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The significance of alternative respiratory pathway (AOXs) during the interaction between soft rot bacteria (Pectobacterium carotovorum subsp. carotovorum, (Pcc.)) and potato tubers is well-defined. However, the role of the AOXs in impaired mitochondrial respiratory chain function during the Pcc. infection is yet to be studied. In this study, the results show that with the aggravation of infection of Pcc., the capacity for alternative respiration in mitochondria of potato tubers increased gradually. The mitochondrial membrane potential increased more significantly after infection with Pcc. when the AOXs in potato tubers was partially blocked using salicylhydroxamic acid (SHAM) beforehand. In addition, the activity of complex III decreased more drastically while the activity of complex IV increased more significantly in the partial absence of the AOXs in the mitochondria. Furthermore, the mitochondrial endogenous respiration, mitochondrial respiratory state 3 and respiratory control rate (RCR) decreased more significantly and the value of RCR reached around 1.0 with the aggravation of infection of Pcc. in the partially absence of AOXs in the mitochondria. [ABSTRACT FROM AUTHOR]

Seed-borne Erwinia persicina can be transmitted globally via alfalfa (Medicago sativa L.) seed trade, but there is limited information about the impact of this plant-pathogenic bacterium on alfalfa plants. In this study, strain Cp2, isolated from alfalfa seeds, was confirmed by whole-genome sequencing to belong to E. persicina. Subsequently, the effects of Cp2 on alfalfa growth and physiology were evaluated by constructing a rhizosphere infection model. Strain Cp2 had a strong inhibitory effect on the elongation and growth of alfalfa roots, which was very unfavorable to these perennial plants. Furthermore, an increased number of leaf spots and yellowing symptoms were observed in plants of the Cp2 group from day 10 to day 21 and the strain Cp2 was re-isolated from these leaves. Correlation between growth and photosynthetic parameters was analyzed and the significant decreases in fresh weight and root and plant lengths in the Cp2 group were related to the marked reduction of chlorophyll b , carotenoid, transpiration rate, and stomatal conductance of leaves (r > 0.75). In addition, nine physiological indicators of root, stem, and leaf were measured in the plants 21 days after treatment with Cp2. The physiological response of root and leaf to Cp2 treatment was stronger than that of stem. The physiological indicators with the greatest response to Cp2 infection were further explored through principal component analysis, and superoxide dismutase, peroxidase, ascorbate peroxidase, and soluble protein showed the greatest changes in roots, stems, and leaves (P < 0.001). Among tissues, the commonality was the change of soluble protein. Therefore, soluble protein is speculated to be a physiological marker during alfalfa– E. persicina interactions. These findings indicate that once E. persicina spreads from alfalfa seeds to the rhizosphere, it can invade alfalfa roots and cause disease. This study demonstrates that this plant pathogenic bacterium may be a potential threat to new environment when they spread via seed trade and these "dangerous hitchhikers" warrant further attention, especially in the study of bacterial diseases in pasture-based production systems. [ABSTRACT FROM AUTHOR]

The plant pathogen Pectobacterium carotovorum subsp. carotovorum (previously Erwinia carotovora subsp. carotovora) causes soft rot and stem rot diseases in a variety of crops, including Chinese cabbage, potato, and tomato. The flagellar-type III secretion systems were used by Pcc's virulence mechanism to export proteins or bacteriocins to the outside of the cell. DGC, a virulence factor that cyclizes c-di-GMP, a common secondary signal in physiological processes and toxin control systems of many bacteria, was discovered in Pcc's genomic DNA. The dgc gene in Pcc was blocked using the method of homologous recombination in our study. In the in vivo setting, the results demonstrated that the dgc knockout strain does not release low molecular weight bacteriocins. The bacteriocin gene (carocin S2, carocin S3, carocin S4) and the flagellar-type III secretion system genes were also unable to be transcribed by the dgc knockout strain in the transcription experiment. We also observed that the amount of bacteriocin expressed changed when the amount of L-glutamine in the environment exceeded a particular level. These data suggested that L-glutamine influenced physiological processes in Pcc strains in some way. We hypothesized a relationship between dgc and the genes involved in Pcc LMWB external export via the flagellar-type secretion system based on these findings. In this study, the current findings led us to propose a mechanism in which DGC's cyclic di-GMP might bind to receptor proteins and positively regulate bacteriocin transcription as well as the synthesis, mobility, and transport of toxins. [ABSTRACT FROM AUTHOR]

The bacterial soft rot and vascular wilt of sugar beet are the major diseases of sugar crops globally induced by Pectobacterium betavasculorum and P. carotovorum subsp. carotovorum (Pcc). The control of this bacterial disease is a severe problem, and only a few copper-based chemical bactericides are available for this disease. Because of the limitations of chemicals to control plant bacterial pathogens, the essential oils and extracts have been considered one of the best alternative strategies for their control. In this study, twenty-seven essential oils and twenty-nine plant extracts were extracted and evaluated for their antibacterial activities against Pectobacterium betavasculorum isolate C3, using the agar diffusion method at 0.01%, 0.1%, and 100% (v/v). Pure Pimpinella anisum L. oil exhibited the most anti-bacterial activity among three different concentrations of essential oils and extracts, followed by Thymus vulgaris L. oil and Rosa multiflora Thunb. extract. The efficacy of effective essential oils and extracts on Ic1 cultivar of sugar beet seeds germination and seedling growth in vivo also were tested. The seed germination of the Ic1 cultivar was inhibited at all the concentrations of essential oils used. Only extracts of Rosa multiflora Thunb., Brassica oleracea L., Lactuca serriola L., Salvia rosmarinus Spenn., Syzygium aromaticum (L.) Merr. and L.M.Perry, Eucalyptus globulus Labill., and essential oils of Ocmium basilicum L., Pimpinella anisum L., and Mentha× piperita L.L. in 0.1% concentration had no inhibition on seed germination and could improve seedling growth. This is the first report of the antibacterial activity of essential oils and extracts on Pectobacterium betavasculorum. [ABSTRACT FROM AUTHOR]

To respond to changing environmental conditions, a population may either shift toward better‐adapted genotypes or adapt on an individual level. The present work aimed to quantify the relevance of these two processes by comparing the responses of defined Drosophila melanogaster populations to different stressors. To do this, we infected two homogeneous populations (isofemale lines), which differ significantly in fitness, and a synthetic heterogeneous population with a specific pathogen and/or exposed them to food restriction. Pectobacterium carotovorum was used to infect Drosophila larvae either fed standard or protein‐restricted diet. In particular, the two homogeneous groups, which diverged in their fitness, showed considerable differences in all parameters assessed (survivorship, protein and lipid contents, phenol‐oxidase (PO) activity, and antibacterial rate). Under fully nutritious conditions, larvae of the homogeneous population with low fitness exhibited lower survivorship and protein levels, as well as higher PO activity and antibacterial rate compared with the fitter population. A protein‐restricted diet and bacterial infection provoked a decrease in survivorship, and antibacterial rate in most populations. Bacterial infection elicited an opposite response in protein and lipid content in both isofemale lines tested. Interestingly, the heterogeneous population showed a complex response pattern. The response of the heterogeneous population followed the fit genotype in terms of survival and antibacterial activity but followed the unfit genotype in terms of PO activity. In conclusion, our results show that defined genotypes exhibit highly divergent responses to varying stressors that are difficult to predict. Furthermore, the responses of heterogeneous populations do not follow a fixed pattern showing a very high degree of plasticity and differences between different genotypes. [ABSTRACT FROM AUTHOR]

