Search

Huanglongbing (HLB) and citrus canker, arising from Candidatus Liberibacter asiaticus (CaLas) and Xanthomonas citri pv. Citri (Xcc), respectively, have been imposing tremendous losses to the global citrus industry. Systemic acquired resistance (SAR) has been shown to be crucial for priming defense against pathogen in citrus. Salicylic acid (SA) binding protein 2 (SABP2), which is responsible for converting methyl salicylate (MeSA) to SA, is essential for full SAR establishment. Here, we characterized the functions of four citrus SABP2 genes (CsSABP2-1, CsSABP2-1V18A, CsSABP2-2 and CsSABP2-3) against HLB and citrus canker. In vitro enzymatic assay revealed that all four proteins had MeSA esterase activities, and CsSABP2-1 and CsSABP2-1V18A has the strongest activity. Their activities were inhibited by SA except for CsSABP2-1V18A. Four genes controlled by a strong promoter 35S were induced into Wanjincheng orange (Citrus sinensis Osbeck) to generate transgenic plants overexpressing CsSABP2. Overexpressing CsSABP2 increased SA and MeSA content and CsSABP2-1V18A had the strongest action on SA. Resistance evaluation demonstrated that only CsSABP2-1V18A had significantly enhanced tolerance to HLB, although all four CsSABP2s had increased tolerance to citrus canker. The data suggested the amino acid Val-18 in the active site of CsSABP2 plays a key role in protein function. Our study emphasized that balancing the levels of SA and MeSA is crucial for regulating SAR and conferring broad-spectrum resistance to HLB and citrus canker. This finding offers valuable insights for enhancing resistance through SAR engineering. [ABSTRACT FROM AUTHOR]

Stilbocrea banihashemiana is a recently described species of ascomycete fungi (Bionectriaceae) that causes canker and dieback diseases on fig and barberry, and dieback on loquat trees. Previous studies suggested this pathogen might affect other trees including economically important fruit crops, such as almonds, apples, and olives. This study assessed the susceptibility of 40 tree species belonging to 20 plant families, including fruit-bearing, ornamental, and edible landscape plants, to S. banihashemiana. The assessment was conducted by wound-inoculating detached shoots and one-year-old saplings with the pathogen. Based on five parameters of induced lesions, we clustered the inoculated tree species using the principal component analysis approach. The results of the approach revealed four distinct groups of tree species based on their susceptibility traits. Almond, apricot, eucalyptus, poplar, and red mulberry clustered in one group as susceptible hosts. The majority of the species examined, which included both tropical and temperate trees, were categorized as moderately susceptible. Platanus and sweet orange were identified as non-host-resistant species, while Arizona cypress, garden privet, and olive were classified as low susceptible. Moreover, our findings suggest a correlation between temperature and the aggressiveness of S. banihashemiana. This study revealed that the emerging pathogen, S. banihashemiana, could have a broader host range than currently recognized. The accidental introduction of this pathogen through the global trade of live host plants may pose a significant threat to economically important trees in temperate and subtropical regions. [ABSTRACT FROM AUTHOR]

Clavibacter michiganensis subsp. michiganensis (Cmm) and C. michiganensis subsp. capsici (Cmc) are phytopathogenic bacteria that cause bacterial canker disease in tomatoes and peppers, respectively. Bacterial canker disease poses serious challenges to solanaceous crops, causing significant yield losses and economic costs. Effective management necessitates the development of sustainable control strategies employing nanobiotechnology. In this study, the antibacterial effects of four Aspergillus sojae -mediated nanoformulations, including cobalt oxide nanoparticles (Co 3 O 4 NPs), zinc oxide nanoparticles (ZnO NPs), cobalt ferrite nanoparticles (CoFe 2 O 4 NPs), and CoFe 2 O 4 /functionalized multi-walled carbon nanotube (fMWCNT) bionanocomposite, were evaluated against Cmm and Cmc. The diameters of the zone of inhibition of A. sojae -mediated Co 3 O 4 NPs, ZnO NPs, CoFe 2 O 4 NPs, and CoFe 2 O 4 /fMWCNT bionanocomposite against Cmm and Cmc were 23.60 mm, 22.09 mm, 27.65 mm, 22.51 mm, and 19.33 mm, 17.66 mm, 21.64 mm, 18.77 mm, respectively. The broth microdilution assay was conducted to determine the minimal inhibitory and bactericidal concentrations. The MICs of Co 3 O 4 NPs, ZnO NPs, CoFe 2 O 4 NPs, and CoFe 2 O 4 /fMWCNT bionanocomposite against Cmm were 2.50 mg/mL, 1.25 mg/mL, 2.50 mg/mL, and 2.50 mg/mL, respectively. While, their respective MBCs against Cmm were 5.00 mg/mL, 2.50 mg/mL, 5.00 mg/mL, and 5.00 mg/mL. The respective MICs of Co 3 O 4 NPs, ZnO NPs, CoFe 2 O 4 NPs, and CoFe 2 O 4 /fMWCNT bionanocomposite against Cmc were 2.50 mg/mL, 1.25 mg/mL, 5.00 mg/mL, and 5.00 mg/mL. While, their respective MBCs against Cmc were 5.00 mg/mL, 2.50 mg/mL, 10.00 mg/mL, and 10.00 mg/mL. The morphological and ultrastructural changes of Cmm and Cmc cells were observed using field-emission scanning and transmission electron microscopy before and after treatment with sub-minimal inhibitory concentrations of the nanoformulations. Nanoformulation-treated bacterial cells became deformed and disrupted, displaying pits, deep cavities, and groove-like structures. The cell membrane detached from the bacterial cell wall, electron-dense particles accumulated in the cytoplasm, cellular components disintegrated, and the cells were lysed. Direct physical interactions between the prepared nanoformulations with Cmm and Cmc cells might be the major mechanism for their antibacterial potency. Further research is required for the in vivo application of the mycosynthesized nanoformulations as countermeasures to combat bacterial phytopathogens. [Display omitted] • A. sojae -mediated nanoformulations exhibited significant antibacterial potential against phytopathogens. • Assessment of morphological and ultrastructural changes using FESEM and TEM. • Elucidation of plausible antibacterial mechanism of A. sojae -mediated nanoformulations. • Mycosynthesis of nanoformulations is a green approach for plant disease management. [ABSTRACT FROM AUTHOR]

Xanthomonas citri subsp. citri (Xcc), the causal agent of citrus canker, elicits canker symptoms in citrus plants because of the transcriptional activator-like (TAL) effector PthA4, which activates the expression of the citrus susceptibility gene CsLOB1. This study reports the regulation of the putative carbohydrate-binding protein gene Cs9g12620 by PthA4-mediated induction of CsLOB1 during Xcc infection. We found that the transcription of Cs9g12620 was induced by infection with Xcc in a PthA4-dependent manner. Even though it specifically bound to a putative TAL effector-binding element in the Cs9g12620 promoter, PthA4 exerted a suppressive effect on the promoter activity. In contrast, CsLOB1 bound to the Cs9g12620 promoter to activate its expression. The silencing of CsLOB1 significantly reduced the level of expression of Cs9g12620, which demonstrated that Cs9g12620 was directly regulated by CsLOB1. Intriguingly, PhtA4 interacted with CsLOB1 and exerted feedback control that suppressed the induction of expression of Cs9g12620 by CsLOB1. Transient overexpression and gene silencing revealed that Cs9g12620 was required for the optimal development of canker symptoms. These results support the hypothesis that the expression of Cs9g12620 is dynamically directed by PthA4 for canker formation through the PthA4-mediated induction of CsLOB1. [ABSTRACT FROM AUTHOR]

Apple canker disease, also named as apple Valsa canker, is one of the most destructive diseases for apples (Malus domestica Borkh.). Cytospora/Valsa spp. are the dominant causal agent of this disease, but many studies have revealed that fungi from some other genus can also cause typical apple canker symptoms. In this study, we performed fungal pathogen isolation from cankered 'Fuji' apple barks. Six representative morphologically different fungi (Strain 1–6) were further subjected to ITS sequencing and evolutionary analysis. Molecular identification results revealed that Strains 1–6 are Cytospora mali, Fusarium cf. solani, Alternaria alternata, C. mali, Diplodia seriata and F. proliferatum, respectively. All these fungi have been reported to be causal agents of apple diseases. By inoculating fungal plugs onto trunks of 'Fuji' apple trees, the pathogenicity of the six fungi were accessed. Only the inoculations of the two C. mali strains (Strain 1 and Strain 4) and the A. alternata strain (Strain 3) resulted in typical apple canker symptoms in trunks. It is worth noting that Strain 1 caused much more severe canker symptoms and higher pathogenicity incidence than the other two fungi. A. alternata has been identified as a pathogen causing diseases on apple fruits and leaves. By further assessing its pathogenicity on apple fruits and leaves, we verified that it can also cause typical fruit rot and leaf spot symptoms. To the best of our knowledge, this is the first report on apple canker disease caused by A. alternata in China. Our present study can provide a theoretical foundation for the prevention and control of apple canker disease. [ABSTRACT FROM AUTHOR]

