The chitinolytic activity of the Curtobacterium sp. isolated from field-grown soybean and analysis of its genome sequence.

Curtobacterium sp. GD1 was isolated from leaves of conventionally grown soybean in Brazil. It was noteworthy that among all bacteria previously isolated from the same origin, only Curtobacterium sp. GD1 showed a strong chitinase activity. The enzyme was secreted and its production was induced by the presence of colloidal chitin in the medium. The chitinase was partially purified and characterized: molecular weight was approximately 37 kDa and specific activity 90.8 U/mg. Furthermore, Curtobacterium sp. GD1 genome was sequenced and analyzed. Our isolate formed a phylogenetic cluster with four other Curtobacterium spp. strains, with ANIb/ANIm � 98%, representing a new, still non described Curtobacterium species. The circular genome visualization and comparison of genome sequences of strains forming new cluster indicated that most regions within their genomes were highly conserved. The gene associated with chitinase production was identified and the distribution pattern of glycosyl hydrolases genes was assessed. Also, genes associated with catabolism of structural carbohydrates such as oligosaccharides, mixed polysaccharides, plant and animal polysaccharides, as well as genes or gene clusters associated with resistance to antibiotics, toxic compounds and auxin biosynthesis subsystem products were identified. The abundance of putative glycosyl hydrolases in the genome of Curtobacterium sp. GD1 suggests that it has the tools for the hydrolysis of different polysaccharides. Therefore, Curtobacterium sp. GD1 isolated from soybean might be a bioremediator, biocontrol agent, an elicitor of the plant defense responses or simply degrader

Chemical Defence in a Millipede: Evaluation and Characterization of Antimicrobial Activity of the Defensive Secretion from Pachyiulus hungaricus (Karsch, 1881) (Diplopoda, Julida, Julidae)

The chemical defence of the millipede Pachyiulus hungaricus is reported in the present paper, in which a chemical characterization is given and antimicrobial activity is determined. In total, independently of sex, 44 compounds were identified. All compounds belong to two groups: quinones and pentyl and hexyl esters of long-chain fatty acids. The relative abundances of quinones and non-quinones were 94.7% vs. 5.3% (males) and 87.3% vs. 12.7% (females), respectively. The two dominant quinones in both sexes were 2-methyl-1,4,-benzoquinone and 2-methoxy-3-methyl-1,4-benzoquinone. Antibacterial and antifungal activity of the defensive secretion was evaluated in vitro against seven bacterial strains and eight fungal species. With the aid of a dilution technique, the antimicrobial potential of the secretion and high sensitivity of all tested strains were confirmed. The lowest minimum concentrations of these compounds (0.20-0.25 mg/mL) were sufficient for inhibition of Aeromonas hydrophila, Listeria monocytogenes and Methicillin resistant Staphylococcus aureus (MRSA). The growth of eight tested fungal species was inhibited by slightly lower concentrations of the secretion, with Fusarium equisetias the most sensitive fungus and Aspergillus flavus as the most resistant. Values of MIC and MFC in the employed microdilution assay ranged from 0.10 to above 0.35 mg/m L. The given extract contains antimicrobial components potentially useful as therapeutic agents in the pharmaceutical and agricultural industries

Genetic diversity and pathogenicity of Pseudomonas syringae pv. aptata isolated from sugar beet

