Arakere Chunchegowda Udayashankar

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.

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.

Endophytic fungi from orchid plants are reported to secrete secondary metabolites which include bioactive antimicrobial siderophores. In this study endophytic fungi capable of secreting siderophores were isolated from Cymbidium aloifolium, a medicinal orchid plant. The isolated extracellular siderophores from orchidaceous fungi act as chelating agents forming soluble complexes with Fe³⁺. The 60% endophytic fungi of Cymbidium aloifolium produced hydroxamate siderophore on CAS agar. The highest siderophore percentage was 57% in Penicillium chrysogenum (CAL1), 49% in Aspergillus sydowii (CAR12), 46% in Aspergillus terreus (CAR14) by CAS liquid assay. The optimum culture parameters for siderophore production were 30 °C, pH 6.5, maltose and ammonium nitrate and the highest resulting siderophore content was 73% in P. chrysogenum. The total protein content of solvent-purified siderophore increased four-fold compared with crude filtrate. The percent Fe³⁺ scavenged was detected by atomic absorption spectra analysis and the highest scavenging value was 83% by P. chrysogenum. Thin layer chromatography of purified P. chrysogenum siderophore showed a wine-colored spot with R_f value of 0.54. HPLC peaks with R_ts of 10.5 and 12.5 min were obtained for iron-free and iron-bound P. chrysogenum siderophore, respectively. The iron-free P. chrysogenum siderophore revealed an exact mass-to-charge ratio (m/z) of 400.46 and iron-bound P. chrysogenum siderophore revealed a m/z of 453.35. The solvent-extracted siderophores inhibited the virulent plant pathogens Ralstonia solanacearum, that causes bacterial wilt in groundnut and Xanthomonas oryzae pv. oryzae which causes bacterial blight disease in rice. Thus, bioactive siderophore-producing endophytic P. chrysogenum can be exploited in the form of formulations for development of resistance against other phytopathogens in crop plants.

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.

Cinnamomum verum plant extract mediated propellant chemistry route was used for the green synthesis of zinc oxide nanoparticles. Prepared samples were confirmed for their nano regime using advanced characterization techniques such as powder X-ray diffraction and microscopic techniques such as scanning electron microscopy and transmission electron microscopy. The energy band gap of the green synthesized zinc oxide (ZnO)-nanoparticles (NPs) were found between 3.25–3.28 eV. Fourier transmission infrared spectroscopy shows the presence of Zn-O bond within the wave number of 500 cm⁻¹. SEM images show the specific agglomeration of particles which was also confirmed by TEM studies. The green synthesized ZnO-NPs inhibited the growth of Escherichia coli and Staphylococcus aureus with a minimum inhibitory concentration (MIC) of 125 µg mL⁻¹ and 62.5 µg mL⁻¹, respectively. The results indicate the prepared ZnO-NPs can be used as a potential antimicrobial agent against harmful pathogens.

Unscientific use of synthetic fungicides in plant disease management has environmental ramifications, such as disease resurgence and serious health problems due to their carcinogenicity. This has prompted the identification and development of eco-friendly greener alternatives. Eclipta alba extract was evaluated for its antifungal activity in in vitro and in vivo against sorghum fungal pathogens Fusarium thapsinum, Alternaria alternata, Epicoccum sorghinum, and Curvularia lunata. The column purified methanolic extract of E. alba exhibited good antifungal activity against the target pathogens. The MIC was observed at 80 mg/mL for all tested pathogenic fungi, whereas MFC was 80 mg/mL for E. sorghinum, 100 mg/mL for F. thapsinum, A. alternata, and C. lunata. In vitro germination percentage was significantly high in seeds treated with E. alba extract (98%) over untreated control (91%). Significant disease protection of 95% was observed in greenhouse and 66% disease protection was noticed in field experiments. The efficacy of E. alba extract in field conditions was improved with the use of E. alba extract formulation. The profile of phytochemicals in E. alba methanol fractions was obtained by ultra-performance liquid chromatography (UPLC) mass spectroscopy. The [M-H]⁻ at m/z 313.3, m/z 797.9, and m/z 269.0 revealed the presence of wedelolactone, eclalbasaponin II, and apigenin, respectively. The H-nuclear magnetic resonance spectroscopy (¹H-NMR) chemical shift value supported the findings of the mass spectrometry. The results highlighted the possible use of E. alba methanolic extract as alternative to chemical fungicide in sorghum disease management.