolysaccharide alginate, a copolymer of d-mannuronic and l-guluronic acids. Alginate production in P. syringae has been associated with increased fitness and virulence in planta. Alginate biosynthesis is tightly controlled by proteins encoded by the algT-muc regulatory gene cluster in P. aeruginosa and A. vinelandii. These genes encode the alternative sigma factor AlgT (σ22), its anti-sigma factors MucA and MucB, MucC, a protein with a controversial function that is absent in P. syringae, and MucD, a periplasmic serine protease and homolog of HtrA in Escherichia coli. We compared an alginate-deficient algT mutant of P. syringae pv. glycinea with an alginate-producing derivative in which algT is intact. The alginate-producing derivative grew significantly slower in vitro growth but showed increased epiphytic fitness and better symptom development in planta. Evaluation of expression levels for algT, mucA, mucB, mucD, and algD, which encodes an alginate biosynthesis gene, showed that mucD transcription is not dependent on AlgT in P. syringae in vitro. Promoter mapping using primer extension experiments confirmed this finding. Results of reverse transcription-PCR demonstrated that algT, mucA, and mucB are cotranscribed as an operon in P. syringae. Northern blot analysis revealed that mucD was expressed as a 1.75-kb monocistronic mRNA in P. syringae.

scholarly journals Apis andreniformis associated Actinomycetes show antimicrobial activity against black rot pathogen (Xanthomonas campestris pv. campestris)

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e12097
Author(s):  
Yaowanoot Promnuan ◽  
Saran Promsai ◽  
Wasu Pathom-aree ◽  
Sujinan Meelai
Keyword(s):  
Antimicrobial Activity ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Honey Bee ◽  
Pseudomonas Syringae ◽  
Xanthomonas Campestris ◽  
Pathogenic Bacteria ◽  
Rrna Gene ◽  
Apis Andreniformis

This study aimed to investigate cultivable actinomycetes associated with rare honey bee species in Thailand and their antagonistic activity against plant pathogenic bacteria. Actinomycetes were selectively isolated from the black dwarf honey bee (Apis andreniformis). A total of 64 actinomycete isolates were obtained with Streptomyces as the predominant genus (84.4%) followed by Micromonospora (7.8%), Nonomuraea (4.7%) and Actinomadura (3.1%). All isolates were screened for antimicrobial activity against Xanthomonas campestris pv. campestris, Pectobacterium carotovorum and Pseudomonas syringae pv. sesame. Three isolates inhibited the growth of X. campestris pv. campestris during in vitro screening. The crude extracts of two isolates (ASC3-2 and ASC5-7P) had a minimum inhibitory concentration (MIC) of 128 mg L−1against X. campestris pv. campestris. For isolate ACZ2-27, its crude extract showed stronger inhibitory effect with a lower MIC value of 64 mg L−1 against X. campestris pv. campestris. These three active isolates were identified as members of the genus Streptomyces based on their 16S rRNA gene sequences. Phylogenetic analysis based on the maximum likelihood algorithm showed that isolate ACZ2-27, ASC3-2 and ASC5-7P were closely related to Streptomyces misionensis NBRC 13063T (99.71%), Streptomyces cacaoi subsp. cacaoi NBRC 12748T (100%) and Streptomyces puniceus NBRC 12811T (100%), respectively. In addition, representative isolates from non-Streptomyces groups were identified by 16S rRNA gene sequence analysis. High similarities were found with members of the genera Actinomadura, Micromonospora and Nonomuraea. Our study provides evidence of actinomycetes associated with the black dwarf honey bee including members of rare genera. Antimicrobial potential of these insect associated Streptomyces was also demonstrated especially the antibacterial activity against phytopathogenic bacteria.

scholarly journals EFFICACY OF BIOCHEMICAL PREPARATIONS AND EXTRACT FROM Hypericum perforatum AGAINST BACTERIAL DISEASES