Pseudomonas syringae pv. aptata is the causal agent of bacterial leaf spot disease of sugar beet (Beta vulgaris). During 2013, 250 samples were collected from leaf lesions with typical symptoms of bacterial leaf spot in commercial fields of sugar beet in Serbia, and 104 isolates of P.syringae pv. aptata were obtained. Identification and characterization was performed using biochemical, molecular and pathogenicity tests. Identification included LOPAT tests and positive reactions using primers Papt2F and Papt1R specific for P.syringae pv. aptata. Repetitive (rep) sequence-based PCR typing with ERIC, REP and BOX primers revealed high genetic variability among isolates and distinguished 25 groups of different fingerprinting profiles. Pulse-field gel electrophoresis (PFGE) and multilocus sequence analysis (MLSA) of representative isolates showed higher genetic variability than in rep-PCR analysis and distinguished three and four major genetic clusters, respectively. A pathogenicity test performed with 25 representative isolates on four cultivars of sugar beet confirmed the occurrence of leaf spot disease and showed correlation between the most aggressive isolates and the genetic clusters obtained in MLSA. All these findings point to the existence of several lines of P.syringae pv. aptata infection in Serbia that are genetically and pathologically different

Biological control of Pseudomonas syringae pv. aptata on sugar beet with Bacillus pumilus SS-10.7 and Bacillus amyloliquefaciens (SS-12.6 and SS-38.4) strains

Aim Assessment of biological control of Pseudomonas syringae pv. aptata using crude lipopeptide extracts (CLEs) of two Bacillus amyloliquefaciens strains (SS-12.6 and SS-38.4) and one Bacillus pumilus strain (SS-10.7). Methods and Results The minimum inhibitory concentration (MIC) of CLEs and their combinations against the pathogen and potential interaction between the extracts were determined in vitro. The most effective antibacterial activity was achieved with the CLE from B. amyloliquefaciens SS-12.6, with an MIC value of 0 center dot 63 mg ml(-1). Interactions between CLE combinations were mostly indifferent. The biocontrol potential of CLEs, mixtures of CLEs, and cell culture of B. amyloliquefaciens SS-12.6 was tested on sugar beet plants inoculated with P. syringae pv. aptata P53. The best result in inhibiting the appearance of tissue necrosis (up to 92%) was achieved with B. amyloliquefaciens SS-12.6 cell culture. Conclusion This work demonstrated significant biocontrol potential of the CLE and cell culture of B. amyloliquefaciens SS-12.6 which successfully suppress leaf spot disease severity on sugar beet plants. Significance and Impact of the Study The findings of biocontrol of sugar beet emerging pathogen will contribute to growers in terms of alternative disease control management. This study represents first assessment of biological control of P. syringae pv. aptata

Screening for the presence of biosynthetic genes for antimicrobial lipopeptides in natural isolates of Bacillus sp.

A collection of 205 natural isolates of Bacillus was tested for the presence of genes for biosynthesis of antimicrobial lipopeptides, iturin, surfactin, fengycin and bacillomycin D. For the detection of iturin producers by PCR screening, we used forward ITUP1-F and reverse ITUP2-R primers which are capable of detecting a 2-kb region that includes the intergenic sequence between the ituA and ituB genes. A 675-bp fragment from the gene sfp from B. subtilis encoding 4’-phosphopantetheinyl transferase involved in the biosynthesis of surfactin was targeted for amplification by using primers P17 and P18. Other two pairs of primers were BACC1F and BACC1R for bacillomycin D and FEND1F and FEND1R for potential fengycin producers, respectively. The results of the screening showed that the majority of tested strains had more than one biosynthetic operon, since 81% possessed the genes for bacillomycin D production, 54% for surfactin, 38% for iturin and 25% for fengycin production. [Projekat Ministarstva nauke Republike Srbije, br. 173026

Sezonska raznovrsnost biodeteriogenih, patogenih i toksigenih gljiva u zraku sakralnoga objekta