Aims: Bacterial soft rot caused by Pectobacterium brasiliense (Pbr) has resulted in severe economic losses of cucumber production in northern China. Quantitative reverse transcription PCR (RT‐qPCR) is widely used to determine the fold change in the expression of genes of interest, and an appropriate reference gene played a critical role in the evaluation of genes expression. However, the suitable reference genes for transcript normalization during the interaction between cucumber and Pbr have not yet been systematically validated. In this study, we aimed to identify the suitable reference genes for accurate and reliable normalization of cucumber and Pbr RT‐qPCR data. Methods and Results: We selected 14 candidate reference genes for cucumber and 10 candidate reference genes for Pbr were analysed by using four algorithms (the deltaCt method, BestKeeper, NormFinder and geNorm). Furthermore, five genes in cucumber involved in plant resistance and five genes in Pbr related to the virulence were selected to confirm the reliability of the reference genes by RT‐qPCR. CsARF (ADP‐ribosylation factor 1) and pgi (glucose‐6‐phosphate isomerase) were suggested as the most suitable reference genes for cucumber and Pbr respectively. Conclusion: Our results suggested that CsARF (ADP‐ribosylation factor 1) and pgi (glucose‐6‐phosphate isomerase) could be as the reference genes to normalize expression data for cucumber and Pbr during the process of pathogen–host interaction respectively. Significance and Impact of the Study: To our knowledge, this is the first systematic study of the optimal reference genes specific to cucumber and Pbr, which could help advance the molecular interactions research in Cucurbitaceae vegetables and Pectobacterium species pathosystems. [ABSTRACT FROM AUTHOR]

Since 2002, papaya black rot has been spreading over several islands of Okinawa Prefecture. The pathogen of the disease was identified as Erwinia sp., genetically close to E. mallotivora and E. papayae. In terms of the disease transmission, it is probably carried by the wind or rain. In order to devise a prevention strategy for the disease, in vitro pesticides susceptibility of the pathogen and tests with papaya pot seedlings were conducted. A minimum inhibitory concentration assay demonstrated that copper (II) hydroxide, basic copper sulfate, and captan present in the papaya-registered pesticides inhibited the growth of the pathogen on nutrient agar plates. In addition, mancozeb that is non-papaya-registered pesticide also showed an inhibitory effect on the pathogen. Thus, there is a high possibility that even the existing papaya-registered or non-papaya-registered pesticides can prevent papaya black rot. In the test with papaya pot seedlings, copper (II) hydroxide exerted a relatively lower pesticide effect; however, captan exhibited a pesticide effect, although it is one of the fungicides not registered for use in the treatment of bacterial diseases of plants in Japan till date. Indeed, based on the result of the present study, the official registration of legal expansion for use of captan to control papaya black rot was approved in Japan on December 22, 2021. [ABSTRACT FROM AUTHOR]

Global crop production depends on strategies to counteract the ever-increasing spread of plant pathogens. Antibiotics are often used for large-scale treatments. As a result, Erwinia amylovora, causal agent of the contagious fire blight disease, has already evolved resistance to streptomycin (Sm). Photodynamic Inactivation (PDI) of microorganisms has been introduced as innovative method for plant protection. The aim of this study is to demonstrate that E. amylovora resistant to Sm (E. amylovoraSmR) can be killed by PDI. Two photosensitizers, the synthetic B17-0024, and the natural derived anionic sodium magnesium chlorophyllin (Chl) with cell-wall-permeabilizing agents are compared in terms of their photo-killing efficiency in liquid culture with or without 100 µg/mL Sm. In vitro experiments were performed at photosensitizer concentrations of 1, 10 or 100 µM and 5 or 30 min incubation in the dark, followed by illumination at 395 nm (radiant exposure 26.6 J/cm2). The highest inactivation of seven log steps was achieved at 100 µM B17-0024 after 30 min incubation. Shorter incubation (5 min), likely to represent field conditions, reduced the photo-killing to 5 log steps. Chlorophyllin at 100 µM in combination with 1.2% polyaspartic acid (PASA) reduced the number of bacteria by 6 log steps. While PASA itself caused some light independent toxicity, an antibacterial effect (3 log reduction) was achieved only in combination with Chl, even at concentrations as low as 10 µM. Addition of 100 µg/mL Sm to media did not significantly increase the efficacy of the photodynamic treatment. This study proves principle that PDI can be used to treat plant diseases even if causative bacteria are resistant to conventional treatment. Therefore, PDI based on natural photosensitizers might represent an eco-friendly treatment strategy especially in organic farming. [ABSTRACT FROM AUTHOR]

Aims: Pectobacterium polaris is a novel species of Pectobacterium taxa, which was reported in different regions of the world and has been recently detected in Turkey. In this study, in order to determine the host range of this novel species, two representative strains were evaluated in terms of their pathogenicity on several host plants, and it is aimed to determine plant species under the risk of the pathogen. Methods and Results: P. polaris strains NV3 and NV19, isolated from potato plants showing blackleg symptoms in Nevşehir province during the vegetation period of 2019, were used for artificial inoculation. Bacterial suspension of both strains was inoculated by injecting into the red cabbage, broccoli, cauliflower, onion, garlic, carrot, tomato, pepper, eggplant, cucumber, squash, melon, watermelon, lettuce, sunflower and sugar beet plants. The appearance of the disease symptoms indicated that both strains of P. polaris caused disease in all the plant species inoculated artificially. Viable bacterial colonies were re-isolated from infected plant tissue. Conclusions: Pectobacterium species are very destructive bacterial plant pathogens that cause soft rot on numerous plant species. It has been determined that P. polaris, which has been reported in different regions of the world and in Turkey as a new emerging pathogen within in a wide host range. Disease protection precautions should be taken in the regions where the pathogen is isolated for the first time, and especially pathogen transmission should be prevented to the clean production areas by potato tubers. Significance and Impact of the Study: All tested plant species were determined as potential hosts for P. polaris. The risk status of the pathogen on these plants, which have economic importance in Turkey, was determined for the first time with this study. [ABSTRACT FROM AUTHOR]

Bacterial soft rot is one of the most destructive diseases of taro (Colocasia esculenta) worldwide. In recent years, frequent outbreaks of soft rot disease have seriously affected taro production and became a major constraint to the development of taro planting in China. However, little is known about the causal agents of this disease, and the only reported pathogens are two Dickeya species and P. carotovorum. In this study, we report taro soft rot caused by two novel Pectobacterium strains, LJ1 and LJ2, isolated from taro corms in Ruyuan County, Shaoguan City, Guangdong Province, China. We showed that LJ1 and LJ2 fulfill Koch's postulates for taro soft rot. The two pathogens can infect taro both individually and simultaneously, and neither synergistic nor antagonistic interaction was observed between the two pathogens. Genome sequencing of the two strains indicated that LJ1 represents a novel species of the genus Pectobacterium , for which the name " Pectobacterium colocasium sp. nov." is proposed, while LJ2 belongs to Pectobacterium aroidearum. Pan-genome analysis revealed multiple pathogenicity-related differences between LJ1, LJ2, and other Pectobacterium species, including unique virulence factors, variation in the copy number and organization of Type III, IV, and VI secretion systems, and differential production of plant cell wall degrading enzymes. This study identifies two new soft rot Pectobacteriaceae (SRP) pathogens causing taro soft rot in China, reports a new case of co-infection of plant pathogens, and provides valuable resources for further investigation of the pathogenic mechanisms of SRP. [ABSTRACT FROM AUTHOR]

