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Verticillium dahliae is a notorious soil-borne pathogen that enters hosts through the roots and proliferates in the plant water-conducting elements to cause Verticillium wilt. Historically, Verticillium wilt symptoms have been explained by vascular occlusion, due to the accumulation of mycelia and plant biomacromolecule aggregation, and also by phytotoxicity caused by pathogen-secreted toxins. Beyond the direct cytotoxicity of some members of the secretome, this review systematically discusses the roles of the V. dahliae secretome in vascular occlusion, including the deposition of polysaccharides as an outcome of plant cell wall destruction, the accumulation of fungal mycelia, and modulation of plant defence responses. By modulating plant defences and hormone levels, the secretome manipulates the vascular environment to induce Verticillium wilt. Thus, the secretome of V. dahliae colludes with plant defence responses to modulate Verticillium wilt symptoms, and thereby bridges the historical concepts of both toxin production by the pathogen and vascular occlusion as the cause of wilting symptoms.

This is the first worldwide report of X. fastidiosa (Xf) subsp. pauca on Castanea sativa and the first characterization of Xf infection on this species. Plants located in three sites (in a long-term affected area in Apulia) were monitored for symptoms and bacterial concentrations in spring and summer, while microscopic analyses were performed to evaluate the pathogen distribution in the xylem vessels. All chestnut plants appeared asymptomatic but the Xf subsp. pauca strain "De Donno" was present at low concentrations (1.14 × 104 and 1.56 × 103 cfu mL−1 in April and June) and with a low incidence (respectively, 38% and 30%). The FISH-CLSM (Fluorescent In Situ Hybridization—Confocal Laser Scanning Microscope) analysis showed evident Xf occlusions but in a low percentage (9.2 ± 3.4%); these data can explain the lack of symptoms on the canopy. Furthermore, through a bibliographic analysis it emerged that Philaenus spumarius and Neophilaenus campestris, two Xf vectors present in Europe, are suckling feeding insects on chestnut trees and could be involved in the bacterial transmission to this species. Asymptomatic Xf host species can play a considerable role in new outbreak emergence or in the expansion of existing ones. So, it is essential to identify them to plan more effective monitoring activities. [ABSTRACT FROM AUTHOR]

Propiconazole has fungistatic and fungicidal properties against Harringtonia lauricola, the causal agent of laurel wilt disease. Propiconazole injections are used by Florida, USA, farmers as a prophylactic method to manage the disease in avocado (Persea americana) trees, but its efficacy has remained questionable for more than a decade due to documented restricted mobility within the tree vascular system. This study was conducted to evaluate the absorption of propiconazole when using soil drenching as an alternative application method, assess the efficacy of propiconazole in controlling disease development when drenched or injected, and its synergistic effect on common cultural management practices used by the local farmers, such as branch removal and trunk cutting (“stumping”). To determine if propiconazole soil-drenching can provide better xylem coverage, potted and mature orchard trees were treated with different concentrations and artificially inoculated with the pathogen. Propiconazole translocation from the roots to above-the-ground tissue was confirmed in potted and orchard trees, but the concentrations in orchard trees were below the fungicidal threshold (1 ppm). Although none of the potted trees developed laurel wilt symptoms, all inoculated branches of the orchard trees did. Furthermore, noninoculated branches in more than 80% of the inoculated and propiconazole-treated orchard trees developed symptoms, even though the inoculated branch was cut at the early stages of disease development. To elucidate if propiconazole injections effectively control the disease, trees from a commercial orchard that were injected five times were challenged by artificial inoculation. Propiconazole concentration in trees was highly variable (ranging from < 0.01 to 294 ppm), but even in trees with a high concentration of propiconazole, inoculated and noninoculated branches developed symptoms. Even though drenched and injected trees were “stumped” soon after symptoms appeared in the noninoculated branches (4 to 5 months after inoculation), all of the stumps in the drenched plot and 80% of the injected trees, showed internal symptoms 5 and 4 months after the cut, respectively. Results demonstrate that the soil-drenching of propiconazole is an ineffective application method in orchard trees, and that the conventional injection does not prevent disease development after artificial inoculation. Moreover, because propiconazole does not prevent the movement of the pathogen to the trunk, the “stumping” of infected trees to reduce the disease in the orchard is an inadequate practice. This study highlights the critical need for other active ingredients with lower fungicidal thresholds, longer half-life, and higher xylem mobility.

This is the first worldwide report of X. fastidiosa (Xf) subsp. pauca on Castanea sativa and the first characterization of Xf infection on this species. Plants located in three sites (in a long-term affected area in Apulia) were monitored for symptoms and bacterial concentrations in spring and summer, while microscopic analyses were performed to evaluate the pathogen distribution in the xylem vessels. All chestnut plants appeared asymptomatic but the Xf subsp. pauca strain “De Donno” was present at low concentrations (1.14 × 104 and 1.56 × 103 cfu mL−1 in April and June) and with a low incidence (respectively, 38% and 30%). The FISH-CLSM (Fluorescent In Situ Hybridization—Confocal Laser Scanning Microscope) analysis showed evident Xf occlusions but in a low percentage (9.2 ± 3.4%); these data can explain the lack of symptoms on the canopy. Furthermore, through a bibliographic analysis it emerged that Philaenus spumarius and Neophilaenus campestris, two Xf vectors present in Europe, are suckling feeding insects on chestnut trees and could be involved in the bacterial transmission to this species. Asymptomatic Xf host species can play a considerable role in new outbreak emergence or in the expansion of existing ones. So, it is essential to identify them to plan more effective monitoring activities.

