Important Metabolites Research Articles

Identification of Key Post‐modification Enzymes Involved in the Biosynthesis of Lanostane‐type Triterpenoids in the Medicinal Mushroom Antrodia camphorata

Lanostane‐type triterpenoids are important bioactive secondary metabolites of mushrooms, though their biosynthetic study has been challenging due to scattered genes. Herein, the strategies of combining metabolomics and transcriptomics analyses, functional motif blast, and KEGG (Kyoto Encyclopedia of Genes and Genomes) annotation were used to discover three key post‐modification enzymes involved in the biosynthesis of lanostanoids in the medicinal mushroom Antrodia camphorata. The cytochrome P450 enzyme AcCYP4 could generate a Δ7,9(11) diene structure and introduce a 15α‐hydroxy group to the triterpene skeleton. The short‐chain dehydrogenase AcSDR6 could regio‐ and stereo‐ selectively catalyze the dehydrogenation of 3β‐OH to produce 3‐keto triterpenoids, and the catalytic mechanisms were interpreted by crystal structure analysis. AcSMT1 could introduce the methyl group at C‐24 to produce a unique 31‐carbon triterpene skeleton. This work elucidated the major biosynthetic pathway of Antrodia lanostanoids in vitro, and the discovered enzymes could be used to synthesize a series of bioactive triterpenoids.

Anthocyanin accumulation enhances drought tolerance in purple-leaf Brassica napus: transcriptomic, metabolomic, and physiological evidence

Anthocyanins are flavonoid compounds that provide plants with various advantages, including resistance to abiotic stress, pollinator attraction, and antioxidant properties. The present study aimed to investigate the impact of anthocyanin accumulation on drought tolerance in two genotypes of Brassica napus; purple-leaf (PL) and green-leaf (GL). Transcriptomic, metabolomic, and physiological analyses were conducted under drought conditions induced by polyethylene glycol (PEG-6000) for 4 and 8 days, with an additional evaluation after 4 days of re-watering. After drought stress, the PL genotype showed higher anthocyanin content, relative water content, and total soluble sugar compared to the GL genotype. Furthermore, the PL genotype exhibited reduced levels of reactive oxygen species, higher antioxidant enzyme activities, and less damage to the chloroplast ultrastructure than the GL genotype. These results indicate that the PL genotype shows significant advantages over GL, demonstrating improved water holding capability and stress tolerance. Moreover, under mild drought and re-watering conditions, the PL genotype of B. napus exhibited significant upregulation of structural genes expression associated with the biosynthesis of flavonoids. These correlate with increased anthocyanin content, particularly cyanidin glycosides and flavanols. The PL genotype showed greater gene expression related to flavonoid biosynthesis, particularly anthocyanin production, compared to GL. Alongside, PL accumulated higher levels of anthocyanin-related metabolites. These results suggest that enhanced flavonoid production contributes to greater drought resistance in PL. Additionally, by identifying important genes and metabolites related to anthocyanin accumulation and drought response, this integrated study provides insights into the intricate regulatory network that underlies these processes. These findings emphasize the potential of PL as a valuable genetic resource for developing drought-resistant rapeseed cultivars.

Macamides as Potential Therapeutic Agents in Neurological Disorders.

Therapeutic treatment of nervous system disorders has represented one of the significant challenges in medicine for the past several decades. Technological and medical advances have made it possible to recognize different neurological disorders, which has led to more precise identification of potential therapeutic targets, in turn leading to research into developing drugs aimed at these disorders. In this sense, recent years have seen an increase in exploration of the therapeutic effects of various metabolites extracted from Maca (Lepidium meyenii), a plant native to the central alpine region of Peru. Among the most important secondary metabolites contained in this plant are macamides, molecules derived from N-benzylamides of long-chain fatty acids. Macamides have been proposed as active drugs to treat some neurological disorders. Their excellent human tolerance and low toxicity along with neuroprotective, immune-enhancing, and and antioxidant properties make them ideal for exploration as therapeutic agents. In this review, we have compiled information from various studies on macamides, along with theories about the metabolic pathways on which they act.