Idesia polycarpa is a woody oil plant with great development and application prospects. However, stem canker disease of I. polycarpa was frequently observed in Henan and Hubei provinces of China in 2020–2022. The seriousness of the disease caused a large number of trees deaths, which affected seedling production and fruit yield, restricting the development of the I. polycarpa industry. We performed isolation, purification and pathogenicity analysis of canker samples. Pathogenicity tests reproduced typical canker disease symptoms on detached branches of I. polycarpa. Based on morphological observations, conidial morphology and phylogenetic analysis of isolates with high similarity to the rDNA internal transcribed spacer (ITS), EF‐1α and TUB2 sequences screened in GenBank by BLAST, the pathogen of canker disease on I. polycarpa was identified as Botryosphaeria dothidea. This is the first report of B. dothidea as the pathogen of stem canker of I. polycarpa. [ABSTRACT FROM AUTHOR]

This article provides a genomic analysis of Seiridium unicorne, a fungus that causes Cypress Canker Disease (CCD). The study aims to understand the evolutionary relationships among different Seiridium species responsible for CCD in order to improve disease management. The researchers isolated the S. unicorne strain from Cryptomeria japonica in New Zealand and conducted DNA extraction and sequencing. The assembled genome consisted of 581 contigs, and gene prediction revealed 13,576 protein-coding genes and 3,155 predicted effector proteins. The study also compared S. unicorne with other Seiridium species and found a higher number of predicted effectors in S. unicorne, possibly due to its wider host range. The genomic data is publicly available in GenBank. [Extracted from the article]

Kiwifruit bacterial canker, caused by Pseudomonas syringae pv. actinidiae (PSA), poses a grave threat to the global kiwifruit industry. In this study, we examined the role of microRNAs (miRNAs) in kiwifruit's response to PSA. Kiwifruit seedlings subjected to PSA treatment showed significant changes in both miRNA and gene expression compared to the control group. We identified 364 differentially expressed miRNAs (DEMs) and 7170 differentially expressed genes (DEGs). Further analysis revealed 180 miRNAs negatively regulating 641 mRNAs. Notably, two miRNAs from the miRNA482 family, miRNA-215-3p and miRNA-29-3p, were found to increase kiwifruit's sensitivity to PSA when overexpressed. These miRNAs were linked to the regulation of NBS-LRR target genes, shedding light on their role in kiwifruit's defence against PSA. This study offers insights into the miRNA482-NBS-LRR network as a crucial component in enhancing kiwifruit bioresistance to PSA infestation and provides promising candidate genes for further research. [ABSTRACT FROM AUTHOR]

The most effective strategy for managing wheat bacterial blight caused by Pseudomonas syringae pv. syringae is believed to be the use of resistant cultivars. Researching the correlation between molecular markers and stress resistance can expedite the plant breeding process. The current study aims to evaluate the response of 27 bread wheat cultivars to bacterial blight disease in order to identify resistant and susceptible cultivars and to pinpoint ISSR molecular markers associated with bacterial blight resistance genes. ISSR markers are recommended for assessing a plant's disease resistance. This experiment is focused on identifying ISSR molecular markers linked to bacterial blight resistance. After applying the bacterial solution to the leaves, we performed sampling to determine the infection percentage in the leaves at different intervals (7, 14, and 18 days after spraying). In most cultivars, the average leaf infection percentage decreased 18 days after spraying on young leaves. However, in some cultivars such as Niknegad, Darab2, and Zarin, leaf infection increased in older leaves and reached up to 100% necrosis. In our study, 12 ISSR primers generated a total of 170 bands, with 156 being polymorphic. The primers F10 and F5 showed the highest polymorphism, while the F7 primer exhibited the lowest polymorphism. Cluster analysis grouped these cultivars into four categories. The resistant group included Qods, Omid, and Atrak cultivars, while the semi-resistant and susceptible groups comprised the rest of the cultivars. Through binary logistic analysis, we identified three Super oxide dismutase-related genes that contribute to plant resistance to bacterial blight. These genes were linked to the F3, F5, and F12 primers in regions I (1500 bp), T (1000 bp), and G (850 bp), respectively. We also identified seven susceptibility-associated genes. Atrak, Omid, and Qods cultivars exhibited resistance against bacterial blight, and three genes associated with this resistance were linked to the F3, F5, and F12 primers. These markers can be used for screening or transferring tolerance to other wheat cultivars in breeding programs. [ABSTRACT FROM AUTHOR]

Mexico ranks second in the world for Persian lime (Citrus latifolia) exports, making it the principal citrus exporter within the national citrus industry, exporting over 600,000 tons per year. However, diseases are the main factor reducing production, resulting in significant economic losses. Among these diseases, fungal diseases like dieback, caused by species of Lasiodiplodia, are an emerging issue in Persian lime. Symptoms include gummosis, twig and branch dieback, cankers, the necrosis of bark and wood, fruit mummification, and tree decline. The aim of this study was to investigate the occurrence and pathogenicity of the fungal species associated with twig and branch dieback, cankers, and decline of Persian lime trees in southern Mexico, and to elucidate the current status of the Lasiodiplodia species causing the disease in Mexico. During June, July, and August of 2023, a total of the 9229 Persian lime trees were inspected across 230 hectares of Persian lime orchards in southern Mexico, and symptoms of the disease were detected in 48.78% of the trees. Branches from 30 of these Persian lime trees were collected. Fungal isolates were obtained, resulting in a collection of 40 strains. The isolates were characterized molecularly and phylogenetically through the partial regions of four loci: the internal transcribed spacer region (ITS), the β-tubulin gene (tub2), the translation elongation factor 1-alpha gene (tef1-α), and the DNA-directed RNA polymerase II second largest subunit (rpb2). Additionally, pathogenicity was assessed, successfully completing Koch's postulates on both detached Persian lime branches and certified 18-month-old Persian lime plants. Through multilocus molecular phylogenetic identification, pathogenicity, and virulence tests, five species were identified as causal agents: L. iraniensis, L. lignicola, L. mexicanensis, L. pseudotheobromae, and L. theobromae. This study demonstrates that in southern Mexico, at least five species of the genus Lasiodiplodia are responsible for dieback in Persian lime. Additionally, this is the first report of L. lignicola and L. mexicanensis as causal agents of the disease in citrus, indicating novel host interactions between species of Lasiodiplodia and C. latifolia. [ABSTRACT FROM AUTHOR]

Torreya taxifolia Arn. (Florida torreya) is a critically endangered (International Union for Conservation of Nature 3.1) conifer with a very limited native range in the USA, occurring in two counties in Florida and one in Georgia where it persists in ravines along the Apalachicola River. The species suffered a major decline, ∼ 99% loss, beginning in the 1930s, with a total population estimated at 357,500 in the early 1900s decreasing to 1,000 individuals currently remaining in the wild. The initial decline was attributed to an unknown fungal disease with Fusarium torreyae (T. Aoki, J. A. Smith, L. Mount, Geiser and O'Donnell), identified in 2011 as the causal agent of a canker disease. The objectives of this work were to develop genetic markers to uniquely identify individuals, examine the structure of the in situ population, and use an ex situ germplasm resource to determine if reproduction without fertilization (apomixis) occurs. We developed a panel of microsatellite markers, sampled individuals from three natural in situ populations, and found population structure at all levels tested. The markers were next applied to ex situ mothers, potential fathers, and progeny to investigate the occurrence of apomixis. One of 29 progeny seedlings tested and its mother had identical multilocus genotypes. The probability of observing this genotype by chance, given the mother and the two possible fathers' genotypes, is approximately one in ten. Based on these data, we cannot reject the hypothesis that apomixis occurs. Our finding of population structure suggests genotypes from as wide an area as possible are required to capture the diversity of the species in conservation efforts. The multiplexed marker panel can be used to identify individual accessions and assist with managing ex situ collections; however, they are not informative enough to clearly differentiate closely related individuals in the ex situ population. [ABSTRACT FROM AUTHOR]

