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Apple scab, caused by the pathogenic fungus Venturia inaequalis, can result in significant economic losses. The frequent use of fungicidal products has led to the emergence of isolates resistant to commonly used active substances. Therefore, biological control offers a sustainable alternative for managing apple scab. In this study, eight Trichoderma isolates were evaluated against five different isolates of V. inaequalis isolated from the Fes-Meknes region. The biocontrol potential of these Trichoderma isolates had previously been demonstrated against other pathogens. The results indicated that the inhibition rate of mycelial growth of V. inaequalis obtained with Trichoderma spp. isolates ranged from 50% to 81%, with significant differences observed among the pathogenic isolates after 5 and 12 days of incubation. In addition, the in vitro tests with Trichoderma cell-free filtrates showed inhibition rates ranging from 2% to 79%, while inhibition rates ranged from 5% to 78% for volatile compound tests. Interestingly, the inhibition of spore germination and elongation was approximately 40–50%, suggesting the involvement of antifungal metabolites in their biocontrol activities. The in vivo bioassay on detached apple leaves confirmed the biocontrol potential of these Trichoderma isolates and demonstrated their ability to preventively control apple scab disease. However, their efficacies were still lower than those of the fungicidal product difenoconazole. These findings could contribute to the development of an effective biofungicide based on these Trichoderma isolates for reliable and efficient apple scab control. [ABSTRACT FROM AUTHOR]

Apple scab incited by the ascomycete Venturia inaequalis poses a significant threat to apple cultivation, necessitating a reassessment of existing disease management strategies. Attempts to manage apple scab include diverse approaches like developing disease forecasting models and the extensive application of synthetic chemical fungicides. However, the efficacy of these methods is compromised by inconsistencies, environmental concerns, and the pathogen's resistance, necessitating the exploration of alternative sustainable strategies. Addressing the challenges associated with apple scab management, this review strongly supports a shift towards the integration of biological control agents (BCAs). Emphasising the transformative synergy between BCAs and their bioactive secondary metabolites, we highlight their efficacy in advancing precision disease control through innovative and sustainable solutions. The review effectively presents a strong justification for the integration of BCAs and their by-products into apple scab management, offering insights into associated benefits, risks, and challenges while outlining promising prospects. Ultimately, it is expected to drive the adoption of environmentally conscious practices for effective apple scab management. [ABSTRACT FROM AUTHOR]

Adequate mineral fertilization helps to ensure optimal tree growth, fruit development, and predictable yield of apple trees. This 7-year study (2016–2022) aims to investigate the effect of nitrogen (N), phosphorus (P), potassium (K), and magnesium (Mg) fertilizer combinations (NP, NPK, NPKMg, and control) on eight parameters (trunk cross-sectional area—TCSA; fruit yield—FY; number of fruit per tree—FNT; crop load—CL; fruit diameter—FD; fruit weight—FW; fruit scab incidence—FSI; and powdery mildew incidence on shoot—PMIS) on the cultivars (cvs) 'Golden Reinders' (disease susceptible) and 'Pinova' (scab and mildew tolerant). In the 7-year period, TCSA values continuously increased for both cultivars over the years. Fertilizer treatments showed significant differences on TCSA but the effect varied greatly annually among fertilizer treatments. Fertilizer treatments had increasing effects on FY and FNT in 2018 and 2022, on CL in 2018, on FD in 2018 and 2019, and on FW in 2016 and 2018 in both cultivars compared to the control treatment. FSI values were the lowest in the NPKMg treatment for cv. 'Golden Reinders' in 2016, 2017, and 2022; for cv. 'Pinova' in 2016; PMIS values for cv. 'Golden Reinders' in 2017, 2018, 2021, and 2022; and for cv. 'Pinova' in 2018. Correlation and regression analyses revealed strong and significant (p = 0.05) relationships between FNT versus (vs.) TCSA, FNT vs. FY, FW vs. TCSA, CL vs. FY, FW vs. FD, and FSI vs. FW. In conclusion, our study showed that multiyear application of fertilizer combinations can successfully increase TCSA and yield parameters as well as reduce fungal disease incidences, especially on the disease-susceptible cultivar in sandy soil with moderate fertility, under Central-European continental climate conditions. [ABSTRACT FROM AUTHOR]

Succinate dehydrogenase inhibitors (SDHIs) are an important group of fungicides (FRAC Group 7) for the control of Venturia inaequalis. Mechanisms which lead to reduced SDHI sensitivity in fungi are based on mutations in the SDH-subunits B, C, or D of the target gene. Monitoring studies have shown that most of the European population is still sensitive to SDHIs. Single cases of SDHI resistance have been detected shortly after market introduction although mainly at trial sites. The underlying resistance mechanisms have been identified as target site mutations (B-T258I, C-N85S and C-H151R). However, these were single findings which did not increase in frequency or even disappeared when the same field was analyzed one or more seasons later. This is in contrast to findings on other resistance cases which are stable over several seasons even without selection pressure, such as QoI and benzimidazole resistance. High frequencies of target site mutations for QoIs and benzimidazoles (G143A and E198A, respectively) have been detected in a 2022 survey, whereas no SDHI target site mutation exceeded the detection limit of our molecular detection assay in any sample. [ABSTRACT FROM AUTHOR]

Apple scab incited by the ascomycete Venturia inaequalis poses a significant threat to apple cultivation, necessitating a reassessment of existing disease management strategies. Attempts to manage apple scab include diverse approaches like developing disease forecasting models and the extensive application of synthetic chemical fungicides. However, the efficacy of these methods is compromised by inconsistencies, environmental concerns, and the pathogen’s resistance, necessitating the exploration of alternative sustainable strategies. Addressing the challenges associated with apple scab management, this review strongly supports a shift towards the integration of biological control agents (BCAs). Emphasising the transformative synergy between BCAs and their bioactive secondary metabolites, we highlight their efficacy in advancing precision disease control through innovative and sustainable solutions. The review effectively presents a strong justification for the integration of BCAs and their by-products into apple scab management, offering insights into associated benefits, risks, and challenges while outlining promising prospects. Ultimately, it is expected to drive the adoption of environmentally conscious practices for effective apple scab management.

Societal Impact statement Reduced fungicide use lowers environmental pollution and enables safer food production. The usage of fungicides in apple orchards can be reduced through the application of sanitation practices which decrease the inoculum sources of apple scab disease on fallen leaves. This study found two non‐chemical sanitation practices, namely the collection of fallen leaves (CFL) and CFL combined with straw mulch in tree rows, were beneficial. These two practices are not only biologically and environmentally valuable, as they reduce disease levels and can replace chemical fungicides, but they are also economically efficient options for integrated and organic orchards compared to non‐sanitized ones. Summary Severe fungicide use can be reduced by applications of sanitation practices in order to reduce scab incidence, yield and fruit quality losses in apple orchards. In a 5‐year study, we aimed to investigate the effect of sanitation practices on biological and cost–benefit parameters in two sustainable apple management systems, and to find significant correlations among the parameters. We investigated the effect of five sanitation treatments (lime sulphur, leaf collection, mulching, lime sulphur + leaf collection, leaf collection + mulching) on four biological (scab incidence, fruit parameters: total yield, yield class I and II) and seven cost–benefit (three cost types, three annual revenue types, income surplus/deficit) parameters in integrated and organic apple orchards. Correlation, linear regression and principal component analyses (PCA) were performed to find correlations among biological and cost–benefit parameters. Results showed that fruit scab incidence was 3.4–8.1 times higher, while total yield was 1.4–1.8 times lower in the organic management system than in the integrated one. The treatment of leaf collection and/or leaf collection + mulching showed higher total cost (180.3 and 675.2 EUR ha−1) but lower scab incidence (5.3 and 27.3%; 4.8 and 26.7%, integrated and organic, respectively) and higher yield with greater total revenues (10,235 and 10,329 EUR ha−1; 8,136 and 8,230 EUR ha−1, integrated and organic, respectively) and income surpluses (851 and 451 EUR ha−1; 897 and 496 EUR ha−1, integrated and organic, respectively) compared to non‐sanitized control treatments in most cases. Other sanitation treatments provided fewer biological and/or no financial benefits. Results from correlation and linear regression analyses indicated strong relationships among the factors of total yield vs surplus, class I vs surplus, and fruit scab vs class II) in both management systems. Further relationships were detected among almost all parameters in the PCA. Overall, our study demonstrated that two non‐chemical sanitation treatments could not only reduce scab incidence and increase fruit yield, but could show positive cost–benefit outcomes in both management systems.

