Bacterial chemotaxis of herbicide atrazine provides an insight into the degradation mechanism through intermediates hydroxyatrazine, N-N-isopropylammelide, and cyanuric acid compounds.

In recent times, the herbicide atrazine (ATZ) has been commonly used before and after the cultivation of crop plants to manage grassy weeds. Despite its effect, the toxic residues of ATZ affect soil fertility and crop yield. Hence, the current study is focused on providing insight into the degradation mechanism of the herbicide atrazine through bacterial chemotaxis involving intermediates responsive to degradation. A bacterium was isolated from ATZ-contaminated soil and identified as Pseudomonas stutzeri based on its morphology, biochemical and molecular characterization. Upon ultra-performance liquid chromatography analysis, the free cells of isolated bacterium strain was found to utilize 174 µg/L of ATZ after 3-days of incubation on a mineral salt medium containing 200 µg/L of ATZ as a sole carbon source. It was observed that immobilized based degradation of ATZ yielded 198 µg/L and 190 µg/L by the cells entrapped with silica beads and sponge, respectively. Furthermore, the liquid chromatography-mass spectroscopy revealed that the secretion of three significant metabolites, namely, cyanuric acid, hydroxyatrazine and N- N-Isopropylammelide is responsive to the biodegradation of ATZ by the bacterium. Collectively, this research demonstrated that bacterium strains are the most potent agent for removing toxic pollutants from the environment, thereby enhancing crop yield and soil fertility with long-term environmental benefits.

Genome-Wide Characterization of Effector Protein-Encoding Genes in Sclerospora graminicola and Its Validation in Response to Pearl Millet Downy Mildew Disease Stress.

Pearl millet [Pennisetum glaucum (L.) R. Br.] is the essential food crop for over ninety million people living in drier parts of India and South Africa. Pearl millet crop production is harshly hindered by numerous biotic stresses. Sclerospora graminicola causes downy mildew disease in pearl millet. Effectors are the proteins secreted by several fungi and bacteria that manipulate the host cell structure and function. This current study aims to identify genes encoding effector proteins from the S. graminicola genome and validate them through molecular techniques. In silico analyses were employed for candidate effector prediction. A total of 845 secretory transmembrane proteins were predicted, out of which 35 proteins carrying LxLFLAK (Leucine-any amino acid-Phenylalanine-Leucine-Alanine-Lysine) motif were crinkler, 52 RxLR (Arginine, any amino acid, Leucine, Arginine), and 17 RxLR-dEER putative effector proteins. Gene validation analysis of 17 RxLR-dEER effector protein-producing genes was carried out, of which 5genes were amplified on the gel. These novel gene sequences were submitted to NCBI. This study is the first report on the identification and characterization of effector genes in Sclerospora graminicola. This dataset will aid in the integration of effector classes that act independently, paving the way to investigate how pearl millet responds to effector protein interactions. These results will assist in identifying functional effector proteins involving the omic approach using newer bioinformatics tools to protect pearl millet plants against downy mildew stress. Considered together, the identified effector protein-encoding functional genes can be utilized in screening oomycetes downy mildew diseases in other crops across the globe.

Biofabricated yeast: super-soldier for detoxification of heavy metals.

The advances in nanotechnology have shown enormous impacts in environmental technology as a potent weapon for degradation of toxic organic pollutants and detoxification of heavy metals. It is either by in-situ or ex-situ adaptive strategies. Mycoremediation of environmental pollutants has been a success story of the past decade, by employing the wide arsenal of biological capabilities of fungi. Recently, the proficiency and uniqueness of yeast cell surface alterations have encouraged the generation of engineered yeast cells as dye degraders, heavy metal reduction and its recovery, and also as detoxifiers of various hazardous xenobiotic compounds. As a step forward, recent trends in research are towards developing biologically engineered living materials as potent, biocompatible and reusable hybrid nanomaterials. They include chitosan-yeast nanofibers, nanomats, nanopaper, biosilica hybrids, and TiO2-yeast nanocomposites. The nano-hybrid materials contribute significantly as supportive stabilizer, and entrappers, which enhances the biofabricated yeast cells' functionality. This field serves as an eco-friendly cutting-edge cocktail research area. In this review, we highlight recent research on biofabricated yeast cells and yeast-based biofabricated molecules, as potent heavy metals, toxic chemical detoxifiers, and their probable mechanistic properties with future application perspectives.

