Biogenic carbon quantum dots from marine endophytic fungi (Aspergillus flavus) to enhance the curcumin production and growth in Curcuma longa L.

In this study, we have investigated the effect of carbon quantum dots (FM-CQDs) synthesized from marine fungal extract on Curcuma longa to improve the plant growth and curcumin production. The isolated fungus, Aspergillus flavus has produced a high amount of indole-3-acetic acid (IAA) (0.025 mg g-1), when treated with tryptophan. CQDs were synthesized from the A. flavus extract and it was characterized using ultraviolet visible spectrophotometer (UV-Vis) and high-resolution transmission electron microscopy (HR-TEM). The synthesized CQDs were excited at 365 nm in an UV-Vis and the HR-TEM analysis showed approximately 7.4 nm in size with a spherical shape. Both fungal crude extract (FCE) at 0-100 mg L-1 and FM-CQDs 0-5 mg L-1 concentrations were tested on C. longa. About 80 mg L-1 concentration FCE treated plants has shown a maximum height of 21 cm and FM-CQDs at 4 mg L-1 exhibited a maximum height of 25 cm compared to control. The FM-CQDs significantly increased the photosynthetic pigments such as total chlorophyll (1.08 mg g-1 FW) and carotenoids (17.32 mg g-1 FW) in C. longa. Further, antioxidant enzyme analysis confirmed that the optimum concentrations of both extracts did not have any toxic effects on the plants. FM-CQDs treated plants increased the curcumin content up to 0.060 mg g-1 by HPLC analysis. Semi quantitative analysis revealed that FCE and FM-CQDs significantly upregulated ClCURS1 gene expression in curcumin production.

Molecular Insights into Abiotic Stresses in Mango.

Mango (Mangifera indica L.) is one of the most economically important fruit crops across the world, mainly in the tropics and subtropics of Asia, Africa, and Central and South America. Abiotic stresses are the prominent hindrance that can adversely affect the growth, development, and significant yield loss of mango trees. Understanding the molecular physiological mechanisms underlying abiotic stress responses in mango is highly intricate. Therefore, to gain insights into the molecular basis and to alleviate the abiotic stress responses to enhance the yield in the mere future, the use of high-throughput frontier approaches should be tied along with the baseline investigations. Taking these gaps into account, this comprehensive review mainly speculates to provide detailed mechanisms and impacts on physiological and biochemical alterations in mango under abiotic stress responses. In addition, the review emphasizes the promising omics approaches in unraveling the candidate genes and transcription factors (TFs) responsible for abiotic stresses. Furthermore, this review also summarizes the role of different types of biostimulants in improving the abiotic stress responses in mango. These studies can be undertaken to recognize the roadblocks and avenues for enhancing abiotic stress tolerance in mango cultivars. Potential investigations pointed out the implementation of powerful and essential tools to uncover novel insights and approaches to integrate the existing literature and advancements to decipher the abiotic stress mechanisms in mango. Furthermore, this review serves as a notable pioneer for researchers working on mango stress physiology using integrative approaches.

Integrated omics profiling and network pharmacology uncovers the prognostic genes and multi-targeted therapeutic bioactives to combat lung cancer.

Non-small cell lung cancer (NSCLC) is the frequent subtype of lung cancer and the currently used treatment methods, diagnosis, and chemoresistance are relatively ineffective. Determining the pharmacological targets from active biomolecules of medicinal plants has become a frontiers era for biomedical research to develop novel therapies. In view of these scenarios, this pilot study, network pharmacology, cheminformatics, integrative omics, molecular docking and in vitro anti-cancer analysis were performed to unveil the multi-targeted treatment mechanisms of novel plant bioactives to treat lung cancer. Bioactive molecules from medicinal plants were compiled from PubChem. Network pharmacology approach revealed that 29 compounds efficiently target the 390 human and lung cancer associated genes. In addition, comparative analysis was performed and identified the 7 bioactive molecules significantly targeting 8 lung cancer genes. The integrative omics analysis discovered unique genes between the lung cancer and normal lung tissues. These genes were further validated through protein-protein interaction, gene ontology, gene functional and pathway enrichment, boxplot and overall survival analyses to understand the function of unique genes and their involvement in cancer signaling pathways. Survival heatmap analyses identified the significant prognostic genes. Docking results revealed that, lupeol and p-coumaric acid displayed high binding affinities with MIF, CCNB1, FABP4. Hence, we selected these two bioactives for in vitro analysis. Furthermore, these selected bioactives were showed concentration dependent cytotoxicity against the lung adenocarcinoma cells (A549). This holistic study has opened up novel avenues and unravels the cancer prognostic genes which could serve as druggable target and bioactives with anti-cancerous efficacy. Further functional validations are prerequisites to deciphering these bioactives as commercial drug candidates.

Recent Updates on Source, Biosynthesis, and Therapeutic Potential of Natural Flavonoid Luteolin: A Review.

Nature gives immense resources that are beneficial to humankind. The natural compounds present in plants provide primary nutritional values to our diet. Apart from food, plants also provide chemical compounds with therapeutic values. The importance of these plant secondary metabolites is increasing due to more studies revealing their beneficial properties in treating and managing various diseases and their symptoms. Among them, flavonoids are crucial secondary metabolite compounds present in most plants. Of the reported 8000 flavonoid compounds, luteolin is an essential dietary compound. This review discusses the source of the essential flavonoid luteolin in various plants and its biosynthesis. Furthermore, the potential health benefits of luteolins such as anti-cancer, anti-microbial, anti-inflammatory, antioxidant, and anti-diabetic effects and their mechanisms are discussed in detail. The activity of luteolin and its derivatives are diverse, as they help to prevent and control many diseases and their life-threatening effects. This review will enhance the knowledge and recent findings regarding luteolin and its therapeutic effects, which are certainly useful in potentially utilizing this natural metabolite.

Bioactive Components and Health Potential of Endophytic Micro-Fungal Diversity in Medicinal Plants.

The endophytic fungi that reside inside medicinal plants have the potential to produce various pharmaco-potential bioactive compounds. The endophytic fungi Graminicolous helminthosporium, Bipolaris australiensis and Cladosporium cladosporioides were isolated from different medicinal plants. The GC-MS analysis of intra- and extracellular products of endophytic fungi revealed the presence of various bioactive metabolites, such as Anthracene, Brallobarbital, Benzo [h] quinolone, Ethylacridine, 2-Ethylacridine, Cyclotrisiloxane, 5 methyl 2 phenylindolizine, and 1,4-Cyclohexadien-1-one, etc. The phytochemical composition analysis of endophytic fungus extracts also revealed the presence of flavonoids, phenols, saponins, carbohydrates, glycosides, and proteins. The intra- and extracellular endophytic extracts exhibited strong antibacterial and antioxidant activity, which was screened with the agar-well diffusion method and DPPH, H2O2, and nitric oxide scavenging activity, respectively. The bioactive compounds identified in the endophytic extracts from GC-MS profiling served as ligands for molecular-docking analysis to investigate the anticancer potential against non-small cell lung carcinoma receptor EGFR. Molecular docking results showed that compounds, such as Brallobarbital, and 5 methyl 2 phenylindolizine had the lowest E- min values, which suggests that these compounds could be used in anticancer drug development. Thus, the isolated endophytic fungal species can be used to produce various bioactive compounds that could be used in novel drug development from natural sources and reduce the environmental burden of synthetic chemical drugs.

Biosynthesis of ZnONP Using Chamaecostus cuspidatus and Their Evolution of Anticancer Property in MCF-7 and A549 Cell Lines.

The ZnO nanoparticle synthesis using the leaf part of Chamaecostus cuspidatus was characterized using UV-Vis spectrophotometry, IR, XRD, DLS, FESEM, EDX, TEM, AFM and XPS. The MTT assay was used to examine the cytotoxicity activity against lung epithelial and breast cell lines, and the IC50 value was determined. The presence of ZnO nanoparticles, which range in size from 200 to 800 nm, was confirmed by the absorption peak at 350 nm. The median particle size was 145.1 nm, and the ζ -the potential was -19.45 mV, showing that ZnONP is stable. Zinc, carbon, and oxygen contribute to the elemental composition of ZnONP, as determined by EDX analysis. MTT assay was used to investigate in vitro cytotoxicity in MCF-7 and A549 cell lines. The cytotoxicity activity IC50 value was determined to be 30 µg/mL for the A549 cell line and 37 µg/mL for the MCF-7 cell line.

Multi-Omics and Integrative Approach towards Understanding Salinity Tolerance in Rice: A Review.

Rice (Oryza sativa L.) plants are simultaneously encountered by environmental stressors, most importantly salinity stress. Salinity is the major hurdle that can negatively impact growth and crop yield. Understanding the salt stress and its associated complex trait mechanisms for enhancing salt tolerance in rice plants would ensure future food security. The main aim of this review is to provide insights and impacts of molecular-physiological responses, biochemical alterations, and plant hormonal signal transduction pathways in rice under saline stress. Furthermore, the review highlights the emerging breakthrough in multi-omics and computational biology in identifying the saline stress-responsive candidate genes and transcription factors (TFs). In addition, the review also summarizes the biotechnological tools, genetic engineering, breeding, and agricultural practicing factors that can be implemented to realize the bottlenecks and opportunities to enhance salt tolerance and develop salinity tolerant rice varieties. Future studies pinpointed the augmentation of powerful tools to dissect the salinity stress-related novel players, reveal in-depth mechanisms and ways to incorporate the available literature, and recent advancements to throw more light on salinity responsive transduction pathways in plants. Particularly, this review unravels the whole picture of salinity stress tolerance in rice by expanding knowledge that focuses on molecular aspects.

Transcriptomics, Cheminformatics, and Systems Pharmacology Strategies Unveil the Potential Bioactives to Combat COVID-19.

Coronavirus disease (COVID-19) is a viral disease caused by the SARS-CoV-2 virus and is becoming a global threat again because of the higher transmission rate and lack of proper therapeutics as well as the rapid mutations in the genetic pattern of SARS-CoV-2. Despite vaccinations, the prevalence and recurrence of this infection are still on the rise, which urges the identification of potential global therapeutics for a complete cure. Plant-based alternative medicine is becoming popular worldwide because of its higher efficiency and minimal side effects. Yet, identifying the potential medicinal plants and formulating a plant-based medicine is still a bottleneck. Hence, in this study, the systems pharmacology, transcriptomics, and cheminformatics approaches were employed to uncover the multi-targeted mechanisms and to screen the potential phytocompounds from significant medicinal plants to treat COVID-19. These approaches have identified 30 unique COVID-19 human immune genes targeted by the 25 phytocompounds present in four selected ethnobotanical plants. Differential and co-expression profiling and pathway enrichment analyses delineate the molecular signaling and immune functional regulations of the COVID-19 unique genes. In addition, the credibility of these compounds was analyzed by the pharmacological features. The current holistic finding is the first to explore whether the identified potential bioactives could reform into a drug candidate to treat COVID-19. Furthermore, the molecular docking analysis was employed to identify the important bioactive compounds; thus, an ultimately significant medicinal plant was also determined. However, further laboratory evaluation and clinical validation are required to determine the efficiency of a therapeutic formulation against COVID-19.

Untargeted Metabolomic Approach to Determine the Regulatory Pathways on Salicylic Acid-Mediated Stress Response in Aphanamixis polystachya Seedlings.

Plants thrive under abiotic and biotic stress conditions with the changes in phytohormones like salicylic acid (SA), resulting in the synthesis of secondary metabolites. The present study determines the response of plants in producing secondary metabolites towards different SA concentrations at varying time intervals. Liquid chromatography-mass spectrometry-based metabolomics studies in Aphanamixis polystachya (Wall.) Parker seedlings are grown at 10 mM, 50 mM, and 100 mM SA concentrations, showed the differential expression of metabolites towards the stress. Alkaloids like amaranthin showed a 15-fold increase on the second day, and analog of androvinblastin showed a 20-fold increase on the sixth day in 10 mM SA compared with other groups. Flavanoid cyanidin 3-3 glucosyl was found to be with a 22-fold increment along with terpenoids betavulgaroside (18-fold), asiaticoside (17-fold), mubenin B (20-fold), and deslanoside (22-fold) increment in 50 mM SA on the sixth day. The shock exerted by 100 mM was too harsh, and the lowered metabolite production level was insufficient for the seedlings to survive at this higher SA condition. Arrangement of stressed groups using Pearson correlation studies, principal component analysis, and partial least square analysis placed 10 mM SA and controlled group closer and 50 mM SA and 100 mM SA groups closer to each other. The study observed that SA regulates metabolites that mediate biotic stress responses at low concentrations, and higher concentrations regulate abiotic stress regulating metabolites.

Enhanced thermo-tolerance in transgenic potato (Solanum tuberosum L.) overexpressing hydrogen peroxide-producing germin-like protein (GLP).

Germins and germin-like proteins (GLPs) were reported to participate in plant response to biotic and abiotic stresses involving hydrogen peroxide (H2O2) production, but their role in mitigating heat stress is poorly understood. Here, we investigated the ability of a Solanum tuberosum L. GLP (StGLP) gene isolated from the yeast cDNA library generated from heat-stressed potato plants and characterized its role in generating innate and/or acquired thermo-tolerance to potato via genetic transformation. The transgenic plants exhibited enhanced tolerance to gradual heat stress (GHS) compared with sudden heat shock (SHS) in terms of maximal cell viability, minimal ion leakage and reduced chlorophyll breakdown. Further, three StGLP transgenic lines (G9, G12 and G15) exhibited enhanced production of H2O2, which was either reduced or blocked by inhibitors of H2O2 under normal and heat stress conditions. This tolerance was mediated by up-regulation of antioxidant enzymes (catalase, ascorbate peroxidase and glutathione reductase) and other heat stress-responsive genes (StHSP70, StHSP20 and StHSP90) in transgenic potato plants. These results demonstrate that H2O2 produced by over-expression of StGLP in transgenic potato plants triggered the reactive oxygen species (ROS) scavenging signaling pathways controlling antioxidant and heat stress-responsive genes in these plants imparting tolerance to heat stress.

Nanotechnologies in Food Science: Applications, Recent Trends, and Future Perspectives.

Nanotechnology is a key advanced technology enabling contribution, development, and sustainable impact on food, medicine, and agriculture sectors. Nanomaterials have potential to lead qualitative and quantitative production of healthier, safer, and high-quality functional foods which are perishable or semi-perishable in nature. Nanotechnologies are superior than conventional food processing technologies with increased shelf life of food products, preventing contamination, and production of enhanced food quality. This comprehensive review on nanotechnologies for functional food development describes the current trends and future perspectives of advanced nanomaterials in food sector considering processing, packaging, security, and storage. Applications of nanotechnologies enhance the food bioavailability, taste, texture, and consistency, achieved through modification of particle size, possible cluster formation, and surface charge of food nanomaterials. In addition, the nanodelivery-mediated nutraceuticals, synergistic action of nanomaterials in food protection, and the application of nanosensors in smart food packaging for monitoring the quality of the stored foods and the common methods employed for assessing the impact of nanomaterials in biological systems are also discussed.

Subcritical water extraction of withanosides and withanolides from ashwagandha (Withania somnifera L) and their biological activities.

Subcritical water extraction (SWE) applied to analyses the bioactives from ashwagandha (W. somnifera) at varying temperature (100-200 °C) and extraction time (10-30 min). The effect of temperature and time has been investigated in terms of extraction yield (EY), total phenolic content (TPC), cytotoxicity, antioxidant, and enzyme inhibitory activities. The withanosides and withanolides responsible for various biological effects were quantified using high performance liquid chromatography (HPLC). The HPLC analysis revealed Withanoside V, Withanoside IV, 12-Deoxywithastramonolide, Withanolide A, and Withaferin A as a principle bioactive compounds in SWE, with high in concentration compared to microwave-assisted extraction (MAE), Soxhlet extraction (SE) and maceration (MC). For SWE the highest EY (65.6%; 200 °C for 30 min), TPC (82.5 mg GAE/g DE), antioxidant activity (DPPH: 80.3%, FRAP: 60.5% and ABTS: 78.9), and potent enzyme inhibitory effects were observed. The SWE and Withaferin A showed significant reduction in cell viability of cervical cancer (HeLa) cells, with IC50 values 10 mg/ml and 8.5 µM/ml, respectively but no cytotoxic effect for normal cells (MDCK). Thus, SWE can provide effective extraction for ashwagandha withanosides and withanolides compared MAE, SE and MC to conventional methods, which could be used for extraction of pharmacologically active fractions with therapeutic applications.

Assessment of the effects of metal oxide nanoparticles on the growth, physiology and metabolic responses in in vitro grown eggplant (Solanum melongena).

Nanoparticles (NPs) are widely used in various domestic products and their usage is constantly increasing which in turn can raise several environmental health issues. Like other abiotic stresses, nanomaterials also affect the growth of crop plants. Solanum melongena is a common vegetable crop grown in the tropics and subtropics regions with medicinal properties. In this study, S. melongena was analyzed for its response to three commercially important metallic nanoparticles, namely NiO, CuO, and ZnO, at four different concentrations (100, 250, 500 and 1000 mg/L). The growth of the eggplant seedlings was suppressed by all the NPs in a concentration-dependent manner and among them, NiO was shown to be more toxic as it suppressed the root and shoot growth effectively. Total chlorophyll contents were decreased in the NP-treated plants compared to control plants. Significant changes were found in the secondary metabolites such as anthocyanins, total phenolic and total flavonoid contents in the NP-treated plants. A dose-dependent increase in the reactive oxygen species (ROS) generation was noticed in the NP-treated plants which are evidenced by the 4-nitro blue tetrazolium chloride (NBT) and 3,3'-diamiobenzidine (DAB) histochemical staining. The DNA damage imposed by the NP in the seedlings of eggplants may be due to the elevated ROS and MDA (malondialdehyde) production. NiO NP was found to be more toxic comparable to CuO and ZnO NPs in the present study. Apart from the toxic effects, nanoparticles also showed profound effects on the production of important secondary metabolites such as phenolics and flavonoid compounds.

Enhanced Tolerance of Transgenic Potato Plants Over-Expressing Non-specific Lipid Transfer Protein-1 (StnsLTP1) against Multiple Abiotic Stresses.

Abiotic stresses such as heat, drought, and salinity are major environmental constraints that limit potato (Solanum tuberosum L.) production worldwide. Previously, we found a potential thermo-tolerance gene, named StnsLTP1 from potato using yeast functional screening. Here, we report the functional characterization of StnsLTP1 and its role in multiple abiotic stresses in potato plants. Computational analysis of StnsLTP1 with other plant LTPs showed eight conserved cysteine residues, and four α-helices stabilized by four disulfide bridges. Expression analysis of StnsLTP1 gene showed differential expression under heat, water-deficit and salt stresses. Transgenic potato lines over-expressing StnsLTP1 gene displayed enhanced cell membrane integrity under stress conditions, as indicated by reduced membrane lipid per-oxidation, and hydrogen peroxide content relative to untransformed (UT) control plants. In addition, transgenic lines over-expressing StLTP1 also exhibited increased antioxidant enzyme activity with enhanced accumulation of ascorbates, and up-regulation of stress-related genes including StAPX, StCAT, StSOD, StHsfA3, StHSP70, and StsHSP20 compared with the UT plants. These results suggests that StnsLTP1 transgenic plants acquired improved tolerance to multiple abiotic stresses through enhanced activation of antioxidative defense mechanisms via cyclic scavenging of reactive oxygen species and regulated expression of stress-related genes.

An Update on Potential Perspectives of Glucosinolates on Protection against Microbial Pathogens and Endocrine Dysfunctions in Humans.

Glucosinolates are the major bioactive secondary metabolites found in the Brassicaceae family and studied extensively in biosynthetic and application perspectives. Because of their potential applications in the welfare of plants (protection against plant pathogens) and human life (prevention of cancer and other diseases), these compounds attracted much interest in the scientific community. In this review, we presented updates on glucosinolate derivatives in protection against microbial pathogens and endocrine related diseases in human. Further, the mechanism of action of glucosinolate derivatives and the strategies to improve their efficiency through modern approaches were discussed. Finally, the genetic enrichment of their contents in plant systems has also been discussed.

A simple and efficient Agrobacterium tumefaciens-mediated plant transformation of Brassica rapa ssp. pekinensis.

The present study aims to investigate the numerous factors influencing Agrobacterium tumefaciens-mediated genetic transformation of Chinese cabbage (Brassica rapa ssp. pekinensis). Factors affecting transformation efficiency, such as age of explants, Agrobacterium concentration, and effect of acetosyringone, pre-cultivation, infection and co-cultivation time of Agrobacterium were examined. The pre-cultured hypocotyls from young seedlings prior to exposure to Agrobacterium showed higher shoot regeneration. The plant transformation with the modest A. tumefaciens concentrations (0.8 OD) and the 3 days co-cultivation periods increased transformation efficiency. Plant growth hormones [1-naphthyl acetic acid (NAA) and 6-benzyl amino purine (BAP)] were essential for callus and shoot formation. Root formation was effective in half strength MS medium without supplementation of root-inducing hormones. To maintain selection pressure, plant subculture was carried out every 2 weeks with selective antibiotics. The putative transgenic plants were acclimatized in the greenhouse. Polymerase chain reaction was performed to confirm the integration of T-DNA into the genome of transgenic plants. A transformation efficiency of 15 % was obtained. This protocol allows effective transformation and indirect regeneration of Brassica rapa.

Impact of biologically synthesized silver nanoparticles on the growth and physiological responses in Brassica rapa ssp. pekinensis.

Silver nanoparticles (AgNPs) were extensively used in various fields, particularly in medicine as an antimicrobial agent. The unavoidable and extensive usage of AgNPs in turn accumulates in the environment. Plants are the essential base of ecosystem and are ready to disturb by environmental pollutants. Therefore, in the present study, we have planned to evaluate the impact of biologically synthesized AgNPs on the essential food crop Chinese cabbage (Brassica rapa ssp. pekinensis). The effects of AgNP-induced plant morphological and physiological changes were investigated in different concentrations (100, 250, and 500 mg/L). The results of morphological features showed that AgNPs at lower concentrations (100 mg/L) exhibit growth-stimulating activity, whereas at higher concentrations (250 and 500 mg/L), particularly, 500 mg/L exhibited growth-suppressing activities which are in terms of reduced root, shoot growth, and fresh biomass. The increased reactive oxygen species (ROS) generation, malondialdehyde production, anthocyanin biosynthesis, and decreased chlorophyll content were also more obviously present at higher concentrations of AgNPs. The concentration-dependent DNA damage was observed in the AgNP-treated plants. The molecular responses of AgNPs indicate that most of the genes related to secondary metabolism (glucosinolates, anthocyanin) and antioxidant activities were induced at higher concentrations of AgNP treatment. The dose-dependent phytotoxicity effects of AgNPs were also observed. Taken together, the highest concentration of AgNPs (500 mg/L) could induce growth-suppressing activities via the induction of ROS generation and other molecular changes in B. rapa seedlings.

Molecular characterization of BrMYB28 and BrMYB29 paralogous transcription factors involved in the regulation of aliphatic glucosinolate profiles in Brassica rapa ssp. pekinensis.

Glucosinolates (GSL) are one of the major secondary metabolites of the Brassicaceae family. In the present study, we aim at characterizing the multiple paralogs of aliphatic GSL regulators, such as BrMYB28 and BrMYB29 genes in Brassica rapa ssp. pekinensis, by quantitative real-time PCR (qRT-PCR) analysis in different tissues and at various developmental stages. An overlapping gene expression pattern between the BrMYBs as well as their downstream genes (DSGs) was found at different developmental stages. Among the BrMYB28 and BrMYB29 paralogous genes, the BrMYB28.3 and BrMYB29.1 genes were dominantly expressed in most of the developmental stages, compared to the other paralogs of the BrMYB genes. Furthermore, the differential expression pattern of the BrMYBs was observed under various stress treatments. Interestingly, BrMYB28.2 showed the least expression in most developmental stages, while its expression was remarkably high in different stress conditions. More specifically, the BrMYB28.2, BrMYB28.3, and BrMYB29.1 genes were highly responsive to various abiotic and biotic stresses, further indicating their possible role in stress tolerance. Moreover, the in silico cis motif analysis in the upstream regulatory regions of BrMYBs showed the presence of various putative stress-specific motifs, which further indicated their responsiveness to biotic and abiotic stresses. These observations suggest that the dominantly expressed BrMYBs, both in different developmental stages and under various stress treatments (BrMYB28.3 and BrMYB29.1), may be potential candidate genes for altering the GSL level through genetic modification studies in B. rapa ssp. pekinensis.