Pierce's disease is a major threat to grapevines caused by the bacterium Xylella fastidiosa. Although devoid of a type 3 secretion system commonly employed by bacterial pathogens to deliver effectors inside host cells, this pathogen is able to influence host parenchymal cells from the xylem lumen by secreting a battery of hydrolytic enzymes. Defining the cellular and biochemical changes induced during disease can foster the development of novel therapeutic strategies aimed at reducing the pathogen fitness and increasing plant health. To this end, we investigated the transcriptional, proteomic, and metabolomic responses of diseased Vitis vinifera compared to healthy plants. We found that several antioxidant strategies were induced, including the accumulation of gamma-aminobutyric acid (GABA) and polyamine metabolism, as well as iron and copper chelation, but these were insufficient to protect the plant from chronic oxidative stress and disease symptom development. Notable upregulation of phytoalexins, pathogenesis-related proteins, and various aromatic acid metabolites was part of the host responses observed. Moreover, upregulation of various cell wall modification enzymes followed the proliferation of the pathogen within xylem vessels, consistent with the intensive thickening of vessels' secondary walls observed by magnetic resonance imaging. By interpreting the molecular profile changes taking place in symptomatic tissues, we report a set of molecular markers that can be further explored to aid in disease detection, breeding for resistance, and developing therapeutics.

Soil-borne pathogens such as Verticillium species, invade into the roots of many herbaceous and woody hosts. The xylem environment supplies these pathogens with a continuous flow of nitrogen-rich nutrition. Detailed quantitative increases in amino acids in the stems, petioles, leaflets and roots of young tomato plants infected with Verticillium. albo-atrum the causal agent of wilt disease, are described in this paper for the first time. Results focus in particular on the vascular environment prior to the emergence of visual symptoms. Total amino acid concentrations in infected stems and petioles increased substantially at 144 and 216 h after inoculation. This effect was evident in leaflets at 216 h after inoculation. By 216 h most amino acid concentrations were substantially increased in stems, petioles and leaflets of infected plants relative to healthy controls. Earlier at 144 h in stems substantial increases were recorded for aspartic acid, threonine, serine, glutamic acid, glycine and ethanolamine. A similar picture emerged for petioles with the addition of increases in proline but not glycine. Amino acids increasing substantially in infected leaflets at 216 h were aspartic acid, glutamic acid and ethanolamine. In the infected roots there was relatively little difference in amino acid concentrations relative to healthy controls with the particular exceptions of proline and ethanolamine. By 18 days (432h), when visual symptoms were well advanced marked increases in amino acid concentrations were found for threonine, serine, α-alanine, valine, methionine, iso-leucine, leucine, tyrosine, ethanolamine, ornithine, lysine, histidine and arginine.

Summary: Xylella fastidiosa is the causal agent of important crop diseases and is transmitted by xylem‐sap‐feeding insects. The bacterium colonizes xylem vessels and can persist with a commensal or pathogen lifestyle in more than 500 plant species. In the past decade, reports of X. fastidiosa across the globe have dramatically increased its known occurrence. This raises important questions: How does X. fastidiosa interact with the different host plants? How does the bacterium interact with the plant immune system? How does it influence the host's microbiome? We discuss recent strain genetic typing and plant transcriptome and microbiome analyses, which have advanced our understanding of factors that are important for X. fastidiosa plant infection. [ABSTRACT FROM AUTHOR]

The effect of olive (Olea europaea) stem extract (OSE) on the viability of conidia of Verticillium dahliae, the causal agent of Verticillium wilt of olive (VWO), is not yet well understood. Thus, the aim of this study was to determine the influence of the olive genotype (cultivar resistance) and the interaction between olive cultivars and biocontrol treatments on the effect of OSE on conidial germination of V. dahliae by in vitro sensitivity tests. To this end, OSE from cultivars Frantoio, Arbequina, and Picual, respectively tolerant, moderately susceptible, and highly susceptible to V. dahliae, were tested alone or after treatments with biological control agents (BCAs) and commercial products efficient at reducing the progress of VWO. Aureobasidium pullulans strain AP08, Phoma sp. strain ColPat-375, and Bacillus amyloliquefaciens strain PAB-24 were considered as BCAs. Aluminium lignosulfonate (IDAI Brotaverd®), copper phosphite (Phoscuprico®), potassium phosphite (Naturfos®), and salicylic acid were selected as commercial products. Our results indicate that the influence of biological treatments against the pathogen depends on the genotype, since the higher the resistance of the cultivar, the lower the effect of the treatments on the ability of OSE to inhibit the germination of conidia. In 'Picual', the BCA B. amyloliquefaciens PAB024 and copper phosphite were the most effective treatments in inhibiting conidia germination by the OSE. This work represents a first approach to elucidate the role of cultivar and biological treatments in modifying the effect on the pathogen of the endosphere content of olive plants. [ABSTRACT FROM AUTHOR]

Quorum sensing (QS) helps the Xanthomonas group of phytopathogens to infect several crop plants. The vascular phytopathogen Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot disease on Brassicaceae leaves, where a typical v-shaped lesion spans both vascular and mesophyll regions with progressive leaf chlorosis. Recently, the role of QS has been elucidated during Xcc early infection stages. However, a detailed insight into the possible role of QS-regulated bacterial invasion in host chlorophagy during late infection stages remains elusive. In this study, using QS-responsive whole-cell bioreporters of Xcc, we present a detailed chronology of QS-facilitated Xcc colonization in the mesophyll region of cabbage (Brassica oleracea) leaves. We report that QS-enabled localization of Xcc to parenchymal chloroplasts triggers leaf chlorosis and promotion of systemic infection. Our results indicate that the QS response in the Xanthomonas group of vascular phytopathogens maximizes their population fitness across host tissues to trigger stage-specific host chlorophagy and establish a systemic infection. [ABSTRACT FROM AUTHOR]