Extracellular vesicles (EVs) were isolated from Pectobacterium zantedeschiae culturing media using direct ultracentrifugation (UC), iodixanol cushion ultracentrifugation (ICUC), and iodixanol density gradient ultracentrifugation (IDGUC) techniques. The isolates were characterized with total protein content assay (bicinchoninic acid assay, BCA), nanoparticles tracking analysis (NTA), and capillary electrophoresis (CE). A satisfactory correlation (R2 > 0.94) between quantitative results obtained with BCA, NTA and CE was achieved only for isolates obtained with the IDGUC. The correlation between protein content and CE was proved to be related to the isolates' purity. The NTA was found unable to provide reliable information on EVs quantity in samples isolated with UC and ICUC, due to the co-isolated particulate impurities. Moreover, the work reports polysaccharides, used as culturing media components, as a potential source of bias of quantitation with total protein content assay and NTA. The study demonstrates the advantageous selectivity of CE in quality control of EVs and its ability to differentiate subpopulations of EVs of Pectobacterium. [ABSTRACT FROM AUTHOR]

Aim: The newly defined species Pectobacterium parmentieri has emerged as an aggressive pathogen that causes soft rot and blackleg diseases on potato and has been widely disseminated across the globe, jeopardizing the productivity and potato food safety. The implementation of a fast and accurate detection tool is imperative to control, monitor and prevent further spread of these pathogens. The objective of this work was to develop a specific and sensitive multiplex TaqMan qPCR to detect P. parmentieri and distinguish it from all known Pectobacterium species. A universal internal control was included to enhance the reliability of the assay. Methods and Results: A comparative genomics approach was used to identify O‐acetyltransferase and the XRE family transcriptional regulator as specific targets for primers/probe design for the detection of the Pectobacterium genus and P. parmentieri, respectively. Specificity was assessed with 35 and 25 strains included in the inclusivity and exclusivity panels, respectively, isolated from different geographical locations and sources. The assay specifically detected all 35 strains of Pectobacterium sp. and all 15 P. parmentieri strains. No cross‐reactivity was detected during assay validation. Our assay detected up to 10 fg genomic DNA and 1 CFU ml−1 bacterial culture. No change in the detection threshold (1 CFU ml−1) was observed in spiked assays after adding host tissue to the reactions. The assay was validated with naturally and artificially infected host tissues and soil rhizosphere samples. All infected plant samples containing the target pathogens were accurately amplified. Conclusion: The presented multiplex TaqMan qPCR diagnostic assay is highly specific, sensitive, reliable for the detection of Pectobacterium species and P. parmentieri with no false positives or false negatives. Significance and Impact of the Study: The developed assay can be adopted for multiple purposes such as seed certification programmes, surveillance, biosecurity, microbial forensics, quarantine, border protection, inspections and epidemiology. [ABSTRACT FROM AUTHOR]

This study aimed to detect soft rot caused by Pectobacterium carotovorum subsp. carotovorum in lettuce using images obtained by multispectral sensors mounted on an unmanned aerial vehicle (UAV). A secondary objective was to identify the best sensor and determine the optimal stage after inoculation to detect infected plants. In the field, soft rot lesions and the agronomic traits of lettuce plants inoculated or not with the bacteria were assessed on different days after inoculation (DAI). Classifications were made using the Support Vector Machine (SVM) and Naive Bayes (NB) algorithms to analyze data groups consisting of spectral bands, vegetation indices and a combination of bands and indices obtained from a conventional visible camera and Mapir Survey3W multispectral camera, as well as agronomic parameters. The results confirmed the possibility of pre-symptomatic detection of P. carotovorum subsp. carotovorum in lettuce at the canopy level. With respect to identifying healthy and infected lettuce plants by supervised classification, the best results were obtained at 4 and 8 DAI, especially when using the subsets derived from the Mapir Survey3W camera (RGN sensor), for both classifiers. The subsets obtained with the conventional visible sensor (RGB sensor) produced the best results at 20 and 24 DAI. [ABSTRACT FROM AUTHOR]

Soft rot caused by Pectobacterium carotovorum is the most serious disease of postharvest green pepper during storage and transportation. The food-grade lactic acid bacteria (LAB) can be promising candidates for development as an antagonist to control soft rot of postharvest green pepper. In this study, a total of 320 isolates were isolated from 52 traditional fermented food samples of China. Among them, 29 isolates showed good antibacterial activity against P. carotovorum in vitro and significantly reduced lesion diameter of green pepper soft rot in vivo. They were identified to be Leuconostoc mesenteroides , L. holzapfelii , L. citreum , Companilactobacillus alimentarius , Lactobacillus plantarum , L. sakei , Staphylococcus edaphicus , S. saprophyticus , Enterococcus faecium and E. hirae. The antagonistic L. mesenteroides SNP037 was focused on as its superior properties. L. mesenteroides SNP037 had a significant higher effectiveness on green pepper soft rot control than the B. subtilis R31, with lower disease incidence and smaller lesion diameter. L. mesenteroides SNP037 treatment resulted in less P. carotovorum cells on the green pepper tissue than B. subtilis R31 treatment. The quality of green pepper in L. mesenteroides SNP037 treatment group was consistently better than that in B. subtilis R31 treatment group during the whole storage period. Then, biocontrol mode of L. mesenteroides SNP037 was investigated. Cell-free supernatant (CFS) and crude bacteriocin of L. mesenteroides SNP037 inhibited the growth of P. carotovorum. A bacteriocin cluster was found on the whole genome of L. mesenteroides SNP037 after sequencing on Illumina HiSeq platform. It was a novel bacteriocin with low identity with known bacteriocins and named FP-41. The bacteriocin gene was ligated to the expression vector pET-30a (+) and transformed into E. coli BL21 (DE3) plysS for heterologous expression. Result showed that the bacteriocin FP-41 had antibacterial activity against P. carotovorum. These results suggest that L. mesenteroides SNP037 has great potentials as an antagonist to control soft rot of green pepper through producing bacteriocin. • 320 strains were isolated from 52 traditional fermented foods of China. • 29 isolates showed good antibacterial activity against soft rot pathogen. • L. mesenteroides SNP037 efficiently controlled soft rot of postharvest green pepper. • It had a higher soft rot control ability than the commercial B. subtilis R31. • The novel bacteriocin FP-41 had antibacterial activity against soft rot pathogen. [ABSTRACT FROM AUTHOR]

• The genus Prodigiosinella belongs to the Pectobacteriaceae family. • Prodigiosinella aquatilis members are pectinolytic bacteria. • Prodigiosinella members have been isolated in four different continents. • They were isolated from water in coastal brackish wetlands. • Colonies are pink due to the production of the red pigment prodigiosin. The Pectobacteriaceae family comprises plant pathogens able to provoke diverse diseases, including plant maceration due to the production of pectinases disrupting the plant cell wall. To better understand their diversity, a survey of pectinolytic bacteria was performed in brackish lakes of the French region La Camargue near the Mediterranean Sea. The genome of six atypical isolates was sequenced; their size is around 4.8 to 5.0 Mb, including a plasmid of 59 to 61 kb; their G+C values range from 49.1 to 49.3 mol%. Phylogenetic analyses indicated that the novel strains form a new clade of Pectobacteriaceae that branches at the basis of the group encompassing the genera Lonsdalea , Musicola , and Dickeya. Based on phenotypic, genomic and phylogenetic characteristics, we propose the creation of a new genus with the name Prodigiosinella gen. nov. Both the phenotypic and phylogenetic analyses separated the strains into two distinct subgroups, G1 and G2. The type strain LS101T (CFBP 8826T = LMG 32072T) and strain CE70 (CFBP 9054 = LMG 32867) are representative G1 and G2 members, respectively. Three genomic methods were used to analyze DNA–DNA relatedness: digital DNA–DNA hybridization (isDDH), average nucleotide identity (ANI), and genome alignment fraction (AF). They revealed a close relationship between genomes of the two groups, supporting their appurtenance to a same species for which we propose the name Prodigiosinella aquatilis sp. nov. Four strains previously designated as Serratia sp. (ATCC 39006), Brenneria " ulupoensis" (K61) or Erwinia sp. (MK01 and MK09) belong to the new genus Prodigiosinella. [ABSTRACT FROM AUTHOR]

