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Xanthomonas fragariae is a global quarantine pathogen, which typically inflicts angular leaf spots. In the present study, we report a new 4.11-Mb high-quality genome sequence of X. fragariae YL19. YL19 can cause the typical angular leaf spot symptoms on strawberry plants in China as well as crown infection pocket symptoms. This new symptom has not been reported in other X. fragariae. Compared with typical X. fragariae strains, including PD885, NBC2815, PD5205, Fap21, and Fap29, the genome and plasmid in YL19 were smaller in size, lacking 109 coding genes, and have more carbohydrate-active enzyme and secondary metabolism genes. The YL19 genome ought to clarify the molecular mechanisms of genome evolution, host adaptation, and pathological process of X. fragariae and help improve strawberry management strategies. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Strawberry production is challenged by several abiotic and biotic stresses, such as drought, soil salinity, and the angular leaf spot (ALS) disease caused by Xanthomonas fragariae. In recent decades, the development of commercial products containing combinations of different Plant-Growth-Promoting (PGP) microorganisms has been one of the main focuses of agricultural research. However, their results are often erratic depending on crop species, environmental conditions, and competition among the different strains or indigenous plant microbiota. The use of beneficial microorganisms selected from the crop-specific microbiota may help overcome this limitation, promoting their utilization for sustainable agriculture. The culturable bacteriota of strawberry plants was screened to identify PGP activities in vitro. Bacterial isolates were tested in vivo on strawberry plants in both optimal and stress (X. fragariae infection or salinity) conditions, allowing the selection of strains of Pseudomonas fluorescens, Stenotrophomonas rhizophila, and Agrobacterium rubi whose application showed a significant increase in plant growth and fruit production (up to seven-fold), even under stress conditions, and the ability to control ALS by over 50%. Potential synergistic effects among PGP isolates were tested by coordinated inoculation. However, plant growth and fruit quality were not promoted, except for fruit weight and size, by coordinate inoculation in comparison to m23 and m27 single-strain treatment.

Strawberry leaf spot caused by Xanthomonas fragariae is one of the main limiting diseases of crops in Brazil, but there are not efficient control strategies. Additionally, the global demand for pesticide-free foods has stimulated research to find low toxicity products to control plant pathogens. Thus, strawberry plants were sprayed with Lentinula edodes polysaccharides (PSHII), propolis extracts (GP1), acibenzolar-s-methyl (ASM) or distilled water (control) and inoculated with the bacterium. Leaf samples were collected to determine phenylalanine ammonia lyase (PAL) and superoxide dismutase (SOD) activities. Antimicrobial activity was checked, based on the effect of the products on CFU and bacterial biofilm. In the cv. Albion, the propolis source GP1 reduced the AUDPC by more than 95%, compared to control, when applied 3 days before inoculation, but PSHII did not differ from control. For cv. San Andreas, GP1 significantly reduced the disease only at 7 days after inoculation. PSHII or ASM promoted significant reduction in all evaluations. Only GP1 inhibited the CFU and biofilm formation of the bacterium and promoted increases in PAL and SOD activities, while PSHII and ASM increased SOD activity. The data suggest that such products have potential for use in programs for control of strawberry diseases.

Xanthomonas fragariae, the bacterium causing angular leaf spot (ALS) of strawberry, is found routinely on strawberry nursery stock. Although ALS can be a serious disease in production fields, it can be particularly problematic in the sale and trade of commercial nursery stock because of international trade regulations. Heat treatment has been shown to be an effective treatment for managing ALS on nursery stock in small-scale experimental trials. The objective of this research was to design, build, and trial precision thermotherapy units for managing ALS for both research and commercial applications and to test a new thermotherapeutic protocol on strawberry nursery stock that combines a conditioning thermal treatment with an eradicative thermal treatment. Small-plot trials were conducted to evaluate the efficacy of precision thermotherapy on control of ALS using known sources of infected nursery stock. Additionally, trials were conducted in cooperation with commercial nurseries to determine the impact of thermotherapy on plant health and yield and on the natural development of ALS. In the small-plot trials, ALS incidence was significantly lower in plots treated with precision thermotherapy. In the commercial trials, precision thermotherapy had a variable, but negligible, effect on plant growth and yield. ALS, when it occurred, was always lower in thermotherapy-treated plots. Heat is a near-universal biocide. Thus, in addition to managing ALS, the commercial application of precision thermal therapy to strawberry nursery stock may be effective for managing a wide range of pest and disease threats to strawberry while simultaneously reducing pesticide usage. [Formula: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 “No Rights Reserved” license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law. 2021