Biodegradable hybrid biopolymer film based on carboxy methyl cellulose and selenium nanoparticles with antifungal properties to enhance grapes shelf life.

In the current study, cellulose was extracted from sugarcane bagasse and further converted into carboxy methyl cellulose. The morphological, chemical, and structural characterization of synthesizeed carboxy methyl cellulose was performed. Further, the biopolymer was fabricated with mycogenic selenium nanoparticles and used to develop the biopolymer films. The developed biopolymer films were examined for the fruit shelf life stability, antifungal activity, and biodegradation potential. The results revealed that grapes wrapped with biofilms showed enhanced shelf life of fruit at all storage time intervals. The study also witnesses the antifungal activity of biopolymer films with a remarkable inhibitory action on the spores of Fusarium oxysporum and Sclerospora graminicola phytopathogens. Lastly, the biopolymer films were significantly degradable in the soil within two weeks of incubation. Thus, the developed biopolymer films exhibit multifaceted properties that can be used as an alternative to synthetic plastics for fruit packaging and also helps in protecting against fungal contaminants during storage with naturally degradable potential.

Chitosan and chitosan-derived nanoparticles modulate enhanced immune response in tomato against bacterial wilt disease.

The present study evaluated the priming efficacy of chitosan and chitosan-derived nanoparticles (CNPs) against bacterial wilt of tomato. In the current study, seed-treated CNPs plus pathogen-inoculated tomato seedlings recorded significant protection of 62 % against pathogen-induced wilt disease and subsequently better growth. The induced resistance was witnessed by a prominent increase in lignin, callose and H2O2 deposition, followed by superoxide radical accumulation in leaves. Additionally, chitosan and CNPs-treated tomato plants recorded a remarkable increase in the upregulation of phenylalanine ammonia-lyase (PAL), peroxidase (POX), polyphenol oxidase (PPO), catalase (CAT) and ß-1, 3 glucanase (GLU) in comparison with untreated plants. The chitosan and CNPs-induced antioxidant enzymes were positively correlated with the stimulation of corresponding gene expression in CNPs treated plants related to pathogen-inoculated ones. The results of this study describe that how the application of chitosan and CNPs elicit defense responses at the cellular, biochemical and gene expression in tomato plants against bacterial wilt disease, thereby improve growth and yield.

Priming with fungal elicitor elicits early signaling defense against leaf spot of broccoli underlying cellular, biochemical and gene expression.

The phytopathogen Alternaria, which causes leaf spot disease in broccoli plants, has developed the ability to adapt to changing climatic conditions with the failure of conventional fungicides. To restrict its infection and further spread, the application of novel molecules that have the ability to trigger innate immune responses of the plants to concurrent pathogenic invasions is essential. In this context, the current study was set out to assess Trichoderma-derived trehalose molecules' beneficial effects from inciting resistance mechanisms in broccoli plants against Alternaria brassicicola at histochemical, biochemical and genetic levels. From the results of the greenhouse experiment, it was evident that trehalose proved to be an exceptional elicitor in combating broccoli leaf spot disease. The induced resistance is positively associated with an early hypersensitive reaction expressed in the primed seedlings. Additionally, the deposition of histochemical such as callose and phenols was found to be accelerated in the tripartite system at 24 and 48 h after post-inoculation (hpi). At the biochemical level, the activities of the enzymes chitinase (CHI), catalase (CAT) and allene oxide synthase (AOS) were significantly enhanced in trehalose-treated broccoli plants inoculated with A. brassicicola. The enzymatic activities were further validated by quantitative real-time polymerase chain reaction (qRT-PCR) analyses wherein the relative expressions of the corresponding gene transcripts were up-regulated in trehalose primed plants. Conclusively, the investigation results have provided vital insights into the histochemical, biochemical and gene expression underlying the enhanced induced systemic resistance against broccoli leaf spot disease.