Erwinia mallotivora, the causal agent of papaya dieback disease, is a devastating pathogen that has caused a tremendous decrease in Malaysian papaya export and affected papaya crops in neighbouring countries. A few studies on bacterial species capable of suppressing E. mallotivora have been reported, but the availability of antagonistic fungi remains unknown. In this study, mycelial suspensions from five rhizospheric Trichoderma isolates of Malaysian origin were found to exhibit notable antagonisms against E. mallotivora during co-cultivation. We further characterised three isolates, Trichoderma koningiopsis UKM-M-UW RA5, UKM-M-UW RA6, and UKM-M-UW RA3a, that showed significant growth inhibition zones on plate-based inhibition assays. A study of the genomes of the three strains through a combination of Oxford nanopore and Illumina sequencing technologies highlighted potential secondary metabolite pathways that might underpin their antimicrobial properties. Based on these findings, the fungal isolates are proven to be useful as potential biological control agents against E. mallotivora, and the genomic data opens possibilities to further explore the underlying molecular mechanisms behind their antimicrobial activity, with potential synthetic biology applications. [ABSTRACT FROM AUTHOR]

L-asparaginases (EC 3.5.1.1) are a family of enzymes that catalyze the hydrolysis of L-asparagine to L-aspartic acid and ammonia. These proteins with different biochemical, physicochemical and pharmacological properties are found in many organisms, including bacteria, fungi, algae, plants and mammals. To date, asparaginases from E. coli and Dickeya dadantii (formerly known as Erwinia chrysanthemi) are widely used in hematology for the treatment of lymphoblastic leukemias. However, their medical use is limited by side effects associated with the ability of these enzymes to hydrolyze L-glutamine, as well as the development of immune reactions. To solve these issues, gene-editing methods to introduce amino-acid substitutions of the enzyme are implemented. In this review, we focused on molecular analysis of the mechanism of enzyme action and to optimize the antitumor activity. [ABSTRACT FROM AUTHOR]

This work introduces an alternative workflow for the discovery of novel bacterial biocontrol agents in wheat against Fusarium head blight (FHB). Unlike the mass testing of isolate collections, we started from a diverse inoculum by extracting microbiomes from ears of field-grown plants at grain filling stage. Four distinct microbial communities were generated which were exposed to 3 14-day culture-independent experimental enrichments on detached wheat spikes infected with F. graminearum PH1. We found that one bacterial community reduced infection symptoms after 3 cycles, which was chosen to subsequently isolate bacteria through limiting dilution. All 94 isolates were tested in an in vitro and in planta assay, and a selection of 14 isolates was further tested on detached ears. The results seem to indicate that our enrichment approach resulted in bacteria with different modes-of-action in regard to FHB control. Erwinia persicina isolate C3 showed a significant reduction in disease severity (Fv/Fm), and Erwinia persicina C3 and Pseudomonas sp. B3 showed a significant reduction in fungal biomass (cGFP). However, the mycotoxin analysis of both these treatments showed no reduction in DON levels. Nevertheless, Pantoea ananatis H3 and H11 and Erwinia persicina H2 were able to reduce DON concentrations by more than 50%, although these effects were not statistically significant. Lastly, Erwinia persicina H2 also showed a significantly greater glucosylation of DON to the less phytotoxic DON-3G. The bacterial genera isolated through the enrichment cycles have been reported to dominate microbial communities that develop in open habitats, showing strong indications that the isolated bacteria can reduce the infection pressure of F. graminearum on the spike phyllosphere. [ABSTRACT FROM AUTHOR]

[Display omitted] • High concentrations of isomaltulose can be obtained by immobilized Erwinia sp. D12 cells. • Algaroba gum is a potential alternative to immobilize Erwinia sp. D12 cells. • Immobilized cells produced 167.52 g/L isomaltulose after 72 h of continuous reaction. • 50 % of alginate can be replaced by incorporating algaroba gum. Isomaltulose is a non-cariogenic sugar widely used in sweets and soft drinks that can be obtained from sucrose via enzymatic reactions catalyzed by microbial glucosyltransferases. In this study, an efficient novel system composed of algaroba gum, cashew gum, and alginate was used on the immobilization of Erwinia sp. D12 aiming improve its capability for conversion of sucrose into isomaltulose. The microbial growth of Erwinia sp. D12 cells was carried out in a bioreactor for obtaining the cell mass of microorganisms. The beads containing the microorganisms were characterized by diameter size, encapsulation efficiency, and FTIR spectroscopy; isomaltulose quantification was performed by high-performance liquid chromatography. The system composed of algaroba gum (1.5 % w/v) and alginate (1.5 % w/v) possessed the highest encapsulation efficiency (91.81 %) of immobilized cells and high isomaltulose production (323.38 g/L) at 12 h of reaction. Additionally, the stability of the immobilized cells was kept until 72 h of continuous reaction resulting in an isomaltulose production of 167.52 g/L; demonstrating that up to 50 % of alginate can be replaced by incorporating algaroba gum. The combination of algaroba gum and alginate is promising and recommended when immobilizing Erwinia sp. D12 cells, as results showed an increase in conversion of sucrose into isomaltulose. [ABSTRACT FROM AUTHOR]

Climatic conditions play a crucial part regarding the development of blackleg caused by Pectobacterium atrocepticum (Pa), which is one of the major threats the potato industry faced. These factors affect the stage of growth, host susceptibility, succulence, vigor, survival, rate of multiplication, direction of pathogen dispersion, rate of spore penetration and germination. In the present experiment, effect of these environmental variables like maximum and minimum temperature (max and min T), relative humidity (RH), rainfall (RF), and wind speed (WS) on disease development were studied. Significant positive correlation was observed for all twenty-five varieties and a multiple regression model (Y = + 24.382 + 0.3592X1 + .0970X2-0.2551X3 + 1.982X4) based on the two-year study was developed to find out the relationship between environmental parameters and disease projection. Goodness of model was indicated by coefficient determination value (97.5%). Correlation of these environmental factors on the development of disease on twenty-five potato varieties during 2018, 2019 was done. Significant positive correlation was observed on all twenty-five varieties. In conclusion, it was established that environmental factors, including max. T (28 °C), min. T (15 °C), RH 70%, and WS 3.5 Km/h were conducive for the development of blackleg. Current study would be helpful for researchers in designing better disease management strategies under changing climatic conditions in the future. [ABSTRACT FROM AUTHOR]

Background and Aims Plant diseases caused by Pectobacterium atrosepticum are often accompanied by extensive rot symptoms. In addition, these bacteria are able to interact with host plants without causing disease for long periods, even throughout several host plant generations. There is, to date, no information on the comparative physiology/biochemistry of symptomatic and asymptomatic plant– P. atrosepticum interactions. Typical (symptomatic) P. atrosepticum infections are associated with the induction of plant responses mediated by jasmonates, which are one of the products of the lipoxygenase cascade that gives origin to many other oxylipins with physiological activities. In this study, we compared the functioning of the lipoxygenase cascade following typical and latent (asymptomatic) infections to gain better insight into the physiological basis of the asymptomatic and antagonistic coexistence of plants and pectobacteria. Methods Tobacco plants were mock-inoculated (control) or infected with the wild type P. atrosepticum (typical infection) or its coronafacic acid-deficient mutant (latent infection). The expression levels of the target lipoxygenase cascade-related genes were assessed by Illumina RNA sequencing. Oxylipin profiles were analysed by GC-MS. With the aim of revising the incorrect annotation of one of the target genes, its open reading frame was cloned to obtain the recombinant protein, which was further purified and characterized using biochemical approaches. Key Results The obtained data demonstrate that when compared to the typical infection, latent asymptomatic P. atrosepticum infection is associated with (and possibly maintained due to) decreased levels of 9-lipoxygenase branch products and jasmonic acid and increased level of cis -12-oxo-10,15-phytodienoic acid. The formation of 9-oxononanoic acid and epoxyalcohols in tobacco plants was based on the identification of the first tobacco hydroperoxide lyase (HPL) with additional epoxyalcohol synthase (EAS) activity. Conclusions Our results contribute to the hypothesis of the oxylipin signature, indicating that different types of plant interactions with a particular pathogen are characterized by the different oxylipin profiles of the host plant. In addition, the tobacco LOC107825278 gene was demonstrated to encode an NtHPL (CYP74C43) enzyme yielding volatile aldehydes and aldoacids (HPL products) as well as oxiranyl carbinols (EAS products). [ABSTRACT FROM AUTHOR]