Soil-borne pathogens such as Verticillium species, invade into the roots of many herbaceous and woody hosts. The xylem environment supplies these pathogens with a continuous flow of nitrogen-rich nutrition. Detailed quantitative increases in amino acids in the stems, petioles, leaflets and roots of young tomato plants infected with Verticillium. albo-atrum the causal agent of wilt disease, are described in this paper for the first time. Results focus in particular on the vascular environment prior to the emergence of visual symptoms. Total amino acid concentrations in infected stems and petioles increased substantially at 144 and 216 h after inoculation. This effect was evident in leaflets at 216 h after inoculation. By 216 h most amino acid concentrations were substantially increased in stems, petioles and leaflets of infected plants relative to healthy controls. Earlier at 144 h in stems substantial increases were recorded for aspartic acid, threonine, serine, glutamic acid, glycine and ethanolamine. A similar picture emerged for petioles with the addition of increases in proline but not glycine. Amino acids increasing substantially in infected leaflets at 216 h were aspartic acid, glutamic acid and ethanolamine. In the infected roots there was relatively little difference in amino acid concentrations relative to healthy controls with the particular exceptions of proline and ethanolamine. By 18 days (432h), when visual symptoms were well advanced marked increases in amino acid concentrations were found for threonine, serine, a-alanine, valine, methionine, iso-leucine, leucine, tyrosine, ethanolamine, ornithine, lysine, histidine and arginine. [ABSTRACT FROM AUTHOR]

Fusarium stalk rot (FSR), caused by Fusarium verticillioides , is one of the most destructive diseases impacting maize yield worldwide. In this study, net carbon assimilation rate (A), stomatal conductance to water vapor (g s), transpiration rate (E), and internal CO 2 concentration (C i) were evaluated on leaves and the activities of enzymes (chitinase (CHI), β -1-3-glucanase (GLU), phenylalanine ammonia-lyase (PAL), polyphenoloxidase (PPO), catalase (CAT), ascorbate peroxidase (APX), peroxidase (POX)) as well the concentrations of total soluble phenolics (TSP), lignin-thioglycolic acid (LTGA) derivatives, and malondialdehyde (MDA) were evaluated in the internodes and nodes of plants from maize hybrids moderately resistant (BRS 1035) and susceptible (30F35Y) to FSR. The upward relative lesion length (URLL) and radial fungal colonization (RFC) were 46 and 29% lower for the BRS 1035 hybrid in comparison to 30F35Y hybrid, respectively, at 30 after inoculation (dai). For both hybrids, A , g s , and E values significantly decreased while the C i values increased on infected leaves compared to noninoculated plants. Inoculated plants from BRS 1035 hybrid showed an increase in A compared to inoculated plants from 30F35Y hybrid, and the increase in C i values was greater for plants from 30F35Y hybrid at 30 dai compared to plants from BRS 1035 hybrid. The CHI, GLU, PPO, CAT, APX, and POX activities increased for inoculated plants from both hybrids compared to the noninoculated plants. In the internodes region, the increase in the activities of CHI (during the infection process of F. verticillioides) and GLU (at earlier stages of F. verticillioides infection) was more pronounced for plants from BRS 1035 hybrid than for plants from 30F35Y hybrid. In the region of the nodes, activities of CHI (during the infection process of F. verticillioides), PAL (at 20 dai), PPO (at 30 dai), and CAT and POX (both at three dai) were more pronounced for plants from BRS 1035 hybrid than for plants from 30F35Y hybrid. In the internodes region, the lower TSP concentration at 30 dai was linked to a high concentration of LTGA derivatives for inoculated plants from BRS 1035 hybrid compared to inoculated plants from 30F35Y hybrid. Taking together, the results of the present study allowed to conclude that the infection by F. verticillioides triggered physiological and biochemical changes on the stalk of maize plants influencing photosynthesis on leaves. A more robust antioxidative metabolism for reactive oxygen species removal in association with an efficient and strong activity of defense enzymes helped to minimize the cellular damage caused by F. verticillioides infection resulting, therefore, in an increase in maize resistance to FSR. • The infection by Fusarium verticillioides on the stalk impaired the photosynthesis of maize plants. • High defense enzymes activities in the stalk was associated with maize resistance to Fusarium stalk rot. • High lignin concentration occurred in the internodes of plants from the more resistant hybrid. • A more robust antioxidative metabolism minimized the cellular damage caused by F. verticillioides infection. [ABSTRACT FROM AUTHOR]

Key message: A histopathological study using one resistant and one susceptible clone ofEucalyptus urophylla × Eucalyptus grandishybrid showed that the colonization ofCeratocystis fimbriatawas limited by rapid and intense host defense responses such as closure of the vessel pits; formation of tyloses and gels; accumulation of amorphous material, starch, phenolic compounds, and calcium oxalate; and tissue lignification. The defense mechanisms of the resistant clone were not lethal to the pathogen because the fungus was reisolated from the diseased tissue. Context: The use of resistant genotypes of eucalypt is widely used to control Ceratocystis wilt caused by Ceratocystis fimbriata. However, little is known regarding the fungal infection process and the host defense responses. Aims: Thus, the objectives of this study were to compare the histopathological responses of one resistant and one susceptible clone of Eucalyptus urophylla × Eucalyptus grandis to artificial inoculation with C. fimbriata and to identify possible host defense responses against fungal infection. Methods: Fungal colonization was analyzed by light and scanning electron microscopy. The host defense responses to artificial fungal inoculation were evaluated through histochemical analysis and determining of the lignin concentration and lesion lengths, whereas the pathogen viability was confirmed by reisolations. Results: Both tested clones showed similar responses against fungal infection but presented defense responses with different speeds and intensities. Fungal colonization was not restricted in the tissue of plants from the susceptible clone, whereas in the resistant clone, fungal colonization was limited to the xylem vessels and parenchyma around the vessels due to closure of the vessel pits; intense formation of tyloses and gels; accumulation of amorphous material, starch, phenolic compounds, and calcium oxalate; and tissue lignification. However, the fungus was reisolated from the inoculated tissues of both clones. Conclusion: The resistance of eucalypt cuttings was found to be based on the rapid and intense defense responses shaped by biochemical and structural mechanisms that contained fungal colonization in the xylem vessels and parenchyma tissues. [ABSTRACT FROM AUTHOR]