Societal Impact statement: Reduced fungicide use lowers environmental pollution and enables safer food production. The usage of fungicides in apple orchards can be reduced through the application of sanitation practices which decrease the inoculum sources of apple scab disease on fallen leaves. This study found two non‐chemical sanitation practices, namely the collection of fallen leaves (CFL) and CFL combined with straw mulch in tree rows, were beneficial. These two practices are not only biologically and environmentally valuable, as they reduce disease levels and can replace chemical fungicides, but they are also economically efficient options for integrated and organic orchards compared to non‐sanitized ones. Summary: Severe fungicide use can be reduced by applications of sanitation practices in order to reduce scab incidence, yield and fruit quality losses in apple orchards. In a 5‐year study, we aimed to investigate the effect of sanitation practices on biological and cost–benefit parameters in two sustainable apple management systems, and to find significant correlations among the parameters.We investigated the effect of five sanitation treatments (lime sulphur, leaf collection, mulching, lime sulphur + leaf collection, leaf collection + mulching) on four biological (scab incidence, fruit parameters: total yield, yield class I and II) and seven cost–benefit (three cost types, three annual revenue types, income surplus/deficit) parameters in integrated and organic apple orchards. Correlation, linear regression and principal component analyses (PCA) were performed to find correlations among biological and cost–benefit parameters.Results showed that fruit scab incidence was 3.4–8.1 times higher, while total yield was 1.4–1.8 times lower in the organic management system than in the integrated one. The treatment of leaf collection and/or leaf collection + mulching showed higher total cost (180.3 and 675.2 EUR ha−1) but lower scab incidence (5.3 and 27.3%; 4.8 and 26.7%, integrated and organic, respectively) and higher yield with greater total revenues (10,235 and 10,329 EUR ha−1; 8,136 and 8,230 EUR ha−1, integrated and organic, respectively) and income surpluses (851 and 451 EUR ha−1; 897 and 496 EUR ha−1, integrated and organic, respectively) compared to non‐sanitized control treatments in most cases. Other sanitation treatments provided fewer biological and/or no financial benefits. Results from correlation and linear regression analyses indicated strong relationships among the factors of total yield vs surplus, class I vs surplus, and fruit scab vs class II) in both management systems. Further relationships were detected among almost all parameters in the PCA.Overall, our study demonstrated that two non‐chemical sanitation treatments could not only reduce scab incidence and increase fruit yield, but could show positive cost–benefit outcomes in both management systems. [ABSTRACT FROM AUTHOR]

Scab, caused by Venturia inaequalis, is the most destructive fungal disease of apple worldwide. Apple scab incidence was studied in apple orchards in the south and southeast of Kazakhstan, including the Almaty, Zhambyl, and Turkestan regions, during 2022 and 2023. Disease incidence was higher in the Zhambyl region than in the Turkestan and Almaty regions in both years. The field evaluation suggested that 19 genotypes showed resistance to apple scab. Molecular screening was carried out using eight gene-specific molecular markers (AM19, CH05e03, OPL19, Hi07f02, AL07, K08, HB09, and CH02f06). The results of the molecular screening revealed that in 38 of the 45 studied cultivars, which included 11 Kazakh cultivars and 34 foreign cultivars, the Rvi (Rvi2, Rvi4, Rvi5, Rvi6, Rvi8, Rvi9, Rvi11, Rvi14, and Rvi15) resistance genes were amplified. Resistance genes such as Rvi2, Rvi4, Rvi6, and Rvi9 are still useful for breeding, but their use is recommended only in extended pyramids of multiple resistance genes. Several cultivars will be strong candidates for further breeding programs against apple scab and for the pyramiding of scab resistance genes in new cultivars. [ABSTRACT FROM AUTHOR]

The outermost hydrophobic layer of plants, i.e. the cuticle, is mainly composed of cutin, a polyester of hydroxy fatty acids with reported eliciting and/or antimicrobial activities for some of them. By-products of the fruit processing industry (fruit pomaces), often strongly enriched in cuticular material, are therefore a potential source of bioactive compounds for crop protection against pathogen attack. We investigated the utilization of tomato and apple pomaces in the development of a cutin-based biocontrol solution against apple scab, a major apple disease caused by Venturia inaequalis. Several cutin monomer extracts obtained through different strategies of depolymerization and purification were first compared for their ability to induce a targeted set of defense genes in apple seedlings after foliar application. After a step of formulation, some extracts were chosen for further investigation in planta and in vitro. Our results show that formulated cutin monomers could trigger a significant transcriptome reprogramming in apple plants and exhibit an antifungal effect on V. inaequalis. Cutin monomers-treated apple seedlings were significantly protected against infection by the apple scab agent. Altogether, our findings suggest that water-dispersed cutin monomers extracted from pomaces are potential new bio-based solutions for the control of apple scab. [ABSTRACT FROM AUTHOR]

Apple scab, caused by the pathogenic fungus Venturia inaequalis, can result in significant economic losses. The frequent use of fungicidal products has led to the emergence of isolates resistant to commonly used active substances. Therefore, biological control offers a sustainable alternative for managing apple scab. In this study, eight Trichoderma isolates were evaluated against five different isolates of V. inaequalis isolated from the Fes-Meknes region. The biocontrol potential of these Trichoderma isolates had previously been demonstrated against other pathogens. The results indicated that the inhibition rate of mycelial growth of V. inaequalis obtained with Trichoderma spp. isolates ranged from 50% to 81%, with significant differences observed among the pathogenic isolates after 5 and 12 days of incubation. In addition, the in vitro tests with Trichoderma cell-free filtrates showed inhibition rates ranging from 2% to 79%, while inhibition rates ranged from 5% to 78% for volatile compound tests. Interestingly, the inhibition of spore germination and elongation was approximately 40–50%, suggesting the involvement of antifungal metabolites in their biocontrol activities. The in vivo bioassay on detached apple leaves confirmed the biocontrol potential of these Trichoderma isolates and demonstrated their ability to preventively control apple scab disease. However, their efficacies were still lower than those of the fungicidal product difenoconazole. These findings could contribute to the development of an effective biofungicide based on these Trichoderma isolates for reliable and efficient apple scab control.

Venturia inaequalis, the causal fungal pathogen of apple scab, has evolved with its Malus hosts during apple domestication. This co‐evolution has resulted in a genetic structure in V. inaequalis populations from Europe and central Asia based on host species and geographic isolation. However, it is not yet clear if geographic isolation has led to population differentiation in North American isolates. We resequenced the genomes of 54 V. inaequalis isolates from North America and analysed them with publicly available genome sequences of 90 European and central Asian isolates for variant discovery and population structure. A total of 204,566 high‐quality single‐nucleotide polymorphisms (SNPs) were identified and used to assess population genetic structure and genomic diversity across 123 Venturia spp. isolates from around the world. Population genetic analysis identified four clusters based on Malus and non‐Malus hosts and Venturia species; this differentiation was supported by genetic diversity parameters including FST, Nei's π and Tajima's D. Genetic structure analysis did not reveal a distinct subpopulation of North American V. inaequalis isolates within the global isolates. Additionally, structure was not observed in V. inaequalis isolates collected on M. × domestica between North America and Europe. These observations indicate that geographical isolation has not contributed to the population differentiation between North America and Europe/central Asia. These results will enhance our understanding of the evolution of V. inaequalis, the emergence of virulent isolates and their implications for managing apple scab effectively. [ABSTRACT FROM AUTHOR]