Bacterial canker disease caused by Clavibacter michiganensis is a substantial threat to the cultivation of tomatoes, leading to considerable economic losses and global food insecurity. Infection is characterized by white raised lesions on leaves, stem, and fruits with yellow to tan patches between veins, and marginal necrosis. Several agrochemical substances have been reported in previous studies to manage this disease but these were not ecofriendly. Thus present study was designed to control the bacterial canker disease in tomato using green fabricated silver nanoparticles (AgNps). Nanosilver particles (AgNPs) were synthesized utilizing Moringa oleifera leaf extract as a reducing and stabilizing agent. Synthesized AgNPs were characterized using UV–visible spectroscopy, scanning electron microscopy (SEM), X-ray diffraction (XRD), energy-dispersive X-ray (EDX), and Fourier transform infrared spectrometry (FTIR). FTIR showed presence of bioactive compounds in green fabricated AgNPs and UV-visible spectroscopy confirmed the surface plasmon resonance (SPR) band in the range of 350 nm to 355 nm. SEM showed the rectangular segments fused together, and XRD confirmed the crystalline nature of the synthesized AgNPs. The presence of metallic silver ions was confirmed by an EDX detector. Different concentrations (10, 20, 30, and 40 ppm) of the green fabricated AgNPs were exogenously applied on tomato before applying an inoculum of Clavibacter michigensis to record the bacterial canker disease incidence at different day intervals. The optimal concentration of AgNPs was found to be 30 µg/mg that exhibited the most favorable impact on morphological (shoot length, root length, plant fresh and dry weights, root fresh and dry weights) and physiological parameters (chlorophyll contents, membrane stability index, and relative water content) as well as biochemical parameters (proline, total soluble sugar and catalase activity). These findings indicated a noteworthy reduction in biotic stress through the increase of both enzymatic and non-enzymatic activities by the green fabricated AgNPs. This study marks a first biocompatible approach in assessing the potential of green fabricated AgNPs in enhancing the well-being of tomato plants that affected with bacterial canker and establishing an effective management strategy against Clavibacter michiganensis. This is the first study suggests that low concentration of green fabricated nanosilvers (AgNPs) from leaf extract of Moringa oleifera against Clavibacter michiganensis is a promisingly efficient and eco-friendly alternative approach for management of bacterial canker disease in tomato crop. [ABSTRACT FROM AUTHOR]

Biotic stresses caused by bacterial and fungal pathogens damage crops; identifying treatments that enhance disease resistance provides important information for understanding plant defenses and sustainable agriculture. Salt stress affects crop yields worldwide; however, studies have focused on the toxic sodium ion, leaving the effects of the chloride ion unclear. In this study, we found that irrigation with a combination of chloride salts (MgCl2, CaCl2, and KCl) suppressed the cell death phenotype of the ceramide kinase mutant acd5. Chloride salt pre-irrigation also significantly limited the cell death caused by Pseudomonas syringae pv maculicola infection and inhibited the multiplication of this bacterial pathogen in a mechanism partially dependent on the salicylic acid pathway. Moreover, chloride salt preirrigation improved plant defenses against the fungal pathogen challenge, confining the lesion area caused by Botrytis cinerea infection. Furthermore, the growth of herbivorous larvae of Spodoptera exigua was retarded by feeding on chloride salt irrigated plants. Thus, our data suggest that treatment with Clincreases broad spectrum resistance to biotic challenges. [ABSTRACT FROM AUTHOR]

The nuclear-encoded mitochondrial protein Tu translation elongation factor, mitochondrial (TUFM) is well-known for its role in mitochondrial protein translation. Originally discovered in yeast, TUFM demonstrates significant evolutionary conservation from prokaryotes to eukaryotes. Dysregulation of TUFM has been associated with mitochondrial disorders. Although early hypothesis suggests that TUFM is localized within mitochondria, recent studies identify its presence in the cytoplasm, with this subcellular distribution being linked to distinct functions of TUFM. Significantly, in addition to its established function in mitochondrial protein quality control, recent research indicates a broader involvement of TUFM in the regulation of programmed cell death processes (e.g., autophagy, apoptosis, necroptosis, and pyroptosis) and its diverse roles in viral infection, cancer, and other disease conditions. This review seeks to offer a current summary of TUFM's biological functions and its complex regulatory mechanisms in human health and disease. Insight into these intricate pathways controlled by TUFM may lead to the potential development of targeted therapies for a range of human diseases. [ABSTRACT FROM AUTHOR]

Both Ceratocystis lukuohia and C. huliohia have been associated with Rapid ʻŌhiʻa Death (ROD), an emerging threat to ʻōhiʻa (Metrosideros polymorpha), a keystone forest tree species. The vascular wilt disease caused by C. lukuohia has been recently described and is responsible for the widespread ROD epidemic on Hawai'i Island. However, the role of C. huliohia in ROD development and tree death is not clear. Artificial inoculation of field‐grown ʻōhiʻa with C. huliohia and dissections of naturally infected, early symptomatic forest trees were conducted to confirm pathogenicity on field grown trees and the pattern of internal colonization. In two trials, crowns of trees with main stems inoculated with C. huliohia were visually healthy at the time of tree harvest after 43–55 days in the first trial, and after 91 days in the second trial. However, elliptical inner bark cankers underlain by reddish‐brown xylem were associated with the inoculation points. Similar canker and stain symptoms were found on stems and branches of ʻōhiʻa (24–26 cm trunk diameter) naturally infected by C. huliohia. This xylem stain manifested as multiple distinct elliptical cankers or the coalescing of multiple cankers. The pathogen was commonly isolated from the perimeter of the stained outer sapwood and to a depth of 4 cm. The coalescence of multiple cankers was associated with the crown symptoms observed on the naturally infected forest trees that were dissected. Multiple C. huliohia infections that lead to coalescing cankers which subsequently girdle stems likely occurs over one or more years compared to the shorter time (e.g., months) required for C. lukuohia‐caused death to occur. [ABSTRACT FROM AUTHOR]

Fusarium circinatum is an important fungal pathogen of Pinus species utilized in commercial forestry worldwide. In Colombia, it was first found on nursery plants and later in plantations associated with basal cankers on young trees. In this study, we explored the population diversity of the pathogen in Colombia by analyzing 136 isolates collected from diseased nursery plants (2005–2007) and plantation trees (2017 and 2020). These were sourced from different geographical regions and Pinus species. Genotyping was performed using 10 microsatellite markers, while mating types were identified with PCRs targeting the MAT1 locus. Using microsatellites, a total of 33 multilocus haplotypes were detected. Genetic diversity indices showed low levels of diversity in both the overall collection and in specific collection groupings. The data also suggested that a small number of isolates had unique origins in the country (p <.05), and relatively low levels of population differentiation were detected between the nursery and plantation collections. All the isolates were scored as having the MAT1‐1 mating type, and no evidence for the random association among microsatellite alleles (p =.0001) was found. Overall, these data suggest that F. circinatum was introduced into Colombia a small number of times, likely on seed for nursery production. Furthermore, the data also indicate that the pathogen has spread from nurseries to the plantations via asexual reproduction and on asymptomatic plants. This has resulted in a highly clonal F. circinatum population in Colombia that has resulted from accidental introductions of the pathogen into a production nursery. [ABSTRACT FROM AUTHOR]

The cultivated apple (Malus × domestica) is one of Türkiye's most important economic crops. Over the growing seasons of 2020–2022, typical symptoms of branch cankers and scaling-off of the bark on apple trees were observed in the Eğirdir district of Isparta province. Botryosphaeriaceae fungi were consistently isolated from symptomatic branches. The fungal pathogen was identified as Diplodia bulgarica based on cultural and morphological characteristics and ITS region of DNA sequence data comparisons. Pathogenicity tests were performed on detached shoots by inoculating them with mycelial plugs. Diplodia bulgarica isolates caused necrotic lesions on Scarlet Spur apple detached shoots. To aid in the development of control strategies for this fungus, nine fungicides were examined in vitro for their antifungal effects against the pathogen at different concentrations. Chlorothalonil, tebuconazole, thiophanate-methyl, and fluazinam were the most efficient fungicides, with EC50 values were <1.0 µg/mL. [ABSTRACT FROM AUTHOR]

Treating plant bacterial diseases is notoriously difficult because of the lack of available antimicrobials. Pseudomonas syringae pathovar syringae (Pss) is a major pathogen of cherry (Prunus avium) causing bacterial canker of the stem, leaf and fruit, impacting productivity and leading to a loss of trees. In an attempt to find a treatment for this disease, naturally occurring bacteriophage (phage) that specifically target Pss is being investigated as a biocontrol strategy. However, before using them as a biocontrol treatment, it is important to both understand their efficacy in reducing the bacterial population and determine if the bacterial pathogens can evolve resistance to evade phage infection. To investigate this, killing curve assays of five MR phages targeting Pss showed that phage resistance rapidly emerges in vitro, even when using a cocktail of the five phages together. To gain insight to the changes occurring, Pss colonies were collected three times during a 66‐h killing curve assay and separately, Pss and phage were also coevolved over 10 generations, enabling the measurement of genomic and fitness changes in bacterial populations. Pss evolved resistance to phages through modifications in lipopolysaccharide (LPS) synthesis pathways. Bacterial fitness (growth) and virulence were affected in only a few mutants. Deletion of LPS‐associated genes suggested that LPS was the main target receptor for all five MR phages. Later generations of coevolved phages from the coevolution experiment were more potent at reducing the bacterial density and when used with wild‐type phages could reduce the emergence of phage‐resistant mutants. This study shows that understanding the genetic mechanisms of bacterial pathogen resistance to phages is important for helping to design a more effective approach to kill the bacteria while minimizing the opportunity for phage resistance to manifest. [ABSTRACT FROM AUTHOR]

