Identification of a Chitooligosaccharide Mechanism against Bacterial Leaf Blight on Rice by In Vitro and In Silico Studies.

This study focuses on a commercial plant elicitor based on chitooligosaccharides (BIG®), which aids in rice plant growth and disease resistance to bacterial leaf blight (BLB). When the pathogen (Xoo) vigorously attacks rice that has suffered yield losses, it can cause damage in up to 20% of the plant. Furthermore, Xoo is a seed-borne pathogen that can survive in rice seeds for an extended period. In this study, when rice seeds were soaked and sprayed with BIG®, there was a significant increase in shoot and root length, as well as plant biomass. Furthermore, BIG®-treated rice plants showed a significant reduction in BLB severity of more than 33%. Synchrotron radiation-based Fourier transform infrared (SR-FTIR) analysis was used to characterize BIG®'s mechanism in the chemical structure of rice leaves. The SR-FTIR results at 1650, 1735, and 1114 cm-1 indicated changes in biochemical components such as pectins, lignins, proteins, and celluloses. These findings demonstrated that commercial BIG® not only increased rice growth but also induced resistance to BLB. The drug's target enzyme, Xoo 1075 from Xanthomonas oryzae (PDB ID: 5CY8), was analyzed for its interactions with polymer ingredients, specifically chitooligosaccharides, to gain molecular insights down to the atomic level. The results are intriguing, with a strong binding of the chitooligosaccharide polymer with the drug target, revealing 10 hydrogen bonds between the protein and polymer. Overall, the computational analysis supported the experimentally demonstrated strong binding of chitooligosaccharides to the drug target.

Rhizosphere Microorganisms Supply Availability of Soil Nutrients and Induce Plant Defense.

Plant health is necessary for food security, which is a key determinant of secure and sustainable food production systems. Deficiency of soil nutrients and invasion of plant pathogens or insects are the main destroyers of the world's food production. Synthetic fertilizers and chemical-based pesticides are frequently employed to combat the problems. However, these have negative impacts on microbial ecosystems and ecosystem functioning. Rhizosphere microorganisms have demonstrated their potency to improve or manage plant nutrients to encourage plant growth, resulting in increased yield and quality by converting organic and inorganic substances around the rhizosphere zone into available plant nutrients. Besides regulating nutrient availability and plant growth enhancement, rhizobacteria or fungi can restrict plant pathogens that cause disease by secreting inhibitory chemicals and boosting plant immunity to combat pests or pathogens. Thus, rhizosphere microorganisms are viewed as viable, alluring economic approaches for sustainable agriculture as biofertilizers and biopesticides. This review provides an overview of the role of rhizosphere microorganisms in soil nutrients and inducing of plant defenses. Moreover, a discussion is presented surrounding the recent consequences of employing these microorganisms and a sustainable strategy towards improving fertilization effectiveness, and encouraging stronger, more pest-resistant plants.

Bio-synthesis and characterization of silver nanoparticles from Trichoderma species against cassava root rot disease.

Cassava root rot disease caused by the fungal pathogens Fusarium solani and Lasiodiplodia theobromae produces severe damages on cassava production. This research was conducted to produce and assess silver nanoparticles (AgNPs) synthesized by Trichoderma harzianum for reducing root rot disease. The results revealed that using the supernatants of T. harzianum on a silver nitrate solution changed it to reddish color at 48 h, indicating the formation of AgNPs. Further characterization was identified using dynamic light scattering (DLS) and scanning electron microscope (SEM). DLS supported that the Z-average size is at 39.79 nm and the mean zeta potential is at - 36.5 mV. SEM revealed the formation of monodispersed spherical shape with a diameter between 60-75 nm. The antibacterial action of AgNPs as an antifungal agent was demonstrated by an observed decrease in the size of the fungal colonies using an increasing concentration of AgNPs until the complete inhibition growth of L. theobromae and F. solani at > 58 µg mL-1 and at ≥ 50 µg mL-1, respectively. At in vitro conditions, the applied AgNPs caused a decrease in the percentage of healthy aerial hyphae of L. theobromae (32.5%) and of F. solani (70.0%) compared to control (100%). The SR-FTIR spectra showed the highest peaks in the first region (3000-2800 cm-1) associated with lipids and fatty acids located at 2962, 2927, and 2854 cm-1 in the AgNPs treated samples. The second region (1700-1450 cm-1) consisting of proteins and peptides revealed the highest peaks at 1658, 1641, and 1548 cm-1 in the AgNPs treated samples. The third region (1300-900 cm-1), which involves nucleic acid, phospholipids, polysaccharides, and carbohydrates, revealed the highest peaks at 1155, 1079, and 1027 cm-1 in the readings from the untreated samples. Finally, the observed root rot severity on cassava roots treated with AgNPs (1.75 ± 0.50) was significantly lower than the control samples (5.00 ± 0.00).