N6-methylated adenine (m6A) is the most prevalent modification of mRNA methylation and can regulate many biological processes in plants, such as mRNA processing, development, and stress response. Some studies have increased our understanding of its various roles in model plants in recent years. Nevertheless, the distribution of m6A and the impact of m6A on the regulation of plant defense responses against pathogen inoculation are virtually unknown in pear. In this study, MeRIP-seq and RNA-seq data from healthy and inoculated plants were analyzed to assess the changes in the transcript levels and posttranscriptional modification of pear in response to the fire blight pathogen Erwinia amylovora. Following the analysis of 97,261 m6A peaks, we found that m6A preferred to modify duplicate genes rather than singleton genes and that m6A-methylated genes underwent stronger purifying selection. A total of 2,935 specific m6A sites were detected at the transcriptome level after inoculation, which may increase defense-related transcript abundance to enhance pear resistance. In addition, 1,850 transcripts were detected only in the mock-inoculated groups. The hypomethylated transcripts were mainly related to transcriptional regulation and various biological processes, such as chloroplast organization and sucrose biosynthetic processes. In addition, we found that the extent of m6A methylation was significantly positively correlated with the transcript level, suggesting a regulatory role for m6A in the plant response. [ABSTRACT FROM AUTHOR]

Bacteria belonging to the Pectobacterium and Dickeya genera are globally distributed phytopathogens that are responsible for economically important soft rot and blackleg diseases of potatoes. Since 2014, there have been increased outbreaks of blackleg disease in the Eastern US, with many cases caused by an especially virulent, nearly clonal strain of Dickeya dianthicola. This disease is thought to be spread via commercial trade of seed tubers with latent infections of these bacteria. There is an urgent need to develop resistant potato varieties to help reduce the accidental spread and damage caused by these diseases. In this study, we conducted an iterative screen of US Potato Genebank (Sturgeon Bay, WI) collections to find wild potato relatives with resistance to tuber soft rot. We found several Solanum microdontum lines with high-level resistance that may be useful as source germplasm for breeding soft rot resistance into commercial potato varieties. [ABSTRACT FROM AUTHOR]

The resistant phenotype of pear cultivars includes either complete resistance or tolerance to the attack by the causative agent of fire blight. The effect of ferritin and active iron (Fe2+) concentrations was compared in resistant (Dargazi), tolerant (Harrow Sweet), and susceptible (Bartlett) pear cultivars inoculated with a wild type strain of Erwinia amylovora (Ea273). The examination of necrosis revealed that resistance to the disease manifested itself as a delay in the appearance of symptoms and progression of the disease. Despite a rise in ferritin levels in all cultivars following disease agent inoculation, resistant and tolerant cultivars showed greater ferritin levels than susceptible cultivar. In addition, only resistant and tolerant cultivars displayed a drop in Fe2+ levels. The rate of decrease in Fe2+ content in Dargazi and Harrow sweet cultivars was 28 and 33%, respectively. Hydrogen peroxide (H2O2) and hydroxyl radical (OH•−) accumulation and intensities were considerably distinctive. Furthermore, tolerant and sensitive cultivars started producing OH•− radicals faster than resistant cultivar. According to these findings, different pathways are employed by pear cultivars to respond to the causative agent of fire blight. [ABSTRACT FROM AUTHOR]

BACKGROUND: Copper agents have been widely used in crop protection because of their unique mechanism against resistant pathogenic bacteria; however, their application brings environmental pollution and biosafety problems. Therefore, environmentally friendly copper agents have attracted attention. In this study, copper sulfide nanoparticles (CuS NPs) were prepared, characterized, analyzed for antibacterial activity and safety. RESULTS: Characterization results showed that the prepared pure CuS NPs have flake nanostructures, hexagonal crystal system, and size range from 40 to 60 nm. These CuS NPs exerted excellent antibacterial effects [median effective concentration (EC50) = 17 mg L−1] against Pectobacterium carotovorum subsp. carotovorum (Pcc) in vitro and can effectively delay and reduce bacterial infection in vivo. Antibacterial mechanism analysis revealed that CuS NPs can increase the levels of reactive oxygen species (ROS) and lipid peroxidation and destroy the structure of bacterial cells as observed through scanning electron microscopy (SEM) and Fourier‐transform infrared (FTIR) spectroscopy. These NPs can also inhibit the motility of Pcc. At 7 and 14 days, the 50% lethal concentrations (LC50) of CuS NPs against earthworms were 1136 and 783 mg kg−1, respectively, indicating their low acute toxicity to earthworms and environmental friendliness. Furthermore, the cells (L02) treated by CuS NPs showed relatively high cell viability (> 96%) and low apoptosis rate (only 5.2%), proving that CuS NPs had low cytotoxicity. CONCLUSION: Compared with commercial dicopper chloride trihydroxide (Cu2(OH)3Cl), CuS NPs could be used as a highly effective, lowly toxic, and environmentally friendly antibacterial agent. © 2021 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

To better understand the taxonomy of Erwinia in the context of the Erwiniaceae family, we carried out a taxogenomic analysis of the Erwiniaceae, a family that was created following the taxonomic revision of the family, Enterobacteriaceae. There has been no systematic analysis of this family, including the agriculturally relevant genus, Erwinia. Our analyses focused on 80 strains of Erwinia along with 37 strains representing 7 other genera in the family. We identified 308 common proteins, generated a genome-level phylogeny and carried out Average Nucleotide Identity, Average Amino Acid Identity and Percentage of Conserved Protein analyses. We show that multiple strains of Erwinia cannot be assigned to established species groups and that both Erwinia gerundensis and "Erwinia mediterraneensis" are not members of Erwinia. We propose the creation of the genus Duffyella gen. nov. and the reclassification of Erwinia gerundensis to this genus as the type species, Duffyella gerundensis comb. nov. Furthermore, divergence between other species within Erwinia as measured by Average Amino Acid Identity is greater than the divergence between Erwinia and other genera, supporting the possible subdivision of the genus Erwinia into at least two genera. Our analyses also suggest that there is no basis for the establishment of the genus Kalamiella within the Erwiniaceae or the taxonomic revision of the Pantoea septica lineage. Therefore, we propose reclassifying Kalamiella piersonii as Pantoea piersonii comb. nov. Our study provides new insight into the diversity of the Erwiniaceae and provides a solid foundation for advancing taxonomic revision of this broadly relevant family. [ABSTRACT FROM AUTHOR]