Background: Venturia inaequalis is an apple scab causing fungal pathogen. It is a highly contagious and destructive pathogen which rapidly spreads infection in the surrounding orchards if not managed. The management and control of disease require multiple fungicides to be sprayed at different development stages of the apple. Persistent applications of fungicides also raises environmental concerns. Here, we demonstrate the potential of using spray induced gene silencing (SIGS) by developing target specific gene constructs for the synthesis of corresponding double-stranded RNA (dsRNA). Methods and Results: The exogenous application of dsRNAs was found to reduce mycelial growth and spore formation of V. inaequalis on culture plates. Four genes of V. inaequalis viz. CIN1, CE5, VICE12 and VICE16 which get upregulated during infection, were selected as targets for the development of gene construct expressing the corresponding dsRNA. The effect of exogenously supplied in vitro synthesized dsRNA on V. inaequalis was assessed in culture bioassay experiments with respect to growth, and spore formation. The expression level of the target genes in treated and control fungus was evaluated using quantitative PCR. Fungus treated with VICE12 targeted dsRNA showed maximum reduction in colony size (~ 55%), conidia formation (~ 93%) and expression level of the corresponding gene (2.2 fold), which was followed by CIN1-dsRNA. VICE16-dsRNA treatment was least effective with 32% reduction in growth, the non-significant effect of conidial spore formation and 1.13 fold down regulation of corresponding target gene expression level. Conclusion: The result of this investigation validates the hypothesis that RNAi is evoked in V. inaequalis by exogenously supplied dsRNA and spray induced gene silencing (SIGS) based solutions may reduce burden of fungicide usage on apple crop against apple scab disease in future. [ABSTRACT FROM AUTHOR]

Adequate mineral fertilization helps to ensure optimal tree growth, fruit development, and predictable yield of apple trees. This 7-year study (2016–2022) aims to investigate the effect of nitrogen (N), phosphorus (P), potassium (K), and magnesium (Mg) fertilizer combinations (NP, NPK, NPKMg, and control) on eight parameters (trunk cross-sectional area—TCSA; fruit yield—FY; number of fruit per tree—FNT; crop load—CL; fruit diameter—FD; fruit weight—FW; fruit scab incidence—FSI; and powdery mildew incidence on shoot—PMIS) on the cultivars (cvs) ‘Golden Reinders’ (disease susceptible) and ‘Pinova’ (scab and mildew tolerant). In the 7-year period, TCSA values continuously increased for both cultivars over the years. Fertilizer treatments showed significant differences on TCSA but the effect varied greatly annually among fertilizer treatments. Fertilizer treatments had increasing effects on FY and FNT in 2018 and 2022, on CL in 2018, on FD in 2018 and 2019, and on FW in 2016 and 2018 in both cultivars compared to the control treatment. FSI values were the lowest in the NPKMg treatment for cv. ‘Golden Reinders’ in 2016, 2017, and 2022; for cv. ‘Pinova’ in 2016; PMIS values for cv. ‘Golden Reinders’ in 2017, 2018, 2021, and 2022; and for cv. ‘Pinova’ in 2018. Correlation and regression analyses revealed strong and significant (p = 0.05) relationships between FNT versus (vs.) TCSA, FNT vs. FY, FW vs. TCSA, CL vs. FY, FW vs. FD, and FSI vs. FW. In conclusion, our study showed that multiyear application of fertilizer combinations can successfully increase TCSA and yield parameters as well as reduce fungal disease incidences, especially on the disease-susceptible cultivar in sandy soil with moderate fertility, under Central-European continental climate conditions.

Apple scab, caused by the fungus Venturia inaequalis, is one of the most damaging diseases of cultivated apples (Malus x domestica) worldwide. It results in huge losses as it diminishes fruit quality and impacts tree growth. Current management revolves around the application of fungicides, however the number of sprays required per season is unsustainable and expensive. Further to this, populations of V. inaequalis have developed fungicide resistance. Breeding new cultivars with higher levels of resistance to scab is a priority, however, this process is long, so introducing mixed cultivar orchards may be a faster solution. We reviewed the general principles of using mixtures to manage plant diseases, and then considered specifically using mixed cultivars to manage apple scab in commercial production. Limited field studies have demonstrated the potential of using mixture to suppress apple scab development; but scab super-races that could emerge from mixture can pose a significant risk in commercial production. However, recent research on population genetics of apple scab in monoculture and mixed orchards suggests that the risk of super-race emergence is probably over-stated, because assortative mating among lesions on the same leaves is likely to occur, rather than commonly assumed random mating. Thus, we conclude that cultivar mixtures can contribute towards sustainable scab management, particularly in commercial cider apple production.

Apple scab, compared to other fungal diseases, consistently causes the most significant economic losses for growers. Venturia inaequalis infection not only reduce actual crop quality and quantity. Impaired tree vitality due to defoliation and a reduced number of buds in the following season are also the consequences of uncontrolled attack of apple trees by this fungal pathogen. Therefore the control of apple scab using fungicides is an essential part of the orchard life cycle. Knowing the sensitivity of local populations of V. inaequalis to active substances used in fungicidal sprays is a key factor for the sustainable protection of apple plantations against scab. In the years 2020–2022, V. inaequalis populations isolated from experimental and production orchards in the Czech Republic were tested for sensitivity to the active substances cyprodinil and difenoconazole. The assessment was carried out using in vivo tests on seedlings of the cultivar ‘Golden Delicious’. The monitored orchards showed the sensitivity to cyprodinil, where only two populations had reduced sensitivity. The majority of tested populations were also sensitive to difenoconazole. Only one population was found to be resistant to difenoconazole, and two populations were found to have reduced sensitivity to this agent. The results are consistent with similar foreign studies.

The natural product chitosan has been shown to reduce plant disease severity and enhance the efficacy of microbial biocontrol agents in several crops. However, little is known about the potential synergisms between chitosan and biopesticides and best use practices in apple production. The objectives of this study were to evaluate the effect of pre-harvest applications of chitosan alone and in combination with a commercial biopesticide to suppress fungal diseases of apple and to investigate the potential for chitosan to reduce the quantity of overwintering Venturia inaequalis spores in orchard leaf litter. Chitosan products, Tidal Grow and ARMOUR-Zen 15, and a commercial biopesticide, Serenade ASO, were tested in a research orchard in Pennsylvania and commercial orchards in New Hampshire. Chitosan applications reduced apple scab incidence and severity by up to 55% on fruit compared to the water control. Chitosan also reduced sooty blotch, flyspeck, and rust incidence on fruit. Furthermore, a chitosan + biopesticide treatment overlayed onto a grower standard spray program reduced diseases more effectively than the grower standard alone. However, this efficacy was dependent on the cultivar and pathogen. Chitosan did not reduce overwintering V. inaequalis ascospores. This research provides evidence that pre-harvest chitosan applications have the potential for disease management in apple production. [ABSTRACT FROM AUTHOR]

The wetting behaviour of the spray and the biological efficacy of Cu2+ active ingredients in agrochemical formulations may be enhanced by tank‐mix additives. We investigated how three BREAK‐THRU® additives (BT301: biodegradable, BT133 and BT420: bio‐based and biodegradable) tank‐mixed with commercial copper preparations influenced the spray distribution, leaf uptake and biological efficacy of copper additive mixtures against apple scab and apple powdery mildew under controlled conditions. We quantified the synergetic effects of these additives in foliar applications. In addition, we determined the phytotoxic potential and evaluated their impacts on photosynthetic activity, non‐photochemical quenching and ROS activity. The additives BT301 and BT420 strongly reduced surface tension and contact angle of copper treatments. The fluorescence observations revealed that BT301 achieved better spreading of copper formulation with more complete coverage of the leaf surface than BT420 and BT133, whereas 'coffee‐ring' spreading was observed with BT133. The additive BT301 showed an increase in relative fluorescence area, indicating higher ROS production as a signal of intra‐cellular tissue activity. The photochemical efficiency of photosystem II (Fv/Fm) was not negatively influenced by copper or additive treatment. Thus, we observed no phytotoxic effects of copper‐additive mixtures on apple leaves at treatment doses of 4 g Cu2+ L−1. All copper treatments reduced apple scab infections significantly, by 53%–76%. Interestingly, addition of BT301 to copper preparations showed the strongest biological efficacy (83% reduction) against Venturia inaequalis, whereas addition of BT420 showed the strongest effect against Podosphaera leucotricha (89% infection reduction). The synergetic effects of additives on the biological efficacy without phytotoxic effects on plants may have potential for reducing copper loads in horticultural production systems. [ABSTRACT FROM AUTHOR]