Dieback and wilt caused by Erwinia psidii is an emerging disease that has been causing considerable damage in eucalypt plantations. Because it is a recently emerged disease, several aspects of the bacterial interaction with its host still remain to be elucidated. In this work, we studied the E. psidii colonization and biofilm formation in eucalypt tissues by specific detection using PCR and scanning electron microscopy (SEM). The results indicate that the bacterium is able to translocate in stem tissue mainly acropetally, although movement in the basipetal direction was also observed to a lesser extent, always through the xylem. No colonization of phloem tissues was observed. In addition to colonizing the xylem, E. psidii colonized the parenchymatous tissue. The bacterium formed cell aggregates enveloped by fibrillar material that evolved into complex, well‐structured biofilms in stem and leaf tissues. In contrast, no biofilm formation was observed on abiotic surfaces. These observations suggest that biofilm formation plays an important role in the elicitation of dieback and wilt symptoms caused by E. psidii on eucalypt plants. This study not only shows ultrastructural aspects of the E. psidii communities but also tissue damage in eucalypt plants that was associated with the presence of bacterial aggregates and formation of tyloses. [ABSTRACT FROM AUTHOR]

Ralstonia solanacearum is a bacterial vascular pathogen causing devastating bacterial wilt. In the field, resistance against this pathogen is quantitative and is available for breeders only in tomato and eggplant. To understand the basis of resistance to R. solanacearum in tomato, we investigated the spatio-temporal dynamics of bacterial colonization using non-invasive live monitoring techniques coupled to grafting of susceptible and resistant varieties. We found four 'bottlenecks' that limit the bacterium in resistant tomato: root colonization, vertical movement from roots to shoots, circular vascular bundle invasion, and radial apoplastic spread in the cortex. Radial invasion of cortical extracellular spaces occurred mostly at late disease stages but was observed throughout plant infection. This study shows that resistance is expressed in both root and shoot tissues, and highlights the importance of structural constraints to bacterial spread as a resistance mechanism. It also shows that R. solanacearum is not only a vascular pathogen but spreads out of the xylem, occupying the plant apoplast niche. Our work will help elucidate the complex genetic determinants of resistance, setting the foundations to decipher the molecular mechanisms that limit pathogen colonization, which may provide new precision tools to fight bacterial wilt in the field. [ABSTRACT FROM AUTHOR]

The effect of olive (Olea europaea) stem extract (OSE) on the viability of conidia of Verticillium dahliae, the causal agent of Verticillium wilt of olive (VWO), is not yet well understood. Thus, the aim of this study was to determine the influence of the olive genotype (cultivar resistance) and the interaction between olive cultivars and biocontrol treatments on the effect of OSE on conidial germination of V. dahliae by in vitro sensitivity tests. To this end, OSE from cultivars Frantoio, Arbequina, and Picual, respectively tolerant, moderately susceptible, and highly susceptible to V. dahliae, were tested alone or after treatments with biological control agents (BCAs) and commercial products efficient at reducing the progress of VWO. Aureobasidium pullulans strain AP08, Phoma sp. strain ColPat-375, and Bacillus amyloliquefaciens strain PAB-24 were considered as BCAs. Aluminium lignosulfonate (IDAI Brotaverd®), copper phosphite (Phoscuprico®), potassium phosphite (Naturfos®), and salicylic acid were selected as commercial products. Our results indicate that the influence of biological treatments against the pathogen depends on the genotype, since the higher the resistance of the cultivar, the lower the effect of the treatments on the ability of OSE to inhibit the germination of conidia. In ‘Picual’, the BCA B. amyloliquefaciens PAB024 and copper phosphite were the most effective treatments in inhibiting conidia germination by the OSE. This work represents a first approach to elucidate the role of cultivar and biological treatments in modifying the effect on the pathogen of the endosphere content of olive plants.

Mango (Mangifera indica) is an economically significant crop, and is affected by dieback in nearly all commercial production areas. Due to the wide range of organisms previously associated with these disease symptoms in Florida, isolations and pathogenicity tests were carried out to determine the causal organism. The pathogen was identified as Neofusicoccum batangarum based on genetic sequences from three loci (internal transcribed spacer of the rDNA (ITS), β-tubulin (BT), and translation elongation factor 1-α (EF)), recommended for members of the Botryosphaeriaceae family. Possible infection routes were determined by inoculating wounded and unwounded stems with N. batangarum. Trees wounded prior to pathogen inoculation developed larger lesions (5.85 cm ± 1.51) than unwounded trees (0.51 cm ± 0.48), p < 0.0003. In addition, lesions only developed at a small number of inoculation sites in the absence of wounds (14.3%), compared to 93% when stems were wounded. No necrosis was observed in the negative controls. This study provides molecular data on N. batangarum, and evidence of its role causing mango dieback in Florida.

Pierce’s disease is a major threat to grapevines caused by the bacterium Xylella fastidiosa. Although devoid of a type 3 secretion system commonly employed by bacterial pathogens to deliver effectors inside host cells, this pathogen is able to influence host parenchymal cells from the xylem lumen by secreting a battery of hydrolytic enzymes. Defining the cellular and biochemical changes induced during disease can foster the development of novel therapeutic strategies aimed at reducing the pathogen fitness and increasing plant health. To this end, we investigated the transcriptional, proteomic, and metabolomic responses of diseased Vitis vinifera compared to healthy plants. We found that several antioxidant strategies were induced, including the accumulation of gamma-aminobutyric acid (GABA) and polyamine metabolism, as well as iron and copper chelation, but these were insufficient to protect the plant from chronic oxidative stress and disease symptom development. Notable upregulation of phytoalexins, pathogenesis-related proteins, and various aromatic acid metabolites was part of the host responses observed. Moreover, upregulation of various cell wall modification enzymes followed the proliferation of the pathogen within xylem vessels, consistent with the intensive thickening of vessels’ secondary walls observed by magnetic resonance imaging. By interpreting the molecular profile changes taking place in symptomatic tissues, we report a set of molecular markers that can be further explored to aid in disease detection, breeding for resistance, and developing therapeutics.