2019 ◽  
Vol 18 (3) ◽  
Author(s):  
Małgorzata Schollenberger ◽  
Sylwia Pudło ◽  
Elżbieta Paduch-Cichal ◽  
Ewa Mirzwa-Mróz
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Pseudomonas Syringae ◽  
Pathogenic Bacteria ◽  
Hypericum Perforatum ◽  
Agar Plate ◽  
Nutrient Agar ◽  
Bacterial Strains ◽  
Bacterial Diseases ◽  
Nutrient Agar Medium

The biotechnical preparations: Biosept Active (based on a grapefruit extract) and BioZell (based on thyme oil) as well as Hypericum perforatum extract, streptomycin solution and fungicide Champion 50WP (active ingredient substance – e.i. 50% copper hydroxide) were investigated for antimicrobial effects against plant pathogenic bacteria: Agrobacterium tumefaciens, Pseudomonas syringae pv. syringae and Xanthomonas ar- boricola pv. corylina. The screening was carried out in vitro on three media: Nutrient Agar (NA Difco), Pseudomonas Agar F (Merck) – analogue of King B and 523. In the experiments, the agar plate method was applied. There were no statistically significant differences in the effect of streptomycin and Champion 50WP on the growth inhibition of three bacteria strains for medium 523 and Nutrient Agar and of P. syringae pv. syringae and X. arboricola pv. corylina for medium King B. It was determined that the antibacterial activity of Biosept Active and BioZell biopreparations and H. perforatum extract against Agrobacterium tumefaciens (strain C58), Pseudomonas syringae pv. syringae (strain 760) and Xanthomonas arboricola pv. corylina (strain RIPF-x13) were dependent on the strain of pathogen as well as the growth medium used. According to the research results obtained, the Biosept Active preparation and H. perforatum extract demonstrated high bacteriostatic activity against three bacterial strains grown on the Nutrient Agar medium.

scholarly journals Appraising Endophyte–Plant Symbiosis for Improved Growth, Nodulation, Nitrogen Fixation and Abiotic Stress Tolerance: An Experimental Investigation with Chickpea (Cicer arietinum L.)

Agronomy ◽  
2019 ◽  
Vol 9 (10) ◽  
pp. 621 ◽  
Author(s):  
Ahmad ◽  
Naseer ◽  
Hussain ◽  
Zahid Mumtaz ◽  
Mustafa ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Plant Growth ◽  
Root Nodules ◽  
Abiotic Stress Tolerance ◽  
Paenibacillus Polymyxa ◽  
Rrna Gene ◽  
Bacterial Strains ◽  
Pgp Traits ◽  
Paenibacillus Sp

Chickpea is an important leguminous crop that improves soil fertility through atmospheric nitrogen fixation with the help of rhizobia present in nodules. Non-rhizobia endophytes are also capable of inducing nodulation and nitrogen fixation in leguminous crops. The aim of the current study was to isolate, characterize and identify the non-rhizobia endophytic bacterial strains from root nodules of chickpea. For this purpose, more than one hundred isolates were isolated from chickpea root nodules under aseptic conditions and were confirmed as endophytes through re-isolating them from root nodules of chickpea after their inoculation. Nineteen confirmed endophytic bacterial strains revealed significant production of indole acetic acid (IAA) both in presence and absence of L-tryptophan and showed their ability to grow under salt, pH and heavy metal stresses. These strains were evaluated for in vitro plant growth promoting (PGP) traits and results revealed that seven strains showed solubilization of P and colloidal chitin along with possessing catalase, oxidase, urease and chitinase activities. Seven P-solubilizing strains were further evaluated in a jar trial to explore their potential for promoting plant growth and induction of nodulation in chickpea roots. Two endophytic strains identified as Paenibacillus polymyxa ANM59 and Paenibacillus sp. ANM76 through partial sequencing of the 16S rRNA gene showed the maximum potential during in vitro PGP activities and improved plant growth and nodulation in chickpea under the jar trial. Use of these endophytic strains as a potential biofertilizer can help to reduce the dependence on chemical fertilizers while improving crop growth and soil health simultaneously.

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