This review gives a perspective of selected advances made since the middle of the 20th century in plant disease modeling, and the associated increase in the number of models published during that time frame. This progress can be mainly atti-ibuted to advances in (i) sensors and automatic environmental data collection technology, (ii) instrumentation and methods for studying botanical epidemiology, and (iii) data analytics and computer science. We review the evolution of techniques for developing data-based (empirical) models and process-based (mechanistic) models using the wheat rusts as a case study. We also describe the increased importance of knowledge about biological processes for plant disease modeling by using apple scab as a second case study. For both wheat rusts and apple scab. we describe how the models have evolved over the last 50 years by considering certain milestones that have been achieved in disease modeling. Finally. we describe how plant disease models are used as part of a multi-modeling approach to develop decision-making tools in the application of integrated pest management. [ABSTRACT FROM AUTHOR]

Scab, caused by Venturia inaequalis, is the most destructive fungal disease of apple worldwide. Apple scab incidence was studied in apple orchards in the south and southeast of Kazakhstan, including the Almaty, Zhambyl, and Turkestan regions, during 2022 and 2023. Disease incidence was higher in the Zhambyl region than in the Turkestan and Almaty regions in both years. The field evaluation suggested that 19 genotypes showed resistance to apple scab. Molecular screening was carried out using eight gene-specific molecular markers (AM19, CH05e03, OPL19, Hi07f02, AL07, K08, HB09, and CH02f06). The results of the molecular screening revealed that in 38 of the 45 studied cultivars, which included 11 Kazakh cultivars and 34 foreign cultivars, the Rvi (Rvi2, Rvi4, Rvi5, Rvi6, Rvi8, Rvi9, Rvi11, Rvi14, and Rvi15) resistance genes were amplified. Resistance genes such as Rvi2, Rvi4, Rvi6, and Rvi9 are still useful for breeding, but their use is recommended only in extended pyramids of multiple resistance genes. Several cultivars will be strong candidates for further breeding programs against apple scab and for the pyramiding of scab resistance genes in new cultivars.

ABSTRACT Scab, caused by the biotrophic fungal pathogen Venturia inaequalis, is the most economically important disease of apples. During infection, V. inaequalis colonizes the subcuticular host environment, where it develops specialized infection structures called runner hyphae and stromata. These structures are thought to be involved in nutrient acquisition and effector (virulence factor) delivery, but also give rise to conidia that further the infection cycle. Despite their importance, very little is known about how these structures are differentiated. Likewise, nothing is known about how these structures are protected from host defenses or recognition by the host immune system. To better understand these processes, we first performed a glycosidic linkage analysis of sporulating tubular hyphae from V. inaequalis developed in culture. This analysis revealed that the V. inaequalis cell wall is mostly composed of glucans (44%) and mannans (37%), whereas chitin represents a much smaller proportion (4%). Next, we used transcriptomics and confocal laser scanning microscopy to provide insights into the cell wall carbohydrate composition of runner hyphae and stromata. These analyses revealed that, during subcuticular host colonization, genes of V. inaequalis putatively associated with the biosynthesis of immunogenic carbohydrates, such as chitin and β-1,6-glucan, are downregulated relative to growth in culture, while on the surface of runner hyphae and stromata, chitin is deacetylated to the less-immunogenic carbohydrate chitosan. These changes are anticipated to enable the subcuticular differentiation of runner hyphae and stromata by V. inaequalis, as well as to protect these structures from host defenses and recognition by the host immune system. IMPORTANCE Plant-pathogenic fungi are a major threat to food security. Among these are subcuticular pathogens, which often cause latent asymptomatic infections, making them difficult to control. A key feature of these pathogens is their ability to differentiate specialized subcuticular infection structures that, to date, remain largely understudied. This is typified by Venturia inaequalis, which causes scab, the most economically important disease of apples. In this study, we show that, during subcuticular host colonization, V. inaequalis downregulates genes associated with the biosynthesis of two immunogenic cell wall carbohydrates, chitin and β-1,6-glucan, and coats its subcuticular infection structures with a less-immunogenic carbohydrate, chitosan. These changes are anticipated to enable host colonization by V. inaequalis and provide a foundation for understanding subcuticular host colonization by other plant-pathogenic fungi. Such an understanding is important, as it may inform the development of novel control strategies against subcuticular plant-pathogenic fungi.

Apple breeding is a laborious and long-lasting process that requires qualified resources, land, time, and funds. In this study, more than 5000 F1 apple hybrids from direct and testcrosses were analyzed. The results revealed how the phenotypic expression of the main quantitative traits of interest assessed in five half-sib families was controlled by the additive genetic effects and by non-additive effects of dominance and epistasis. The statistical number of hybrids required to ensure efficient selection increased exponentially with the number of desirable traits. The minimum number of progenies required to obtain a hybrid with associated quantitative traits of agronomic interest was highly variable. For two independent traits essential in selection (fruit size and quality), but incorporated together in the same hybrid, the statistical number was between about 30 and 300. If three more cumulative traits were added (a large number of fruits per tree, resistance/tolerance to apple scab, and powdery mildew attack), the limits increased to between 1500 and 18,000. The study highlighted the need for new apple varieties due to the narrowing of the genetic diversity of the cultivated species and how the choice of parents used in hybridizations (as well as the objectives pursued in the selection) can increase the efficiency of apple breeding. [ABSTRACT FROM AUTHOR]

Demethylation inhibitor fungicides (DMIs) are widely used against Venturia inaequalis in north western Himalayas. The sensitivity of pathogen populations to DMIs viz., myclobutanil, difenconazole and flusilazole was evaluated. Thirty baseline isolates of V. inaequalis were collected from an orchard never exposed to fungicides and 200 other isolates from fungicide-exposed orchards. To establish the baseline sensitivity, the thirty baseline isolates were tested at seven concentrations viz., 0.001, 0.01, 0.03, 0.1, 0.3, 1.0, and 10.0 µg ml−1 of each fungicide using mycelial growth assays. The mean EC50 values of myclobutanil, difenoconazole and flusilazole for baseline population were 0.17, 0.02 and 0.022 µg ml−1, respectively, and accordingly the respective discriminatory doses of 0.20, 0.03 and 0.03 µg ml−1 were selected. To monitor the shift in fungicide-exposed populations, all the two hundred isolates were tested for their sensitivity at the respective discriminatory doses of different fungicides and the frequency distributions of their relative growth (RG) values were compared to that of the baseline isolates. A major shift towards reduced sensitivity was observed in myclobutanil, flusilazole and difenoconazole. The highest proportion of isolates (35.5%) were 'resistant' to myclobutanil followed by 19.5 per cent isolates resistant to flusilazole while only 7.5 per cent of isolates were resistant to difenoconazole. [ABSTRACT FROM AUTHOR]

Apple orchards are perennially planted where pesticides are applied to control numerous pests and diseases. The extensive long-term use of fungicides can lead to overall environmental load and resistance risk. This study aims to assess which fungicide-active substances have been used more intensively in the last decade in Latvia, evaluating the overall environmental load using the Pesticide Load Indicator (PLI). It was essential to see whether the amount of active substance usage rises, how it correlates with the total changes of the PLI and which substances are with the highest scores. The other issue was to test the sensitivity of Venturia inaequalis populations to systemic fungicides. Six full-bearing apple orchards that reflected local plant protection practices were selected from the different growing regions of Latvia to analyze fungicide use from 2012 to 2021 and test V. inaequalis populations' sensitivity to systemic substances difenoconazole and cyprodinil. The PLI demonstrated that the protective fungicides were the most crucial group overall, with the highest potential impact on the environment and human health. Systemic fungicides had a relatively lower environmental impact, but after long-term use, the pathogen population's sensitivity to difenoconazole and cyprodinil was reduced. Introducing new fungicide classes and biological control agents could help growers improve plant protection strategies against V. inaequalis, reducing the risk of resistance and environmental load. [ABSTRACT FROM AUTHOR]