Tree decline is described as the loss in tree vigor and increased mortality initiated by climate change events, and also involves pathogens and pests. Stem bleeding and bark canker of oak (Quercus castaneifolia) were observed in Mazandaran and Golestan forests during summer 2020–2021. Symptoms included cracks in the outer bark, stem tissue necrosis, bleeding, dark exudate and dark brown lesions in the inner bark. Eighty-nine strains with a metallic green sheen pigment were isolated on eosin methylene blue agar. The pathogenicity of all strains recovered in this study was assessed on oak seedlings and acorns. Forty-four strains produced rotting on oak acorns 2 weeks after inoculation. Inoculation of six representative strains on oak seedlings resulted in twig dieback of the plants after 4 weeks. Strains were negative for Gram reaction, oxidase and levan formation from sucrose. The gyrB and infB gene sequence similarity values of strains were 98.87–99.57% with the type strain of Gibbsiella quercinecans, 98.66–98.86% with the type strain of Gibbsiella greigii and 99.46–99.64% with the type strain of Dryocola boscaweniae. In the phylogenetic tree based on concatenated sequences of gyrB and infB genes or each gene individually, the strains were divided into three clusters containing the type strains of G. quercinecans, G. greigii and D. boscaweniae, each with high bootstrap support and confirming their identity as belonging to these three species. To the best of our knowledge, this is the first report of oak bacterial canker caused by D. boscaweniae, G. greigii and G. quercinecans in Iran and the first report of D. boscaweniae associated with oak decline symptoms. [ABSTRACT FROM AUTHOR]

Fungal endophytes inhabit a similar ecological niche to that occupied by many phytopathogens, with several pathogens isolated from healthy tissues in their latent phase. This study aimed to evaluate the pathogenicity, the colonisation ability, and the enzyme activity of 37 endophytic fungal isolates recovered from apparently healthy apple shoot and leaf tissues. The pathogenicity of the isolates was assessed on 'Royal Gala' and 'Braeburn' fruit and detached 'Royal Gala' shoots. For the non-pathogenic isolates, their ability to endophytically colonise detached 'Royal Gala' shoots was evaluated. Enzyme activity assays were undertaken to determine whether the pathogenicity of the endophytes was related to the production of the extracellular enzymes, amylase, cellulase, pectinase, protease, and xylanase. Of the 37 isolates studied, eight isolates, representing the genera Colletotrichum , Diaporthe , Fusarium, and Penicillium, were shown to be pathogenic on both apple shoots and fruit. Two isolates identified as Trichoderma atroviride , were pathogenic only on shoots, and three isolates, representing the genus Diaporthe , were pathogenic only on fruit. Of the remaining 24 isolates, 22 (Biscogniauxia (n = 8), Chaetomium (n = 4), Trichoderma (n = 3), Epicoccum (n = 2), Neosetophoma (n = 2), Xylaria (n = 1), Daldinia (n = 1), and Paraphaeosphaeria (n = 1)) were recovered from the inoculated apple shoots but two failed to colonise the shoot tissues. Of the isolates tested, 20 produced amylase, 15 cellulase, 25 pectinase, 26 protease, and 13 xylanase. There was no correlation between the range and type of enzymes produced by the isolates and their pathogenicity or ability to endophytically colonise the shoot tissue. The study showed that approximately one-third (13/37) of the isolates recovered from the apparently healthy apple shoot tissues were observed as latent pathogens. The isolates that did not cause disease symptoms may have the ability to reduce colonisation of apple tissues by pathogens including Neonectria ditissima associated with European canker of apple. • Latent plant pathogens isolated from healthy plant tissue identified as endophytes. • Pathogenicity and enzyme production of 37 fungal apple endophyte isolates assessed. • Thirteen isolates produced disease symptoms on apple fruit and/or detached shoots. • Twenty-two isolates endophytically colonised apple shoots, with no symptoms. • Enzyme production did not differentiate between pathogenic and endophytic isolates. [ABSTRACT FROM AUTHOR]

BACKGROUND: Citrus canker caused by Xanthomonas citri subsp. citri (Xcc) is a devastating bacterial disease that reduces citrus yield and quality, posing a serious threat to the citrus industry. Several conventional chemicals have been used to control citrus canker. However, this approach often leads to the excessive use of chemical agents, can exacerbate environmental pollution and promotes the development of resistant Xcc. Therefore, there is significant interest in the development of efficient and environmentally friendly technologies to control citrus canker. RESULTS: In this study, water‐soluble ZnO quantum dots (ZnO QDs) were synthesised as an efficient nanopesticide against Xcc. The results showed that the antibacterial activity of ZnO QDs irradiated with visible light [half‐maximal effective concentration (EC50) = 33.18 μg mL−1] was ~3.5 times higher than that of the dark‐treated group (EC50 = 114.80 μg mL−1). ZnO QDs induced the generation of reactive oxygen species (•OH, •O−2 and 1O2) under light irradiation, resulting in DNA damage, cytoplasmic destruction, and decreased catalase and superoxide dismutase activities. Transcription analysis showed downregulation of Xcc genes related to 'biofilms, virulence, adhesion' and 'DNA transfer' exposure to ZnO QDs. More importantly, ZnO QDs also promoted the growth of citrus. CONCLUSION: This research provides new insights into the photocatalytic antibacterial mechanisms of ZnO QDs and supports the development of more efficient and safer ZnO QDs‐based nanopesticides to control citrus canker. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

BACKGROUND: Valsa canker caused by Valsa pyri is one of the most destructive diseases of pear, leading to severe yield and economic losses. Volatile organic compounds (VOCs) from endophytes have important roles in the regulation of plant disease. In this study, we investigated the biocontrol activity of the endophytic fungus Aspergillus niger strain La2 and its antagonistic VOCs against pear Valsa canker. RESULTS: Strain La2 exhibited an obvious inhibitory effect against V. pyri. A colonization assay suggested that strain La2 could complete its life cycle on pear twigs. The symptoms of pear Valsa canker were weakened on detached pear twigs after treatment with strain La2. In addition, VOCs from strain La2 also significantly suppressed mycelial growth in V. pyri. Based on the results of headspace solid‐phase microextraction/gas chromatography–mass spectrometry analysis, six possible VOCs produced by strain La2 were detected, of which 2,4‐di‐tert‐butylphenol and 4‐methyl‐1‐pentanol were the main antagonistic VOCs in terms of their effect on pear Valsa canker in vitro and in vivo. Further results showed that 4‐methyl‐1‐pentanol could destroy the V. pyri hyphal structure and cell membrane integrity. Importantly, the activities of pear defense‐related enzymes (polyphenol oxidase, phenylalanine ammonia lyase and superoxide dismutase) were enhanced after 4‐methyl‐1‐pentanol treatment in pear twigs, suggesting that 4‐methyl‐1‐pentanol might induce a plant disease resistance response. CONCLUSION: Aspergillus niger strain La2 and its VOCs 2,4‐di‐tert‐butylphenol and 4‐methyl‐1‐pentanol have potential as novel biocontrol agents of pear Valsa canker. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Bacterial canker is an important disease of sweet cherry plants mainly caused by Pseudomonas syringae pv. syringae (Pss). Water deficit profoundly impairs the yield of this crop. Nitric oxide (NO) is a molecule that plays an important role in the plant defense mechanisms. To evaluate the protection exerted by NO against Pss infection under normal or water-restricted conditions, sodium nitroprusside (SNP), a NO donor, was applied to sweet cherry plants cv. Lapins, before they were exposed to Pss infection under normal or water-restricted conditions throughout two seasons. Well-watered plants treated with exogenous NO presented a lower susceptibility to Pss. A lower susceptibility to Pss was also induced in plants by water stress and this effect was increased when water stress was accompanied by exogenous NO. The lower susceptibility to Pss induced either by exogenous NO or water stress was accompanied by a decrease in the internal bacterial population. In well-watered plants, exogenous NO increased the stomatal conductance and the net CO2 assimilation. In water-stressed plants, NO induced an increase in the leaf membranes stability and proline content, but not an increase in the CO2 assimilation or the stomatal conductance. [ABSTRACT FROM AUTHOR]