Xanthomonas fragariae is the causal agent of an important bacterial disease in strawberry production regions worldwide and a quarantine plant pathogen in many countries including New Zealand. Xanthomonas fragariae mainly infects the foliage of strawberry plants but can also infect the calyx tissue associated with strawberry fruit. Fresh strawberries are a high‐value internationally traded commodity that has a short shelf‐life. When making biosecurity decisions based on the finding of a quarantine organism such as X. fragariae by PCR, one of the major challenges is the inability to differentiate positive results originating from viable or dead cells. Viability PCR (vPCR) is a technique that selectively inhibits PCR amplification of DNA derived from dead cells through the use of a nucleic acid intercalating dye, for example, PEMAX™. A vPCR protocol has been optimized to enable rapid detection of viable X. fragariae in a tissue sample. PEMAX™ treatment resulted in complete inhibition of PCR amplification of 10⁸–10³ cfu/ml dead X. fragariae cells in strawberry host tissue. The most important parameters for optimization were the dilution of the sample, amplicon length and choice of nucleic acid intercalating dye. This study provides a rapid protocol to discriminate between viable and dead X. fragariae in strawberry in a phytosanitary environment. This test will help timely decisions to be made at the border on imported fresh strawberry consignments that test positive for X. fragariae.

Xanthomonas fragariae causes angular leaf spot in strawberry. The pathogen’s association with its host tissue is thought to be a condition for its survival. Consequently, transmission of the pathogen to field production sites occurs almost exclusively through the movement of contaminated planting stock. The aim of this study was to develop a propidium monoazide (PMA)-quantitative PCR (qPCR) protocol for specific detection of viable X. fragariae cells. The qPCR procedure was developed for two different primer pairs: one producing a long amplicon (863 bp) and the other a short amplicon (61 bp). Both pairs were tested on mixtures of viable and heat-killed bacteria cells, bacteria-spiked strawberry petiole samples, and petioles collected from symptomatic, inoculated plants. The results showed that long-amplicon PMA-qPCR enabled specific and sensitive detection of X. fragariae with a detection limit of 103 CFU/ml, and it significantly improved PMA efficiency in differentiating viable from dead bacterial cells relative to short-amplicon PMA-qPCR. Based on the delta threshold cycle (Ct) values (i.e., the difference in Ct values between PMA-treated and nontreated samples), the long-amplicon PMA-qPCR was able to suppress the detection of dead X. fragariae cells 1.9- to 3.1-fold across all petiole samples tested. The quantification results from PMA-qPCR for mixtures of viable and dead cells were highly correlated with the predicted bacterial concentrations in a linear relationship (R2 = 0.981). This assay can be useful for identifying inoculum sources in the strawberry production cycle, which may lead to improved disease management strategies.

This is the first study which describes a unique procedure of isolating of high-quality, intact RNA from strawberry leaves of Xanthomonas fragariae, three most suitable reference genes, crucial for the normalization of RT-qPCR data for this pathogen and accurate expression analysis of target genes. In our study, various mathematic algorithms: NormFinder geNorm, BestKeeper, the delta CT method, RefFinder were adopted for validation of most stable reference genes from nine candidate genes (ffh, glyA, gyrA, gyrB, proC, pykA, recA, rpoB, rpoD). The analyses allowing to select three most suitable pioneer reference genes, gyrB, ffh, and pykA, that we recommend for the normalization of RT-qPCR data and for the study of the expression of target genes in Xf. Moreover, their combination as references allowed for an accurate expression analysis and computation of the fold change of the flhF and iroN2 genes in Xf. These two genes are important for the success of the colonization of plant tissue and pathogenicity and sequences of primers designed to study these genes, are presented.