Tomato (Lycopersicon esculentum L) is one of the most important vegetable crops widely grown around the world. However, the global economic productivity of this crop is limited to a number of phytopathogens. Bacterial spotting and mild rot of tomatoes caused by Pseudomonas syringae pv. tomato and Pectobacterium carotovorum subsp. carotovorum, are one of the most common bacterial diseases that cause significant losses and reduce both the quality and quantity of fruits and impair the nutritional value of tomatoes. Tomato plants are affected by these pathogens throughout the growing season, the lesion is characterized by chlorosis, necrosis and rot. The aim of the study was to analyze the impact on the physiological and biochemical parameters of tomato sprouts affected by pathogens of P. syringae pv. tomato and P. carotovorum subsp. carotovorum. It was found that strains of P. syringae pv. tomato IZ202014 and P. carotovorum subsp. carotovorum IZ207 cause a 30-40% decrease in germination energy and seed germination of Black Prince, Northern Queen, Riddle and Liana tomatoes compared to the control. According to the analysis of physiological and biochemical parameters, the most resistant to pathogens P. syringae pv. tomato, P. carotovorum subsp. carotovorum variety Northern Queen. Treatment of tomato plants with a suspension of the studied strains causes a decrease in chlorophyll content in seedlings by 58-93%. According to the chlorophyll a/b ratio, the Northern Queen and Black Prince varieties have the highest resistance to bacterial damage. Under the influence of cell suspensions of P. syringae pv. tomato IZ202014 and P. carotovorum subsp. carotovorum IZ207 showed changes in the activity of enzymes of the antioxidant system in tomato seedlings, which indicates a decrease in the oxidative processes of cells. [ABSTRACT FROM AUTHOR]

The paper presents data on the useful properties of quinoa seeds, which are rapidly gaining popularity due to their rich chemical composition. The results of studying the changes in the organoleptic, chemical, and microbiological characteristics of quinoa grain during storage under different conditions are presented. The quinoa seeds considered in the research were stored for 12 months at the temperature +(5–30)°С and relative humidity 20–80%. It has been shown that after storing the quinoa seeds for 12 months at +(5–30)°C, their organoleptic characteristics changed: a noticeable rancid and unpleasant aftertaste appeared, accompanied by a foreign smell and a slight change in the colour. When the quinoa grain was stored at +5°С, its acidity remained within the standards of good grain quality throughout the whole storage period. At +15°C, the acidity remained normal for up to 9 months of storage, and at +30°C, up to 6 months. Analysis of the obtained results on the microbiological characteristics has shown that the predominant component of the bacterial microflora of quinoa grain (70.5%) is non-spore-forming bacterium Erwinia herbicola, a representative of the epiphytic microflora. The micromycetes found are mould fungi of the genus Aspergillus and field fungi of the genus Mucor. It has been shown that when quinoa grain is stored at +(5–30)°С, the vital activity of microorganisms is significantly reduced, the development of bacteria and even moulds is retarded, which has a positive effect on maintaining the grain quality. The studies have made it possible to recommend storing quinoa grain for up to 12 months at +5°C, up to 9 months at +15°C, and not more than 6 months at +30°C. [ABSTRACT FROM AUTHOR]

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The purpose of the study in this paper was to detect and identify pathogens in some dendrofloric plants in different green spaces of Craiova. The detection and identification of the attack of different phytopathogens consisted both in periodic visual observations in the experimental area on certain organs of the host plants that showed symptoms of disease, and in performing laboratory tests. Regarding the detection of certain bacterial diseases, in the monitored period, following bacteria were identified: Agrobacterium tumefaciens, Erwinia carotovora, Pseudomonas marginalis, Xanthomonas hyacinthi, Xanthomonas campestris pv. campestris, Xanthomonas hortorum pv. pelargoni. For proper identification, not only macroscopic observations, but also laboratory tests are required to certify the initial identification in the field and, where possible, a pathogenicity test on susceptible plants was performed, obtaining the reproduction of the disease in the test plants. [ABSTRACT FROM AUTHOR]

BACKGROUND: The treatment of acute lymphoblastic leukemia (ALL) uses the biopharmaceutical l‐asparaginase (ASNase) as the main medication. This drug, from bacterial origin (Escherichia coli or Erwinia chrysanthemi), depletes l‐asparagine (Asn) and secondarily l‐glutamine (Gln – GLNase activity) from the bloodstream, leading leukemic cells to die by deprivation of Asn. The use of ASNase is limited by the high incidence of adverse effects, which collectively can specifically impair quality of life of patients. Its high toxicity caused by the product of the hydrolysis of amino acids and the formation of anti‐ASNase antibodies often required treatment interruption, thus reducing the chances of cure and increasing the rates of disease relapse. RESULTS: In order to improve enzymatic activity, while reducing toxicity, we developed through directed evolution a double‐mutant ASNase from Erwinia chrysanthemi (Erw_DM), which has specific activity for Asn 46% higher than the wild‐type enzyme (Erw_WT). This makes it possible to reduce the amount of protein necessary for depletion of this amino acid and, consequently, the reduction of GLNase activity, considered toxic. In silico analysis showed that a lower number of epitopes was exposed, resulting in reduced recognition of the recombinant protein by antibody anti‐ASNase observed in vitro assay. Furthermore, we observed the same cytotoxic profile for the MOLT‐4 and REH ALL cell lines using 40% lower protein mass of Erw_DM to achieve the minimum enzyme activity required in the bloodstream during treatment. CONCLUSION: Altogether, our findings describe a potent and less immunogenic ASNase, an improvement that may alleviate treatment adverse effects developed in anti‐ALL therapy. © 2021 Society of Chemical Industry (SCI). [ABSTRACT FROM AUTHOR]

Acute lymphoblastic leukemia is the most common childhood malignancy. One of the drugs used in the treatment is Asparaginase, and monitoring of its activity levels enables better outcomes. Since 2018, our laboratory has been working to establish a regular analysis of activity. This implementation allowed to qualify care by detecting silent inactivation and also establishing desensitization as a safe way to overcome the lack of Erwinia. We were able to monitor children aged 0 to 18 years who were being treated with PEG-ASNase. The activity was assessed on days 7 (90 samples) and 14 (52 samples) after ASNase infusions. 142 samples were analyzed. 95.7% reached an adequate activity level (≥ 0.1 IU/mL). Patients treated with ASNase can develop allergic reactions. With the activity monitoring, is possible to circumvent situations like these and implement desensitization protocols for patients who had clinical hypersensitivity without inactivation. Desensitization induces temporary unresponsiveness to drug antigens, allowing the patients to proceed with the prescribed chemotherapy. We have received samples from four patients being treated with different desensitization protocols. Patients tolerated the protocols well. Only one had a grade 2 reaction during the infusion and activity < 0.1 IU/mL, which resulted in the switch to Erwinia. The dose adaptation is a possible and more recent use of ASNase monitoring and we were able to confirm the feasibility of PEG-ASNase desensitization protocols. [ABSTRACT FROM AUTHOR]

Pectinolytic enzymes or pectinases are synthesized naturally by numerous microbes and plants. These enzymes degrade various kinds of pectin which exist as the major component of the cell wall in plants. A pectinase gene encoding endo-polygalacturonase (endo-PGase) enzyme was isolated from Pectobacterium carotovorum a plant pathogenic strain of bacteria and successfully cloned into a secretion vector pHT43 having σA-dependent promoter for heterologous expression in Bacillus subtilis (WB800N).The desired PCR product was 1209bp which encoded an open reading frame of 402 amino acids. Recombinant proteins showed an estimated molecular weight of 48 kDa confirmed by sodium dodecyl sulphate–polyacrylamide-gel electrophoresis. Transformed B. subtilis competent cells harbouring the engineered pHT43 vector with the foreign endo-PGase gene were cultured in 2X-yeast extract tryptone medium and subsequently screened for enzyme activity at various temperatures and pH ranges. Optimal activity of recombinant endo-PGase was found at 40°C and pH 5.0. To assay the catalytic effect of metal ions, the recombinant enzyme was incubated with 1 mM concentration of various metal ions. Potassium chloride increased the enzyme activity while EDTA, Zn++ and Ca++, strongly inhibited the activity. The chromatographic analysis of enzymatic hydrolysates of polygalacturonic acid (PGA) and pectin substrates using HPLC and TLC revealed tri and tetra-galacturonates as the end products of recombinant endo-PGase hydrolysis. Conclusively, endo-PGase gene from the plant pathogenic strain was successfully expressed in Bacillus subtilis for the first time using pHT43 expression vector and could be assessed for enzyme production using a very simple medium with IPTG induction. These findings proposed that the Bacillus expression system might be safer to escape endotoxins for commercial enzyme production as compared to yeast and fungi. Additionally, the hydrolysis products generated by the recombinant endo-PGase activity offer their useful applications in food and beverage industry for quality products. [ABSTRACT FROM AUTHOR]