Background: Pseudomonas syringae pv. actinidiae (Psa) is an important bacterial plant pathogen that causes severe damage to the kiwifruit industry worldwide. Three Psa strains were recently obtained from different kiwifruit orchards in Anhui Province, China. The present study mainly focused on the variations in virulence and genome characteristics of these strains based on the pathogenicity assays and comparative genomic analyses. Results: Three strains were identified as biovar 3 (Psa3), along with strain QSY6 showing higher virulence than JZY2 and YXH1 in pathogenicity assays. The whole genome assembly revealed that each of the three strains had a circular chromosome and a complete plasmid. The chromosome sizes ranged from 6.5 to 6.6 Mb with a GC content of approximately 58.39 to 58.46%, and a predicted number of protein-coding sequences ranging from 5,884 to 6,019. The three strains clustered tightly with 8 Psa3 reference strains in terms of average nucleotide identity (ANI), whole-genome-based phylogenetic analysis, and pangenome analysis, while they were evolutionarily distinct from other biovars (Psa1 and Psa5). Variations were observed in the repertoire of effectors of the type III secretion system among all 15 strains. Moreover, synteny analysis of the three sequenced strains revealed eight genomic regions containing 308 genes exclusively present in the highly virulent strain QSY6. Further investigation of these genes showed that 16 virulence-related genes highlight several key factors, such as effector delivery systems (type III secretion systems) and adherence (type IV pilus), which might be crucial for the virulence of QSY6. Conclusion: Three Psa strains were identified and showed variant virulence in kiwifruit plant. Complete genome sequences and comparative genomic analyses further provided a theoretical basis for the potential pathogenic factors responsible for kiwifruit bacterial canker. [ABSTRACT FROM AUTHOR]

Pakistan's economy greatly benefits from citrus production since these fruits are sold and consumed all over the world. Although citrus fruits are easy to cultivate, they are susceptible to diseases caused by bacteria, viruses, and fungi. These challenges, as well as difficulties in obtaining the proper nutrients, might negatively impact fruit yields and quality. Citrus canker is another complicated problem caused by the germ Xanthomonas axonopodis. This germ affects many types of citrus fruits all over the world. This study looked closely at how citrus canker affects the leaves and the quality of the fruit in places like Sargodha, Bhalwal, Kotmomin, and Silanwali, which are big areas for growing citrus in the Sargodha district. What we found was that plants without the disease had more chlorophyll in their leaves compared to the sick plants. Also, the healthy plants had better amounts of important minerals like calcium, magnesium, potassium, and phosphorus in their fruits. But the fruits with the disease had too much sodium, and the iron levels were a bit different. The fruits with the disease also didn't have as much of something that protects them called antioxidants, which made them more likely to get sick. This study helps us understand how citrus canker affects plants and fruit, so we can think of ways to deal with it. [ABSTRACT FROM AUTHOR]

Background: Citrus canker is a significant bacterial disease caused by Xanthomonas citri subsp. citri (Xcc) that severely impedes the healthy development of the citrus industry. Especially when citrus fruit is infected by Xcc, it will reduce or even lost its commercial value. However, due to the prolonged fruiting cycle and intricate structure, much less research progress had been made in canker disease on fruit than on leaf. In fact, limited understanding has been achieved on canker development and the response to Xcc infection in fruit. Results: Herein, the progression of canker disease on sweet orange fruit was tracked in the field. Results indicated that typical lesions initially appear on the sepal, style residue, nectary disk, epicarp, and peduncle of young fruits after petal fall. The susceptibility of fruits to Xcc infection diminished as the fruit developed, with no new lesions forming at the ripening stage. The establishment of an efficient method for inoculating Xcc on fruit as well as the artificial inoculation throughout the fruit's developmental cycle clarified this infection pattern. Additionally, microscopic observations during the infection process revealed that Xcc invasion caused structural changes on the surface and cross-section of the fruit. Conclusions: An efficient system for inoculation on citrus fruit with Xcc was established, by which it can serve for the evaluation of citrus germplasm for canker disease resistance and systematic research on the interactions between Xcc and citrus fruits. [ABSTRACT FROM AUTHOR]

Introduction: Valsa canker, caused by Cytospora mali, is a destructive disease in apple production. However, the mechanism by which apple defend against C. mali infection remains unclear. Methods: In this study, the integrative transcriptional and metabolic analysis were used to investigate the responses of the 'Jin Hong' apple branches to the invasion of C. mali. Results and Discussion: Results showed that the differentially expressed genes were mainly enriched in the pathways of carbon metabolism, photosynthesisantenna proteins, and biosynthesis of amino acids pathways. Additionally, the differentially accumulated metabolites were significantly enriched in aminoacyltRNA biosynthesis, fructose and mannose metabolism, and alanine, aspartate, and glutamate metabolism pathways. Conjoint analysis revealed that C. mali infection significantly altered 5 metabolic pathways, 8 highly relevant metabolites and 15 genes of apples. Among which the transcription factors WRKY and basic domain leucine zipper transcription family were induced, the α-linolenic acid and betaine were significantly accumulated in C. mali infected apple stems. This work presents an overview of the changes in gene expression and metabolic profiles in apple under the inoculation of C. mali, which may help to further screen out the mechanism of plant-pathogen interaction at the molecular level. [ABSTRACT FROM AUTHOR]

Wall‐associated kinases (WAKs) have been determined to recognize pathogenic signals and initiate plant immune responses. However, the roles of the family members in host resistance against Valsa canker, a serious fungal disease of apples and pears, are largely unknown. Here, we identified MbWAK1 in Malus baccata, a resistant germplasm differentially expressed during infection by Valsa mali (Vm). Over‐expression of MbWAK1 enhanced the Valsa canker resistance of apple and pear fruits and 'Duli‐G03' (Pyrus betulifolia) suspension cells. A large number of phloem, cell wall, and lipid metabolic process‐related genes were differentially expressed in overexpressed suspension cell lines in response to Valsa pyri (Vp) signals. Among these, the expression of xyloglucan endotransglucosylase/hydrolase (XTH) gene PbeXTH1 and sieve element occlusion B‐like (SEOB) gene PbeSEOB1 were significantly inhibited. Transient expression of PbeXTH1 or PbeSEOB1 compromised the expressional induction of MbWAK1 and the resistance contributed by MbWAK1. In addition, PbeXTH1 and PbeSEOB1 suppressed the immune response induced by MbWAK1. Our results enriched the molecular mechanisms for MbWAK1 against Valsa canker and resistant breeding. [ABSTRACT FROM AUTHOR]

Highly virulent Pseudomonas syringae pv. actinidiae biovar 3 (Psa3) causes kiwifruit bacterial canker, which seriously damages kiwifruits worldwide. We previously screened Psa3-transposon-inserted mutants and obtained reduced-virulence mutants with genes encoding the flagellar protein. Flagella are involved in the motility of bacteria and contribute to P. syringae invasion of plant leaves. However, whether flagella are required for the ability to enter stomata and then multiply in the plant apoplast is not known. Here, we sprayed kiwifruit leaves with Psa3-flagellar-defective mutants and found that their virulence was reduced compared to that of the Psa3 wild type, but their virulence was not reduced when the leaves were infiltrated with Psa3-flagellar-defective mutants using a syringe. Motility and bacterial entry into apoplastic space of the flagellar-defective mutants were also significantly reduced. These results indicate that flagella contribute to Psa3 motility and stomata-mediated entry, leading to disease development, but not multiplication in apoplast space after Psa3 entry. This study provides new insight into bacterial motility in host plants. [ABSTRACT FROM AUTHOR]

The control of kiwifruit canker, caused by Pseudomonas syringae pv. actinidiae (Psa), largely relies on copper pesticides. After epidemics of kiwifruit canker in Ehime Prefecture, Japan, we comprehensively surveyed isolates of Psa biovars 1 (Psa 1) and 3 (Psa 3) for copper resistance from 2002 to 2019. In Psa 1, copper-resistant isolates drastically increased in 2006 and onward during the survey from 2002 to 2008 and were frequently found in 2017. By contrast, copper-resistant Psa 3 isolates emerged soon after its first identification in 2014 and spread to different orchards with an isolation ratio of about 10%. Identification of copper resistance-related genes by whole-genome resequencing revealed that cop genes from Psa 1 isolates in 2017 were almost identical to those from Psa 1 isolates from the late 1980s. On the contrary, in Psa 3 isolates, we found two different sets of cop genes. One of them was closely related to those from other Pseudomonas species, and the other to those from P. syringae pv. tomato. Notably, copper-sensitive Psa1 and 3 isolates had copAB and copRS homologs but no homologs of copCD. Despite the diversity in cop gene sequences, PCR detection of copCD sequences from different lineages matched perfectly with their copper resistance. [ABSTRACT FROM AUTHOR]