The ascomycete fungus Venturia inaequalis causes scab disease, a major problem in apple production. The utilization of resistant cultivars is hindered by emerging new pathogen races, which erode their resistance. Increasing our knowledge on the population genetic processes of the fungus can contribute to the development of resistance gene deployment strategies and more durable resistance. We investigated the effect of four scab race indicator cultivars, 'Gala' (no R-gene), 'Golden Delicious' (Rvi1), 'Geneva' (Rvi3, complex), and OR45t132 (Rvi5), on the V. inaequalis population genetic structure and diversity. Sixty-six monosporic fungal isolates from the four cultivars were genotyped with seven simple sequence repeat primers. Furthermore, the partial resistance of the indicators and the pathogenicity profile of the conidia from each host were assessed. The genetic diversity and structure of the investigated V. inaequalis subpopulations correspond to the partial resistance of the original hosts as well as the subpopulations' pathogenicity profiles. Indicators carrying Rvi3 and Rvi5 had strongly diverged and specialized V. inaequalis populations on them and fewer symptoms on the field. In line with the complete breakdown of the Rvi1 gene, the population from 'Golden Delicious' did not segregate from the susceptible 'Gala', and virulence towards Rvi1 was present in all subpopulations. [ABSTRACT FROM AUTHOR]

Apple scab is caused by an ascomycete fungus, Venturia inaequalis (Cke.) Wint., which is one of the most severe disease of apple (Malus × Domestica Borkh.) worldwide. The disease results in 30–40% fruit loss annually and even complete loss in some places. Owing to the evolving susceptibility of resistant apple genotypes harboring R-genes to new variants of V. inaequalis, a comparative transcriptome analysis using Illumina (HiSeq) platform of three scab-resistant (Florina, Prima, and White Dotted Red) and three susceptible (Ambri, Vista Bella, and Red Delicious) apple genotypes was carried out to mine new scab resistance genes. The study led to the identification of 822 differentially expressed genes in the tested scab-resistant and scab-susceptible apple genotypes. The most upregulated genes uniformly expressed in resistant varieties compared to susceptible ones were those coding for 17.3 kDa class II heat shock protein-like, chaperone protein ClpB1, glutathione S-transferase L3-like protein, B3 domain-containing protein At3g18960-like, transcription factor bHLH7, zinc finger MYM-type protein 1-like, and nine uncharacterized proteins, besides three lncRNAs. The genes that were downregulated in susceptible and upregulated in resistant cultivars were those coding for non-specific lipid transfer protein GPI-anchored 1, rust resistance kinase Lr10-like, disease resistance protein RPS6-like, and many uncharacterized proteins. DESeq2 analysis too revealed 20 DEGs that were upregulated in scab-resistant cultivars. Furthermore, a total of 361 genes were significantly upregulated in scab-susceptible variety, while 461 were found downregulated (P value < 0.05 and Log2 (FC) > 1). The differentially expressed genes (DEGs) were related to various pathways, i.e., metabolic, protein processing, biosynthesis of secondary metabolites, plant hormone signal transduction, autophagy, ubiquitin-mediated proteolysis, plant-pathogen interaction, lipid metabolism, and protein modification pathways. Real-time expression of a set of selected twelve DEGs further validated the results obtained from RNA-seq. Overall, these findings lay the foundation for investigating the genetic basis of apple scab resistance and defense pathways that might have a plausible role in governing scab resistance in apple against V. inaequalis. [ABSTRACT FROM AUTHOR]

The apple scab induced by Venturia inaequalis is an economically significant disease of apples worldwide and is predominantly controlled by multiple fungicide applications. Therefore, resistant apple cultivation is important for long-standing disease control. The knowledge about cultivar resistance is mainly founded on their evaluation in orchard conditions and testing in a greenhouse that is laborious and requires a large space. This study evaluated apple cultivar resistance and the virulence of V. inaequalis strains by inoculating detached leaves and immature fruits. Nine V. inaequalis strains originating from different apple-growing regions and host genotypes were tested on eight apple genotypes. Microscopic and macroscopic symptom development and host tissue reactions were monitored during the experiments. The tested V. inaequalis strains and cultivars showed different levels of virulence and resistance, respectively. Cultivar ‘Lobo’ was scored as partially susceptible, with almost all strains tested. The incompatible interaction with or without host tissue reactions was observed only on the cultivar ‘Priscilla’ with all the strains and field populations tested. The results of this study using detached leaves and immature fruits were concordant with the data obtained in the cultivar evaluation trials in the orchards. [ABSTRACT FROM AUTHOR]

Difenoconazole sensitivity was assessed in three populations of the apple scab agent Venturia inaequalis of the Krasnodar region. One of the populations was fungicide naïve population; its sample was collected in natural habitats of the eastern crabapple Malus orientalis in foothills of the Severskii raion. The two other populations were from commercial orchards of the apple variety Reinette Simirenko (the Krasnoarmeiskii and Dinskoi raions), where fungicide treatments with various agents, including those with difenoconazole as an active ingredient, were performed on a regular basis. Single-spore V. inaequalis isolates were isolated from fresh leaves with signs of the disease or fallen leaves with fungal fruiting bodies. The median effective concentration (EC50) was defined as the concentration that halves the colony growth and was estimated in a series of in vitro experiments with 120 isolates. Difenoconazole (Score EC, 250 mg/L, Syngenta) was used at six concentrations: 0.005, 0.01, 0.025, 0.05, 0.5, and 1 mg a.i./L. Mean EC50 values were 0.0078 mg a.i./L in the natural population and 0.12 and 0.25 mg a.i./L in the orchard populations. Fungicide sensitivity was lower in both of the orchard populations; their resistance factors were estimated at 16 and 32. The proportion of sensitive and low-sensitive isolates differed between the two orchard populations. A discriminatory dose, or single-assessment concentration (SAC), of 0.01 mg a.i./L was proposed to simplify the laboratory monitoring of the difenoconazole sensitivity in V. inaequalis by using a test for relative growth (RG) of the mycelium. Comparable results were obtained with EC50 and RG at the discriminatory dose. The portion of isolates with RGs exceeding the threshold (RG > 70%) was 97% in one of the orchard populations. The results indicate that difenoconazole resistance develops in V. inaequalis populations from commercial orchards of the Krasnodar region. [ABSTRACT FROM AUTHOR]

Increasing the reliability of fruit crops in unstable weather conditions of a temperate continental climate has become particularly relevant. This research was carried out based on a bioresource collection from the Russian Research Institute of Fruit Crop Breeding (VNIISPK BRC). Apple cultivars (diploids and triploids) of different maturation periods were studied. Triploid cultivars (3×) of summer maturation were developed using diploid gametes of the 'Papirovka tetraploid' cultivar as a donor (2-4-4-4×), triploids of winter maturation were developed using 13-6-106 (Suvorovetz: open pollination), and 'Wealthy tetraploid' (2-4-4-4×) or 'Giant Spy' (2-4-4-4×) were used as donors of diploid gametes. 'Antonovka' and 'Melba' were taken as control cultivars. The aim of this work was to evaluate the apple breeding cultivars according to the main economic and biological indicators and to determine the threshold of resistance to unfavorable conditions of the winter period for possible cultivation in specific conditions of a temperate continental climate. As a result of using the method of modeling damaging factors, apple cultivars that withstood not only the critical temperatures of January, but also especially dangerous frosts after thaws at the end of winter were identified, with the stability of vital tissues at the level of 'Antonovka' (a control cultivar), scab immune cultivars (RVi6) 'Ivanovskoye', 'Zdorovie' and 'Pamyati Hitrovo', and immune triploids (3×) 'Vavilovskoye', 'Aleksandr Boyko', Rozhdestvenskoye' and 'Academic Saveliev'. During the whole winter, the winter resistance of summer scab immune triploids, 'Zhilinskoye', 'Maslovskoye', 'Yablochny Spas' and 'Spasskoye', was at the level of 'Antonovka', but was higher than 'Melba'. The high precocity of scab-immune cultivars, 'Afrodita', 'Ivanovskoye', 'Veniaminovskoye', and 'Yubiley Moskvy', and triploids 'Patriot' and 'Rozhdestvenskoye', have been identified. Triploid cultivars are superior to diploid cultivars and control 'Antonovka' in fruit size. According to the complex of characteristics, promising apple cultivars were identified for planting in intensive orchards of the temperate continental climate zones, namely 'Avgusta' and 'Solnyshko' (summer cultivars), and 'Aleksandr Boyko', 'Vavilovskoye', 'Venyaminovskoye', 'Ivanovskoye' and 'Rozhdestvenskoye' (winter cultivars). [ABSTRACT FROM AUTHOR]