Pectobacterium carotovorum subsp. carotovorum (Pcc) causes soft-rot disease in a wide variety of plants resulting in economic losses worldwide. It produces various types of bacteriocin to compete against related plant pathogens. Studies on how bacteriocins are extracellularly secreted are conducted to understand the mechanism of interbacterial competition. In this study, the secretion of the low-molecular-weight bacteriocins (LMWB) Carocin S1 and Carocin S3 produced by a multiple-bacteriocin producing strain of Pcc, 89-H-4, was investigated. Tn5 insertional mutagenesis was used to generate a mutant, TH22–6, incapable of LMWBs secretion. Sequence and homology analyses of the gene disrupted by transposon Tn5 insertion revealed that the gene sctT, an essential component of the injectisome type III secretion machinery (T3aSS), is required for the secretion of the bacteriocins. This result raised a question regarding the nature of the secretion mechanism of Pcc bacteriocins which was previously discovered to be secreted via T3bSS, a system that utilizes the bacterial flagellum for extracellular secretions. Our previous report has shown that bacteriocin Carocin S1 cannot be secreted by mutants that are defective of T3bSS-related genes such as flhA, flhC, flhD and fliC. We knocked out several genes making up the significant structural components of both T3aSS and T3bSS. The findings led us to hypothesize the potential roles of the T3aSS-related proteins, SctT, SctU and SctV, as flagellar T3SS chaperones in the secretion of Pcc bacteriocins. This current discovery and the findings of our previous study helped us to conceptualize a unique Type III secretion system for bacteriocin extracellular export which is a hybrid of the injectisome and flagellar secretion systems. [ABSTRACT FROM AUTHOR]

Background: The native potatoes (Solanum tuberosum subsp. tuberosum L.) grown in Chile (Chiloé) represent a new, unexplored source of endophytes to find potential biological control agents for the prevention of bacterial diseases, like blackleg and soft rot, in potato crops. Result : The objective of this study was the selection of endophytic actinobacteria from native potatoes for antagonistic activity against Pectobacterium carotovorum subsp. carotovorum and Pectobacterium atrosepticum, and their potential to suppress tissue maceration symptoms in potato tubers. This potential was determined through the quorum quenching activity using a Chromobacterium violaceaum ATCC 12472 Wild type (WT) bioassay and its colonization behavior of the potato plant root system (S. tuberosum) by means of the Double labeling of oligonucleotide probes for fluorescence in situ hybridization (DOPE-FISH) targeting technique. The results showed that although Streptomyces sp. TP199 and Streptomyces sp. A2R31 were able to inhibit the growth of the pathogens, only the Streptomyces sp. TP199 isolate inhibited Pectobacterium sp. growth and diminished tissue maceration in tubers (p ≤ 0.05). Streptomyces sp. TP199 had metal-dependent acyl homoserine lactones (AHL) quorum quenching activity in vitro and was able to colonize the root endosphere 10 days after inoculation. Conclusions: We concluded that native potatoes from southern Chile possess endophyte actinobacteria that are potential agents for the disease management of soft rot and blackleg. [ABSTRACT FROM AUTHOR]

JZP‐458 is a recombinant Erwinia asparaginase produced using a novel Pseudomonas fluorescens expression platform that yields an enzyme expected to lack immunologic cross‐reactivity to Escherichia coli–derived asparaginases. It is being developed as part of a multiagent chemotherapeutic regimen to treat acute lymphoblastic leukemia or lymphoblastic lymphoma patients who develop E coli–derived asparaginase hypersensitivity. A population pharmacokinetic (PopPK) model was developed for JZP‐458 using serum asparaginase activity (SAA) data from a phase 1, single‐dose study (JZP458‐101) in healthy adults. Effects of intrinsic covariates (body weight, body surface area, age, sex, and race) on JZP‐458 PK were evaluated. The model included SAA data from 24 healthy adult participants from the phase 1 study who received JZP‐458: intramuscular (IM) data at 12.5 mg/m2 (N = 6) and 25 mg/m2 (N = 6), and intravenous (IV) data at 25 mg/m2 (N = 6) and 37.5 mg/m2 (N = 6). Model simulations of adult and pediatric SAA profiles were performed to explore the likelihood of achieving a therapeutic target nadir SAA (NSAA) level ≥0.1 IU/mL based on different administration strategies. PopPK modeling and simulation suggest JZP‐458 is expected to achieve 72‐hour NSAA levels ≥0.1 IU/mL in 100% of adult or pediatric populations receiving IM administration at 25 mg/m2, and in 80.9% of adult and 94.5% of pediatric populations receiving IV administration at 37.5 mg/m2 on a Monday/Wednesday/Friday (M/W/F) dosing schedule. Based on these results, the recommended starting dose for the phase 2/3 pivotal study is 25 mg/m2 IM or 37.5 mg/m2 IV on a M/W/F dosing schedule in pediatric and adult patients. [ABSTRACT FROM AUTHOR]

Pectobacterium brasiliense (Pbr) infects a wide range of crops worldwide, causing potato blackleg and soft rot and vegetable soft rots. This study aimed to characterize the genetic diversity and virulence variability among 68 Pbr strains isolated from either symptomless potato progeny tubers, diseased potato plants, ware potatoes wash water, or vegetables grown in Israel, as well as strains isolated from symptomless seed tubers grown in Europe, or diseased potato plants grown in France. The collection was typed using PCR and TaqMan real‐time PCR analyses, dnaX sequence analysis, pulsed‐field gel electrophoresis (PFGE), and pectolytic activity. dnaX phylogeny grouped almost all strains in a common genetic clade related to Pbr, which was distinct from the other Pectobacterium species. PFGE analysis identified two main clusters, including one major group of 47 strains with 95%–100% similarity. Maceration assays on two potato cultivars showed significant differences between strains but with no correlations with the source of the strains nor the status of the host (with/without symptoms). Molecular (dnaX sequences and PFGE profiles) and phenotypic analyses (tuber maceration tests) showed that the tested Pbr strains are not a homogeneous group. Analysis of the tested Pbr strains isolated from potato and vegetables grown in fields with a history of potato cultivation suggests that seed tubers imported from Europe may be the main source for Pbr in Israel. To the best of our knowledge, this is the first study that describes biodiversity and population structure of P. brasiliense isolated from potato and vegetables under hot climate conditions. [ABSTRACT FROM AUTHOR]