Citrus canker is a disease caused by the gram-negative bacterium Xanthomonas citri subp. citri (X. citri), which affects all commercially important varieties of citrus and can lead to significant losses. Fruit sanitization with products such as chlorine-based ones can reduce the spread of the disease. While effective, their use raises concerns about safety of the workers. This work proposes essential oils (EOs) as viable alternatives for fruit sanitization. EOs from Cymbopogon species were evaluated as to their antibacterial activity, their effect on the bacterial membrane, and their ability to sanitize citrus fruit. The in vitro assays revealed that the EOs from C. schoenanthus and C. citratus had a lower bactericidal concentration at 312 mg L−1, followed by 625 mg L−1 for C. martini and C. winterianus. Microscopy assay revealed that the bacterial cell membranes were disrupted after 15 min of contact with all EOs tested. Regarding the sanitizing potential, the EOs with higher proportions of geraniol were more effective in sanitizing acid limes. Fruit treated with C. shoenanthus and C. martini showed a reduction of ∼68% in the recovery of viable bacterial cells. Therefore, these EOs can be used as viable natural alternatives in citrus fruit disinfection. [ABSTRACT FROM AUTHOR]

Radiata pine is one of the most commonly planted tree species in Chile due to its fast growth and desirable wood and pulp properties. However, its productivity is hampered by several diseases. Pitch canker disease (PCC) caused by Fusarium circinatum, is considered the most damaging disease to the pine forest industry. Several control measures have been established, with biological control emerging as an environmentally friendly and effective way for F. circinatum control. Previous studies support the value of Clonostachys rosea in reducing PCC damage, with evidence suggesting a potential induced systemic resistance (ISR) triggered in radiata pines by this agent. Ten-month-old radiata pine plants were pre-treated with C. rosea on a substrate at 8 and 1 days before inoculation with F. circinatum on the stem tip, and expression levels were determined for DXS1, LOX, PAL, and PR3 genes 24 h later. Lesion length was 45% lower on plants pre-treated with C. rosea and infected with F. circinatum compared to non-pre-treated and infected plants. Additionally, LOX and PR3 were induced 23 and 62 times more, respectively, in comparison to untreated plants. Our results indicate that C. rosea causes an ISR response in pre-treated plants, significantly increasing the expression of resistance genes and reducing lesion length. [ABSTRACT FROM AUTHOR]

The glyphosate‐tolerant soybean [Glycine max (L.) Merr.] line 'TN16‐520R1' (Reg. no. CV‐560, PI 703273) was released as a high‐yielding cultivar by University of Tennessee Agricultural Research in 2018. TN16‐520R1 is a BC4F2 cultivar derived from the recurrent parent 'Ellis', with drought tolerance and a maturity group (MG) of V‐early. It has white flowers, gray pubescence, tan podwall, and a determinate growth habit. The seed has buff hila, is yellow with smooth seed coat, and contains 40% protein and 22% oil on a dry weight basis. TN16‐520R1 has good resistance to lodging and good seed quality (1.8 and 1.6, respectively, on 1–5 scale), and a seed size of 11.8 g per 100 seed. Like Ellis, TN16‐520R1 has performed competitively throughout much of the mid‐South United States over several years, with testing occurring over a 6‐year period in 100 environments. Further, TN16‐520R1 is remarkably like the recurrent parent Ellis for yield, maturity, lodging, height, protein, and oil. Also, like Ellis, TN16‐520R1 is resistant to stem canker [caused by Diaporthe phaseolorum (Cooke & Ellis) Sacc. var. caulivora K.L. Athow & R.M. Caldwell] and southern root‐knot nematode [Meloidogyne incognita (Kofoid and White) Chitwood] and has field tolerance to frogeye leafspot [caused by Cercospora sojina (Hara)]. This combination of beneficial traits and broad adaptability make TN16‐520R1 an excellent choice as a soybean cultivar for producers or a crossing parent for breeders. Core Ideas: This study describes the development of a high‐yielding soybean cultivar through backcross breeding.Drought tolerance is a distinguishing feature of TN16‐520R1.TN16‐520R1 possesses glyphosate resistance, which differs from the conventional recurrent parent. [ABSTRACT FROM AUTHOR]

BACKGROUND: Phoma stem canker is a damaging disease of oilseed rape caused by two related fungal species, Leptosphaeria maculans and L. biglobosa. However, previous work has mainly focused on L. maculans and there has been little work on L. biglobosa. This work provides evidence of the importance of L. biglobosa to stem canker epidemics in the UK. RESULTS: Quantification of L. maculans and L. biglobosa DNA using species‐specific quantitative PCR showed that L. biglobosa caused both upper stem lesions and stem base cankers on nine oilseed rape cultivars in the UK. Upper stem lesions were mainly caused by L. biglobosa. For stem base cankers, there was more L. maculans DNA than L. biglobosa DNA in the susceptible cultivar Drakkar, while there was more L. biglobosa DNA than L. maculans DNA in cultivars with the resistance gene Rlm7 against L. maculans. The frequency of L. biglobosa detected in stem base cankers increased from 14% in 2000 to 95% in 2013. Ascospores of L. biglobosa and L. maculans were mostly released on the same days and the number of L. biglobosa ascospores in air samples increased from the 2010/2011 to 2012/2013 growing seasons. CONCLUSION: Effective control of L. maculans increased infection by L. biglobosa, causing severe upper stem lesions and stem base cankers, leading to yield losses. The importance of L. biglobosa to phoma stem canker epidemics can no longer be ignored. Effective control of phoma stem canker epidemics needs to target both L. maculans and L. biglobosa. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Introduction: Pear Valsa canker, caused by Valsa pyri (V. pyri), poses a major threat to pear production. We aimed to assess the effectiveness of the cell-free supernatant (CFS) produced by Trichoderma virens (T. virens) to control the development of pear Valsa canker and reveal the inhibitory mechanism against the pathogenic fungi. Results: Using morphological characteristics and phylogenetic analysis, the pathogen G1H was identified as V. pyri, and the biocontrol fungus WJ561 was identified as Trichoderma virens. CFS derived from WJ561 exhibited strong inhibition of mycelial growth and was capable of reducing the pathogenicity of V. pyri on pear leaves and twigs. Scanning electron microscopy (SEM) observations revealed deformations and shrinkages in the fungal hyphae treated with CFS. The CFS also destroyed the hyphal membranes leading to the leakage of cellular contents and an increase in the malondialdehyde (MDA) content. Additionally, CFS significantly inhibited the activities of catalase (CAT) and superoxide dismutase (SOD), and downregulated the expression of antioxidant defense-related genes in V. pyri, causing the accumulation of reactive oxygen species (ROS). Artesunate, identified as the main component in CFS by liquid chromatograph-mass spectrometry (LC--MS), exhibited antifungal activity against V. pyri. Conclusion: Our findings demonstrate the promising potential of T. virens and its CFS in controlling pear Valsa canker. The primary inhibitory mechanism of CFS involves multiple processes, including membrane damage and negatively affecting enzymatic detoxification pathways, consequently leading to hyphal oxidative damage of V. pyri. This study lays a theoretical foundation for the utilization of T. virens to control V. pyri in practical production. [ABSTRACT FROM AUTHOR]

In ascomycetous fungi, sexual mate recognition requires interaction of the Ste2 receptor protein produced by one partner with the α-factor peptide pheromone produced by the other partner. In some fungi, Ste2 is further needed for chemotropism towards plant roots to allow for subsequent infection and colonization. Here, we investigated whether this is also true for the pine pitch canker fungus, Fusarium circinatum, which is a devastating pathogen of pine globally. Ste2 knockout mutants were generated for two opposite mating-type isolates, after which all strains were subjected to chemotropism assays involving exudates from pine seedling roots and synthetic α-factor pheromone, as well as a range of other compounds for comparison. Our data show that Ste2 is not required for chemotropism towards any of these other compounds, but, in both wild-type strains, Ste2 deletion resulted in the loss of chemotropism towards pine root exudate. Also, irrespective of mating type, both wild-type strains displayed positive chemotropism towards α-factor pheromone, which was substantially reduced in the deletion mutants and not the complementation mutants. Taken together, these findings suggest that Ste2 likely has a key role during the infection of pine roots in production nurseries. Our study also provides a strong foundation for exploring the role of self-produced and mate-produced α-factor pheromone in the growth and overall biology of the pitch canker pathogen. [ABSTRACT FROM AUTHOR]

During our biodiversity investigations in Tibet, China, typical Cytospora canker symptoms were observed on branches of hosts Myricaria paniculate, Prunus cerasifera and Sibiraea angustata. Samples were studied, based on morphological features coupled with multigene phylogenetic analyses of ITS , act , rpb2 , tef1 and tub2 sequence data, which revealed two new species (Cytospora myricicola sp. nov. and C. sibiraeicola sp. nov.) and a known species (C. populina). In addition, Cytospora populina is newly discovered on the host Prunus cerasifera and in Tibet. [ABSTRACT FROM AUTHOR]

