Evaluation of the effect of pre-sowing electron irradiation of barley seeds on plant development and disease incidence

The effect of electron irradiation of barley seeds of the Vladimir variety on the development of plants, disease incidence and productivity in controlled greenhouse conditions has been studied. It was found that the effectiveness of the effect of seed treatment with low-energy electronic radiation in the dose range of 30–150 kGy on the morphometric parameters of barley largely depends on the accelerating voltage – 130 kV (mode 1) and 160 kV (mode 2). It was noted that irradiation suppressed the formation of chlorophyll a and b and carotenoids in barley leaves at all doses and irradiation regimes and, at the tendency level, reduced the infestation of Bipolaris sorokiniana plants in the tillering and heading phases. It was shown that irradiation of seeds caused an increase in the total tillering of plants (mode 2), but at the same time reduced the productive tillering. Electron irradiation caused an increase in the mass of 1000 grains and did not affect the yield of grain and straw.

Extracellular vesicles isolated from dsRNA sprayed barley plants exhibit no growth inhibition or gene silencing in Fusarium graminearum

Incorporating a double-stranded RNA (dsRNA)-expressing transgene into plants or applying dsRNA by spraying it onto plant leaves successfully protects plants against invading pathogens with RNA interference (RNAi). How dsRNAs or siRNAs are transferred between donor host cells and recipient fungal cells is largely unknown It is speculated that plant extracellular vesicles (EVs) function as RNA shuttles between plants and their interacting pathogens. Recently, we found that EVs isolated from HIGS or SIGS plants contained dsRNA-derived siRNAs. In this study, we evaluated whether isolated EVs from RNA-sprayed barley ( Hordeum vulgare ) plants affected the growth of the phytopathogenic ascomycete Fusarium graminearum ( Fg ). Encouraged by our previous finding that dropping barley-derived EVs on Fg cultures caused fungal stress phenotypes, we conducted an in vitro growth experiment in microtiter plates where we co-cultivated Fg with plant EVs isolated from dsRNA-sprayed barley leaves. We observed that co-cultivation of Fg macroconidia with barley EVs did not affect fungal growth. Furthermore, plant EVs containing SIGS-derived siRNA appeared not to affect Fg growth and showed no gene silencing activity on FgCYP51 genes. We conclude that either the amount of spray-derived sRNA was insufficient to induce target gene silencing (SIGS) in Fg or Fg uptake of plant EVs from liquid cultures was inefficient or impossible.

MoGLN2 Is Important for Vegetative Growth, Conidiogenesis, Maintenance of Cell Wall Integrity and Pathogenesis of Magnaporthe oryzae

Glutamine is a non-essential amino acid that acts as a principal source of nitrogen and nucleic acid biosynthesis in living organisms. In Saccharomyces cerevisiae, glutamine synthetase catalyzes the synthesis of glutamine. To determine the role of glutamine synthetase in the development and pathogenicity of plant fungal pathogens, we used S. cerevisiae Gln1 amino acid sequence to identify its orthologs in Magnaporthe oryzae and named them MoGln1, MoGln2, and MoGln3. Deletion of MoGLN1 and MoGLN3 showed that they are not involved in the development and pathogenesis of M. oryzae. Conversely, ∆Mogln2 was reduced in vegetative growth, experienced attenuated growth on Minimal Medium (MM), and exhibited hyphal autolysis on oatmeal and straw decoction and corn media. Exogenous l-glutamine rescued the growth of ∆Mogln2 on MM. The ∆Mogln2 mutant failed to produce spores and was nonpathogenic on barley leaves, as it was unable to form an appressorium-like structure from its hyphal tips. Furthermore, deletion of MoGLN2 altered the fungal cell wall integrity, with the ∆Mogln2 mutant being hypersensitive to H2O2. MoGln1, MoGln2, and MoGln3 are located in the cytoplasm. Taken together, our results shows that MoGLN2 is important for vegetative growth, conidiation, appressorium formation, maintenance of cell wall integrity, oxidative stress tolerance and pathogenesis of M. oryzae.

Proteinous pancreatic lipase inhibitor is responsible for the antiobesity effect of young barley (Hordeum vulgare L.) leaf extract

Young barley leaves (Hordeum vulgare L.) have various health effects and are employed as an ingredient in the production of health-promoting foods. Promoting antiobesity is one such health effect; however, the mechanism and bioactive compounds are unclear. In this research, young barley leaf extract (YB) was demonstrated to possess pancreatic lipase inhibitory activity. The addition of YB to a high-fat diet in mice increased fecal lipid content, indicating reduced absorption of lipids as the mechanism underlying antiobesity effect. The investigation of bioactive compounds in YB resulted in the identification of fructose–bisphosphate aldolase as a proteinous lipase inhibitor. Maximum inhibition of the protein was 45%, but inhibition was displayed at a concentration as low as 16 ng/mL, which is a characteristic inhibition compared with other reported proteinous lipase inhibitors.

Evaluation of dsRNA delivery methods for targeting macrophage migration inhibitory factor MIF in RNAi-based aphid control

Macrophage migration inhibitory factors (MIFs) are multifunctional proteins regulating major processes in mammals, including activation of innate immune responses. In invertebrates, MIF proteins participate in the modulation of host immune responses when secreted by parasitic organisms, such as aphids. In this study, we assessed the possibility to use MIF genes as targets for RNA interference (RNAi)-based control of the grain aphid Sitobion avenae (Sa) on barley (Hordeum vulgare). When nymphs were fed on artificial diet containing double-stranded (ds)RNAs (SaMIF-dsRNAs) that target sequences of the three MIF genes SaMIF1, SaMIF2 and SaMIF3, they showed higher mortality rates and these rates correlated with reduced MIF transcript levels as compared to the aphids feeding on artificial diet containing a control dsRNA (GFP-dsRNA). Comparison of different feeding strategies showed that nymphs’ survival was not altered when they fed from barley seedlings sprayed with naked SaMIF-dsRNAs, suggesting they did not effectively take up dsRNA from the sieve tubes of these plants. Furthermore, aphids’ survival was also not affected when the nymphs fed on leaves supplied with dsRNA via basal cut ends of barley leaves. Consistent with this finding, the use of sieve tube-specific YFP-labeled Arabidopsis reporter lines confirmed that fluorescent 21 nt dsRNACy3, when supplied via petioles or spraying, co-localized with xylem structures, but not with phloem tissue. Our results suggest that MIF genes are a potential target for insect control and also imply that application of naked dsRNA to plants for aphid control is inefficient. More efforts should be put into the development of effective dsRNA formulations.

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.

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.

Light and CO2 Modulate the Accumulation and Localization of Phenolic Compounds in Barley Leaves

Barley (Hordeum vulgare) accumulates phenolic compounds (PhCs), which play a key role in plant defense against environmental stressors as antioxidants or UV screening compounds. The influence of light and atmospheric CO2 concentration ([CO2]) on the accumulation and localization of PhCs in barley leaves was examined for two varieties with different tolerances to oxidative stress. PhC localization was visualized in vivo using fluorescence microscopy. Close relationships were found between fluorescence-determined localization of PhCs in barley leaves and PhC content estimated using liquid chromatography coupled with mass spectroscopy detection. Light intensity had the strongest effect on the accumulation of PhCs, but the total PhC content was similar at elevated [CO2], minimizing the differences between high and low light. PhCs localized preferentially near the surfaces of leaves, but under low light, an increasing allocation of PhCs in deeper mesophyll layers was observed. The PhC profile was significantly different between barley varieties. The relatively tolerant variety accumulated significantly more hydroxycinnamic acids, indicating that these PhCs may play a more prominent role in oxidative stress prevention. Our research presents novel evidence that [CO2] modulates the accumulation of PhCs in barley leaves. Mesophyll cells, rather than epidermal cells, were most responsive to environmental stimuli in terms of PhC accumulation.

Evaluation of dsRNA delivery methods for targeting macrophage migration inhibitory factor MIF in RNAi-based aphid control

Macrophage migration inhibitory factors (MIF) are multifunctional proteins regulating major processes in mammals, including activation of innate immune responses. In invertebrates, MIF proteins participate in the modulation of host immune responses when secreted by parasitic organisms, such as aphids. In this study, we assessed the possibility to use MIF genes as targets for RNA interference (RNAi)-based control of the grain aphid Sitobion avenae (Sa) on barley (Hordeum vulgare). When nymphs were fed on artificial diet containing double-stranded (ds)RNAs (SaMIF-dsRNAs) that target sequences of the three MIF genes SaMIF1, SaMIF2 and SaMIF3, they showed higher mortality rates and these rates correlated with reduced MIF transcript levels as compared to the aphids feeding on artificial diet containing a control dsRNA (GFP-dsRNA). Comparison of different feeding strategies showed that nymphs’ survival was not altered when they fed from barley seedlings sprayed with SaMIF-dsRNAs, suggesting they did not effectively take up dsRNA from the sieve tubes of these plants. Furthermore, aphids’ survival was also not affected when the nymphs fed on leaves supplied with dsRNA via basal cut ends of barley leaves. Consistent with this finding, the use of sieve-tube-specific YFP-labeled Arabidopsis reporter lines confirmed that fluorescent 21 nt dsRNACy3 supplied via petioles co-localized with xylem structures, but not with phloem tissue. Our results suggest that MIF genes are a potential target for insect control and also imply that application of naked dsRNA to plants for aphid control is inefficient. More efforts should be put into the development of effective dsRNA formulations.