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 postinfection on seedling leaves using Cascade R-CNN (region-based convolutional neural network) 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 postinfection, 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.

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.

Gene expression and metabolite analysis in barley inoculated with net blotch fungus and plant growth-promoting rhizobacteria.

Net blotch, caused by the ascomycete Drechslera teres, can compromise barley production. Beneficial bacteria strains are of substantial interest as biological agents for plant protection in agriculture. Belonging to the genus Paraburkholderia, a bacterium, referred to as strain B25, has been identified as protective for barley against net blotch. The strain Paraburkholderia phytofirmans (strain PsJN), which has no effect on the pathogen's growth, has been used as control. In this study, the expression of target genes involved in cell wall-related processes, defense responses, carbohydrate and phenylpropanoid pathways was studied under various conditions (with or without pathogen and/or with or without bacterial strains) at different time-points (0-6-12-48 h). The results show that specific genes were subjected to a circadian regulation and that the expression of most of them increased in barley infected with D. teres and/or bacterized with the strain PsJN. On the contrary, a decreased gene expression was observed in the presence of strain B25. To complement and enrich the gene expression analysis, untargeted metabolomics was carried out on the same samples. The data obtained show an increase in the production of lipid compounds in barley in the presence of the pathogen. In addition, the presence of strain B25 leads to a decrease in the production of defense compounds in this crop. The results contribute to advance the knowledge on the mechanisms occurring at the onset of D. teres infection and in the presence of a biocontrol agent limiting the severity of net blotch in barley.

Expression Analysis of Cell Wall-Related Genes in the Plant Pathogenic Fungus Drechslera teres.

Drechslera teres (D. teres) is an ascomycete, responsible for net blotch, the most serious barley disease causing an important economic impact. The cell wall is a crucial structure for the growth and development of fungi. Thus, understanding cell wall structure, composition and biosynthesis can help in designing new strategies for pest management. Despite the severity and economic impact of net blotch, this is the first study analyzing the cell wall-related genes in D. teres. We have identified key genes involved in the synthesis/remodeling of cell wall polysaccharides, namely chitin, ß-(1,3)-glucan and mixed-linkage glucan synthases, as well as endo/exoglucanases and a mitogen-activated protein kinase. We have also analyzed the differential expression of these genes in D. teres spores and in the mycelium after cultivation on different media, as well as in the presence of Paraburkholderia phytofirmans strain PsJN, a plant growth-promoting bacterium (PGPB). The targeted gene expression analysis shows higher gene expression in the spores and in the mycelium with the application of PGPB. Besides analyzing key cell-wall-related genes, this study also identifies the most suitable reference genes to normalize qPCR results in D. teres, thus serving as a basis for future molecular studies on this ascomycete.

Cell wall composition and transcriptomics in stem tissues of stinging nettle (Urtica dioica L.): Spotlight on a neglected fibre crop.

Stinging nettle (Urtica dioica L.) produces silky cellulosic fibres, as well as bioactive molecules. To improve the knowledge on nettle and enhance its opportunities of exploitation, a draft transcriptome of the "clone 13" (a fibre clone) is here presented. The transcriptome of whole internodes sampled at the top and middle of the stem is then compared with the core and cortical tissues sampled at the bottom. Young internodes show an enrichment in genes involved in the biosynthesis of phytohormones (auxins and jasmonic acid) and secondary metabolites (flavonoids). The core of internodes collected at the bottom of the stem is enriched in genes partaking in different aspects of secondary cell wall formation (cellulose, hemicellulose, lignin biosynthesis), while the cortical tissues reveal the presence of a C starvation signal probably due to the UDP-glucose demand necessary for the thickening phase of bast fibres. Cell wall analysis indicates a difference in rhamnogalacturonan structure/composition of mature bast fibres, as evidenced by the higher levels of galactose measured, as well as the occurrence of more water-soluble pectins in elongating internodes. The targeted quantification of phenolics shows that the middle internode and the cortical tissues at the bottom have higher contents than top internodes. Ultrastructural analyses reveal the presence of a gelatinous layer in bast fibres with a lamellar structure. The data presented will be an important resource and reference for future molecular studies on a neglected fibre crop.