In-silico analysis of ribosome inactivating protein (RIP) of the Cucurbitaceae family.

Ribosome-inactivating proteins (RIPs) are highly active N-glycosidases that depurinate both bacterial and eukaryotic rRNAs, halting protein synthesis during translation. Found in a diverse spectrum of plant species and tissues, RIPs possess antifungal, antibacterial, antiviral, and insecticidal properties linked to plant defense. In this study, we investigated the physiochemical properties of RIP peptides from the Cucurbitaceae family through bioinformatics approaches. Molecular weight, isoelectric point, aliphatic index, extinction coefficient, and secondary structures were analyzed, revealing their hydrophobic nature. The novelty of this work lies in the comprehensive examination of RIPs from the Cucurbitaceae family and their potential therapeutic applications. The study also elucidated the binding interactions of Cucurbitaceae RIPs with key biological targets, including Interleukin-6 (IL-6). Strong hydrogen bond interactions between RIPs and these targets suggest potential for innovative insilico drug design and therapeutic applications, particularly in cancer treatment. Comprehensive analysis of bond lengths using Ligpolt + software provides insights for optimizing molecular interactions, offering a valuable tool for drug design and structural biology studies.

In-silico characterization of LSDV132 protein divulged its BCL-2-like nature.

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.

Toxicity assessment of heavy metal (Pb) and its bioremediation by potential bacterial isolates.

Lead (Pb) is a non-essential metal with high toxicity, is persistent, is not biodegradable, and has no known biological function. It is responsible for severe health and environmental issues that need appropriate remediation. Therefore, microbes have thrived in a lead-contaminated environment without exhibiting any negative impacts. The present study aimed to examine the toxic effects of lead on animals and the isolation, identification, and characterization of lead-resistant bacterial strains and their biodegradation potential. After oral administration of lead for 4 weeks, mice showed an elevated level of leukocytes and a decrease in TEC, Hb, PCV, MCV, MCH, and MCHC levels. However, a decline in body weight and inflammation and oxidative stress was observed in liver tissues. To remediate toxic heavy metal, lead-resistant bacterial strains were isolated, among which Enterobacter exhibited maximum degradation potential at high lead concentrations. It was identified by molecular basis and after 16S rRNA sequencing, and 99% resemblance was observed with Enterobacter cloacae. FT-IR analysis of the bacteria illustrated the presence of functional groups, including hydroxyl, carboxyl group, sulfide, and amino groups, on the bacterial cell surface involved in the adsorption of lead. Moreover, electron microscopy (SEM) revealed the morphological and physiochemical changes in the bacterial cell after biosorption, indicating the interaction of Cu ions with functional groups. To summarize, the findings show the highly toxic effects of lead on animals and humans and its effective biodegradation by the bacterial strains in the lead-contaminated environment. This biological strategy can be an ideal alternative to remediate heavy metals from contaminated sites to clean up the environment.

Molecular and Biochemical Mechanisms of Elicitors in Pest Resistance.

Insect herbivores have a variety of life cycles and feeding habits, making them extremely diverse. With their host plants, they form close relationships and suppress their defense mechanisms. Molecular elicitors are the key bio-elements in the detection and recognition of attacking enemies in tissue consumption. Insect oral secretion, frass, and fluid of egg deposition contain biologically active molecules called herbivore-associated elicitors (HAEs) that are recognized by pattern-recognition receptors (PRRs). Many plants distinguish insect feeding from wounding by HAEs present in their oral secretions (OS) and induce local and/or systemic responses against arthropod feeding. PRRs perceive HAEs in the oral secretion of caterpillars in a species-specific manner to elicit exclusive defense responses. HAEs-PRRs interactions induce plant resistance by reprogramming plant metabolism and transcriptional machinery. Quantitative, timely, and coordinated plant response initiate early signaling events, including Ca2+, reactive oxygen species (ROS), and mitogen-activated protein kinases (MAPKs). However, in insect herbivory, little is known about the molecular basis of signal transduction and regulation of plant resistance. We discuss here how early signaling cascades converge into the accumulation of phytohormones that regulate downstream special metabolites against herbivores. In this review, we propose a hypothetical model of PPRs-HAEs-mediated-induced responses in plants and discuss how PRRs-HAEs interactions elicit short- and long-term induced defenses in plants. The understanding of PRRs-HAEs interactions will help to explore the fundamental molecular mechanisms of host manipulation and may generate prospects to develop novel pest-resistance strategies.

Mythimna separata herbivory primes maize resistance in systemic leaves.

Biotic and abiotic cues can trigger priming in plants, which enables plants to respond to subsequent challenge with stronger and/or faster responses. It is well known that herbivory activates defense-related responses in systemic leaves. However, little is known about whether insect feeding activates priming in systemic leaves. To determine whether and how herbivory induces priming in maize systemic leaves, a combination of insect bioassays, phytohormone and defense metabolite quantification, and genetic and transcriptome analyses were performed. Actual and simulated Mythimna separata herbivory in maize local leaves primed the systemic leaves for enhanced accumulation of jasmonic acid and benzoxazinoids and increased resistance to M. separata. Activation of priming in maize systemic leaves depends on both the duration of simulated herbivory and perception of M. separata oral secretions in the local leaves, and genetic analysis indicated that jasmonic acid and benzoxazinoids mediate the primed defenses in systemic leaves. Consistently, in response to simulated herbivory, the primed systemic leaves exhibited a large number of genes that were uniquely regulated or showed further up- or down-regulation compared with the non-primed systemic leaves. This study provides new insight into the regulation and ecological function of priming in maize.