Xanthomonas fragariae, the causal agent of angular leaf spot (ALS) of strawberry, is a quarantine pathogen in some export markets, causing trade restrictions and economic loss to the California fresh-market strawberry industry. Preharvest chemical management options are limited to copper, and there are no postharvest treatments available that reduce populations of the pathogen if ALS is detected at an export destination. Here, we report high preharvest efficacy for the experimental bactericide amino thiadiazole and the commercial product zinc thiadiazole, alone and in mixtures with low rates of copper or the antibiotic kasugamycin, with average disease incidence reduction of up to 92.8% compared with the control. Although effective against quarantine insect pests of strawberry, postharvest methyl bromide fumigation was ineffective against X. fragariae in diseased plant tissue at a standard commercial rate. Postharvest propylene oxide fumigation, used for decades by the California nut industries for insect and microbial disinfestation, significantly reduced X. fragariae populations in infected leaflet tissues by at least 2.5-log compared with controls at a dose of ≥142 µg/ml for 2 h at 15 to 20°C. Fumigated leaflets showed little to no phytotoxicity at effective rates, and fumigated fruit were not significantly affected in appearance or susceptibility to postharvest gray mold or Rhizopus rot following storage at 2°C for 3 days and at 15°C for an additional 5 days. Together, these new treatments offer potential strategies for establishing a systems approach with preharvest treatments significantly reducing the risk of ALS on plants and, in response to quarantine detections, a postharvest fumigation treatment that reduces viable pathogen populations in existing lesions.

Xanthomonas fragariae is the causal agent of angular leaf spot of strawberry. Short-read sequences were generated for two X. fragariae strains with different virulence phenotypes on the Illumina HiSeq 2000 platform. The genome sequences will contribute to a better understanding of pathogen evolution and the genes contributing to virulence in X. fragariae., Xanthomonas fragariae is the causal agent of angular leaf spot of strawberry. Short-read sequences were generated for two X. fragariae strains with different virulence phenotypes on the Illumina HiSeq 2000 platform. These genome sequences will contribute to a better understanding of pathogen evolution and the genes contributing to virulence in X. fragariae.

Strawberry is economically important and widely grown, but susceptible to a large variety of phytopathogenic organisms. Among them, Xanthomonas fragariae is a quarantine bacterial pathogen threatening strawberry productions by causing angular leaf spots. Using whole transcriptome sequencing, the gene expression of both plant and bacteria in planta was analyzed at two time points, 12 and 29 days post inoculation, in order to compare the pathogen and host response between the stages of early visible and of well-developed symptoms. Among 28,588 known genes in strawberry and 4046 known genes in X. fragariae expressed at both time points, a total of 361 plant and 144 bacterial genes were significantly differentially expressed, respectively. The identified higher expressed genes in the plants were pathogen-associated molecular pattern receptors and pathogenesis-related thaumatin encoding genes, whereas the more expressed early genes were related to chloroplast metabolism as well as photosynthesis related coding genes. Most X. fragariae genes involved in host interaction, recognition, and pathogenesis were lower expressed at late-phase infection. This study gives a first insight into the interaction of X. fragariae with its host. The strawberry plant changed gene expression in order to consistently adapt its metabolism with the progression of infection.

The quarantine bacterium Xanthomonas fragariae causes angular leaf spots on strawberry. Its population structure was recently found to be divided into four (sub)groups resulting from two distinct main groups. Xanthomonas arboricola pv. fragariae causes bacterial leaf blight, but the bacterium has an unclear virulence status on strawberry. In this study, we use comparative genomics to provide an overview of the genomic variations of a set of 58 X. fragariae and five X. arboricola pv. fragariae genomes with a focus on virulence-related proteins. Structural differences within X. fragariae such as differential plasmid presence and large-scale genomic rearrangements were observed. On the other hand, the virulence-related protein repertoire was found to vary greatly at the interspecies level. In three out of five sequenced X. arboricola pv. fragariae strains, the major part of the Hrp type III secretion system was lacking. An inoculation test with strains from all four X. fragariae (sub)groups and X. arboricola pv. fragariae resulted in an interspecies difference in symptom induction since no symptoms were observed on the plants inoculated with X. arboricola pv. fragariae. Our analysis suggests that all X. fragariae (sub)groups are pathogenic on strawberry plants. On the other hand, the first genomic investigations of X. arboricola pv. fragariae revealed a potential lack of certain key virulence-related factors which may be related to the difficulties to reproduce symptoms on strawberry and could question the plant-host interaction of the pathovar.

