Powdery mildew (PM) is one of the most devastating and widespread foliar diseases globally. Despite the critical need for developing a durable PM resistance, the number of cloned genes remains limited, along with a shortage of Mildew Locus O (MLO) resistance-conferring genes in wheat breeding programs. Here, utilizing the latest wheat reference genome data, we comprehensively identified and characterized 47 MLO genes through a genome-wide search approach. These genes are randomly distributed among 21 wheat chromosomes, harbor seven transmembrane domains, and are predicted to be primarily localized in the plasma membrane. Comparative phylogenetic analysis with model plants classified wheat MLOs into four clades (I–IV) harboring 6, 28, 6, and 7 genes, respectively. The phylogenetic grouping was strongly supported by gene structures and motif distribution among members of different clades. Evolution analysis revealed that the MLO gene arsenal expanded through segmental duplications, and purifying selection is potentially conserving their stress-associated functions. In-silico expression analysis highlighted at least 10 genes with overlapping expression patterns among different growth and development stages and under abiotic and biotic stress conditions. The quantitative real-time polymerase chain reaction (qRT-PCR) validated the differential expression patterns of these 10 overlapping genes in PM-resistant and susceptible wheat genotypes after challenging these with a PM pathogen strain at different time intervals. The identified wheat MLO genes, especially the 10 overlapping genes, highlight untapped genetic diversity for engineering a durable and broad-spectrum tolerance/resistance against abiotic and biotic stresses, especially the PM resistance. Collectively, this study provides a compendium of wheat MLO genes, which could be functionally characterized to confirm their roles in PM resistance and further exploited in wheat breeding programs for the development of climate-resilient cultivars for sustainable wheat production.

Lumpy skin disease virus (LSDV) belongs to Poxviridae family. This virus possesses various proteins which impart potential functions to it including assembly of newly synthesized viruses in the replication cycle and forming their structure. LSDV132 protein is also one of such proteins. Its key characteristics were unknown because, no any relevant study was reported about it. This study aimed to investigate its characteristic features and essential functions using several bioinformatics techniques. These analyses included physiochemical characterization and exploring the crucial functional and structural perspectives. Upon analysis of the physiochemical properties, the instability index was computed to be 30.89% which proposed LSDV132 protein to be a stable protein. Afterwards, the phosphorylation sites were explored. Several sites were found in this regard which led to the hypothesis that it might be involved in the regulation of apoptosis and cell signaling, among other cellular processes. Furthermore, the KEGG analysis and the analysis of protein family classification confirmed that the LSDV132 protein possessed Poxvirus-BCL-2-like motifs, indicating that it might be responsible in modulating the apoptosis of host cells. This crucial finding suggested that the protein under study possessed BCL-2-like features. Proceeding this very important finding, the molecular docking analysis was performed. In this context, various viral BCL-2 inhibitors were retrieved from the ChEMBL database for docking purpose. The docking results revealed that pelcitoclax exhibited best docking scores i.e., −9.1841 kcal/mol, among all of the other docked complexes. This fact signified that this compound might serve as an inhibitor of LSDV132 protein.

Brassica rapa (turnip) belongs to the Brassicaceae family. Due to the diseases caused by fungal pathogens such as Erysiphe cruciferarum, Alternaria japonica, and Alternaria brassicae, the yield of B. rapa was reduced in many Asian and European countries. Plant defensins (PDFs) are proteins that have antifungal properties. This research aimed to comprehend the key attributes of B. rapa PDF 1.1s by incorporating a wide range of in silico approaches. Initially, we explored orthologues of PDF 1.1 in B. rapa through Blastp in Phytozome v.13 which highlighted fourteen different orthologues. Afterward, by using Mega X, the homology of B. rapa PDF1.1 proteins with Arabidopsis thaliana PDF1.1 proteins was also inferred. While, the results of NCBI CD show that these proteins possessed a common domain, i.e., gamma-thionin. The major findings of this study included the crucial characteristics of PDF1.1 proteins that were explored by utilizing the PANZZER2 web server and BlastKOALA, which proposed that these proteins possessed degradation ability of pathogen’s cells and were also found to be involved in a mitogen-activated protein kinase signaling pathway. Overall, this research provided a better understanding of the crucial contribution of B. rapa PDFs 1.1 under biotic stress specifically, the pathogenic fungal attack.