Metabolomic and transcriptomic analyses reveal the regulation mechanism of postharvest light-induced phenolics accumulation in mango peel

Phenolics are important secondary metabolites and antioxidants in plants. Mango fruit accumulate abundant phenolic compounds, while the effect of light on the accumulation of different phenolic components in mango peel and the relevant molecular mechanism are still unknown. In this study, mature ‘Guifei’ mango fruit was subjected to postharvest UV-B/white light treatment, fruit peel was sampled for metabolomic and transcriptomic analyses. The results showed that light induced the accumulation of anthocyanins, flavonoids, and phenolics, thus increasing the antioxidant capacity in mango peel. A total of 1223 phenolic metabolites were detected in mango peel, 36 phenolic compounds were defined as key differentially accumulated metabolites (DAMs) regulated by light. Among the DAMs, the accumulation of 33 compounds was promoted by light, and 30 of these were flavonoids. Light up-regulated most phenolics biosynthesis and light signaling pathway genes and also regulated expression of plant hormone signaling pathway genes. Transcription factors (TFs) such as MYB, C2H2 and HSF were identified as candidates regulating phenolics biosynthesis in mango. Our findings not only provide information for commercial application of light in promoting mango fruit appearance and increasing phenolics content and antioxidant capacity, but also reveal important aspects of the molecular regulation of light-induced phenolics accumulation in mango peel.

Apigeninidin chloride disrupts Toxoplasma gondii Mitochondrial membrane potential and induce reactive oxygen species and metabolites production.

Apigeninidin chloride (APi) is a form of 3-deoxyanthrocyanidins (3-DAs) abundantly produced by the red Sorghum bicolor plant. It has been previously reported to be effective against Toxoplasma gondii (T. gondii) tachyzoites grown in vitro with less cytotoxic effect. However, its possible mechanism(s) of action has not been elucidated. Biochemically, we discovered that APi induced high reactive oxygen species (ROS) and mitochondria superoxide (MitoSOX) productions in tachyzoites, leading to mitochondrial membrane potential (MMP) disruption in vitro. To confirm our biochemical results at the molecular level, we performed a liquid chromatography-mass spectrometry (LC-MS) analysis on APi-treated parasites to assess any metabolite and lipid alterations often associated with high ROS/MitoSOX production in cells. Noteworthy is that we detected several important oxidative stress-induced metabolites such as hexanal, aldehydes, methyl undeo10-enoate, butadiynyl phenyl ketone, 16-hydroxyhexadecanoic acid (16-OH, 16:0), 2-hydroxytricosanoic acid (C23:0; O), 3-oxodecanosanoic acid (C22:1; O), 2-hydroxypropylsterate, and furan fatty acids F6 (19FU-FA). These metabolites are associated with lipid, protein, and nucleic acid disruptions. Using atovaquone (Atov) as a control, we observed that it disrupted intracellular tachyzoites' mitochondrial membrane potential, increased ROS and MitoSOX production, and altered metabolite and lipid production similar to what was observed with our experimental compound APi. Overall, our results indicated that APi targets T. gondii tachyzoite growth through inducing oxidative stress, mitochondrial dysfunction, and eventually parasite death.

Effect of Ag+ and Cd2+ Elicitation on Polyphenol Production in Shoot Culture of Dracocephalum ruyschiana L.