Genome-Wide Identification, Diversification, and Expression Analysis of Lectin Receptor-Like Kinase (LecRLK) Gene Family in Cucumber under Biotic Stress.

Members of the lectin receptor-like kinase (LecRLKs) family play a vital role in innate plant immunity. Few members of the LecRLKs family have been characterized in rice and Arabidopsis, respectively. However, little literature is available about LecRLKs and their role against fungal infection in cucumber. In this study, 60 putative cucumber LecRLK (CsLecRLK) proteins were identified using genome-wide analysis and further characterized into L-type LecRLKs (24) and G-type LecRLKs (36) based on domain composition and phylogenetic analysis. These proteins were allocated to seven cucumber chromosomes and found to be involved in the expansion of the CsLecRLK gene family. Subcellular localization of CsaLecRLK9 and CsaLecRLK12 showed green fluorescence signals in the plasma membrane of leaves. The transcriptional profiling of CsLecRLK genes showed that L-type LecRLKs exhibited functional redundancy as compared to G-type LecRLKs. The qRT-PCR results indicated that both L- and G-type LecRLKs showed significant response against plant growth-promoting fungi (PGPF-Trichoderma harzianum Rifai), powdery mildew pathogen (PPM-Golovinomyces orontii (Castagne) V.P. Heluta), and combined (PGPF+PPM) treatments. The findings of this study contribute to a better understanding of the role of cucumber CsLecRLK genes in response to PGPF, PPM, and PGPF+PPM treatments and lay the basis for the characterization of this important functional gene family.

Ameliorated Antibacterial and Antioxidant Properties by Trichoderma harzianum Mediated Green Synthesis of Silver Nanoparticles.

Biosynthesis of silver nanoparticles using beneficial Trichoderma harzianum is a simple, eco-friendly and cost-effective route. Secondary metabolites secreted by T. harzianum act as capping and reducing agents that can offer constancy and can contribute to biological activity. The present study aimed to synthesize silver nanoparticles using T. harzianum cell filtrate and investigate different bioactive metabolites based on LC-MS/MS analysis. The synthesized silver nanoparticles (AgNPs) from T. harzianum were characterized by ultraviolet-visible spectrophotometry, Fourier transform infrared spectrometry (FT-IR), energy-dispersive spectroscopy (EDS), dynamic light scattering (DLS), X-ray powder diffraction (XRD) and scanning electron microscopy (SEM). The surface plasmon resonance of synthesized particles formed a peak centered near 438 nm. The DLS study determined the average size of AgNPs to be 21.49 nm. The average size of AgNPs was measured to be 72 nm by SEM. The cubic crystal structure from XRD analysis confirmed the synthesized particles as silver nanoparticles. The AgNPs exhibited remarkable antioxidant properties, as determined by DPPH and ferric reducing antioxidant power (FRAP) assay. The AgNPs also exhibited broad-spectrum antibacterial activity against two Gram-positive bacteria (S. aureus and B. subtilis) and two Gram-negative bacteria (E. coli and R. solanacearum). The minimum inhibitory concentration (MIC) of AgNPs towards bacterial growth was evaluated. The antibacterial activity of AgNPs was further confirmed by fluorescence microscopy and SEM analysis.

Trehalose: A mycogenic cell wall elicitor elicit resistance against leaf spot disease of broccoli and acts as a plant growth regulator.