Xanthomonas fragariae causes angular leaf spot of strawberry, an important disease in strawberry growing regions worldwide. To better understand how X. fragariae multiplies and moves in strawberry plants, a green fluorescent protein (GFP)-labeled strain was constructed and used to monitor the pathogen’s presence in leaf, petiole, and crown tissue with fluorescence microscopy following natural and wound inoculation in three strawberry cultivars. Taqman PCR was used to quantify bacterial densities in these same tissues regardless of the presence of GFP signal. Results showed X. fragariae colonized leaf mesophyll, the top 1 cm portion of the petiole adjacent to the leaf blade, and was occasionally found colonizing xylem vessels down to the middle of the petioles. The colonization of vascular bundles and the limited systemic movement that was observed appeared to be a passive process, of which the frequency increased with wounding and direct infiltration of bacteria into leaf veins. X. fragariae was able to directly enter petioles and colonize the space under the epidermis. Systemic movement of the bacteria into crown and other uninoculated tissues was not detected visually by GFP. However, X. fragariae was occasionally detected in these tissues by qPCR, but at quantities very near the qPCR detection limit. Petiole tissue harboring bacteria introduced either by direct entry through natural openings or wounds, or by systemic movement from infected foliar tissue, likely serves as a main source of initial inoculum in field plantings.

Xanthomonas fragariae is the causative agent of angular leaf spot of strawberry, a quarantine organism in plant propagation material in the European Union. Field experiments were conducted to assess the risks for infection of strawberry plants through dispersal of an aerosolized inoculum. In practice, pathogen aerosols can be formed during mowing of an infected crop or by water splashing on symptomatic plants during overhead irrigation or rain. In our experiments, aerosols were generated by spraying suspensions of X. fragariae with a density of 108 cfu ml−1 or water under pressure vertically up into the air. In strawberry plants (cv Elsanta) placed at 1.3, 5 and 10 m distance downwind from the spray boom, infections were found, as evidenced with a combination of dilution–plating and molecular techniques, but more frequently in plants wetted prior to inoculation than in plants kept dry. A logarithmic decrease in infection incidence was found with the distance to the inoculum source. Symptomatic plants were found up to 5 m distance from the inoculum source. No infected plants were found in plants placed 4 m upwind or treated with water. In glasshouse studies, it was shown that under conditions favorable for disease development, spray-inoculation of strawberry plants with estimated densities of X. fragariae as low as 2000 cfu per plant were able to cause symptoms both in cv Elsanta and cv Sonata. Results indicate that there is a considerable risk on infections of strawberry plants exposed to aerosolized inoculum.

Xanthomonas fragariae es el agente causal de la mancha angular de la hoja en el cultivo de fresa (Fragaria × ananassa Duch) en México. El objetivo de este estudio fue caracterizar e identificar las bacterias aisladas basadas en la caracterización fisiológica, bioquímica, morfológica y en pruebas de patogenicidad y genética por PCR. Se aislaron colonias de bacterias con morfología similar a la descrita para X. fragariae a partir de tejido foliar de diferentes genotipos de fresa en Cd. Guzmán, Jalisco, México con síntomas de manchas angulares acuosas en el área adaxial, exudado en nervaduras primarias y secundarias y manchas necróticas. Los resultados de la caracterización de los aislamientos bacterianos los identificaron como X. fragariae, lo cual fue confirmado por el análisis genético con la amplificación del gen hrp y la amplificación y secuenciación del gen 16S rADN. La inoculación de la cepa X. fragariae-22 produjo lesiones angulares y necrosis adyacentes a la nervadura de las hojas en plantas susceptibles de fresa var. Jacona.

The spread of Xanthomonas fragariae in a strawberry field by mowing and runner cutting machinery was studied in the Netherlands during autumn. The blades of a rotary mower became heavily contaminated after trimming the leaves of symptomatic plants. Use of a contaminated mower resulted in infection of plants that were initially free of X. fragariae, even at a distance of 4 m (i.e. 10 mother plants) from the symptomatic plants. Directly after mowing,the pathogen could be detected on damaged leaves at a density of 103–105 cells g-1.Infections were established, although 3 months after mowing symptoms were still absent. There were indications that the use of the mower resulted in the spread of infected leaf material from symptomatic plants to neighbouring beds. Spread by a runner cutting machine was also studied. Again, after cutting the runners of symptomatic plants, high numbers of X. fragariae cells were recovered from debris on the discs of the machine. During cutting, the cut ends of runners of plants initially free of X. fragariae became contaminated, even ata distance of 4 m from the symptomatic plants, but only at low densities of 103 cfu g-1. Three months after stolon cutting, infections were detected in some of the cut plants.