Citrus fruits, revered for their nutritional value, face significant threats from diseases like citrus canker, particularly impacting global citrus cultivation, notably in Pakistan. This study delves into the critical role of NPR1-like genes, the true receptors for salicylic acid (SA), in the defense mechanisms of citrus against Xanthomonas axonopodis pv. citri (Xcc). By conducting a comprehensive genome-wide analysis and phylogenetic study, the evolutionary dynamics of Citrus limon genes across diverse citrus cultivars are elucidated. Structural predictions unveil conserved domains, such as the BTB domain and ankyrin repeat domains, crucial for the defense mechanism. Motif analysis reveals essential conserved patterns, while cis-regulatory elements indicate their involvement in transcription, growth, response to phytohormones, and stress. The predominantly cytoplasmic and nuclear localization of NPR1-like genes underscores their pivotal role in conferring resistance to various citrus species. Analysis of the Ks/Ka ratio indicates a purifying selection of NPR1-like genes, emphasizing their importance in different species. Synteny and chromosomal mapping provide insights into duplication events and orthologous links among citrus species. Notably, Xac infection stimulates the expression of NPR1-like genes, revealing their responsiveness to pathogenic challenges. Interestingly, qRT-PCR profiling post-Xac infection reveals cultivar-specific alterations in expression within susceptible and resistant citrus varieties. Beyond genetic factors, physiological parameters like peroxidase, total soluble protein, and secondary metabolites respond to SA-dependent PR genes, influencing plant characteristics. Examining the impact of defense genes (NPR1) and plant characteristics on disease resistance in citrus, this study marks the inaugural investigation into the correlation between NPR1-associated genes and various plant traits in both susceptible and resistant citrus varieties to citrus bacterial canker. [ABSTRACT FROM AUTHOR]

Citrus canker disease affects citrus production. This disease is caused by Xanthomonas citri subsp. citri (Xcc). Previous studies confirmed that during Xcc infection, PthA4, a transcriptional activator like effector (TALE), is translocated from the pathogen to host plant cells. PthA4 binds to the effector binding elements (EBEs) in the promoter region of canker susceptibility gene LOB1 (EBEPthA4-LOBP) to activate its expression and subsequently cause canker symptoms. Previously, the Cas12a/CBE co-editing method was employed to disrupt EBEPthA4-LOBP of pummelo, which is highly homozygous. However, most commercial citrus cultivars are heterozygous hybrids and more difficult to generate homozygous/biallelic mutants. Here, we employed Cas12a/CBE co-editing method to edit EBEPthA4-LOBP of Hamlin (Citrus sinensis), a commercial heterozygous hybrid citrus cultivar grown worldwide. Binary vector GFPp1380N-ttLbCas12a:LOBP1-mPBE:ALS2:ALS1 was constructed and shown to be functional via Xcc-facilitated agroinfiltration in Hamlin leaves. This construct allows the selection of transgene-free regenerants via GFP, edits ALS to generate chlorsulfuron-resistant regenerants as a selection marker for genome editing resulting from transient expression of the T-DNA via nCas9-mPBE:ALS2:ALS1, and edits gene(s) of interest (i.e., EBEPthA4-LOBP in this study) through ttLbCas12a, thus creating transgene-free citrus. Totally, 77 plantlets were produced. Among them, 8 plantlets were transgenic plants (#HamGFP1 - #HamGFP8), 4 plantlets were transgene-free (#HamNoGFP1 - #HamNoGFP4), and the rest were wild type. Among 4 transgene-free plantlets, three lines (#HamNoGFP1, #HamNoGFP2 and #HamNoGFP3) contained biallelic mutations in EBEpthA4, and one line (#HamNoGFP 4) had homozygous mutations in EBEpthA4. We achieved 5.2% transgene-free homozygous/biallelic mutation efficiency for EBEPthA4-LOBP in C. sinensis cv. Hamlin, compared to 1.9% mutation efficiency for pummelo in a previous study. Importantly, the four transgenefree plantlets and 3 transgenic plantlets that survived were resistant against citrus canker. Taken together, Cas12a/CBE co-editing method has been successfully used to generate transgene-free canker-resistant C. sinensis cv. Hamlin in the T0 generation via biallelic/homozygous editing of EBEpthA4 of the canker susceptibility gene LOB1. [ABSTRACT FROM AUTHOR]

Simple Summary: Pathogens of diseases in branches and trunks are commonly inoculated through punching, burning, and toothpick inoculation. At present, there is a lack of comparative analyses of the inoculation outcomes of these three methods. We determined that toothpick inoculation is an optimal option for studying the pathogenesis of Valsa sordida in six-year-old Populus alba var. pyramidalis, providing technical support for research on poplar diseases and offering a theoretical basis for the inoculation of other diseases in branches and trunks. A key step in the study of tree pathology is the identification of an appropriate method for inoculating pathogens of diseases in branches and trunks. Pathogens of diseases in branches and trunks are commonly inoculated through punching, burning, and toothpick inoculation. However, there is a lack of comparative analyses of the inoculation outcomes of these three methods. In this work, six-year-old P. alba var. pyramidalis were inoculated with V. sordida using punching, burning, and toothpick techniques to investigate the differences in the effectiveness of these inoculation methods. Results reveal that the incidence rate was 93.55% in the toothpick inoculation group, significantly higher than the 80.65% in the burning inoculation group (chi-square, n = 90, p = 0.007), while punching inoculation exhibited significant pathological responses in the early stages, with spontaneous healing in the later stage. Additionally, toothpick inoculation was more efficient in inducing Valsa canker when inoculating the pathogen at the bottom of the tree, with lower intra- and inter-row spacing (stand density) providing better outcomes than higher intra- and inter-row spacing. The results of this study demonstrate that toothpick inoculation is an optimal option for studying the artificial inoculation of V. sordida in six-year-old P. alba var. pyramidalis, providing technical support for research on poplar diseases and offering a theoretical basis for the inoculation of other diseases in the branch and trunk. [ABSTRACT FROM AUTHOR]

Aims Citrus canker caused by Xanthomonas citri subsp. citri (X. citri) is a disease of economic importance. Control of this disease includes the use of metallic copper, which is harmful to the environment and human health. Previous studies showed that the crude extract from the fungus Pseudogymnoascus sp. LAMAI 2784 isolated from Antarctic soil had in vitro antibacterial action against X. citri. The aim of the present study was to expand the applications of this extract. Methods and results In greenhouse assays, the crude extract was able to reduce bacterial infection on citrus leaves from 1.55 lesions/cm2 (untreated plants) to 0.04 lesions/cm2. Bisdechlorogeodin was identified as the main compound of the bioactive fraction produced by Pseudogymnoascus sp. LAMAI 2784, which inhibited bacterial growth in vitro (IC90 ≈ 156 µg ml−1) and permeated 80% of X. citri cells, indicating that the membrane is the primary target. Conclusion The present results showed that the bioactive fraction of the extract is mainly composed of the compound bisdechlorogeodin, which is likely responsible for the biological activity against X. citri , and the main mechanism of action is the targeting of the cell membrane. This study indicates that bisdechlorogeodin has valuable potential for the control of X. citri. [ABSTRACT FROM AUTHOR]

Field surveys were conducted in British Columbia fruit tree growing regions to determine the incidence of dieback and to identify the main fungal species causing cankers. Fruit trees showing cankers and dieback symptoms were recorded from 94% of orchards and 5.5% of trees surveyed. Overall, higher dieback incidence was observed in cherry than apple with 33% of cherry blocks showing between 5% and 26% of trees affected. Morphological studies along with DNA sequencing of the internal transcribed spacer (ITS) including the 5.8S rDNA, and parts of the translation elongation factor 1-α (TEF1), beta-tubulin (TUB2), and actin (ACT1) genes, identified seven fungi for the first time in fruit trees in Canada, including Calosphaeria pulchella, Cytospora parasitica, Cytospora populicola, Cytospora sorbicola, Ilyonectria robusta, Nectria dematiosa, and Phaeoacremonium minimum. In addition, this study reports for the first time Diplodia mutila and Diplodia seriata from cankers in sweet cherry in Canada. The already known fungal pathogens Neofabraea perennans and Neonectria ditissima were also identified. Pathogenicity studies showed N. ditissima and C. sorbicola to cause the largest vascular lesions in apple and cherry, respectively. This study identified the main fungal pathogens causing tree fruit cankers and dieback in British Columbia providing important information for the development of effective control strategies. [ABSTRACT FROM AUTHOR]