Elicitors are biochemicals, and the cell wall-derived elicitors from fungi can trigger defence mechanisms in plants by increasing the phytoalexin accumulation when they encounter the pathogens. The main objective of this research was to purify and characterize a cell wall elicitor from Trichoderma atroviride (TaCWE) and evaluate the seed priming effect of that elicitor for inducing systemic resistance in broccoli plants against leaf spot disease. Amongst the tested TaCWE concentrations of the seed priming (5, 10, & 25 mg ml-1), 10.0 mg ml-1 showed significantly (P < 0.05) improved early emergence, the rate of germination at 94%, and observed seedling vigour of 2601. Also, elicitor (10 mg ml-1) treatment alone induced 57% plant protection. On the contrary, the elicitor treated and pathogen inoculated plants induced a notable 72% protection against leaf spot disease of broccoli caused by A. brassicicola. Thus, the primed seeds with elicitor showed induced disease resistance and plant growth promotion. The prominent molecule present in the purified extracted cell wall elicitor is confirmed as trehalose. The AFM analysis indicated the trehalose length and width as 10.16 µm and 2.148 µm, respectively. FTIR chromatogram further confirmed trehalose in abundance with traces of carbon, hydrogen, nitrogen, oxygen, and LC-MS profile with a single peak eluted with a retention time of 3.78 min. The findings of this study contribute to understanding better the role of trehalose, a biogenic cell-wall elicitor that can induce systemic resistance against leaf spot disease and regulate plant growth in the broccoli plants.

Myco-engineered selenium nanoparticles elicit resistance against tomato late blight disease by regulating differential expression of cellular, biochemical and defense responsive genes.

With the advent of rapid evolution of oomycete pathogen lineages, the need for sustainable agriculture practices has become the need of the hour. The late blight of tomato caused by Phytopthora infestans, has recently emerged as one such devastating disease in India that led to huge crop losses. Hence, in the present work seed priming with mycogenic selenium nanoparticles (SeNPs) for elicitation of resistance against tomato late blight disease is investigated. It also aims to understand the defense responses triggered by SeNPs at cellular, biochemical and transcriptomic levels. Enhanced plant growth parameters were observed in bioactive SeNPs-primed tomato plants as compared to control plants. SeNPs-primed and pathogen inoculated plants exhibited a significant protection of 72.9 % against late blight disease. The primed plants also recorded a remarkable accumulation of lignin, callose and hydrogen peroxide that serve as the cellular defense over the control plants. Further, an elevated level of lipoxygenase (LOX), phenylalanine lyase (PAL), ß-1,3-glucanase (GLU), superoxide dismutase (SOD) corroborated the biochemical defense in primed plants, which was also reflected in the corresponding transcriptome profiling of the genes encoding the enzymes. Thus, the present study represents an orchestrated correlation between resistance and defense responses incited by SeNPs against tomato late blight disease, which can be used as nano-biostimulant fungicide in protecting tomato plants.

Exogenous priming of chitosan induces upregulation of phytohormones and resistance against cucumber powdery mildew disease is correlated with localized biosynthesis of defense enzymes.

In recent years, exploration of biopolymer-based materials to avoid hazardous chemicals in agriculture has gained enormous importance for sustainable crop improvement. In the present study, chitosan a biopolymer derived from crab-shell was used in different concentrations as priming agent to cucumber seeds and were evaluated for its effect to enhance plant growth parameters as well as its ability to induce resistance against powdery mildew disease. Among the treatments, seeds-primed with 2.5 mg/mL exhibited early seedling germination of 90% and vigour of 2665 and also remarkably enhanced the cucumber growth parameters which might be fairly attributed to the stimulation of phytohormones content in primed plants over the controls. More importantly, under greenhouse conditions a significant induced disease protection of 66.6% against powdery mildew disease was noticed in chitosan-pretreated plants at 2.5 mg/mL. The induced resistant plants also showed a significant deposition of lignin, callose and H2O2. Notably, polyphenol oxidase, phenylalanine ammonia-lyase, peroxidase and glucanase defense-responsive enzymes were upregulated in chitosan-primed plants. Considered together, these results determine that the susceptible cucumber cultivar elicits immunity after perception of priming with chitosan to upregulate phytohormones and synthesize defense-responsive enzymes, thereby induce resistance against powdery mildew disease and strengthen the growth-promotion of cucumber plants.

Synchronised regulation of disease resistance in primed finger millet plants against the blast disease.