Strawberry (Fragaria × ananassa Duch.) is a kind of fruit with great economic importance and widely cultivated in the world. From 2019 to 2020, a serious crown rot disease was sporadically observed in several strawberry cultivars including 'Zhang Ji', 'Hong Yan' and 'Yue Xiu' in Shanghai, China. Initially, water-soaked rot appeared in inner tissue of strawberry crown, then progressed into browning and hollowing symptoms accompanied with yellow discolorations of young leaves. To isolate and identify the causal agent, small pieces of tissue taken from ten diseased crowns were sterilized by 70% alcohol. The cut-up pieces were macerated and serially diluted. The dilutions were placed on nutrient agar (NA) medium. After incubation at 25°C for 4-5 days, the yellow bacterial colonies were tiny and were streaked on NA plate for purification. The colonies were yellow, mucoid, smooth-margined, and five independent representative colonies were used for further confirmation. To confirm the species identity of the bacterial, genomic DNA was extracted from the five representative isolates, and 16S rRNA gene was amplified and sequenced using universal primers 27F/1492R. The 16S rRNA sequence was deposited in GenBank (MW725235) and showed 99% nucleotide similarity with Xanthomonas fragariae strain LMG 708 (NR_026318). The isolate's identity was further confirmed by X. fragariae-specific primers XF9/XF12 (Roberts et al. 1996). All five isolates could be detected by XF9/XF12 primer. To confirm Koch's postulates, five healthy strawberry plants were placed in 1000 ml glass beakers by submerging the cutting wound in 50 ml the bacterial suspension of 108 CFU/ml. Five additional strawberry plants treated with sterilized water served as a control. The beakers containing inoculated plants were sealed with plastic film at 25°C. Water-soaked rot appeared on internal tissue of crown similar to those observed in the field within 10-12 days after inoculation, while the control samples remained healthy. The bacteria were re-isolated from rot of inoculated crowns, and confirmed by X. fragariae-specific primers XF9/XF12. X. fragariae has been reported to cause angular leaf spot on strawberry in China (Wang et al. 2017; Wu et al., 2020). It's also found that X. fragariae could systematically infect crown tissue (Milholland et al. 1996; Mahuku and Goodwin, 1997). To our knowledge, this is the first report of X. fragariae causing strawberry crown rot in China. This report increased our understanding of X. fragariae, and showed that the spread of this disease might seriously threaten the development of strawberry industry in the future.

Strawberry (Fragaria × ananassa Duch.) is an important fruit crop in China. Typical crown infection pockets symptoms were observed on the infected strawberry in Liaoning province, China (121°60'E, 38°90'N) in the autumn of 2017. The disease incidence was estimated to be around 5 to 10 %, but could reach 30 to 40 % in some heavily affected plastic tunnels. The infected plants early displayed water-soaked symptoms on the abaxial leaf surface and subsequently developed reddish-brown shaped stripes and coalesced lesions on the adaxial leaf surface around the main veins (Fig. 1-A, 1-B). Several variable-size (0.3-0.8 mm in diameter) pockets were observed inside the crown tissues after dissection (Fig. 1-C). The diseased plants rarely reached fruiting and were easily broken between the crown tissue and the stem, and would eventually die. To identify the causal agent of this disease, the several surface-disinfested infected main veins and crown tissues were individually ground in sterile water and plated on sucrose peptone agar(SPA) medium (Hayward 1960) with 10-fold serial dilutions and incubated at 25℃. A number of yellow colonies grew on the medium at the 10-4 dilution 7 days after plating (Fig. 1-D) in all specimens. The colonies were aerobic, yellow, viscous, smooth, and gram-negative, which is a typical characteristic of Xanthomonas. To confirm identity of the causal bacteria, 18 colonies selected randomly were subjected to polymerase chain reactions (PCR) for the amplification of the cpn60 (Sahin et al. 2010), gyrB, rpoD, and fyuA (C Manceau et al. 2011), respectively. The results showed that the 18 colonies are identical. The cpn60, gyrB, rpoD, and fyuA sequences of this isolate were deposited in GenBank with accession numbers MT513132.1, MW233896, MW233897, and MW233895, respectively. BLAST searches with sequences of this isolate cpn60, gyrB, rpoD, and fyuA revealed 97.7%, 96.4%, 97.8%, and 97.3% similarity with the corresponding sequences of X. fragariae strain NBC2815 (LT853880.1), respectively. The resulting concatenated data set of cpn60-gyrB-rpoD-fyuA was used to build a Multilocus Sequence Analysis (MLSA) by maximum likelihood criteria (Fig. 2). The cpn60-gyrB-rpoD-fyuA sequences of the isolate from Liaoning clustered in the clade containing the type strain of X. fragariae NBC2815, indicating that it belongs to X. fragariae. Thus, the bacterial strain from Liaoning was designated as X. fragariae strain YL19. To fulfill Koch's postulates, the base of leaf petioles of disease-free strawberry plants were syringe-infiltrated inoculated with bacterial suspension (2×108 CFU) prepared from colonies of X. fragariae YL19 washed from SPA plates. The inoculated and control (treated with sterile water) were placed in a chamber (25/20℃day/night,≥90% relative humidity(RH), 12/12 h photoperiod) for three months. After one month, water-soaked symptoms were observed in the crown tissues of all X. fragariae YL19-inoculated plants. Two months after inoculation, a significant crown pocket similarly to initial symptoms observed in the field was developed on all inoculated plants. No symptoms were observed in the control plants. The bacteria were re-isolated from the symptomatic leaves, petioles and crowns, and confirmed as X. fragariae YL19 by the above mentioned morphological and molecular analyses. Pathogenicity tests were conducted three times and the same results were obtained. It was reported that X. fragariae usually causes angular leaf spot, a serious bacterial disease in many strawberry production regions worldwide. The typical symptoms of angular leaf spot caused by X. fragariae include reddish-brown, irregular spots on the upper leaf surface, water-soaked lesions developed along leaf veins. Although angular leaf spot caused by X. fragariae has been reported in Tianjin and Taiwan province, China (Wang et al. 2017; Wu et al. 2020), there is no report about the symptoms that infection pockets on crowns caused by X. fragariae strain YL19 as described above. This result indicated that YL19 is different from the other two X. fragariae strains reported in China or the disease caused by YL19 could be a severe case of angular leaf spot and vascular decline or collapse in strawberry (Bradbury, 1977). . To the best of our knowledge, these results showed a previously unreported new strain YL19 of X. fragariae is the causal agent of crown infecton pocket in strawberry in China, it may lead to serious losses to the local strawberry industry. This report will assist in developing management measures for this disease promptly.

In this work it was evaluated the bactericidal effect of silver and silver-gold nanoparticles coated with silicon dioxide on Xanthomonas fragariae. Nanoparticles were synthesized by combining laser ablation and chemical synthesis techniques. Irradiating the samples at the same wavelength where nanoparticles exhibit its maximum absorbance is possible to photo induce the bactericidal effect by detonating the surface plasmon resonance. The results showed that the minimum bactericidal concentration (MBC) of silver nanoparticles was 16 µM (referred to [AgNO3]) and for silver-gold nanoparticles were 32 µM (referred to [AgNO3-HAuCl4]) at 60 minutes of irradiation.

Bacterial angular leaf spot disease (Xanthomonas fragariae) is an important strawberry disease and responsible for significant yield losses. The efficacy of emulsions containing tea tree or palmarosa essential oils (EO) against angular leaf spot, seeking to elucidate their mode of action, was evaluated in this study. Tea tree or palmarosa EO were added in an aqueous suspension of aloe polysaccharide (AP) and tea tree EO emulsion (TAP) and palmarosa EO microemulsion (PAP) were obtained. The antimicrobial potential of AP, TAP and PAP were evaluated in plants of two cultivars (Albion and San Andreas). The ability of these preparations to reduce disease severity and to activate plant defenses were evaluated. AP, TAP and PAP reduced X. fragariae growth by up to 44.0%, 94.4% and 86.1%, respectively. In addition, TAP and PAP suppressed completely the symptoms in both cultivars. Increased activities of superoxide dismutase, dismutase, catalase and peroxidase were observed in the more susceptible cultivar (Albion) treated with the emulsions, while a higher concentration of total phenolic compounds was verified in the less susceptible cultivar (San Andreas). Thus, emulsions with EO and AP can be used to control angular leaf spot by activating latent defense mechanisms in strawberry.

Xanthomonas phage RiverRider is a novel N4-like bacteriophage and the first phage isolated from the plant pathogen Xanthomonas fragariae. Electron microscopy revealed a Podoviridae morphology consisting of isometric heads and short noncontractile tails. The complete genome of RiverRider is 76,355 bp in length, with 90 open reading frames and seven tRNAs. The genome is characteristic of N4-like bacteriophages in both content and organization, having predicted proteins characterized into the functional groups of transcription, DNA metabolism, DNA replication, lysis, lysis inhibition, structure and DNA packaging. Amino acid sequence comparisons for proteins in these categories showed highest similarities to well-characterized N4-like bacteriophages isolated from Achromobacter xylosoxidans and Erwinia amylovora. However, the tail fiber proteins of RiverRider are clearly distinct from those of other N4-like phages. RiverRider was able to infect seven different strains of X. fragariae and none of the other species of Xanthomonas tested.

The use of lactic acid bacteria (LAB) to control multiple pathogens that affect different crops was studied, namely, Pseudomonas syringae pv. actinidiae in kiwifruit, Xanthomonas arboricola pv. pruni in Prunus and Xanthomonas fragariae in strawberry. A screening procedure based on in vitro and in planta assays of the three bacterial pathogens was successful in selecting potential LAB strains as biological control agents. The antagonistic activity of 55 strains was first tested in vitro and the strains Lactobacillus plantarum CC100, PM411 and TC92, and Leuconostoc mesenteroides CM160 and CM209 were selected because of their broad-spectrum activity. The biocontrol efficacy of the selected strains was assessed using a multiple-pathosystem approach in greenhouse conditions. L. plantarum PM411 and TC92 prevented all three pathogens from infecting their corresponding plant hosts. In addition, the biocontrol performance of PM411 and TC92 was comparable to the reference products (Bacillus amyloliquefaciens D747, Bacillus subtilis QST713, chitosan, acibenzolar-S-methyl, copper and kasugamycin) in semi-field and field experiments. The in vitro inhibitory mechanism of PM411 and TC92 is based, at least in part, on a pH lowering effect and the production of lactic acid. Moreover, both strains showed similar survival rates on leaf surfaces. PM411 and TC92 can easily be distinguished because of their different multilocus sequence typing and random amplified polymorphic DNA profiles Funding was provided by AGL2015‐69876‐C2‐1‐R (Spain Ministerio de Economía y Competitividad and FEDER of the European Union), by FP7‐KBBE.2013.1.2‐04 613678 DROPSA of the European Union, and by the MPCUdG2016 grant (University of Girona). N. Daranas was recipient of the research grant 2015 FI_B00515 (Secretaria d'Universitats i Recerca, Departament d'Economia i Coneixement, Generalitat de Catalunya; ES, EU). The research group is accredited by SGR 2014‐697 and TECNIO net from the Department d'Economia i Coneixement‐ACCIÓ Catalonia

Bakterien Xanthomonas fragariae kan gjøre stor skade på jordbærplanter. Den ødelegger bladene, og kan i tillegg gi stygge, skjemmende flekker på begerbladene slik at kvaliteten på bærene blir dårlig. Sykdommen har før denne sesongen ikke blitt påvist i Norge. I mange land med stor jordbærproduksjon har sykdommen mange ganger ført til store tap. For å dokumentere status for Xanthomonas fragariae i Norge ble det på oppdrag av Mattilsynet gjennomført en landsomfattende kartleggingsundersøkelse i 2013, 2014, 2015, 2016 og 2017. Oppfølging av OK programmet i 2018 bestod av testing hos virksomheter med felt etablert med importerte jordbærplanter. Det ble sendt inn og analysert totalt 258 prøver fra Mattilsynets kontorer for Region Midt og Region Stor Oslo. Alle prøver ble undersøkt med den internasjonalt anbefalte og anerkjente analysemetoden real-time PCR. I august 2018 påviste vi latent smitte av bakterien Xanthomona fragariae i jordbærplanter av sorten 'Malwina' hos en jordbærdyrker på Østlandet. Påvisningen ble gjort i et felt etablert i 2017 med planter importert fra Nederland. Ingen av prøvene som ble tatt ut hos dyrkeren hadde synlige symptomer, men ved oppflgingsuttak av prøver hos dyrkeren var igjen en av prøvene, sort 'Malwina', positiv ved PCR. Det kan ikke utelukkes at også andre dyrkere kan ha fått latent smitte av Xanthomona fragariae i sine jordbærplanter.

Angular leaf spot of strawberry, considered an A2 quarantine pest by the European and Mediterranean Plant Protection Organization (EPPO 2019), is an important bacterial disease in many regions. Since 2017, symptoms similar to angular leaf spot were observed in several strawberry cultivars including 'Taoyuan No. 1' and 'Xiang-Shui'. Early symptoms were angular, water-soaked lesions on the abaxial leaf surface, and later, reddish-brown irregular spots and coalesced lesions developed on the adaxial surface. In the humid conditions, sticky bacterial ooze exuding from lesions was observed. To isolate the causal agent, leaves showing water-soaked lesions were surface sterilized, cut into small pieces and soaked in 5 ml sterile water for at least 15 min. The supernatant from the cut-up pieces was serially diluted followed by spreading on sucrose peptone agar (SPA) (Hayward 1960). After incubating at 20°C for 4-5 days, single colonies grown on SPA were transferred to a new SPA plate and cultured at 20°C until colonies appeared. The yellow, glossy and mucoid colonies, which resembled the colony morphology of Xanthomonas fragariae, were selected as candidates for further confirmation. First, bacterial DNA of four candidate isolates, B001, B003 and B005 from Miaoli County and B004 from Taoyuan City, was PCR amplified with X. fragariae-specific primers: XF9/XF12 (Roberts et al. 1996) and 245A/B and 295A/B (Pooler et al. 1996). All four isolates could be detected by XF9/XF12 primer. Furthermore, isolates B003 and B004 could be detected by both 245A/B and 295A/B primers, while B001 and B005 could be detected by 295A/B only. Next, DNA gyrase subunit B (gyrB) was PCR amplified with the primers XgyrB1F/XgyrB1R (Young et al. 2008). The gyrB sequences of these four isolates were deposited in GenBank with accession numbers MT754942 to MT754945. The gyrB phylogenetic tree was constructed based on Bayesian inference analysis and maximum likelihood analysis. The gyrB sequences of the four isolates from Taiwan clustered in the clade containing the type strain of X. fragariae ICMP5715, indicating that they belong to X. fragariae. B001 and B005 formed a sub-group separated from B003 and B004, suggesting genetic differences between these isolates. To fulfill Koch's postulates, the abaxial surface of strawberry leaves were syringe infiltrated (KJP Silva et al., 2017) or wounded inoculated (Wang et al., 2017) with bacterial suspensions (final OD600 = 1.0-2.0) prepared from colonies of B001 and B003 washed from SPA plates. Inoculated plants were enclosed in a plastic bag (> 90% RH) at 25/20°C (day/night) under a 12-h/12-h photoperiod. After 7-14 days, water-soaked lesions similar to those observed in the field were developed on all inoculated leaves. The bacteria were successfully re-isolated from lesions of inoculated leaves and confirmed by specific primers XF9/XF12, 245A/B and 295A/B. We also found that the disease commonly occurs in the strawberry fields/nurseries with sprinkler irrigation during winter or early spring, and was particularly serious in the windward side or near riverside. To our knowledge, this is the first report of X. fragariae causing angular leaf spot on strawberry in Taiwan. Currently, the disease only occurs severely in certain regions, but establishment of effective management strategies will be needed to prevent spreading of this disease and potential economic loss in the future.

Strawberry bacterial angular leaf spot (ALS) disease, caused by Xanthomonas fragariae has become increasingly problematic in the strawberry agro-industry. ALS causes small angular water-soaked lesions to develop on the abaxial leaf surface. Studies reported optimum temperature conditions for X. fragariae are 20°C and the pathogen suffers mortality above 32°C. However, at the nursery stage, disease symptoms have been observed under high temperature conditions. In the present study, results showed X. fragariae transmission was via infected maternal plants, precipitation, and sprinkler irrigation systems. Systemic infections were detected using X. fragariae specific primers 245A/B and 295A/B, where 300-bp and 615-bp were respectively amplified. During the nursery stage (from May to August), the pathogen was PCR detected only in maternal plants, but not in soil or irrigation water through the nursery stage. During the cultivation period, from September to March, the pathogen was detected in maternal plants, progeny, and soil, but not in water. Additionally, un-infected plants, when planted with infected plants were positive for X. fragariae via PCR at the late cultivation stage. Chemical control for X. fragariae with oxolinic acid showed 87% control effects against the disease during the nursery period, in contrast to validamycin-A, which exhibited increased efficacy against the disease during the cultivation stage (control effect 95%). To our knowledge, this is the first epidemiological study of X. fragariae in Korean strawberry fields.