Bacillus vallismortis TU-Orga21 blocks rice blast through both direct effect and stimulation of plant defense.

Beneficial microorganisms are an important strategy for sustainable plant production processes such as stimulate root exudation, stress tolerance, and yield improvement. This study investigated various microorganisms isolated from the rhizosphere of Oryza sativa L. in order to inhibit Magnaporthe oryzae cause of rice blast, by direct and indirect mode of action. The results indicated that Bacillus vallismortis strain TU-Orga21 significantly reduced M. oryzae mycelium growth and deformed the hyphal structures. The effects of biosurfactant TU-Orga21 was studied against M. oryzae spore development. The dose of ≥5% v/v biosurfactant significantly inhibited the germ tubes and appressoria formation. The biosurfactants were evaluated as surfactin and iturin A by Matrix-assisted laser desorption ionization dual time-of-flight tandem mass spectrometry. Under greenhouse conditions, priming the biosurfactant three times before M. oryzae infection significantly accumulated endogenous salicylic acid, phenolic compounds, and hydrogen peroxide (H2O2) during the infection process of M. oryzae. The SR-FT-IR spectral changes from the mesophyll revealed higher integral area groups of lipids, pectins, and proteins amide I and amide II in the elicitation sample. Furthermore, scanning electron microscope revealed appressorium and hyphal enlargement in un-elicitation leaves whereas appressorium formation and hyphal invasion were not found in biosurfactant-elicitation at 24 h post inoculation. The biosurfactant treatment significantly mitigated rice blast disease severity. Therefore, B. vallismortis can be a promising novel biocontrol agent which contains the preformed active metabolites for a rapid control of rice blast by a direct action against pathogen and by boosting plant immunity.

Resistance Induction by Salicylic Acid Formulation in Cassava Plant against Fusarium solani.

Fusarium root rot caused by the soil-borne fungus Fusarium solani is one of the most important fungal diseases of cassava in Thailand, resulting in high yield losses of more than 80%. This study aimed to investigate if the exogenous application of salicylic acid formulations (Zacha) can induce resistance in cassava against Fusarium root rot and observe the biochemical changes in induced cassava leaf tissues through synchrotron radiation based on Fourier-transform infrared (SR-FTIR) microspectroscopy. We demonstrated that the application of Zacha11 prototype formulations could induce resistance against Fusarium root rot in cassava. The in vitro experimental results showed that Zacha11 prototype formulations inhibited the growth of F. solani at approximately 34.83%. Furthermore, a significant reduction in the disease severity of Fusarium root rot disease at 60 days after challenge inoculation was observed in cassava plants treated with Zacha11 at a concentration of 500 ppm (9.0%). Population densities of F. solani were determined at 7 days after inoculation. Treatment of the Zacha11 at a concentration of 500 ppm resulted in reduced populations compared with the distilled water control and differences among treatment means at each assay date. Moreover, the SR-FTIR spectral changes of Zacha11-treated epidermal tissues of leaves had higher integral areas of lipids, lignins, and pectins (1,770-1,700/cm), amide I (1,700-1,600/cm), amide II (1,600-1,500/cm), hemicellulose, lignin (1,300-1,200/cm), and cellulose (1,155/cm). Therefore, alteration in defensive carbohydrates, lipids, and proteins contributed to generate barriers against Fusarium invasion in cassava roots, leading to lower the root rot disease severity.

Efficacy of Chitosan Nanoparticle Loaded-Salicylic Acid and -Silver on Management of Cassava Leaf Spot Disease.

Leaf spot is one of the most important cassava diseases. Nanotechnology can be applied to control diseases and improve plant growth. This study was performed to prepare chitosan (CS) nanoparticle (NP)-loaded salicylic acid (SA) or silver (Ag) by the ionic gelation method, and to evaluate their effectiveness on reducing leaf spot disease and enhancing the growth of cassava plants. The CS (0.4 or 0.5%) and Pentasodium triphosphate (0.2 or 0.5%) were mixed with SA varying at 0.05, 0.1, or 0.2% or silver nitrate varying at 1, 2, or 3 mM to prepare three formulations of CS-NP-loaded SA named N1, N2, and N3 or CS-NP-loaded Ag named N4, N5, and N6. The results showed that the six formulations were not toxic to cassava leaves up to 800 ppm. The CS-NP-loaded SA (N3) and CS-NP-loaded Ag (N6) were more effective than the remaining formulations in reducing the disease severity and the disease index of leaf spot. Furthermore, N3 at 400 ppm and N6 at 200, 400, and 800 ppm could reduce disease severity (68.9-73.6% or 37.0-37.7%, depending on the time of treatment and the pathogen density) and enhance plant growth more than or equal to commercial fungicide or nano-fungicide products under net-house conditions. The study indicates the potential to use CS-NP-loaded SA or Ag as elicitors to manage cassava leaf spot disease.

Chitosan Nanoparticles-Based Ionic Gelation Method: A Promising Candidate for Plant Disease Management.

By 2050, population growth and climate change will lead to increased demand for food and water. Nanoparticles (NPs), an advanced technology, can be applied to many areas of agriculture, including crop protection and growth enhancement, to build sustainable agricultural production. Ionic gelation method is a synthesis of microparticles or NPs, based on an electrostatic interaction between opposite charge types that contains at least one polymer under mechanical stirring conditions. NPs, which are commonly based on chitosan (CS), have been applied to many agricultural fields, including nanopesticides, nanofertilizers, and nanoherbicides. The CS-NP or CS-NPs-loaded active ingredients (Cu, saponin, harpin, Zn, hexaconazole, salicylic acid (SA), NPK, thiamine, silicon, and silver (Ag)) are effective in controlling plant diseases and enhancing plant growth, depending on the concentration and application method by direct and indirect mechanisms, and have attracted much attention in the last five years. Many crops have been evaluated in in vivo or in greenhouse conditions but only maize (CS-NP-loaded Cu, Zn, SA, and silicon) and soybean (CS-NP-loaded Cu) were tested for manage post flowering stalk rot, Curvularia leaf spot, and bacterial pustule disease in field condition. Since 2019, five of eight studies have been performed in field conditions that have shown interest in CS-NPs synthesized by the ionic gelation method. In this review, we summarized the current state of research and provided a forward-looking view of the use of CS-NPs in plant disease management.

Investigation of bioactive compounds from Bacillus sp. against protein homologs CDC42 of Colletotrichum gloeosporioides causing anthracnose disease in cassava by using molecular docking and dynamics studies.

Manihot esculenta, commonly called cassava, is an economically valuable crop and important staple food, grown in tropical and subtropical regions of the world. Demand for cassava in the food and fuel industry is growing worldwide. However, anthracnose disease caused by Colletotrichum gloeosporioides severely affects cassava yield and production. The bioactive molecules from Bacillus are widely used to control fungal diseases in several plants. Therefore, in this study, bioactive compounds (erucamide, behenic acid, palmitic acid, phenylacetic acid, and ß-sitosterol) from Bacillus megaterium were assessed against CDC42, a key protein for virulence, from C. gloeosporioides. Structure of the CDC42 protein was generated through the comparative homology modeling method. The binding site of the ligands and the stability of the complex were analyzed through docking and molecular dynamics simulation studies, respectively. Furthermore, a protein interaction network was envisaged through the STRING database, followed by enrichment analysis in the WebGestalt tool. From the enrichment analysis, it is apparent that bioactive from B. megaterium chiefly targets the MAP kinase pathway that is essential for filamentous growth and virulence. Further exploration through experimental studies could be advantageous for cassava improvement as well as to combat against C. gloeosporioides pathogen.

Identification of Salicylic Acid Mechanism against Leaf Blight Disease in Oryza sativa by SR-FTIR Microspectroscopic and Docking Studies.

The present study was to investigate the application and mechanism of salicylic acid (SA) as SA-Ricemate for the control of leaf blight disease using a Synchrotron Radiation-based Fourier-Transform Infra-Red (SR-FTIR) microspectroscopy and docking studies. After treating rice plants cv. KDML 105 with SA-Ricemate, the leaves were inoculated with Xanthomonas oryzae pv. oryzae, the causal agent of leaf blight, and disease severity were assessed. The leaves were also used to detect changes in endogenous SA content. The results indicated that SA-Ricemate, as an activated compound, reduced disease severity by 60% at three weeks post-inoculation and increased endogenous content by 50%. The SR-FTIR analysis of changes in the mesophyll of leaves (treated and untreated) showed that the groups of lipids, pectins, and proteins amide I and amide II occurred at higher values, and polysaccharides were shown at lower values in treated compared to untreated. Besides, docking studies were used to model a three-dimensional structure for Pathogenesis-related (PR1b) protein and further identify its interaction with SA. The results showed that ASP28, ARG31, LEU32, GLN97, and ALA93 are important residues that have strong hydrogen bonds with SA. The docking results showed that SA has a good interaction, confirming its role in expression.