Erwinia tracheiphila, the causal pathogen of bacterial wilt of cucurbit crops, is disseminated by cucumber beetles. A bacteriophage, designated FBB1 (Fu-Beattie-Beetle-1), was isolated from spotted cucumber beetles (Diabrotica undecimpunctata) that were collected from a field in which E. tracheiphila is endemic. FBB1 was classified into the Myoviridae family based on its morphology, which includes an elongated icosahedral head (106 × 82 nm) and a putatively contractile tail (120 nm). FBB1 infected all 62 E. tracheiphila strains examined and three Pantoea spp. strains. FBB1 virions were stable at 55°C for 1 h and tolerated a pH range from 3 to 12. FBB1 has a genome of 175,994 bp with 316 predicted coding sequences and a GC content of 36.5%. The genome contains genes for a major bacterial outer-membrane protein, a putative exopolysaccharide depolymerase, and 22 predicted transfer RNAs. The morphology and genome indicate that FBB1 is a T4-like virus and thus in the Tevenvirinae subfamily. FBB1 is the first virulent phage of E. tracheiphila to be reported and, to date, is one of only two bacteriophages to be isolated from insect vectors of phytopathogens. Collectively, the results support FBB1 as a promising candidate for biocontrol of E. tracheiphila based on its virulent (lytic) rather than lysogenic lifestyle, its infection of all E. tracheiphila strains examined to date, and its infection of a few nonpathogenic bacteria that could be used to support phage populations when pathogen numbers are low. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Agricultural Science & Technology (1008-0864) is the property of Journal of Agricultural Science & Technology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Our previous study identified a new β-galactosidase in Erwinia sp. E602. To further understand the lactose metabolism in this strain, de novo genome assembly was conducted by using a strategy combining Illumina and PacBio sequencing technology. The whole genome of Erwinia sp. E602 includes a 4.8 Mb chromosome and a 326 kb large plasmid. A total of 4,739 genes, including 4,543 protein-coding genes, 25 rRNAs, 82 tRNAs and 7 other ncRNAs genes were annotated. The plasmid was the largest one characterized in genus Erwinia by far, and it contained a number of genes and pathways responsible for lactose metabolism and regulation. Moreover, a new plasmid-borne lac operon that lacked a typical β-galactoside transacetylase (lacA) gene was identified in the strain. Phylogenetic analysis showed that the genes lacY and lacZ in the operon were under positive selection, indicating the adaptation of lactose metabolism to the environment in Erwinia sp. E602. Our current study demonstrated that the hybrid de novo genome assembly using Illumina and PacBio sequencing technologies, as well as the metabolic pathway analysis, provided a useful strategy for better understanding of the evolution of undiscovered microbial species or strains. [ABSTRACT FROM AUTHOR]

Pectobacterium parmentieri (formerly Pectobacterium wasabiae), which causes soft rot disease in potatoes, is a newly established species of pectinolytic bacteria within the family Pectobacteriaceae. Despite serious damage caused to the potato industry worldwide, no field-deployable diagnostic tests are available to detect the pathogen in plant samples. In this study, we aimed to develop a reliable, rapid, field-deployable loop-mediated isothermal amplification (LAMP) assay for the specific detection of P. parmentieri. Specific LAMP primers targeting the petF1 gene region, found in P. parmentieri but no other Pectobacterium spp., were designed and validated in silico and in vitro using extensive inclusivity (15 strains of P. parmentieri) and exclusivity (94 strains including all other species in the genus Pectobacterium and host DNA) panels. No false positives or negatives were detected when the assay was tested directly with bacterial colonies, and with infected plant and soil samples. Sensitivity (analytical) assays using serially diluted bacterial cell lysate and purified genomic DNA established the detection limit at 10 CFU/mL and 100 fg (18–20 genome copies), respectively, even in the presence of host crude DNA. Consistent results obtained by multiple users/operators and field tests suggest the assay's applicability to routine diagnostics, seed certification programs, biosecurity, and epidemiological studies. [ABSTRACT FROM AUTHOR]

Soft rot disease by Pectobacterium carotovorum subsp. carotovorum (Pcc) is one of the causes of post-harvest damage to economically important vegetables such as carrots, cabbage, cucumbers, and others. The study reported growth optimization to increase the antimicrobial activity of the bacteriocin from Pediococcus pentosaceus 2397 to inhibit Pcc. The antimicrobial activity of cell-free supernatant and crude bacteriocin was carried out using the well agar diffusion method. The results showed that the antimicrobial activity of the bacteriocin from strain 2397 could be increased by regulating the growth environment and adding nutrients to the growth medium. The optimum growth conditions for strain 2397 to produce bacteriocins that have high antimicrobial activity were 24 h incubation time, pH 6.3-7.0, and 7.5% starter concentration, the addition of 4% Carbon sources (sucrose, lactose, mannose, fructose, glucose), and Nitrogen sources (peptone, beef extract, yeast extract, ammonium sulfate) and 2% of tween (tween 20, tween 40 and tween 80). Thus, bacteriocin of P. pentosaceus 2397 strain isolated from dadih has the potential to be used as a safe bio-preservative in place of chemical preservatives to avoid the post-harvest damage to the fresh vegetables. [ABSTRACT FROM AUTHOR]

Endophytes have recently garnered importance worldwide and multiple studies are being conducted to understand their important role and mechanism of interaction inside plants. But before we indulge in their functions it is necessary to dig into the microbiome. This will help to get a complete picture of the microbes intrinsic to their host and understand changes in community composition with respect to their habitats. To fulfil this requirement in our study we have attempted to dissect the endophytic diversity in roots of rice plant grown across the various agro-ecological zones of West Bengal by undergoing amplicon analysis of their 16S rRNA gene. Based on the measured environmental parameters agro-ecological zones can be divided into two groups: nutrient dense groups, representing zones like Gangetic, Northern hill and Terai-Teesta zone characterised by soil with higher levels of nitrogen (N) and total organic carbon and nutrient low groups representing Coastal saline, Red-laterite and Vindhyan zone mainly characterised by high electroconductivity and pH. Gammaproteobacteria, Alphaproteobacteria, Bacilli and Bacteroidetes were mostly abundant in nutrient dense sites whereas Clostridia and Planctomycetes were concentrated in nutrient low sites. Few genera (Aeromonas, Sulfurospirillum, Uliginosibacterium and Acidaminococcus) are present in samples cultivated in all the zones representing the core microbiome of rice in West Bengal, while some other genera like Lactococcus, Dickeya, Azonexus and Pectobacterium are unique to specific zone. Hence it can be concluded that this study has provided some insight in to the endophytic status of rice grown across the state of West Bengal. [ABSTRACT FROM AUTHOR]

The native plant microbiome is composed of diverse microbial communities that influence overall plant health, with some species known to promote plant growth and pathogen resistance. Here, we show the antibacterial and growth promoting activities of autoclaved culture metabolites (ACM) from native endophytic bacteria (NEB). These NEB were isolated from a papaya cultivar (var. Cariflora) that is tolerant to bacterial crown rot (BCR) caused by Erwinia mallotivora. In this cultivar, bacterial colonization in tissues recovering from the disease was observed before onset of tissue regeneration or 'regrowth'. We further isolated and characterized these bacteria and were able to identify two culturable stem NEB related to plant endophytic genera Kosakonia sp. (ex. Enterobacter sp., isolate EBW), and to Sphingomonas sp. (isolate EBY). We also identified root NEB under genus Bacillus (isolates BN, BS, and BT). Inhibition assays indicated that ACM from these NEB promptly (within 18-30 h) and efficiently inhibited (60–65% reduction) E. mallotivora proliferation in vitro. When surface-sterilized papaya seeds were soaked in ACM from isolates EBY and EBW, germination was variably retarded (20–60% reduction) depending on plant genotype, but plant biomass accumulation was significantly stimulated, at around two-fold increase. Moreover, greenhouse experiments show that ACM from all isolates, especially isolate EBW, significantly reduced BCR incidence and severity in a susceptible genotype (var. Solo), at around two-fold. In general, our observations of pathogen antagonism and plant growth promotion leading to disease reduction, suggested the influence of native endophytic bacteria to increased fitness in plants, and tolerance against the re-emerging crown rot disease of papaya. [ABSTRACT FROM AUTHOR]