Abiotic elicitation with heavy metals has demonstrated considerable potential to stimulate the production of industrially important secondary metabolites in plant in vitro cultures. The present study investigates the effect of exogenous silver nitrate and cadmium chloride supplementation on flavonoid and phenolic acid production, as well as other indicators of oxidative stress, in shoot cultures of Dracocephalum ruyschiana L. Owing to the presence of bioactive polyphenolic compounds, this Mongolian medicinal plant is traditionally used as an anti-inflammatory, antibacterial and antipyretic agent. The shoots were cultured for three weeks, and then, cadmium (Cd2+) and silver (Ag+) ions (50 or 100 µM) were added to the medium. The maximum proliferation rate was observed in the presence of 100 µM Ag+ (almost 5), the highest chlorophyll content in the presence of 100 µM Cd2+ (0.6 mg/g FW) and the highest biomass was observed with both these treatments (73.4-75.7 g FW and 7.53-7.72 g DW). UPLC-PDA-ESI-MS analysis revealed four phenolic acids and five flavonoid derivatives in the hydromethanolic extract of D. ruyschiana shoots. All treatments stimulated the production of rosmarinic acid (RA), which was the dominant compound in the analyzed culture; the highest level of RA, i.e., about three times higher than the control, was noted in shoots exposed to 50 µM Cd2+ (14.72 mg/g DW), whereas the level of most flavonoids in the culture increased most significantly when exposed to Cd2+ at a concentration of 100 µM. Moreover, the shoots grown in the presence of 100 µM Cd2+ exhibited significantly higher antioxidant potential in comparison to the control. Our findings indicate that heavy metals are able to stimulate phenolic compound biosynthesis in Dracocephalum shoots without any negative impact on their growth. These results could be of significant importance for the medical, nutraceutical and agronomic industries.

Ethnobotany, Phytochemistry, Pharmacology and Therapeutic Potential of Su‘ad Kūfī (Cyperus rotundus L.): An Insight

Background: Cyperus rotundus, an important medicinal plant belonging to the Cyperaceae family, is therapeutically used in many traditional systems of medicine, such as Unani medicine, Ayurveda, Siddha, Homeopathy, Chinese medicine, etc. Aim: The prime goal of this review is to provide a scientific basis and classical references for clinical use and further scientific exploration of Cyperus rotundus. Materials and Methods: This systemic review was carried out after searching Unani classical and other ethnobotanical literature and published research work available on PubMed, SCOPUS, Science Direct, Google Scholar, Research Gate, etc. Results: Cyperus rotundus L. is the accepted botanical name of Su'ad Kūfī, Nagarmotha, or Nutgrass as referred in Unani and other traditional medicines. It is grown in India, China, the Philippines, Thailand, Taiwan, Indonesia, Korea, Vietnam, Malaysia, the Pacific Islands, South America, Africa, the Middle East, North America, Mexico, Australia, New Zealand, and Europe. Usually, the rhizome of this important medicinal plant is therapeutically used for various ailments, including indigestion, constipation, dysentery, neurogenic gastralgia, skin diseases, mental weakness, cardiac weakness, nervine weakness, palpitation, weakness of stomach, flatulence, retention of urine, amenorrhea, dribbling of urine, etc. It contains many important secondary metabolites, including flavonoids, tannins, glycosides, monoterpenes, sesquiterpenes, sitosterol, alkaloids, saponins, terpenoids, essential oils, sugar, protein, amino acids, etc., which have been reported to possess antibacterial, anti-tumour, anticonvulsant, antidiabetic, anti-diarrhoeal, anti-inflammatory, antilipidemic, antimalarial, anti-obesity, antioxidant, hepatoprotective, cardioprotective, and neuroprotective pharmacological activities. Conclusion: It is concluded that abundant bioactive compounds identified and separated from Cyperus rotundus possess potential medicinal values on different systems of the body.

PhAN2 regulated Ph3GT silencing changes the flower color and anthocyanin content in petunias.