Management of scab diseases of apple, pecan and almond is mostly based on fungicide applications according to host phenological stage and wetness events. Resistance to widely used locally systemic fungicides including the methyl benzimidazoles, demethylation inhibitors (DMIs) and quinone outside inhibitors (QoIs) has been reported. We evaluated the activity of a new premixed hybrid fungicide containing the DMI difenoconazole and essential tea tree oil (TTO) against mycelial growth of Venturia inaequalis in vitro and against scab of apple, pecan and almond in field trials. Mycelial growth of 24 isolates of V. inaequalis collected from commercial apple orchards varied in their sensitivity to difenoconazole (EC50 values of 0.0002–1.29 mg/L). Difenoconazole–TTO was significantly more inhibitory to some of the isolates than difenoconazole alone, primarily to those that were less sensitive to difenoconazole. Protective foliar applications of difenoconazole–TTO in field trials were highly effective in controlling scab of apple and generally provided significantly higher disease control than difenoconazole alone. On apple and almond, difenoconazole–TTO treatments were similarly or more effective than applications with other synthetic fungicides belonging to the DMI, QoI and succinate dehydrogenase inhibitor (SDHI) groups, or their mixtures. Difenoconazole–TTO completely prevented scab on pecan trees. Results suggest that difenoconazole–TTO can be integrated into scab control programmes for these crops and be used as a strategic approach in fungicide resistance management in orchards. [ABSTRACT FROM AUTHOR]

Apple scab caused by the ascomycete Venturia inaequalis is an economically significant disease worldwide. The annual sexual reproduction of V. inaequalis leads to high variation, changes in the population's genetic structure and adaptations to the changing environment, including overcoming the host's resistance. The objective of this study is to characterise and assess the genetic diversity of V. inaequalis populations in two main apple-growing regions in Latvia. In total, 143 V. inaequalis isolates were collected from Latvia, six reference strains with known virulence were obtained from other countries, and all strains were genotyped by 12 SSR markers. The SSR markers were highly variable and informative, identifying 158 alleles that ranged from two to 29 per locus. The Bayesian clustering identified three genetic lineages among the Latvian isolates that did not correlate to the geographic origin, host genotype, organ (leaves or fruits) from which the pathogen was isolated, time of collection, and type of isolation (single conidium or ascospore). The possible relatedness to virulence was detected when reference strains with known virulence were included in the analysis. Our findings correspond with previous studies demonstrating that V. inaequalis in Europe has a high genetic diversity within populations, but low diversity among the populations. [ABSTRACT FROM AUTHOR]

Apple scab caused by the pathogenic fungus Venturia inaequalis causes significant damage to apples. The creation of apple-scab resistant varieties is considered an alternative to pesticide-based management. To evaluate R gene inheritance, 862 apple hybrid samples were analyzed and divided into populations depending on the resistance genes (Rvi6 and Rvi5) present in the parents' genotypes and their combinations. The field evaluation was carried out in 2016–2018 using the international VINQUEST scab evaluation methodology, but the genes were identified using specific scab resistance-gene molecular markers. Parent plants were genotyped using 22 SSR markers. The study determined that the field resistance of apple scab is influenced not only by the resistance genes present in the genotype but also by the genetic background of the apple cultivar, the tree's general health status and resistance to other diseases. It was found that the apple scab resistance genes Rvi6 (Vf) and Rvi5 (Vm) are inherited according to Mendelian laws, and when both genes are pyramided in offspring, they are inherited as separate genes. In general, the inheritance of both genes in the offspring is not a determining factor for apple field resistance, as field resistance to scab is influenced by a combination of several factors. [ABSTRACT FROM AUTHOR]

Apple scab is an important disease conventionally controlled by chemical fungicides, which should be replaced by more environmentally friendly alternatives. One of these alternatives could be the use of lipopeptides produced by Bacillus subtilis. The objective of this work is to study the action of the three families of lipopeptides and different mixtures of them in vitro and in vivo against Venturia inaequalis. Firstly, the antifungal activity of mycosubtilin/surfactin and fengycin/surfactin mixtures was determined in vitro by measuring the median inhibitory concentration. Then, the best lipopeptide mixture ratio was produced using Design of Experiment (DoE) to optimize the composition of the culture medium. Finally, the lipopeptides mixtures efficiency against V. inaequalis was assessed in orchards as well as the evaluation of the persistence of lipopeptides on apple. In vitro tests show that the use of fengycin or mycosubtilin alone is as effective as a mixture, with the 50–50% fengycin/surfactin mixture being the most effective. Optimization of culture medium for the production of fengycin/surfactin mixture shows that the best composition is glycerol coupled with glutamic acid. Finally, lipopeptides showed in vivo antifungal efficiency against V. inaequalis regardless of the mixture used with a 70% reduction in the incidence of scab for both mixtures (fengycin/surfactin or mycosubtilin/surfactin). The reproducibility of the results over the two trial campaigns was significantly better with the mycosubtilin/surfactin mixture. The use of B. subtilis lipopeptides to control this disease is very promising. [ABSTRACT FROM AUTHOR]

Apple scab, which is caused by the fungus Venturia inaequalis, is one of the most important diseases of apple trees. It mainly affects leaves, fruits, and sometimes also flowers and shoots. The aim of this work was monitoring the resistance of Venturia inaequalis to pyrimethanil and stobilurins fungicides. Part of our work included evaluation of the monitoring and assignment of the population to the relevant category of resistance to the active substances. A total of 46 populations were tested. Our results show that the population of V. inaequalis was generally more sensitive to pyrimethanil while resistant to strobilurins. A large percentage of the populations (76 %) was sensitive to pyrimethanil and just 1 was moderately resistant, only 31 % of the population was sensitive to strobilurins and 62 % were resistant.

The natural product chitosan has been shown to reduce plant disease severity and enhance the efficacy of microbial biocontrol agents in several crops. However, little is known about the potential synergisms between chitosan and biopesticides and best use practices in apple production. The objectives of this study were to evaluate the effect of pre-harvest applications of chitosan alone and in combination with a commercial biopesticide to suppress fungal diseases of apple and to investigate the potential for chitosan to reduce the quantity of overwintering Venturia inaequalis spores in orchard leaf litter. Chitosan products, Tidal Grow and ARMOUR-Zen 15, and a commercial biopesticide, Serenade ASO, were tested in a research orchard in Pennsylvania and commercial orchards in New Hampshire. Chitosan applications reduced apple scab incidence and severity by up to 55% on fruit compared to the water control. Chitosan also reduced sooty blotch, flyspeck, and rust incidence on fruit. Furthermore, a chitosan + biopesticide treatment overlayed onto a grower standard spray program reduced diseases more effectively than the grower standard alone. However, this efficacy was dependent on the cultivar and pathogen. Chitosan did not reduce overwintering V. inaequalis ascospores. This research provides evidence that pre-harvest chitosan applications have the potential for disease management in apple production.