An efficient system composed of maize protoplast transfection and HPLC-MS for studying the biosynthesis and regulation of maize benzoxazinoids.

BACKGROUND: Insect herbivory poses a major threat to maize. Benzoxazinoids are important anti-insect secondary metabolites in maize, whose biosynthetic pathway has been extensively studied. However, yet little is known about how benzoxazinoids are regulated in maize, partly due to lack of mutant resources and recalcitrance to genetic transformation. Transient systems based on mesophyll- or cultured cell-derived protoplasts have been exploited in several plant species and have become a powerful tool for rapid or high-throughput assays of gene functions. Nevertheless, these systems have not been exploited to study the regulation of secondary metabolites. RESULTS: A protocol for isolation of protoplasts from etiolated maize seedlings and efficient transfection was optimized. Furthermore, a 10-min-run-time and highly sensitive HPLC-MS method was established to rapidly detect and quantify maize benzoxazinoids. Coupling maize protoplast transfection and HPLC-MS, we screened a few genes potentially regulating benzoxazinoid biosynthesis using overexpression or silencing by artificial microRNA technology. CONCLUSIONS: Combining the power of maize protoplast transfection and HPLC-MS analysis, this method allows rapid screening for the regulatory and biosynthetic genes of maize benzoxazinoids in protoplasts, before the candidates are selected for in planta functional analyses. This method can also be applied to study the biosynthesis and regulation of other secondary metabolites in maize and secondary metabolites in other plant species, including those not amenable to transformation.

The oriental armyworm ( Mythimna separata) feeding induces systemic defence responses within and between maize leaves.

Maize ( Zea mays) is a staple cereal crop cultivated all over the world but that is threatened by various insects. Feeding of the lepidopteran insect Mythimna separata triggers defence signalling and increases anti-herbivore benzoxazinoids (Bxs) in the insect-damaged maize leaves. However, the herbivory-elicited within-leaf and leaf-to-leaf systemic signalling in maize remains largely unexplored. Here, we show that simulated M. separata herbivory and mechanical wounding elicited increased levels of jasmonic acid (JA), JA-Ile (JA-isoleucine conjugate) and Bxs in the damaged areas and in specific systemic regions within a leaf. Importantly, increased contents of Bxs were detected in a systemic leaf, and consistently, this leaf exhibited increased defence against M. separata. Increased JA/JA-Ile and altered transcriptome, including Bx biosynthesis genes, were detected in systemic leaves after wounding or simulated herbivory treatments, although only simulated herbivory induced increase of the contents of Bxs systemically. Promoter and co-expression analysis revealed that transcription factors bHLH57 and WRKY34 may regulate Bx biosynthesis genes in systemic leaves. Moreover, leaf ablation experiment indicated that the systemic signal rapidly exited the local leaves within 30 min after elicitation. This study provides new insight into the temporal and spatial regulation of defence responses of maize against lepidopteran insects. This article is part of the theme issue 'Biotic signalling sheds light on smart pest management'.

Elevated CO2 differentially affects tobacco and rice defense against lepidopteran larvae via the jasmonic acid signaling pathway.

Atmospheric CO2 levels are rapidly increasing due to human activities. However, the effects of elevated CO2 (ECO2 ) on plant defense against insects and the underlying mechanisms remain poorly understood. Here we show that ECO2 increased the photosynthetic rates and the biomass of tobacco and rice plants, and the chewing lepidopteran insects Spodoptera litura and Mythimna separata gained less and more mass on tobacco and rice plants, respectively. Consistently, under ECO2 , the levels of jasmonic acid (JA), the main phytohormone controlling plant defense against these lepidopteran insects, as well as the main defense-related metabolites, were increased and decreased in insect-damaged tobacco and rice plants. Importantly, bioassays and quantification of defense-related metabolites in tobacco and rice silenced in JA biosynthesis and perception indicate that ECO2 changes plant resistance mainly by affecting the JA pathway. We further demonstrate that the defensive metabolites, but not total N or protein, are the main factors contributing to the altered defense levels under ECO2 . This study illustrates that ECO2 changes the interplay between plants and insects, and we propose that crops should be studied for their resistance to the major pests under ECO2 to predict the impact of ECO2 on future agroecosystems.

Current understanding of maize and rice defense against insect herbivores.

Plants have sophisticated defense systems to fend off insect herbivores. How plants defend against herbivores in dicotyledonous plants, such as Arabidopsis and tobacco, have been relatively well studied, yet little is known about the defense responses in monocotyledons. Here, we review the current understanding of rice (Oryza sativa) and maize (Zea mays) defense against insects. In rice and maize, elicitors derived from insect herbivore oral secretions or oviposition fluids activate phytohormone signaling, and transcriptomic changes mediated mainly by transcription factors lead to accumulation of defense-related secondary metabolites. Direct defenses, such as trypsin protein inhibitors in rice and benzoxazinoids in maize, have anti-digestive or toxic effects on insect herbivores. Herbivory-induced plant volatiles, such as terpenes, are indirect defenses, which attract the natural enemies of herbivores. R gene-mediated defenses against herbivores are also discussed.