Anthocyanins are important secondary metabolites in plants. After the formation of anthocyanidins, Flavonoid 3-O-glucosyltransferase (3GT) mediated glycosylation first occurs at the C-3 site, forming a stable anthocyanin 3-O-glucoside. Several studies have investigated the function of 3GT using biochemical methods. However, it is necessary to provide further genetic evidence for the role of Ph3GT in petunia (Petunia hybrida). In addition, there is no information regarding the subcellular localization of Ph3GT and the regulation of transcription factors on Ph3GT. In this study, the full-length Ph3GT gene from petunia (Petunia hybrida) was isolated. We found that Ph3GT is localized in the cytoplasm. Ph3GT exhibited high expression levels in the corollas during the coloring period of petunia flowers. VIGS-mediated Ph3GT silencing resulted in a lighter corolla color and a significant decrease in the anthocyanin content in six petunia cultivars. The silencing of Ph3GT affected the expression levels of eight key genes in the anthocyanin synthesis pathway. Additionally, dual luciferase and yeast one-hybrid assays showed that R2R3-MYB transcription factor PhAN2 directly regulates the transcript of Ph3GT.

LncNAT11-GbMYB11-GbF3'H/GbFLS module mediates flavonol biosynthesis to regulate salt stress tolerance in Ginkgo biloba.

Flavonols are important secondary metabolites that enable plants to resist environmental stresses. Although MYB regulation of flavonol biosynthesis has been well studied, the lncRNA-MYB networks involved in regulating flavonol biosynthesis remain unknown. Ginkgo biloba is rich in flavonols, which are the most important medicinal components. Based on multi-omics data and phylogenetic trees, we identified GbMYB11 as a potential key transcription factor regulating flavonol biosynthesis. Overexpression and VIGS experiments confirmed that GbMYB11 acts as a pivotal positive regulator in flavonol biosynthesis. In the transcriptome of calli overexpressing GbMYB11, we identified significant upregulation of GbF3'H and GbFLS in the flavonol biosynthetic pathway. Yeast one-hybrid and dual-luciferase assays demonstrated that GbMYB11 enhances the expression of GbF3'H and GbFLS by binding to their promoters. Interestingly, we identified LncNAT11, an antisense lncRNA complement to GbMYB11, which negatively regulates flavonol biosynthesis by repressing the expression of GbMYB11. Consequently, we established the LncNAT11-GbMYB11-GbF3'H/GbFLS module as a critical regulator of flavonol biosynthesis in G. biloba, and further elucidated that this module can mitigate the accumulation of reactive oxygen species by modulating flavonol biosynthesis during salt stress. These findings unveil a novel mechanism underlying flavonol biosynthesis and a lncRNA-MYB mediated salt stress tolerance strategy in plants.

Comparative Metabolome and Transcriptome Analysis Revealed the Accumulative Mechanism of Rubusoside in Chinese Sweet Tea.

Terpenoids are important secondary metabolites in Rubus. Rubusoside is a relatively specific diterpenoid bioactive component in the leaves of Chinese Sweet Tea (Rubus suavissimus). However, the terpenoid anabolic pathway of Rubus and the molecular mechanism underlying the specific accumulation of rubusoside in R. suavissimus remain unclear. Here, metabolomics and transcriptomics analyses were performed on differences in terpenoid metabolism levels between R. suavissimus (sweet leaves) and Rubus chingii (bitter leaves). Steviol glycosides and goshonosides primarily accumulated in R. suavissimus and R. chingii, respectively. Three pairs of highly homologous glycosyltransferase genes (UGT85A57, UGT75L20, and UGT75T4) associated with rubusoside biosynthesis in the two Rubus species were identified. The three pairs of UGT proteins in both species could glycosylate steviol. Thus, the transcriptional regulation of UGTs in R. suavissimus appears to play a pivotal role in rubusoside accumulation. Our findings provide insights into the differences in terpenoid metabolism between R. suavissimus and R. chingii and reveal the molecular mechanism of rubusoside accumulation in R. suavissimus.

Mechanistic effects of lipid binding pockets within soluble signaling proteins: Lessons from acyl-CoA-binding and START-domain-containing proteins.

While lipids serve as important energy reserves, metabolites, and cellular constituents in all forms of life, these macromolecules also function as unique carriers of information in plant communication given their diverse chemical structures. The signal transduction process involves a sophisticated interplay between messengers, receptors, signal transducers, and downstream effectors. Over the years, an array of plant signaling proteins have been identified for their crucial roles in perceiving lipid signals. However, the mechanistic effects of lipid binding on protein functions remain largely elusive. Recent literature has presented numerous fascinating models that illustrate the significance of protein-lipid interactions in mediating signaling responses. This review focuses on the category of lipophilic signaling proteins that encompass a hydrophobic binding pocket located outside of cellular membranes and provides an update on the lessons learned from two of these structures, namely the acyl-CoA-binding and START domains. It begins with a brief overview of the latest advances in understanding the functions of the two protein families in plant communication. The second part highlights five functional mechanisms of lipid ligands in concert with their target signaling proteins.

Manipulating the light spectrum to increase the biomass production, physiological plasticity and nutritional quality of Eruca sativa L

The extensive development in light-emitting diodes (LEDs) in recent years provides an opportunity to positively influence plant growth and biomass accumulation and to optimize biochemical composition and nutritional quality. This study aimed to assess how different light spectra affect the growth, photosynthesis and biochemical properties of Eruca sativa. Therefore two LED lighting modes - red:blue (RB, 1:1) and red:green:blue (RGB, 2:1:2) were compared to the conventional white light fluorescent tubes (WL). Plant biomass, photosynthetic performance, several antioxidants, polyamines and nitrates contents were analyzed across different treatments. The plant growth was affected by the light quality - the presence of green light in the spectrum resulted in smaller plants and leaves, and correspondingly less biomass. RB spectral mode enhanced the total antioxidant and guaiacol peroxidase activity, pigments, flavonoids, polyphenols, ascorbate and polyamines contents. This effect under RB was combined with better leaf development compared to RGB and less nitrate in the leaves among all treatments. The RB light generated modifications in polyamines, which are interrelated with the nitrate content, further induce important metabolite and antioxidant changes. Both RB and RGB enhanced photosynthesis. The afterglow thermoluminescence band varied according to leaves development, being higher in RB and WL as a consequence of their faster growth. The RB light spectrum was found to be the most efficient for promoting the growth, biochemical composition, and overall quality of Eruca sativa compared to RGB and WL. These findings suggest that RB LEDs can be an effective tool for improving crop production in controlled environments.

ABA deficiency and its impact on ion and metabolite profiles in tomato roots under single and combined stress conditions

Abscisic acid (ABA) plays a crucial role in the stress response of plants. Although the impact of ABA on individual stresses has been extensively studied, there is less research on its role in plants grown under stress combination, such as salinity and high temperature. This study analyzes the response at the ionomic and metabolomic levels of tomato roots to ABA deficiency under single salinity or heat stress, as well as under the combination of both stresses, using ABA-deficient (flacca, flc) tomato plants. ABA was found to be crucial in ionic regulation, particularly for Na, K, Ca, and other ions such as Cu, Mn, Zn, Mo, and Fe. However, its influence depended on the type of stress applied, indicating the complexity of plant responses to these adverse environmental factors. In our study, phenylpropanoids, terpenoids, and nitrogenous compounds were the metabolites most affected by endogenous ABA concentration and environmental treatment. On the other hand, the application of exogenous ABA to flc mutants did not fully restore root dry weight, although it did cause significant changes at the ionomic and metabolomic levels. Salinity and heat applied in combination aggravated the vulnerability of flc mutants compared to single stresses, making the application of exogenous ABA more effective under single stresses than under the combined stresses. Finally, a correlation analysis between the ionome and metabolome revealed that the accumulation or deficiency of some ions (i.e., Na, Zn, and Fe) was correlated with the abundance of important metabolites related to amino acid biosynthesis and terpenoid metabolism, among others.

Distributions of Lanostene-Derived Triterpenoids and Glucan Content in the Fruiting Bodies of the Australian Ganoderma Species.

Lanostene-derived triterpenoids and β-glucans are important metabolites in Ganoderma mushrooms associated with benefits to human health. The medicinal value of the Australian Ganoderma species remains unclear, with no data on triterpenoid distribution or glucan content. In the present study, 22 Australian Ganoderma specimens were analyzed for triterpenoid and glucan contents. Thirty-two triterpenoids were identified in the fruiting bodies of 19 of the specimens. Distinct patterns in triterpenoid distribution between laccate and matte fruiting bodies were observed, leading to the classification of four groups of Ganoderma. Most of the glucans in the Ganoderma fruiting bodies were β-glucans (~99%), with a nominal α-glucan content (~1%). The β-glucan content ranged from 19.5 to 43.5% (w/w). A range of antioxidant activities was observed for methanol extracts using the ABTS (1.8 to 8.4 mg GAE.g-1), DPPH (1.7 to 9.4 mg GAE/g-1) and FRAP (24.7 to 111.6 mmol FeSO4.g-1) assays, with four specimens presenting relatively high radical scavenging and reducing activities. For the first time, we demonstrated that Australian Ganoderma mushrooms contain medicinal triterpenoids, including ganoderic acid A, and we established a link between its distribution and the fruiting body morphology. However, further research is required to isolate diploid clones and determine factors that impact triterpenoid and glucan synthesis in these strains.

Integrated Analysis of Transcriptome and Metabolome Provides Insights into Flavonoid Biosynthesis of Blueberry Leaves in Response to Drought Stress.

Blueberries (Vaccinium spp.) are extremely sensitive to drought stress. Flavonoids are crucial secondary metabolites that possess the ability to withstand drought stress. Therefore, improving the drought resistance of blueberries by increasing the flavonoid content is crucial for the development of the blueberry industry. To explore the underlying molecular mechanism of blueberry in adaptation to drought stress, we performed an integrated analysis of the metabolome and transcriptome of blueberry leaves under drought stress. We found that the most enriched drought-responsive genes are mainly involved in flavonoid biosynthesis and plant hormone signal transduction pathways based on transcriptome data and the main drought-responsive metabolites come from the flavonoid class based on metabolome data. The UDP-glucose flavonoid 3-O-glucosyl transferase (UFGT), flavonol synthase (FLS), and anthocyanidin reductase (ANR-2) genes may be the key genes for the accumulation of anthocyanins, flavonols, and flavans in response to drought stress in blueberry leaves, respectively. Delphinidin 3-glucoside and delphinidin-3-O-glucoside chloride may be the most important drought-responsive flavonoid metabolites. VcMYB1, VcMYBPA1, MYBPA1.2, and MYBPA2.1 might be responsible for drought-induced flavonoid biosynthesis and VcMYB14, MYB14, MYB102, and MYB108 may be responsible for blueberry leaf drought tolerance. ABA responsive elements binding factor (ABF) genes, MYB genes, bHLH genes, and flavonoid biosynthetic genes might form a regulatory network to regulate drought-induced accumulation of flavonoid metabolites in blueberry leaves. Our study provides a useful reference for breeding drought-resistant blueberry varieties.

Identifying Potential Natural Antibiotics from Unani Formulas through Machine Learning Approaches.

The Unani Tibb is a medical system of Greek descent that has undergone substantial dissemination since the 11th century and is currently prevalent in modern South and Central Asia, particularly in primary health care. The ingredients of Unani herbal medicines are primarily derived from plants. Our research aimed to address the pressing issues of antibiotic resistance, multi-drug resistance, and the emergence of superbugs by examining the molecular-level effects of Unani ingredients as potential new natural antibiotic candidates. We utilized a machine learning approach to tackle these challenges, employing decision trees, kernels, neural networks, and probability-based methods. We used 12 machine learning algorithms and several techniques for preprocessing data, such as Synthetic Minority Over-sampling Technique (SMOTE), Feature Selection, and Principal Component Analysis (PCA). To ensure that our model was optimal, we conducted grid-search tuning to tune all the hyperparameters of the machine learning models. The application of Multi-Layer Perceptron (MLP) with SMOTE pre-processing techniques resulted in an impressive accuracy precision and recall values. This analysis identified 20 important metabolites as essential components of the formula, which we predicted as natural antibiotics. In the final stage of our investigation, we verified our prediction by conducting a literature search for journal validation or by analyzing the structural similarity with known antibiotics using asymmetric similarity.

Research Progress on the Mechanism for Improving Glucose and Lipid Metabolism Disorders Using Phenolic Acid Components from Medicinal and Edible Homologous Plants.

Glucose and lipid metabolism disorders are the core pathological mechanism of a variety of metabolic diseases, and the incidence of related diseases is increasing year by year, which seriously threatens human life and health. Traditional Chinese medicine with medicinal and edible properties refers to Chinese medicinal resources that have both medicinal and edible characteristics. Due to its safety and its health-promoting and medicinal functions, traditional Chinese medicine has received increasing attention in the development of functional health foods. Phenolic acids are important secondary metabolites that are ubiquitous in medicinal and edible homologous plants, and the regulation of glycolipid metabolism is an important activity and plays a key role in many diseases. In this paper, we focus on the alleviation of glycolipid disorders using MEHH phenolic acids, which regulate glucose metabolism and lipid metabolism, improve insulin resistance, inhibit inflammatory responses, alleviate oxidative stress, and regulate intestinal flora; additionally, we summarize the mechanism in order to provide a reference for MEHH phenolic acids in the treatment of glycolipid metabolism diseases.

Multitemporal UAV study of phenolic compounds in slash pine canopies

Phenolic compounds (PC) are important secondary metabolites in plants, playing a crucial role in plant defense mechanisms against pathogens and other plants. Monitoring PC levels is important for understanding tree stress and implementing effective breeding programs. However, traditional methods for monitoring PC are time-consuming, prone to altering the phenolic composition, and mostly applicable only on a small scale. In this study, we evaluated the performance of Unoccupied Aerial Vehicles (UAV) multispectral imaging in estimating the canopy phenolic content in slash pine over an 11-month period in 2021 and a seven-month period in 2022. Three machine learning models including Partial least squares regression (PLSR), Random forest (RF) and Support Vector Machine (SVM) were compared to determine the optimal predictive model for canopy PC. The RF model provided the best predictive results, with R2 values of 0.82 for the validation set and 0.94 for the calibration set. Additionally, the study assesses the heritable variation in canopy PC over time, with the monthly heritability (h2) of PC ranging from 0 to 0.26 in 2021 and from 0 to 0.35 in 2022; The highest h2 levels were observed in July and September 2021and July 2022. The findings demonstrate significant genetic control over the variation of PC. Furthermore, we observed higher breeding values and genetic gains in July and November, which further supports the strong correlation between PC and environmental factors such as temperature and light intensity. To the best of our knowledge, this is the first study to employ time-series UAV multispectral imaging to predict secondary metabolites in pine trees and estimate their genetic variation over time. As a proof of concept, these findings provide more reliable information for tree breeding programs, ultimately enhancing their overall performance.

Improving organoleptic and antioxidant properties by inhibition of novel miRstv_7 to target key genes of steviol glycosides biosynthetic pathway in Stevia rebaudiana Bertoni.

Stevioside (5-10%) and rebaudioside-A (2-4%) are well-characterized diterpene glycosides found in leaves of Stevia rebaudiana known to have natural sweetening properties with zero glycaemic index. Stevioside has after-taste bitterness, whereas rebaudioside-A is sweet in taste. The ratio of rebaudioside-A to stevioside needs to be changed in order to increase the effectiveness and palatability of this natural sweetener. Plant-specific miRNAs play a significant role in the regulation of metabolic pathways for the biosynthesis of economically important secondary metabolites. In this study inhibition of miRNA through antisense technology was employed to antagonize the repressive action of miRstv_7 on its target mRNAs involved in the steviol glycosides (SGs) biosynthesis pathway. In transgenic plants expressing anti-miRstv_7, reduced expression level of endogenous miRstv_7 was observed than the non-transformed plants. As a result, enhanced expression of target genes, viz. KO (Kaurene oxidase), KAH (Kaurenoic acid-13-hydroxylase), and UGT76G1 (UDP-glycosyltransferase 76G1) led to a significant increase in the rebaudioside-A to stevioside ratio. Furthermore, metabolome analysis revealed a significant increase in total steviol glycosides content as well as total flavonoids content. Thus, our study can be utilized to generate more palatable varieties of Stevia with improved nutraceutical values including better organoleptic and antioxidant properties.