Alfalfa growers in the Intermountain West of the United States have recently seen an increased incidence in bacterial stem blight (BSB), which can result in significant herbage yield losses from the first harvest. BSB has been attributed to Pseudomonas syringae pv. syringae and P. viridiflava; however, little is known about the genetic diversity and pathogenicity of these bacteria or their interaction with alfalfa plants. Here, we present a comprehensive phylogenetic and phenotypic analysis of P. syringae and P. viridiflava strains causing BSB on alfalfa. A multilocus sequence analysis found that they grouped exclusively with P. syringae PG2b and P. viridiflava PG7a. Alfalfa symptoms caused by both bacterial groups were indistinguishable, although there was a large range in mean disease scores for individual strains. Overall, PG2b strains incited significantly greater disease scores than those caused by PG7a strains. Inoculated plants showed browning in the xylem and collapse of epidermal and pith parenchyma cells. Inoculation with a mixture of PG2b and PG7a strains did not result in synergistic activity. The populations of PG2b and PG7a strains were genetically diverse within their clades and did not group by location or haplotype. The PG2b strains had genes for production of the phytotoxin coronatine, which is unusual in PG2b strains. The results indicate that both pathogens are well established on alfalfa across a wide geographic range and that a recent introduction or evolution of more aggressive strains as the basis for emergence of the disease is unlikely. [ABSTRACT FROM AUTHOR]

Peptidoglycan hydrolases are enzymes responsible for breaking the peptidoglycan present in the bacterial cell wall, facilitating cell growth, cell division and peptidoglycan turnover. Xanthomonas citri subsp. citri (X. citri), the causal agent of citrus canker, encodes an Escherichia coli M23 peptidase EnvC homolog. EnvC is a LytM factor essential for cleaving the septal peptidoglycan, thereby facilitating the separation of daughter cells. In this study, the investigation focused on EnvC contribution to the virulence and cell separation of X. citri. It was observed that disruption of the X. citri envC gene (ΔenvC) led to a reduction in virulence. Upon inoculation into leaves of Rangpur lime (Citrus limonia Osbeck), the X. citri ΔenvC exhibited a delayed onset of citrus canker symptoms compared with the wild-type X. citri. Mutant complementation restored the wild-type phenotype. Sub-cellular localization confirmed that X. citri EnvC is a periplasmic protein. Moreover, the X. citri ΔenvC mutant exhibited elongated cells, indicating a defect in cell division. These findings support the role of EnvC in the regulation of cell wall organization, cell division, and they clarify the role of this peptidase in X. citri virulence. [ABSTRACT FROM AUTHOR]

Pathogenic fungi secrete numerous effectors into host cells to manipulate plants' defense mechanisms. Valsa mali, a necrotrophic fungus, severely impacts apple production in China due to the occurrence of Valsa canker. Here, we predicted 210 candidate effector protein (CEP)-encoding genes from V. mali. The transcriptome analysis revealed that 146 CEP-encoding genes were differentially expressed during the infection of the host, Malus sieversii. Proteome analysis showed that 27 CEPs were differentially regulated during the infection stages. Overall, 25 of the 146 differentially expressed CEP-encoding genes were randomly selected to be transiently expressed in Nicotiana benthamiana. Pathogenicity analysis showed that the transient expression of VM1G-05058 suppressed BAX-triggered cell death while the expression of VM1G-10148 and VM1G-00140 caused cell death in N. benthamiana. In conclusion, by using multi-omics analysis, we identified potential effector candidates for further evaluation in vivo. Our results will provide new insights into the investigation of virulent mechanisms of V. mali. [ABSTRACT FROM AUTHOR]

This article reports on the first documented case of Micrococcus luteus causing bacterial blight in pomegranate trees in India. Pomegranate is an economically important fruit crop in India, and the disease was observed in orchards in Maharashtra. The infected leaves showed symptoms of bacterial leaf blight, including chlorosis, brown spots, and leaf loss. The pathogenicity of the bacteria was confirmed through laboratory tests and molecular identification. This finding is significant for growers as it provides valuable information for disease control measures. [Extracted from the article]

BACKGROUND: Botryosphaeria dieback is a canker disease caused by fungal species of the Botryosphaeriaceae family that threatens almond productivity. The most common control measure to prevent canker development is the application of fungicides which are being phased out by European Union regulations. In the present study, two sets of bacterial strains were evaluated for their antifungal activity against pathogenic Botryosphaeriaceae species through in vitro and in vivo antagonism assays. RESULTS: The rhizospheric bacteria Pseudomonas aeruginosa AC17 and Bacillus velezensis ACH16, as well as the endophytic bacteria Bacillus mobilis Sol 1–2, respectively inhibited 87, 95, and 63% of the mycelial growth of Neofusicoccum parvum, Botryosphaeria dothidea, Diplodia seriata, and Macrophomina phaseolina. Additionally, they significantly reduced the length of lesions caused by N. parvum and B. dothidea in artificially inoculated detached almond twigs. All these bacterial strains produce hydrolytic enzymes that are able to degrade the fungal cell wall. P. aeruginosa AC17 also produces toxic volatile compounds, such as hydrogen cyanide. This strain was the most effective in controlling Botryosphaeria dieback in planta under controlled conditions at a level similar to the biocontrol agent Trichoderma atroviride and standard chemical fungicide treatments. CONCLUSION: Pseudomonas aeruginosa AC17 is the best candidate to be considered as a potential biocontrol agent against Botryosphaeriaceae fungi affecting almond. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Citrus canker is a bacterial disease caused by Xanthomonas citri subsp. citri (Xcc) that affects the citrus industry worldwide. Hrp pili subunits (HrpE), an essential component of Type III secretion system (T3SS) bacteria, play a crucial role in the pathogenesis of Xcc by transporting effector proteins into the host cell and causing canker symptoms. Therefore, development of antibodies that block HrpE can suppress disease progression. In this study, a specific scFv detecting HrpE was developed using phage display technique and characterized using sequencing, ELISA, Western blotting, and molecular docking. In addition, a plant expression vector of pCAMBIA-scFvH6 was constructed and agroinfiltrated into Nicotiana tabacum cv. Samson leaves. The hypersensitive response (HR) in the leaves of transformed and non-transformed plants was evaluated by inoculating leaves with Xcc. After three rounds of biopanning of the phage library, a specific human scFv antibody, named scFvH6, was identified that showed high binding activity against HrpE in ELISA and Western blotting. Molecular docking results showed that five intermolecular hydrogen bonds are involved in HrpE-scFvH6 interaction, confirming the specificity and high binding activity of scFvH6. Successful transient expression of pCAMBIA-scFvH6 in tobacco leaves was verified using immunoassay tests. The binding activity of plant-produced scFvH6 to detect HrpE in Western blotting and ELISA was similar to that of bacterial-produced scFvH6 antibody. Interestingly, tobacco plants expressing scFvH6 showed a remarkable reduction in HR induced by Xcc compared with control plants, so that incidence of necrotic lesions was significantly higher in non-transformed controls (≥ 1.5 lesions/cm2) than in the plants producing scFvH6 (≤ 0.5 lesions/cm2) after infiltration with Xcc inoculum. Our results revealed that the expression of scFvH6 in tobacco leaves can confer resistance to Xcc, indicating that this approach could be considered to provide resistance to citrus bacterial canker disease. [ABSTRACT FROM AUTHOR]

Apple is the most important fruit tree in West Azarbaijan province of Iran. In a survey of apple orchards, a disease with crown and collar canker and necrosis symptoms was observed in three young apple orchards in Urmia, affecting 15% and 1% of 'Red Delicious' and 'Golden Delicious' cultivars, respectively. A fungus with typical characteristics of the asexual morph of Cytospora was regularly isolated from the diseased tissues. Morphological characteristics and phylogenetic analyses inferred from the combined dataset of the ITS-rDNA, parts of LSU, tef1-α, rpb2, and act1 genes revealed that the isolates represent a new species of Cytospora, described herein as Cytospora balanejica sp. nov.. The pathogenicity of all isolates was confirmed on apple cv. 'Red Delicious' based on Koch's postulates. Also, the reaction of 12 other apple cultivars was assessed against five selected isolates with the highest virulence. The results showed that except for cv. 'Braeburn', which did not produce any symptoms of the disease, the other 11 cultivars showed characteristic disease symptoms including sunken and discolored bark and wood. The mean length of the discolored area was different among the 11 so-called susceptible cultivars, hence cvs. 'M4' and 'Golden Delicious' showed the highest and the lowest lesion length, respectively. Moreover, the aggressiveness of the five tested isolates was different, and the isolates BA 2-4 and BA 3-1 had the highest and lowest aggressiveness, respectively. Based on our observations on the potential ability of the fungus to cause disease on young and actively growing apple trees, it will be a serious threat to apple cultivation and industry. [ABSTRACT FROM AUTHOR]