Pierce's disease is a major threat to grapevines caused by the bacterium Xylella fastidiosa. Although devoid of a type 3 secretion system commonly employed by bacterial pathogens to deliver effectors inside host cells, this pathogen is able to influence host parenchymal cells from the xylem lumen by secreting a battery of hydrolytic enzymes. Defining the cellular and biochemical changes induced during disease can foster the development of novel therapeutic strategies aimed at reducing the pathogen fitness and increasing plant health. To this end, we investigated the transcriptional, proteomic, and metabolomic responses of diseased Vitis vinifera compared to healthy plants. We found that several antioxidant strategies were induced, including the accumulation of gamma-aminobutyric acid (GABA) and polyamine metabolism, as well as iron and copper chelation, but these were insufficient to protect the plant from chronic oxidative stress and disease symptom development. Notable upregulation of phytoalexins, pathogenesis-related proteins, and various aromatic acid metabolites was part of the host responses observed. Moreover, upregulation of various cell wall modification enzymes followed the proliferation of the pathogen within xylem vessels, consistent with the intensive thickening of vessels' secondary walls observed by magnetic resonance imaging. By interpreting the molecular profile changes taking place in symptomatic tissues, we report a set of molecular markers that can be further explored to aid in disease detection, breeding for resistance, and developing therapeutics. [ABSTRACT FROM AUTHOR]

Verticillium dahliae is the most overwhelming plant pathogen, causing Verticillium wilt in a number of economic crops. The molecular mechanism is still unclear and identification of the genes involved in the pathogenicity or virulence of this fungus would benefit to uncover such mechanism. STT3 is a catalytic subunit of the multi-subunit oligosaccharyl transferase (OST) and plays an essential role in glycoprotein modification. Here, we characterized STT3 gene (VDAG_03232.1) of V. dahliae to explore its regulatory role in the development and virulence by deletion and complementation of this gene, as well as its silence in transgenic plants. The expression of the STT3 gene increased at the stage of conidia germination and reached its peak level with germ tube formation and elongation. We generated the knockout mutants (Δ STT3) using protoplast transformation. Mycelial growth, sporulation ability and glycoprotein secretion were impaired when Δ STT3 mutants were grown on media supplemented with different carbon sources. Moreover, Δ STT3 mutants exhibited distinctly decreased germination ratio and reduction in virulence compared with the wild type (Vd wt) and complementary (Δ STT3- C) strains. We also generated transgenic Nicotiana benthamiana (Trans-1 and -2) plants by expressing dsRNA against the STT3 gene. Transgenic plants showed significant reduction in the disease index and fungal biomass resulting in elevated resistance to V. dahliae compared with the wild-type plants when inoculated with Vd wt. Our results indicated that STT3 mediates the full virulence through the regulation in fungal development, hyphal growth, glycoprotein secretion of V. dahliae and merits further study as a potential RNAi target to control this fungus. [ABSTRACT FROM AUTHOR]

* Verticillium wilt of olive (VWO) is one of the most serious biotic constraints for this tree crop. Our knowledge of the genetics of the tolerance/resistance to this disease is very limited. Here we show that tolerance of the cv Frantoio relies on both basal and early pathogen-induced differential transcriptomic responses. * A comparative transcriptomic analysis (RNA-seq) was conducted in root tissues of cvs Frantoio (VWO-tolerant) and Picual (VWO-susceptible). RNA samples originated from roots of inoculated olive plants during the early infection stages by Verticillium dahliae (highly virulent, defoliating pathotype). * A huge number of differentially expressed genes (DEGs) were found between 'Frantoio' and 'Picual' (27 312 unigenes) in the absence of the pathogen. Upon infection with V. dahliae, 'Picual' and 'Frantoio' plants responded differently too. In the early infection stages, four clusters of DEGs could be identified according to their time-course expression patterns. Among others, a pathogenesis-related protein of the Bet v I family and a dirigent-like protein involved in lignification, and several BAK1, NHL1, reactive oxygen species stress response and BAM unigenes showed noticeable differences between cultivars. * Tolerance of 'Frantoio' plants to VWO is a consequence of a complex and multifaceted process which involves many plant traits. [ABSTRACT FROM AUTHOR]

Wilt diseases caused by vascular pathogens include some of the most damaging stresses affecting trees. Dutch elm disease (DED), caused by the fungus Ophiostoma novo-ulmi, destroyed most of North American and European elm populations in the 20th century. The highly susceptible English elm, also known as Atinian clone, suffered the highest mortality rates during the last pandemic event, probably due to its lack of genetic diversity. To study the DED pathosystem, we inoculated English elm ramets with O. novo-ulmi and evaluated xylem anatomy, molecular response, and disease symptoms. The high DED susceptibility of the clone was linked to xylem structure. The transcript levels changed significantly for 1,696 genes during O. novo-ulmi invasion. Genes covering different steps of the plant immune system were identified, many of which showed homology with Arabidopsis thaliana genes involved in systemic acquired resistance. Induction of several pathogenesis-related proteins and repression of fasciclin-like arabinogalactan proteins and other cell wall biosynthesis pathways evidence unbalanced costs between growth and defence mechanisms far from the inoculation point. This study sheds light on elm molecular defence mechanisms against DED. [ABSTRACT FROM AUTHOR]

The soil-borne fungus Verticillium causes serious vascular disease in a wide variety of annual crops and woody perennials. Verticillium wilt is notoriously difficult to control by conventional methods, so there is great potential for biocontrol to manage this disease. In this study we aimed to review the research about Verticillium biocontrol to get a better understanding of characteristics that are desirable in a biocontrol agent (BCA) against Verticillium wilt. We only considered studies in which the BCAs were tested on plants. Most biocontrol studies were focused on plants of the Solanaceae, Malvaceae, and Brassicaceae and within these families eggplant, cotton, and oilseed rape were the most studied crops. The list of bacterial BCAs with potential against Verticillium was dominated by endophytic Bacillus and Pseudomonas isolates, while non-pathogenic xylem-colonizing Verticilliumand Fusariumisolates topped the fungal list. Predominant modes of action involved in biocontrol were inhibition of primary inoculum germination, plant growth promotion, competition and induced resistance. Many BCAs showed in vitro antibiosis and mycoparasitism but these traits were not correlated with activity in vivo and there is no evidence that they play a role in planta. Good BCAs were obtained from soils suppressive to Verticillium wilt, disease suppressive composts, and healthy plants in infested fields. Desirable characteristics in a BCA against Verticillium are the ability to (1) affect the survival or germination of microsclerotia, (2) colonize the xylem and/or cortex and compete with the pathogen for nutrients and/or space, (3) induce resistance responses in the plant and/or (4) promote plant growth. Potential BCAs should be screened in conditions that resemble the field situation to increase the chance of successful use in practice. Furthermore, issues such as large scale production, formulation, preservation conditions, shelf life, and applicationmethods should be considered early in the process of selecting BCAs against Verticillium. [ABSTRACT FROM AUTHOR]