Phytochemical Screening, Antioxidant, Cytotoxic, Analgesic, Antidiarrheal Activity and GC-MS Analysis of the Leaves of Stachyphrynium Placentarium

The goal of the present study is to analyze the phytochemicals, antioxidants, cytotoxic, analgesic and anti-diarrheal properties of the leaves of Stachyphrynium placentarium methanol extract as well as its dichloromethane soluble fraction using GC-MS technique. The leaves of the S. placentarium were extracted with methanol and designated as SPM. Phytochemicals investigation exhibited the presence of several important secondary metabolites. Antioxidants such as total phenolic content, flavonoids content, total antioxidant, reducing power capacity and DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging assay SPM revealed non-significant antioxidant properties. The cytotoxicity study of the extract showed significant cytotoxicity (LC50 35.1093 µg/mL) compared to standard vincristine sulfate (LC50 0.482 µg/mL). Analgesic study of S. placentarium by hot plate model indicates both the doses exhibited significant central analgesic activity compared with the control. The maximum analgesic activity was observed at 90 min. SPM (500 mg/kg) is 60.21 %. SPM250 also showed a significant percentage of elongation or latency from 30 to 150 min 57.00, 59.64, 59.88, 58.07 and 53.71 %, respectively 30 min time interval. In the acetic acid writhing test, S. placentarium significantly outperformed the control and standard (SPM250 - 52.27 %, SPM500 - 34.09 and STD - 27.27 %), showing a significant increase in peripheral analgesic activity (p < 0.001). Antidiarrheal activity of SPM showed that SPM250 and SPM500 substantially decreased the feces in castor oil-induced diarrhea (p < 0.05). According to the study, the incidence and severity of diarrhea are significantly reduced by SPM500. Four chemicals are found in the dichloromethane-soluble portion of the methanol extract of S. placentarium leaves, according to GC-MS studies such as Octadecane, 6-methyl- (1.326 %), Dodecane (1.02 %), 1-Iodo-2-methylundecane (0.47 %), tetradecanoic acid 12-methyl, methyl ester (72.64 %). Tetradecanoic acid exhibits anti-inflammatory, anticancer, 5-reductase inhibitory, antioxidant, hemolytic and antifibrinolytic properties. HIGHLIGHTS The phytochemicals, antioxidants, cytotoxic, analgesic and anti-diarrheal properties of Stachyphrynium placentarium methanol extract (SPM) leaves. The dichloromethane soluble fraction is examined using Gas chromatography–mass spectrometry (GC-MS) and identified Octadecane, 6-methyl-, Dodecane, 1-Iodo-2-methylundecane, tetradecanoic acid 12-methyl, methyl ester. The brine shrimp lethality test exhibited that SPM has significant cytotoxicity (LC50 35.1093 µg/mL) compared to vincristine sulphate (LC50 0.482µg/mL), the extract showed significant cytotoxicity (LC50 35.1093 µg/mL). S. placentarium at both dosages (250 and 500 mg) provided significant central analgesia compared to the control in a hot plate model. In the acetic acid writhing test, S. placentarium showed significantly higher peripheral analgesic action (SPM250 - 52.27, SPM500 - 34.09 and STD - 27.27 %) compared to control and standard. In castor oil-induced diarrhoea, SPM250 and SPM500 (250 and 500 mg) showed significantly reduced faeces production (p < 0.05). GRAPHICAL ABSTRACT