The main purpose of this study was to isolate airborne fungi and assess seasonal variations in air contamination with their particulates by determining the levels of their propagules in the nave and exonarthex of a church. We also monitored indoor microclimate as a determining factor for fungal proliferation on wall paintings, spore release, and transmission through the air. The temperature and relative humidity of the nave favoured fungal growth. A total of 33 fungi were isolated, mainly of the phylum Ascomycota, and to the lesser extent of the phyla Zygomycota and Basidiomycota. The most common were the fungi of the genera Penicillium and Aspergillus (23.55 % and 20.58 %, respectively). Sørensen's quotient of similarity (0.37) suggests moderate species overlap and constant exchange of fungal propagules between the nave and exonarthex. The autumn had the highest diversity, with 17 documented taxa, followed by the summer and the winter. The spring had only eight taxa. Quantitative analysis of the airborne mycobiota in the nave (430±84.85 to 1880±106.07 CFU m -3 ) and exonarthex (715±59.62 to 2295±91.92 CFU m -3 ) showed very high contamination throughout the year, with values exceeding the maximum permissible concentrations by most standards. Many of the fungi determined in this study are known for their biodeteriogenic, toxigenic, and allergenic properties, and are a threat not only to occasional visitors and staff, but also to valuable works of art decorating nave walls. © 2018 Sciendo. All rights reserved. All Rights Reserved.Glavni cilj ovoga istraživanja bio je izolirati gljive iz zraka i procijeniti sezonske promjene u onečišćenju zraka gljivičnim propagulama u naosu i egzonarteksu istraživane crkve. Također su praćeni mikroklimatski parametri kao ograničavajući čimbenici za razvoj i rast gljiva na zidnim slikama, za oslobađanje spora i njihovu transmisiju kroz zrak. Zabilježena temperatura i relativna vlažnost zraka u naosu pogodovale su razvoju i rastu gljiva. Dokumentirana su 33 taksona gljiva, uglavnom pripadnika koljena Ascomycota, a manje su zastupljeni oni Zygomycota i Basidiomycota. Najčešće su bile prisutne plijesni rodova Penicillium (23,55 %) i Aspergillus (20,58 %). Sørensenov indeks sličnosti (0,37) upućuje na stalnu i umjerenu razmjenu gljivičnih propagula između naosa i egzonarteksa. Uzorci uzeti u jesen pokazali su najveću raznolikost sa 17 zabilježenih taksona, a oni uzeti u proljeće samo osam taksona. Kvantitativna mikološka analiza u naosu (430±84,85 do 1880±106,07 CFU m-3) i egzonarteksu (715±59,62 do 2295±91,92 CFU m-3) pokazala je visoku godišnju razinu onečišćenja zraka, s vrijednostima koje prema većini standarda prelaze dopuštene koncentracije. Mnoge identificirane gljive mogu dovesti do biodeterioracije, proizvesti mikotoksine i izazvati alergijske reakcije. Stoga su nepoželjne u sakralnim objektima ne samo zbog očuvanja murala nego i zbog zaštite zdravlja zaposlenih, posjetitelja i konzervatora

An Insight into an Olive Scab on the “Istrska Belica” Variety: Host‐Pathogen Interactions and Phyllosphere Mycobiome

The olive tree is one of the most important agricultural plants, affected by several pests and diseases that cause a severe decline in health status leading to crop losses. Olive leaf spot disease caused by the fungus Venturia oleaginea can result in complete tree defoliation and consequently lower yield. The aim of the study was to obtain new knowledge related to plant–pathogen interaction, reveal mechanisms of plant defense against the pathogen, and characterize fungal phyllosphere communities on infected and symptomless leaves that could contribute to the development of new plant breeding strategies and identification of novel biocontrol agents. The highly susceptible olive variety “Istrska Belica”' was selected for a detailed evaluation. Microscopy analyses led to the observation of raphides in the mesophyll and parenchyma cells of infected leaves and gave new insight into the complex V. oleaginea pathogenesis. Culturable and total phyllosphere mycobiota, obtained via metabarcoding approach, highlighted Didymella, Aureobasidium, Cladosporium, and Alternaria species as overlapping between infected and symptomless leaves. Only Venturia and Erythrobasidium in infected and Cladosporium in symptomless samples with higher abundance showed statistically significant differences. Based on the ecological role of identified taxa, it can be suggested that Cladosporium species might have potential antagonistic effects on V. oleaginea. © 2022, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature