Deep Learning Based Barley Disease Quantification for Sustainable Crop Production.

Net blotch disease caused by Drechslera teres is a major fungal disease that affects barley (Hordeum vulgare) plants and can result in significant crop losses. In this study, we developed a deep-learning model to quantify net blotch disease symptoms on different days post-infection on seedling leaves using Cascade R-CNN (Region-Based Convolutional Neural Networks) and U-Net (a convolutional neural network) architectures. We used a dataset of barley leaf images with annotations of net blotch disease to train and evaluate the model. The model achieved an accuracy of 95% for cascade R-CNN in net blotch disease detection and a Jaccard index score of 0.99, indicating high accuracy in disease quantification and location. The combination of Cascade R-CNN and U-Net architectures improved the detection of small and irregularly shaped lesions in the images at 4-days post infection, leading to better disease quantification. To validate the model developed, we compared the results obtained by automated measurement with a classical method (necrosis diameter measurement) and a pathogen detection by real-time PCR. The proposed deep learning model could be used in automated systems for disease quantification and to screen the efficacy of potential biocontrol agents to protect against disease.

Bacillus velezensis BE2 controls wheat and barley diseases by direct antagonism and induced systemic resistance.

Wheat and barley rank among the main crops cultivated on a global scale, providing the essential nutritional foundation for both humans and animals. Nevertheless, these crops are vulnerable to several fungal diseases, such as Septoria tritici blotch and net blotch, which significantly reduce yields by adversely affecting leaves and grain quality. To mitigate the effect of these diseases, chemical fungicides have proven to be genuinely effective; however, they impose a serious environmental burden. Currently, biocontrol agents have attracted attention as a sustainable alternative to fungicides, offering an eco-friendly option. The study aimed to assess the efficacy of Bacillus velezensis BE2 in reducing disease symptoms caused by Zymoseptoria tritici and Pyrenophora teres. This bacterium exhibited significant antagonistic effects in vitro by suppressing fungal development when pathogens and the beneficial strain were in direct confrontation. These findings were subsequently confirmed through microscopic analysis, which illustrated the strain's capacity to inhibit spore germination and mycelial growth in both pathogens. Additionally, the study analysed the cell-free supernatant of the bacterium using UPLC-MS (ultra-performance liquid chromatography-mass spectrometry). The results revealed that strain BE2 produces, among other metabolites, different families of cyclic lipopeptides that may be involved in biocontrol. Furthermore, the beneficial effects of strain BE2 in planta were assessed by quantifying the fungal DNA content directly at the leaf level after bacterization, using two different application methods (foliar and drenching). The results indicated that applying the beneficial bacterium at the root level significantly reduced pathogens pressure. Finally, gene expression analysis of different markers showed that BE2 application induced a priming effect within the first hours after infection. KEY POINTS: • BE2 managed Z. tritici and P. teres by direct antagonism and induced systemic resistance. • Strain BE2 produced seven metabolite families, including three cyclic lipopeptides. • Application of strain BE2 at the root level triggered plant defense mechanisms.

PGPR-Soil Microbial Communities' Interactions and Their Influence on Wheat Growth Promotion and Resistance Induction against Mycosphaerella graminicola.

The efficiency of plant-growth-promoting rhizobacteria (PGPR) may not be consistently maintained under field conditions due to the influence of soil microbial communities. The present study aims to investigate their impact on three PGPR-based biofertilizers in wheat. We used the PGPR Paenibacillus sp. strain B2 (PB2), PB2 in co-inoculation with Arthrobacter agilis 4042 (Mix 2), or with Arthrobacter sp. SSM-004 and Microbacterium sp. SSM-001 (Mix 3). Inoculation of PB2, Mix 2, and Mix 3 into non-sterile field soil had a positive effect on root and aboveground dry biomass, depending on the wheat cultivar. The efficiency of the PGPR was further confirmed by the protection they provided against Mycosphaerella graminicola, the causal agent of Septoria leaf blotch disease. PB2 exhibited protection of ≥37.8%, while Mix 2 showed ≥47.9% protection in the four cultivars tested. These results suggest that the interactions between PGPR and native soil microbial communities are crucial for promoting wheat growth and protection. Additionally, high-throughput sequencing of microbial communities conducted 7 days after PGPR inoculations revealed no negative effects of PB2, Mix 2, and Mix 3 on the soil microbial community structure. Interestingly, the presence of Arthrobacter spp. appeared to mitigate the potential negative effect of PB2 on bacterial community and foster root colonization by other beneficial bacterial strains.

Enhancing Maize Productivity and Soil Health under Salt Stress through Physiological Adaptation and Metabolic Regulation Using Indigenous Biostimulants.

Salinity poses a persistent threat to agricultural land, continuously jeopardizing global food security. This study aimed to enhance sweet corn (SC) fitness under varying levels of salinity using indigenous biostimulants (BioS) and to assess their impacts on plant performance and soil quality. The experiment included control (0 mM NaCl), moderate stress (MS; 50 mM NaCl), and severe stress (SS; 100 mM NaCl) conditions. Indigenous biostimulants, including compost (C), Bacillus sp., Bacillus subtilis (R), and a consortium of arbuscular mycorrhizal fungi (A) were applied either individually or in combination. Growth traits, physiological and biochemical parameters in maize plants, and the physico-chemical properties of their associated soils were assessed. SS negatively affected plant growth and soil quality. The RC combination significantly improved plant growth under SS, increasing aerial (238%) and root (220%) dry weights compared to controls. This treatment reduced hydrogen peroxide by 54% and increased peroxidase activity by 46% compared to controls. The indigenous biostimulants, particularly C and R, enhanced soil structure and mineral composition (K and Mg). Soil organic carbon and available phosphorus increased notably in C-treated soils. Furthermore, RC (437%) and CAR (354%) treatments exhibited a significant increase in glomalin content under SS. Indigenous biostimulants offer a promising strategy to mitigate salinity-related threats to agricultural land. They improve plant fitness, fine-tune metabolism, and reduce oxidative stress. In addition, the biostimulants improved the soil structure and mineral composition, highlighting their potential for reconstitution and sustainability in salt-affected areas. This approach holds promise for addressing salinity-related threats to global food security.

Evaluating Food Additives Based on Organic and Inorganic Salts as Antifungal Agents against Monilinia fructigena and Maintaining Postharvest Quality of Apple Fruit.

A set of commonly used food additives was evaluated for their antifungal activity against the brown rot disease of fruits caused by the fungal pathogen Monilinia fructigena, which is one of the most economically important agents, causing important damage to pome fruits, such as pears and apples. The radial mycelial growth of the fungal pathogen was assessed in PDA amended with different concentrations (0.5, 2, 2.5, and 5%) of each additive. The results underlined that most of the additives displayed a significant inhibition of mycelial growth, with the extent of inhibition varying depending on the specific additive and concentration used. Five food additives showed high inhibition rates (above 88%), of which sodium bicarbonate, sodium carbonate, copper sulphate, and sodium hydroxide were the most effective, whereas ammonium carbonate, magnesium chlorite, and citric acid were the least effective. Interestingly, the coatings containing sodium bicarbonate, copper sulphate, and ammonium bicarbonate significantly reduced the incidence of brown rot disease in apples, but other additives were not effective, such as ammonium carbonate and magnesium sulphate. The anhydrous sodium sulphate used at a concentration of 2%, was found to be one of the least effective additives, with a reduction rate of 20%. Subsequently, food additives showing good growth inhibition rates and reduction in disease severity were then tested in semi-commercial trials at temperatures of 4 °C and 22 °C. The results indicated that these additives demonstrate effectiveness in controlling M. fructigena at specific concentrations, and lower temperatures (4 °C) can improve the efficiency of the control measures. In addition, the selected food additives exhibited significant antimicrobial activity against M. fructigena, suggesting their application as a promising alternative for managing brown rot disease in apple fruits.

Paraburkholderia phytofirmans PsJN delays Botrytis cinerea development on grapevine inflorescences.

Grapevine flowering is an important stage in the epidemiology of Botrytis cinerea, the causal agent of gray mold disease. To prevent infection and to minimize postharvest losses, the control of this necrotrophic fungus is mainly based on chemical fungicides application. However, there is a growing interest in other control alternatives. Among them, the use of beneficial microorganisms appears as an eco-friendly strategy. This study aims to investigate the effect of Paraburkholderia phytofirmans PsJN, root-inoculated or directly sprayed on fruiting cuttings inflorescences to control B. cinerea growth. For this purpose, quantification by real time PCR of Botrytis development, direct effect of PsJN on fungal spore germination and chemotaxis were assayed. Our results showed a significant protective effect of PsJN only by direct spraying on inflorescences. Moreover, we demonstrated an inhibition exerted by PsJN on Botrytis spore germination, effective when there was a direct contact between the two microorganisms. This study showed that PsJN is positively attracted by the pathogenic fungus B. cinerea and forms a biofilm around the fungal hyphae in liquid co-culture. Finally, microscopic observations on fruit cuttings revealed a co-localization of both beneficial and pathogenic microorganisms on grapevine receptacle and stigma that might be correlated with the protective effect induced by PsJN against B. cinerea via a direct antimicrobial effect. Taking together, our findings allowed us to propose PsJN as a biofungicide to control grapevine gray mold disease.

Drivers of Post-Harvest Aflatoxin Contamination: Evidence Gathered from Knowledge Disparities and Field Surveys of Maize Farmers in the Rift Valley Region of Kenya.

Maize-dependent populations in sub-Saharan Africa are continually exposed to aflatoxin poisoning owing to their regular consumption of this dietetic cereal. Being a staple in Kenyan households, consumption of maize-based meals is done almost daily, thereby exposing consumers to aflatoxicoses. This study assessed awareness levels, knowledge disparities, and perceptions regarding aflatoxin contamination at the post-harvest phase among farmers in the Rift Valley Region of Kenya. Households were randomly selected using a geographical positioning system (GPS) overlay of the agro-ecological zones within Uasin Gishu and Elgeyo Marakwet counties. Face-to-face interviews were conducted in 212 smallholder and large-scale farms. The study documented the demographic profiles of farmers and knowledge, awareness, and perceptions of aflatoxin contamination using a pre-designed structured questionnaire. Most farmers were familiar with aflatoxins and the adverse effects they present to health (61.32%). Almost all the farmers (94.37%) were aware of storage molds and food-spoilage fungi. However, few farmers adopted good post-harvest practices (PHPs), such as avoiding premature harvests (49.8%), using well-ventilated storage spaces (44.6%), grain sorting (30.5%), proper drying of maize (17.8%), and using hermetic bags for storage (30.5%). Conclusively, intensified farmer education is required to train farmers on good PHPs to protect their maize from aflatoxigenic fungi and aflatoxin accumulation.

Application of Indigenous Rhizospheric Microorganisms and Local Compost as Enhancers of Lettuce Growth, Development, and Salt Stress Tolerance.

This study aimed to mitigate salt stress effects on lettuce by using native biostimulants (arbuscular mycorrhizal fungi (M, consortium), plant growth-promoting rhizobacteria (R, Z2, and Z4 strains), and compost (C)) applied alone or in combination under salinity stress (0, 50, and 100 mM NaCl). Physiological, biochemical, nutritional, mycorrhizal, growth, and soil characteristics were evaluated. Results revealed that growth and physiological traits were negatively affected by salinity. However, mycorrhizal colonization was enhanced under 100 mM NaCl after compost application. The applied biostimulants, particularly M and/or R improved the salinity tolerance of lettuce by increasing the dry biomass by 119% and 113% under 100 mM NaCl, respectively, for M and MR treatments. Similarly, MR enhanced stomatal conductance (47%), water content (260%), total chlorophyll (130%), phosphorus content (363%), and reduced the malondialdehyde (54%) and hydrogen peroxide (78%) compared to the control. Moreover, peroxidase activity (76%) and sugar content (36%) were enhanced by CM treatment, while protein (111%) and proline (104%) contents were significantly boosted by R treatment under 100 mM NaCl. Furthermore, glomalin content was enhanced by MR treatment under severe salinity. In conclusion, the applied biostimulants alone or in combination might help lettuce to tolerate salt stress and enhance its production in degraded areas.

Plasmopara viticola the Causal Agent of Downy Mildew of Grapevine: From Its Taxonomy to Disease Management.

Plasmopara viticola (P. viticola, Berk. & M. A. Curtis; Berl. & De Toni) causing grapevine downy mildew is one of the most damaging pathogens to viticulture worldwide. Since its recognition in the middle of nineteenth century, this disease has spread from America to Europe and then to all grapevine-growing countries, leading to significant economic losses due to the lack of efficient disease control. In 1885 copper was found to suppress many pathogens, and is still the most effective way to control downy mildews. During the twentieth century, contact and penetrating single-site fungicides have been developed for use against plant pathogens including downy mildews, but wide application has led to the appearance of pathogenic strains resistant to these treatments. Additionally, due to the negative environmental impact of chemical pesticides, the European Union restricted their use, triggering a rush to develop alternative tools such as resistant cultivars breeding, creation of new active ingredients, search for natural products and biocontrol agents that can be applied alone or in combination to kill the pathogen or mitigate its effect. This review summarizes data about the history, distribution, epidemiology, taxonomy, morphology, reproduction and infection mechanisms, symptoms, host-pathogen interactions, host resistance and control of the P. viticola, with a focus on sustainable methods, especially the use of biocontrol agents.

D-Tagatose-Based Product Triggers Sweet Immunity and Resistance of Grapevine to Downy Mildew, but Not to Gray Mold Disease.

The use of natural bio-based compounds becomes an eco-friendly strategy to control plant diseases. Rare sugars would be promising compounds as inducers of plant "sweet immunity". The present study aimed to investigate the induced resistance of grapevine leaves against Plasmopara viticola and Botrytis cinerea by a rare sugar-based product (IFP48) and its active ingredient D-tagatose (TAG), in order to elucidate molecular mechanism involved in defense-related metabolic regulations before and after pathogen challenge. Data showed that spraying leaves with IFP48 and TAG lead to a significant reduction of downy mildew, but not of gray mold disease. The induced protection against P. viticola relies on IFP48's and to a lesser extent TAG's ability to potentiate the activation of salicylic acid- and jasmonic acid/ethylene-responsive genes and stilbene phytoalexin accumulation. Most of defense responses remained upregulated in IFP48-treated plants after infection with P. viticola, but inconsistent following challenge with B. cinerea. The beneficial effects of IFP48 were associated with an enhanced accumulation of tagatose inside leaf tissues compared to TAG treatment. Meanwhile, the amounts of sugars, glucose, fructose, maltose, galactose and trehalose remained unchanged or decreased in IFP48-treated leaves after P. viticola infection, although only a few genes involved in sugar transport and metabolism showed transcriptional regulation. This suggests a contribution of sugar homeostasis to the IFP48-induced sweet immune response and priming plants for enhanced resistance to P. viticola, but not to B. cinerea.

Pseudomonas Lipopeptide-Mediated Biocontrol: Chemotaxonomy and Biological Activity.

Pseudomonas lipopeptides (Ps-LPs) play crucial roles in bacterial physiology, host-microbe interactions and plant disease control. Beneficial LP producers have mainly been isolated from the rhizosphere, phyllosphere and from bulk soils. Despite their wide geographic distribution and host range, emerging evidence suggests that LP-producing pseudomonads and their corresponding molecules display tight specificity and follow a phylogenetic distribution. About a decade ago, biocontrol LPs were mainly reported from the P. fluorescens group, but this has drastically advanced due to increased LP diversity research. On the one hand, the presence of a close-knit relationship between Pseudomonas taxonomy and the molecule produced may provide a startup toolbox for the delineation of unknown LPs into existing (or novel) LP groups. Furthermore, a taxonomy-molecule match may facilitate decisions regarding antimicrobial activity profiling and subsequent agricultural relevance of such LPs. In this review, we highlight and discuss the production of beneficial Ps-LPs by strains situated within unique taxonomic groups and the lineage-specificity and coevolution of this relationship. We also chronicle the antimicrobial activity demonstrated by these biomolecules in limited plant systems compared with multiple in vitro assays. Our review further stresses the need to systematically elucidate the roles of diverse Ps-LP groups in direct plant-pathogen interactions and in the enhancement of plant innate immunity.

Beneficial Microorganisms to Control the Gray Mold of Grapevine: From Screening to Mechanisms.

In many vineyards around the world, Botrytis cinerea (B. cinerea) causes one of the most serious diseases of aerial grapevine (Vitis vinifera L.) organs. The control of the disease relies mainly on the use of chemical products whose use is increasingly challenged. To develop new sustainable methods to better resist B. cinerea, beneficial bacteria were isolated from vineyard soil. Once screened based on their antimicrobial effect through an in vivo test, two bacterial strains, S3 and S6, were able to restrict the development of the pathogen and significantly reduced the Botrytis-related necrosis. The photosynthesis analysis showed that the antagonistic strains also prevent grapevines from considerable irreversible PSII photo-inhibition four days after infection with B. cinerea. The 16S rRNA gene sequences of S3 exhibited 100% similarity to Bacillus velezensis, whereas S6 had 98.5% similarity to Enterobacter cloacae. On the other hand, the in silico analysis of the whole genome of isolated strains has revealed the presence of "biocontrol-related" genes supporting their plant growth and biocontrol activities. The study concludes that those bacteria could be potentially useful as a suitable biocontrol agent in harvested grapevine.

Advances in Wheat Physiology in Response to Drought and the Role of Plant Growth Promoting Rhizobacteria to Trigger Drought Tolerance.

In the coming century, climate change and the increasing human population are likely leading agriculture to face multiple challenges. Agricultural production has to increase while preserving natural resources and protecting the environment. Drought is one of the major abiotic problems, which limits the growth and productivity of crops and impacts 1-3% of all land.To cope with unfavorable water-deficit conditions, plants use through sophisticated and complex mechanisms that help to perceive the stress signal and enable optimal crop yield are required. Among crop production, wheat is estimated to feed about one-fifth of humanity, but faces more and more drought stress periods, partially due to climate change. Plant growth promoting rhizobacteria are a promising and interesting way to develop productive and sustainable agriculture despite environmental stress. The current review focuses on drought stress effects on wheat and how plant growth-promoting rhizobacteria trigger drought stress tolerance of wheat by highlighting several mechanisms. These bacteria can lead to better growth and higher yield through the production of phytohormones, osmolytes, antioxidants, volatile compounds, exopolysaccharides and 1-aminocyclopropane-1-carboxylate deaminase. Based on the available literature, we provide a comprehensive review of mechanisms involved in drought resilience and how bacteria may alleviate this constraint.

Rare Sugars: Recent Advances and Their Potential Role in Sustainable Crop Protection.

Rare sugars are monosaccharides with a limited availability in the nature and almost unknown biological functions. The use of industrial enzymatic and microbial processes greatly reduced their production costs, making research on these molecules more accessible. Since then, the number of studies on their medical/clinical applications grew and rare sugars emerged as potential candidates to replace conventional sugars in human nutrition thanks to their beneficial health effects. More recently, the potential use of rare sugars in agriculture was also highlighted. However, overviews and critical evaluations on this topic are missing. This review aims to provide the current knowledge about the effects of rare sugars on the organisms of the farming ecosystem, with an emphasis on their mode of action and practical use as an innovative tool for sustainable agriculture. Some rare sugars can impact the plant growth and immune responses by affecting metabolic homeostasis and the hormonal signaling pathways. These properties could be used for the development of new herbicides, plant growth regulators and resistance inducers. Other rare sugars also showed antinutritional properties on some phytopathogens and biocidal activity against some plant pests, highlighting their promising potential for the development of new sustainable pesticides. Their low risk for human health also makes them safe and ecofriendly alternatives to agrochemicals.

Pyrenophora teres: Taxonomy, Morphology, Interaction With Barley, and Mode of Control.

Net blotch, induced by the ascomycete Pyrenophora teres, has become among the most important disease of barley (Hordeum vulgare L.). Easily recognizable by brown reticulated stripes on the sensitive barley leaves, net blotch reduces the yield by up to 40% and decreases seed quality. The life cycle, the mode of dispersion and the development of the pathogen, allow a quick contamination of the host. Crop residues, seeds, and wild grass species are the inoculum sources to spread the disease. The interaction between the barley plant and the fungus is complex and involves physiological changes with the emergence of symptoms on barley and genetic changes including the modulation of different genes involved in the defense pathways. The genes of net blotch resistance have been identified and their localizations are distributed on seven barley chromosomes. Considering the importance of this disease, several management approaches have been performed to control net blotch. One of them is the use of beneficial bacteria colonizing the rhizosphere, collectively referred to as Plant Growth Promoting Rhizobacteria. Several studies have reported the protective role of these bacteria and their metabolites against potential pathogens. Based on the available data, we expose a comprehensive review of Pyrenophora teres including its morphology, interaction with the host plant and means of control.

A biological agent modulates the physiology of barley infected with Drechslera teres.

Recognized as the causal agent of net blotch, Drechslera teres is responsible for major losses of barley crop yield. The consequences of this leaf disease are due to the impact of the infection on the photosynthetic performance of barley leaves. To limit the symptoms of this ascomycete, the use of beneficial bacteria known as "Plant Growth Promoting Rhizobacteria" constitutes an innovative and environmentally friendly strategy. A bacterium named as strain B25 belonging to the genus Burkholderia showed a strong antifungal activity against D. teres. The bacterium was able to limit the development of the fungus by 95% in detached leaves of bacterized plants compared to the non-bacterized control. In this study, in-depth analyses of the photosynthetic performance of young barley leaves infected with D. teres and/or in the presence of the strain B25 were carried out both in and close to the necrotic area. In addition, gas exchange measurements were performed only near the necrotic area. Our results showed that the presence of the beneficial bacterium reduced the negative impact of the fungus on the photosynthetic performance and modified only the net carbon assimilation rate close to the necrotic area. Indeed, the presence of the strain B25 decreased the quantum yield of regulated non-photochemical energy loss in PSII noted as Y(NPQ) and allowed to maintain the values stable of maximum quantum yield of PSII photochemistry known as Fv/Fm and close to those of the control in the presence of D. teres. To the best of our knowledge, these data constitute the first study focusing on the impact of net blotch fungus and a beneficial bacterium on photosynthesis and respiratory parameters in barley leaves.

Development of a DNA-Based Real-Time PCR Assay To Quantify Allorhizobium vitis Over Time in Grapevine (Vitis vinifera L.) Plantlets.

Allorhizobium vitis is the primary causal pathogen of grapevine crown gall disease. Because this endophytic bacterium can survive as a systemic latent (symptomless) infection in grapevine, detecting and monitoring its development in planta is of great importance. In plant bacteria studies, plate counting is routinely used as a simple and reliable method to evaluate the bacterial population level in planta. However, isolation techniques are time-consuming and present some disadvantages such as the risk of contamination and the need for fresh samples for research. In this study, we developed a DNA-based real-time PCR assay that can replace the classical method to monitor the development of Allorhizobium vitis in grapevine plantlets. Primers targeting Allorhizobium vitis chromosomic genes and the virulent tumor-inducing plasmid were validated. The proposed quantitative real-time PCR technique is highly reliable and reproducible to assess Allorhizobium vitis numeration at the earliest stage of infection until tumor development in grapevine plantlets. Moreover, this low-cost technique provides rapid and robust in planta quantification of the pathogen and is suitable for fundamental research to monitor bacterial development over time.

The mode of action of plant associated Burkholderia against grey mould disease in grapevine revealed through traits and genomic analyses.

Plant-associated Burkholderia spp. have been shown to offer a promising alternative method that may address concerns with ecological issue associated with pesticide overuse in agriculture. However to date, little work has studied the role of Burkholderia species as biocontrol agents for grapevine pathogens. To this end, two Burkholderia strains, BE17 and BE24 isolated from the maize rhizosphere in France, were investigated to determine their biocontrol potential and their ability to induce systemic resistance against grey mould disease in grapevine. Results showed the capacity of both strains to inhibit spore germination and mycelium growth of Botrytis cinerea. Experimental inoculation with BE17 and BE24 showed a significant protection of bacterized-plantlets against grey mould compared to the non-bacterized control. BE17 and BE24-bacterized plants accumulated more reactive oxygen species and an increased callose deposition was observed in leaves of bacterized plantlets compared to the control plantlets. In bacterized plants, gene expression analysis subsequent to B. cinerea challenge showed that strains BE17 and BE24 significantly increased the relative transcript level of pathogenesis-related (PR) proteins PR5 and PR10, two markers involved in the Salicylic acid (SA)-signaling pathway. Furthermore, in silico analysis of strains revealed the presence of genes involved in plant growth promotion and biocontrol highlighting the attractiveness of these strains for sustainable agricultural applications.