Verticillium wilt of olive (VWO) is caused by the vascular pathogen Verticillium dahliae. One of the best VWO management measures is the use of tolerant cultivars; however, our knowledge on VWO tolerance/resistance genetics is very limited. A transcriptomic analysis was conducted to (i) identify systemic defence responses induced/repressed in aerial tissues of the tolerant cultivar Frantoio upon root colonization by V. dahliae, and (ii) determine the expression pattern of selected defence genes in olive cultivars showing differential susceptibility to VWO. Two suppression subtractive hybridization cDNA libraries, enriched in up-regulated (FU) and down-regulated (FD) genes respectively, were generated from ‘Frantoio’ aerial tissues. Results showed that broad systemic transcriptomic changes are taking place during V. dahliae-‘Frantoio’ interaction. A total of 585 FU and 381 FD unigenes were identified, many of them involved in defence response to (a)biotic stresses. Selected genes were then used to validate libraries and evaluate their temporal expression pattern in ‘Frantoio’. Four defence genes were analyzed in cultivars Changlot Real (tolerant) and Picual (susceptible). An association between GRAS1 and DRR2 gene expression patterns and susceptibility to VWO was observed, suggesting that these transcripts could be further evaluated as markers of the tolerance level of olive cultivars to V. dahliae.

The species Verticillium represents a group of highly destructive fungal pathogens, responsible for vascular wilt in a number of crops. The host response to infection by V. longisporum at the level of secondary plant metabolites has not been well explored. Natural variation in the glucosinolate (GLS) composition of four Arabidopsis thaliana accessions was characterized: the accessions Bur-0 and Hi-0 accumulated alkenyl GLS, while 3-hydroxypropyl GLS predominated in Kn-0 and Ler-0. With respect to GLS degradation products, Hi-0 and Kn-0 generated mainly isothiocyanates, whereas Bur-0 released epithionitriles and Ler-0 nitriles. An analysis of the effect on the composition of both GLS and its breakdown products in the leaf and root following the plants' exposure to V. longisporum revealed a number of organ- and accession-specific alterations. In the less disease susceptible accessions Bur-0 and Ler-0, colonization depressed the accumulation of GLS in the rosette leaves but accentuated it in the roots. In contrast, in the root, the level of GLS breakdown products in three of the four accessions fell, suggestive of their conjugation or binding to a fungal target molecule(s). The plant-pathogen interaction influenced both the organ- and accession-specific formation of GLS degradation products.

Verticillium wilt of olive (VWO) is caused by the vascular pathogen Verticillium dahliae. One of the best VWO management measures is the use of tolerant cultivars; however, our knowledge on VWO tolerance/resistance genetics is very limited. A transcriptomic analysis was conducted to (i) identify systemic defense responses induced/repressed in aerial tissues of the tolerant cultivar Frantoio upon root colonization by V. dahliae, and (ii) determine the expression pattern of selected defense genes in olive cultivars showing differential susceptibility to VWO. Two suppression subtractive hybridization cDNA libraries, enriched in up-regulated (FU) and down-regulated (FD) genes respectively, were generated from "Frantoio" aerial tissues. Results showed that broad systemic transcriptomic changes are taking place during V. dahliae-"Frantoio" interaction. A total of 585 FU and 381 FD unigenes were identified, many of them involved in defense response to (a)biotic stresses. Selected genes were then used to validate libraries and evaluate their temporal expression pattern in "Frantoio." Four defense genes were analyzed in cultivars Changlot Real (tolerant) and Picual (susceptible). An association between GRAS1 and DRR2 gene expression patterns and susceptibility to VWO was observed, suggesting that these transcripts could be further evaluated as markers of the tolerance level of olive cultivars to V. dahliae. [ABSTRACT FROM AUTHOR]

The species Verticillium represents a group of highly destructive fungal pathogens, responsible for vascular wilt in a number of crops. The host response to infection by Verticillium longisporum at the level of secondary plant metabolites has not been well explored. Natural variation in the glucosinolate (GLS) composition of four Arabidopsis thaliana accessions was characterized: the accessions Bur-0 and Hi-0 accumulated alkenyl GLS, while 3-hydroxypropyl GLS predominated in Kn-0 and Ler-0. With respect to GLS degradation products, Hi-0 and Kn-0 generated mainly isothiocyanates, whereas Bur-0 released epithionitriles and Ler-0 nitriles. An analysis of the effect on the composition of both GLS and its breakdown products in the leaf and root following the plants' exposure to V. longisporum revealed a number of organ- and accession-specific alterations. In the less disease susceptible accessions Bur-0 and Ler-0, colonization depressed the accumulation of GLS in the rosette leaves but accentuated it in the roots. In contrast, in the root, the level of GLS breakdown products in three of the four accessions fell, suggestive of their conjugation or binding to a fungal target molecule(s). The plant-pathogen interaction influenced both the organ- and accession-specific formation of GLS degradation products. [ABSTRACT FROM AUTHOR]