Plants, being sessile, are exposed to an array of abiotic and biotic stresses. To adapt towards the changing environments, plants have evolved mechanisms that help in perceiving stress signals wherein phytohormones play a critical role. They have the ability to network enabling them to mediate defense responses. These endogenous signals, functioning at low doses are a part of all the developmental stages of the plant. Phytohormones possess specific functions as they interact with each other positively or negatively through cross-talks. In the present study, variations in the amount of phytohormones produced during biotic stress caused due to Magnoporthe grisea infection was studied through targeted metabolomics in both primed and control finger millet plants. Histochemical studies revealed callose deposition at the site of pathogen entry in the primed plants indicating its role during plant defense. The knowledge on the genetic makeup during infection was obtained by quantification of MAP kinase kinases 1 and 2 (MKK1/2) and lipoxygenase (LOX) genes, wherein the expression levels were high in the primed plants at 6 hours post-inoculation (hpi) compared to mock-control. Studies indicate the pivotal role of mitogen-activated protein kinase (MAPK or MAP kinases) during defense signalling. It is the first report to be studied on MAPK role in finger millet-blast disease response. Temporal accumulation of LOX enzyme along with its activity was also investigated due to its significant role during jasmonate synthesis in the plant cells. Results indicated its highest activity at 12 hpi. This is the first report on the variation in phytohormone levels in fingermillet - M. grisea pathosystem upon priming which were substantiated through salicylic acid (SA) pathway.

Green Synthesis and Characterization of Zinc Oxide Nanoparticles Using Eucalyptus globules and Their Fungicidal Ability Against Pathogenic Fungi of Apple Orchards.

Eucalyptus globules belonging to the Myrtaceae family was explored for the synthesis of zinc oxide nanoparticles and for biological applications. The aqueous extract of the synthesized zinc nanoparticles (ZnNPs) was characterized using UV-visible spectrophotometer, FTIR, SEM and TEM. The aqueous broth was observed to be an efficient reducing agent, leading to the rapid formation of ZnNPs of varied shapes with sizes ranging between 52-70 nm. In addition, antifungal activity of the biosynthesized ZnNPs was evaluated against major phytopathogens of apple orchards. At 100 ppm of ZnNPs, the fungal growth inhibition rate was found to be 76.7% for Alternaria mali, followed by 65.4 and 55.2% inhibition rate for Botryosphaeria dothidea and Diplodia seriata, respectively. The microscopic observations of the treated fungal plates revealed that ZnNPs damages the topography of the fungal hyphal layers leading to a reduced contraction of hyphae. This considerable fungicidal property of ZnNPs against phytopathogenic fungi can have a tremendous impact on exploitation of ZnNPs for fungal pest management and ensure protection in fruit crops.

Mycogenic Selenium Nanoparticles as Potential New Generation Broad Spectrum Antifungal Molecules.

The current challenges of sustainable agricultural development augmented by global climate change have led to the exploration of new technologies like nanotechnology, which has potential in providing novel and improved solutions. Nanotools in the form of nanofertilizers and nanopesticides possess smart delivery mechanisms and controlled release capacity for active ingredients, thus minimizing excess run-off to water bodies. This study aimed to establish the broad spectrum antifungal activity of mycogenic selenium nanoparticles (SeNPs) synthesized from Trichoderma atroviride, and characterize the bioactive nanoparticles using UV-Vis spectroscopy, dynamic light scattering (DLS), Fourier transform infrared (FT-IR), X-ray diffraction (XRD), scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDS), and high-resolution transmission electron microscopy (HR-TEM). The synthesized nanoparticles displayed excellent in vitro antifungal activity against Pyricularia grisea and inhibited the infection of Colletotrichum capsici and Alternaria solani on chili and tomato leaves at concentrations of 50 and 100 ppm, respectively. The SEM-EDS analysis of the bioactive SeNPs revealed a spherical shape with sizes ranging from 60.48 nm to 123.16 nm. The nanoparticles also possessed the unique property of aggregating and binding to the zoospores of P. infestans at a concentration of 100 ppm, which was visualized using light microscope, atomic force microscopy, and electron microscopy. Thus, the present study highlights the practical application of SeNPs to manage plant diseases in an ecofriendly manner, due to their mycogenic synthesis and broad spectrum antifungal activity against different phytopathogens.