Background: Scab caused by Venturia inaequalis (Cke.) Wint. is the most important fungal disease of apple. Fungicide application is a widely practiced method of disease control. However, the use of chemicals is costintensive, tedious, and ecologically unsafe. The development of genetic resistance and the breeding of resistant cultivars is the most reliable and safest option. One such source of scab resistance happens to be the variety 'Shireen', released from SKUAST-Kashmir. However, to date, the nature of resistance and its genetic control have not been characterized. Objective This research aimed to elucidate the genetic basis of scab resistance in Shireen. Methods: Genetic mapping of quantitative trait loci (QTL) for resistance to apple scab disease was performed using an F1 cross developed between the susceptible cultivar 'StarKrimson' and the resistant cultivar 'Shireen'. The population was evaluated for two consecutive years. Further, six candidate genes were analyzed via quantitative real-time PCR, to determine their expression level in response to the pathogen infestation. Results: Genotyping and disease phenotyping of populations led us to identify two quantitative trait loci (QTLs), namely qRVI.SS-LG2.2019 and qRVI.SS-LG8.2019 on chromosomes 2 and 8 with LOD-values of 7.67 and 4.99 respectively, and six potential CDGs for the polygenic resistance in 'Shireen'. The genomic region corresponding to the mapped QTLs in LG 2 and LG 8 of 'Shireen' was examined for candidate genes possibly related to scab resistance using in silico analysis. Conclusion: The QTLs mapped in the genetic background of Shireen are the novel QTLs and may be transferred to desirable genetic backgrounds and provide opportunities for isolation and cloning of genes apart from their utility to achieve durable resistance to scab. [ABSTRACT FROM AUTHOR]

In Agriculture Sciences, detection of diseases is one of the most challenging tasks. The mis-interpretations of plant diseases often lead to wrong pesticide selection, resulting in damage of crops. Hence, the automatic recognition of the diseases at earlier stages is important as well as economical for better quality and quantity of fruits. Computer aided detection (CAD) has proven as a supportive tool for disease detection and classification, thus allowing the identification of diseases and reducing the rate of degradation of fruit quality. In this research work, a model based on convolutional neural network with 19 convolutional layers has been proposed for effective and accurate classification of Marsonina Coronaria and Apple Scab diseases from apple leaves. For this, a database of 50,000 images has been acquired by collecting images of leaves from apple farms of Himachal Pradesh (H.P) and Uttarakhand (India). An augmentation technique has been performed on the dataset to increase the number of images for increasing the accuracy. The performance analysis of the proposed model has been compared with the new two Convolutional Neural Network (CNN) models having 8 and 9 layers respectively. The proposed model has also been compared with the standard machine learning classifiers like support vector machine, k-Nearest Neighbour, Random Forest and Logistic Regression models. From experimental results, it has been observed that the proposed model has outperformed the other CNN based models and machine learning models with an accuracy of 99.2%. [ABSTRACT FROM AUTHOR]

Apple scab, caused by the fungal pathogen Venturia inaequalis , is the most economically important disease of apple (Malus x domestica) worldwide. To develop durable control strategies against this disease, a better understanding of the genetic mechanisms underlying the growth, reproduction, virulence and pathogenicity of V. inaequalis is required. A major bottleneck for the genetic characterization of V. inaequalis is the inability to easily delete or disrupt genes of interest using homologous recombination. Indeed, no gene deletions or disruptions in V. inaequalis have yet been published. Using the melanin biosynthesis pathway gene trihydroxynaphthalene reductase (THN) as a target for inactivation, which has previously been shown to result in a light-brown colony phenotype when transcriptionally silenced using RNA interference, we show, for the first time, that the CRISPR-Cas9 gene editing system can be successfully applied to the apple scab fungus. More specifically, using a CRISPR-Cas9 single guide RNA (sgRNA) targeted to the THN gene, delivered by a single autonomously replicating Golden Gate-compatible plasmid, we were able to identify six of 36 stable transformants with a light-brown phenotype, indicating an ∼16.7% gene inactivation efficiency. Notably, of the six THN mutants, five had an independent mutation. As part of our pipeline, we also report a high-resolution melting (HRM) curve protocol for the rapid detection of CRISPR-Cas9 gene-edited mutants of V. inaequalis. This protocol identified a single base pair deletion mutation in a sample containing only 5% mutant genomic DNA, indicating high sensitivity for mutant screening. In establishing CRISPR-Cas9 as a tool for gene editing in V. inaequalis , we have provided a strong starting point for studies aiming to decipher gene function in this fungus. The associated HRM curve protocol will enable CRISPR-Cas9 transformants to be screened for gene inactivation in a high-throughput and low-cost manner, which will be particularly powerful in cases where the CRISPR-Cas9-mediated gene inactivation efficiency is low. • First report of CRISPR-Cas9 gene editing in Venturia inaequalis. • Golden Gate-compatible plasmid used for delivery of the sgRNA. • First use of HRM to identify CRISPR-Cas9 gene-edited mutants in fungi. [ABSTRACT FROM AUTHOR]

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This paper presents the results recorded following the application in apple culture of different treatment programs with fungicide in order to control Venturia inaequalis, a pathogen recognized for the quantitative and qualitative damages caused. In order to control this pathogen, the chemical measures are necessary in addition to the agro-phytotechnical ones. The efficacy of the new active substance fluopyram, present in Luna Experience, with effect in blocking the pathogen's respiration. The apple variety on which the experiments were carried out is Idared, chosen especially for its sensitivity to the apple scab. The observations took place in 2016 at I.C.D.P. Mărăcineni. The evolution of the meteorological parameters was noted using Watchdog station, the dedicated software and IRFAN VIEW (Freeware). With its help, the dynamics of the infections were graphically highlighted and warnings were elaborated. In the field, observations were made to determine the frequency (F%) and intensity (I%) of the attack, in order to calculate the degree attack (DA%). In the end, the effectiveness of the treatment programs was calculated. [ABSTRACT FROM AUTHOR]

Apple scab caused by Venturia inaequalis has the most destructive effects among other phytopathogens in apple crops all over the world. The integration of resistance genes from local and domestic cultivars is a prerequisite for the efficient control of this disease and is a main target in efficient breeding approaches. Across Greece, many domestic apple cultivars are reported without deep knowledge about the presence and diversity of scab resistance genes. In this study, the presence of five resistance genes (Rvi2, Rvi4, Rvi6, Rvi8 and Rvi11) was evaluated across twenty local and domestic apple genotypes, employing twelve molecular markers closely linked to known apple scab resistance loci. Significant differences and polymorphisms among the tested cultivars were detected suggesting that some of them carry a sufficient number of resistance genes. This observed genetic diversity could be exploited in ongoing breeding approaches as a natural source of polygenic resistance against apple scab.

Apple breeding is a laborious and long-lasting process that requires qualified resources, land, time, and funds. In this study, more than 5000 F1 apple hybrids from direct and testcrosses were analyzed. The results revealed how the phenotypic expression of the main quantitative traits of interest assessed in five half-sib families was controlled by the additive genetic effects and by non-additive effects of dominance and epistasis. The statistical number of hybrids required to ensure efficient selection increased exponentially with the number of desirable traits. The minimum number of progenies required to obtain a hybrid with associated quantitative traits of agronomic interest was highly variable. For two independent traits essential in selection (fruit size and quality), but incorporated together in the same hybrid, the statistical number was between about 30 and 300. If three more cumulative traits were added (a large number of fruits per tree, resistance/tolerance to apple scab, and powdery mildew attack), the limits increased to between 1500 and 18,000. The study highlighted the need for new apple varieties due to the narrowing of the genetic diversity of the cultivated species and how the choice of parents used in hybridizations (as well as the objectives pursued in the selection) can increase the efficiency of apple breeding.

Apple orchards are perennially planted where pesticides are applied to control numerous pests and diseases. The extensive long-term use of fungicides can lead to overall environmental load and resistance risk. This study aims to assess which fungicide-active substances have been used more intensively in the last decade in Latvia, evaluating the overall environmental load using the Pesticide Load Indicator (PLI). It was essential to see whether the amount of active substance usage rises, how it correlates with the total changes of the PLI and which substances are with the highest scores. The other issue was to test the sensitivity of Venturia inaequalis populations to systemic fungicides. Six full-bearing apple orchards that reflected local plant protection practices were selected from the different growing regions of Latvia to analyze fungicide use from 2012 to 2021 and test V. inaequalis populations’ sensitivity to systemic substances difenoconazole and cyprodinil. The PLI demonstrated that the protective fungicides were the most crucial group overall, with the highest potential impact on the environment and human health. Systemic fungicides had a relatively lower environmental impact, but after long-term use, the pathogen population’s sensitivity to difenoconazole and cyprodinil was reduced. Introducing new fungicide classes and biological control agents could help growers improve plant protection strategies against V. inaequalis, reducing the risk of resistance and environmental load.

DMIs (sterol demethylation inhibitors) provide an important mode of action for control of Venturia inaequalis. They target the enzyme 14a-demethylase, encoded by the Cyp51 gene, which acts on an essential precursor for ergosterol biosynthesis. One mechanism, which leads to reduced DMI sensitivity of fungi, is based on mutations in Cyp51. In contrast to other phytopathogenic fungi, only little information is available on changes in the Cyp51 gene of V. inaequalis and their potential influence on DMI sensitivity. In this study, 126 monoconidial isolates were generated from field samples of V. inaequalis collected in apple orchards in different countries in 2018 and 2019. The Cyp51 gene of all isolates was sequenced. Furthermore, a new liquid culture method for V. inaequalis was established, which provides strongly sporulating cultures as a basis for the sensitivity tests. The sequence analysis revealed 15 different point mutations, and each of them resulted in an amino acid exchange in the 14α-demethylase. For some of them, corresponding mutations had been reported in other plant pathogens and shown to influence DMI sensitivity. We measured fungal growth in microtiter-plate tests to evaluate the influence of the amino acid changes in the Cyp51 gene on the sensitivity of the isolates towards the DMIs mefentrifluconazole, difenoconazole, and myclobutanil. While some mutations (e.g. those in the YGYG region 443–446) showed little impact on DMI sensitivity, others caused higher EC50 values, in particular Y133F and M141T. The sensitivity reducing effects differed for the three DMIs and were most pronounced for myclobutanil and less for mefentrifluconazole. The variability in the DMI sensitivity in each Cyp51 haplotype suggests the presence of additional resistance mechanisms caused by genetic background. [ABSTRACT FROM AUTHOR]

The large genetic evolution due to the sexual reproduction-mediated gene assortments and propensities has made Venturia inaequalis (causing apple scab) unique with respect to its management strategies. The resistance in apple germplasm against the scab, being controlled for by more than fifteen genes, has limited gene alteration-based investigations. Therefore, a biological approach of bacterial endophyte community dynamics was envisioned across the apple germplasm in context to the fungistatic behavior against V. inaequalis. A total of 155 colonies of bacterial endophytes were isolated from various plant parts of the apple, comprising 19 varieties, and after screening for antifungal behavior followed by morphological, ARDRA, and sequence analysis, a total of 71 isolates were selected for this study. The alpha diversity indices were seen to fluctuate greatly among the isolation samples in context to microflora with antifungal behavior. As all the isolates were screened for the presence of various metabolites and some relevant genes that directly or indirectly influence the fungistatic behavior of the isolated microflora, a huge variation among the isolated microflora was observed. The outstanding isolates showing highest percentage growth inhibition of V. inaequalis were exploited to raise a bio-formulation, which was tested against the scab prevalence in eight apple varieties under controlled growth conditions. The formulation at all the concentrations caused considerable reductions in both the disease severity and disease incidence in all the tested apple varieties. Red Delicious being most important cultivar of the northwestern Himalayas was further investigated for its biochemical behavior in formulation and the investigation revealed different levels of enzyme production, chlorophyll, and sugars against the non-inoculated control. [ABSTRACT FROM AUTHOR]

The ascomycete fungus Venturia inaequalis causes scab disease, a major problem in apple production. The utilization of resistant cultivars is hindered by emerging new pathogen races, which erode their resistance. Increasing our knowledge on the population genetic processes of the fungus can contribute to the development of resistance gene deployment strategies and more durable resistance. We investigated the effect of four scab race indicator cultivars, ‘Gala’ (no R-gene), ‘Golden Delicious’ (Rvi1), ‘Geneva’ (Rvi3, complex), and OR45t132 (Rvi5), on the V. inaequalis population genetic structure and diversity. Sixty-six monosporic fungal isolates from the four cultivars were genotyped with seven simple sequence repeat primers. Furthermore, the partial resistance of the indicators and the pathogenicity profile of the conidia from each host were assessed. The genetic diversity and structure of the investigated V. inaequalis subpopulations correspond to the partial resistance of the original hosts as well as the subpopulations’ pathogenicity profiles. Indicators carrying Rvi3 and Rvi5 had strongly diverged and specialized V. inaequalis populations on them and fewer symptoms on the field. In line with the complete breakdown of the Rvi1 gene, the population from ‘Golden Delicious’ did not segregate from the susceptible ‘Gala’, and virulence towards Rvi1 was present in all subpopulations.

Apple scab caused by the ascomycete Venturia inaequalis is an economically significant disease worldwide. The annual sexual reproduction of V. inaequalis leads to high variation, changes in the population’s genetic structure and adaptations to the changing environment, including overcoming the host’s resistance. The objective of this study is to characterise and assess the genetic diversity of V. inaequalis populations in two main apple-growing regions in Latvia. In total, 143 V. inaequalis isolates were collected from Latvia, six reference strains with known virulence were obtained from other countries, and all strains were genotyped by 12 SSR markers. The SSR markers were highly variable and informative, identifying 158 alleles that ranged from two to 29 per locus. The Bayesian clustering identified three genetic lineages among the Latvian isolates that did not correlate to the geographic origin, host genotype, organ (leaves or fruits) from which the pathogen was isolated, time of collection, and type of isolation (single conidium or ascospore). The possible relatedness to virulence was detected when reference strains with known virulence were included in the analysis. Our findings correspond with previous studies demonstrating that V. inaequalis in Europe has a high genetic diversity within populations, but low diversity among the populations.

Increasing the reliability of fruit crops in unstable weather conditions of a temperate continental climate has become particularly relevant. This research was carried out based on a bioresource collection from the Russian Research Institute of Fruit Crop Breeding (VNIISPK BRC). Apple cultivars (diploids and triploids) of different maturation periods were studied. Triploid cultivars (3×) of summer maturation were developed using diploid gametes of the ‘Papirovka tetraploid’ cultivar as a donor (2-4-4-4×), triploids of winter maturation were developed using 13-6-106 (Suvorovetz: open pollination), and ‘Wealthy tetraploid’ (2-4-4-4×) or ‘Giant Spy’ (2-4-4-4×) were used as donors of diploid gametes. ‘Antonovka’ and ‘Melba’ were taken as control cultivars. The aim of this work was to evaluate the apple breeding cultivars according to the main economic and biological indicators and to determine the threshold of resistance to unfavorable conditions of the winter period for possible cultivation in specific conditions of a temperate continental climate. As a result of using the method of modeling damaging factors, apple cultivars that withstood not only the critical temperatures of January, but also especially dangerous frosts after thaws at the end of winter were identified, with the stability of vital tissues at the level of ‘Antonovka’ (a control cultivar), scab immune cultivars (RVi6) ‘Ivanovskoye’, ‘Zdorovie’ and ‘Pamyati Hitrovo’, and immune triploids (3×) ‘Vavilovskoye’, ‘Aleksandr Boyko’, Rozhdestvenskoye’ and ‘Academic Saveliev’. During the whole winter, the winter resistance of summer scab immune triploids, ‘Zhilinskoye’, ‘Maslovskoye’, ‘Yablochny Spas’ and ‘Spasskoye’, was at the level of ‘Antonovka’, but was higher than ‘Melba’. The high precocity of scab-immune cultivars, ’Afrodita’, ‘Ivanovskoye’, ‘Veniaminovskoye’, and ‘Yubiley Moskvy’, and triploids ‘Patriot’ and ‘Rozhdestvenskoye’, have been identified. Triploid cultivars are superior to diploid cultivars and control ‘Antonovka’ in fruit size. According to the complex of characteristics, promising apple cultivars were identified for planting in intensive orchards of the temperate continental climate zones, namely ‘Avgusta’ and ‘Solnyshko’ (summer cultivars), and ‘Aleksandr Boyko’, ‘Vavilovskoye’, ‘Venyaminovskoye’, ‘Ivanovskoye’ and ‘Rozhdestvenskoye’ (winter cultivars).