Mango wilt is one of the most important diseases affecting mango production, yet the physiological mechanisms underlying host responses to this fungal infection are poorly known. The disease attenuation displayed by some mango cultivars might be related to higher antioxidant capacities in leaves. To test this hypothesis, the activities of a range of enzymes (e.g. superoxide dismutase and catalase) and metabolites (e.g. ascorbate and glutathione) involved in the antioxidant system, as well as some markers for oxidative stress (e.g. hydrogen peroxide and malondialdehyde equivalents), were evaluated for 30 days after the inoculation of mango plants with Ceratocystis fimbriata. Tommy Atkins plants displayed higher values for the upward, downward and radial fungal colonization of the stem tissues than Ubá plants. In contrast to the working hypothesis, Tommy Atkins plants exhibited more prominent increases in enzyme activities and metabolite concentrations related to oxidative stress responses. These increases were minimal, if any, in Ubá plants. These results suggest that Tommy Atkins plants are less effective at restraining fungal spread through vascular tissues and thus trigger leaf antioxidant defences to cope with the developing stress. [ABSTRACT FROM AUTHOR]

PIRIN (PRN) is a member of the functionally diverse cupin protein superfamily. There are four members of the Arabidopsis thaliana PRN family, but the roles of these proteins are largely unknown. Here we describe a function of the Arabidopsis PIRIN2 (PRN2) that is related to susceptibility to the bacterial plant pathogen Ralstonia solanacearum. Two prn2 mutant alleles displayed decreased disease development and bacterial growth in response to R. solanacearum infection. We elucidated the underlying molecular mechanism by analyzing PRN2 interactions with the papain-like cysteine proteases (PLCPs) XCP2, RD21A, and RD21B, all of which bound to PRN2 in yeast two-hybrid assays and in Arabidopsis protoplast co-immunoprecipitation assays. We show that XCP2 is stabilized by PRN2 through inhibition of its autolysis on the basis of PLCP activity profiling assays and enzymatic assays with recombinant protein. The stabilization of XCP2 by PRN2 was also confirmed in planta. Like prn2 mutants, an xcp2 single knockout mutant and xcp2 prn2 double knockout mutant displayed decreased susceptibility to R. solanacearum, suggesting that stabilization of XCP2 by PRN2 underlies susceptibility to R. solanacearum in Arabidopsis. [ABSTRACT FROM AUTHOR]

Mango wilt, caused by Ceratocystis fimbriata, is one of the most important diseases affecting mango yields in Brazil. Information regarding the infection process of C. fimbriata in the stem tissues of mango from different cultivars and the basis of host resistance to the pathogen is rare in the literature. Thus, the objective of the study was to investigate how infection by two isolates of C. fimbriata can be affected by mango cultivar-specific mechanisms of resistance. Disease progress on the inoculated stem tissues of the mango cultivars was evaluated and stem sections were obtained from the site of inoculation and prepared for histopathological observations using light microscopy. The factors mango cultivars and C. fimbriata isolates and their interaction were significant for all measures of disease development. Plants from the cultivars Espada, Haden and Palmer inoculated with isolates of C. fimbriata were more susceptible, whereas plants from the cultivars Tommy and Ubá were moderately resistant and resistant, respectively. Histopathologically, fungal isolates apparently massively colonized the stem tissues of plants from the susceptible cultivars Espada, Haden and Palmer, starting from the collenchyma and moving in the direction of the cortical parenchyma, xylem vessels and pith parenchyma. By contrast, on stem tissues of plants from the resistant cultivars Tommy Atkins and Ubá, most of the cells reacted to C. fimbriata infection by accumulating amorphous material. The results from the present study strongly indicated the importance of phenolic compounds for mango cultivar resistance against infection by Brazilian C. fimbriata isolates. [ABSTRACT FROM AUTHOR]

Xanthomonas axonopodis pathovar vasculorum strain NCPPB 900 was isolated from sugarcane on Reunion island in 1960. Consistent with its belonging to fatty-acid type D, multi-locus sequence analysis confirmed that NCPPB 900 falls within the species X. axonopodis. This genome harbours sequences similar to plasmids pXCV183 from X. campestris pv. vesicatoria 85-10 and pPHB194 from Burkholderia pseudomallei. Its repertoire of predicted effectors includes homologues of XopAA, XopAD, XopAE, XopB, XopD, XopV, XopZ, XopC and XopI and transcriptional activator-like effectors and it is predicted to encode a novel phosphonate natural product also encoded by the genome of the phylogenetically distant X. vasicola pv. vasculorum. Availability of this novel genome sequence may facilitate the study of interactions between xanthomonads and sugarcane, a host-pathogen system that appears to have evolved several times independently within the genus Xanthomonas and may also provide a source of target sequences for molecular detection and diagnostics. [ABSTRACT FROM AUTHOR]

This study was designed to characterize and describe host cell responses of stem tissue to mango wilt disease caused by the fungus Ceratocystis fimbriata in Brazil. Disease progress was followed, through time, in inoculated stems for two cultivars, 'Ubá' (field resistant) and 'Haden' (field susceptible). Stem sections from inoculated areas were examined using fluorescence light microscopy and transmission and scanning electron microscopy, coupled with energy-dispersive X-ray microanalysis. Tissues from Ubá colonized by C. fimbriata had stronger autofluorescence than those from Haden. The X-ray microanalysis revealed that the tissues of Ubá had higher levels of insoluble sulfur and calcium than those of Haden. Scanning electron microscopy revealed that fungal hyphae, chlamydospores (aleurioconidia), and perithecia-like structures of C. fimbriata were more abundant in Haden relative to Ubá. At the ultrastructural level, pathogen hyphae had grown into the degraded walls of parenchyma, fiber cells, and xylem vessels in the tissue of Haden. However, in Ubá, plant cell walls were rarely degraded and hyphae were often surrounded by dense, amorphous granular materials and hyphae appeared to have died. Taken together, the results of this study characterize the susceptible and resistant basal cell responses of mango stem tissue to infection by C. fimbriata. [ABSTRACT FROM AUTHOR]

Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense, is a disease that causes large reductions in banana yield worldwide. Considering the importance of silicon (Si) to potentiate the resistance of several plant species to pathogen infection, this study aimed to investigate, at the histochemical level, whether this element could enhance the production of phenolics on the roots of banana plants in response to F. oxysporum f. sp. cubense infection. Plants of cultivar Maçā, which is susceptible to F. oxysporum f. sp. cubense, were grown in plastic pots amended with 0 (-Si) or 0.39 g of Si (+Si) per kilogram of soil and inoculated with race 1 of F oxysporum f. sp. cubense. The root Si concentration was increased by 35.6% for +Si plants in comparison to the -Si plants, which contributed to a 27% reduction in the symptoms of Fusarium wilt on roots. There was an absence of fluorescence for the root sections of the -Si plants treated with the Neu and Wilson's reagents. By contrast, for the root sections obtained from the +Si plants treated with Neu's reagent, strong yellow-orange fluorescence was observed in the phloem, and lemon-yellow fluorescence was observed in the sclerenchyma and metaxylem vessels, indicating the presence of flavonoids. For the root sections of the +Si plants treated with Wilson's reagent, orange-yellowish autofluorescence was more pronounced around the phloem vessels, and yellow fluorescence was more pronounced around the metaxylem vessels, also indicating the presence of flavonoids. Lignin was more densely deposited in the cortex of the roots of the +Si plants than for the -Si plants. Dopamine was barely detected in the roots of the -Si plants after using the lactic and glyoxylic acid stain, but was strongly suspected to occur on the phloem and metaxylem vessels of the roots of the +Si plants as confirmed by the intense orange-yellow fluorescence. The present study provides new evidence of the pivotal role of the phenylpropanoid pathway in the resistance of banana plants to F. oxysporum f. sp. cubense infection when supplied with Si. [ABSTRACT FROM AUTHOR]

The predicted repertoire of type III secretion system effectors (T3SEs) in Erwinia tracheiphila , causal agent of cucurbit bacterial wilt, is much larger than in xylem pathogens in the closely related genera Erwinia and Pantoea . The genomes of strains BHKY and SCR3, which represent distinct E. tracheiphila clades, encode at least 6 clade-specific and 12 shared T3SEs. The strains expressed the majority of the T3SE genes examined in planta. Among the shared T3SE genes, eop1 was expressed most highly in both strains in squash ( Cucurbita pepo ) and muskmelon ( Cucumis melo ) but the clade-specific gene avrRpm2 was expressed 40- to 900-fold more than eop1 in BHKY. The T3SEs AvrRpm2, Eop1, SrfC, and DspE contributed to BHKY virulence on squash and muskmelon, as shown using combinatorial mutants involving six T3SEs, whereas OspG and AvrB4 contributed to BHKY virulence only on muskmelon, demonstrating host-specific virulence functions. Moreover, Eop1 was functionally redundant with AvrRpm2, SrfC, OspG, and AvrB4 in BHKY, and BHKY mutants lacking up to five effector genes showed similar virulence to mutants lacking only two genes. The T3SEs OspG, AvrB4, and DspE contributed additively to SCR3 virulence on muskmelon and were not functionally redundant with Eop1. Rather, loss of eop1 and avrB4 restored wild-type virulence to the avrB4 mutant, suggesting that Eop1 suppresses a functionally redundant effector in SCR3. These results highlight functional differences in effector inventories between two E. tracheiphila clades, provide the first evidence of OspG as a phytopathogen effector, and suggest that Eop1 may be a metaeffector influencing virulence. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Inactivation of Arabidopsis WAT1 (Walls Are Thin1), a gene required for secondary cell-wall deposition, conferred broad-spectrum resistance to vascular pathogens, including the bacteria Ralstonia solanacearum and Xanthomonas campestris pv. campestris, and the fungi Verticillium dahliae and Verticillium albo-atrum. Introduction of NahG, the bacterial salicylic acid (SA)-degrading salicylate hydroxylase gene, into the wat1 mutant restored full susceptibility to both R. solanacearum and X. campestris pv. campestris. Moreover, SA content was constitutively higher in wat1 roots, further supporting a role for SA in wat1-mediated resistance to vascular pathogens. By combining transcriptomic and metabolomic data, we demonstrated a general repression of indole metabolism in wat1-1 roots as shown by constitutive down-regulation of several genes encoding proteins of the indole glucosinolate biosynthetic pathway and reduced amounts of tryptophan (Trp), indole-3-acetic acid and neoglucobrassicin, the major form of indole glucosinolate in roots. Furthermore, the susceptibility of the wat1 mutant to R. solanacearum was partially restored when crossed with either the trp5 mutant, an over-accumulator of Trp, or Pro35S:AFB1-myc, in which indole-3-acetic acid signaling is constitutively activated. Our original hypothesis placed cell-wall modifications at the heart of the wat1 resistance phenotype. However, the results presented here suggest a mechanism involving rootlocalized metabolic channeling away from indole metabolites to SA as a central feature of wat1 resistance to R. solanacearum. [ABSTRACT FROM AUTHOR]

Mango (Mangifera indica) is an economically significant crop, and is affected by dieback in nearly all commercial production areas. Due to the wide range of organisms previously associated with these disease symptoms in Florida, isolations and pathogenicity tests were carried out to determine the causal organism. The pathogen was identified as Neofusicoccum batangarum based on genetic sequences from three loci (internal transcribed spacer of the rDNA (ITS), β-tubulin (BT), and translation elongation factor 1-α (EF)), recommended for members of the Botryosphaeriaceae family. Possible infection routes were determined by inoculating wounded and unwounded stems with N. batangarum. Trees wounded prior to pathogen inoculation developed larger lesions (5.85 cm ± 1.51) than unwounded trees (0.51 cm ± 0.48), p < 0.0003. In addition, lesions only developed at a small number of inoculation sites in the absence of wounds (14.3%), compared to 93% when stems were wounded. No necrosis was observed in the negative controls. This study provides molecular data on N. batangarum, and evidence of its role causing mango dieback in Florida. [ABSTRACT FROM AUTHOR]