<div style="text-align: justify;">The Indian perfumery industry is shifting towards natural product. In India including West Bengal betel leaves produces high quality essential oil as well contribute to Indian fresh vegetable export. The crop is cultivated from stem cutting and suffers from authenticity problem of cultivars with redundant names. The genetic screening and characterization of cultivars were not initiated due to unavailability of reliable markers. The essential oil metabolomic study identified some polar and non-polar volatile signature compounds. Metabolomic profiling of cultivars is not consistent due to seasonal variation in the production of secondary metabolites and ignorance in marking of unique trace discriminatory compounds. In this paper gene ontogeny study was made on major signature compounds to obtain the complete coding sequence (CDS) of the aroma-genes. The CDS information of aroma-genes could be utilized to construct robust DNA markers to eradicate authentication problem and germplasm management of Piper. The direct genomic analysis could supersede the metabolome profiling. Information available in NCBI, DDBJ and EMBL database were searched for gene ontogeny study utilizing available metabolomic data. The information and method depicted could be efficiently utilized for Piper genomics. Aroma-scientists could apply this technique to validate promising cultivars and competent germplasm management. &nbsp;</div>

<div style="text-align: justify;">Conversion of Metabolomic Data to Genomic Marker for Genetic Characterization of Piper betle L. Chemotypes: A Review</div>

<div>Background: Solanaceae plants have the potential to be a source of food, medicine, industrial materials and antioxidants.&nbsp;Antioxidants are compounds that can delay or prevent oxidation caused by free radicals significantly. Metabolomic analysis is a method used to study metabolite profiles. It is necessary to use elicitors to optimize the synthesis of metabolomic compounds and antioxidant potential: methyl jasmonate and salicylic acid as elicitors. 
 
Methods: This research aims to analyze the Solanaceae plant&rsquo;s metabolomic compounds and antioxidant potential using GCMS after the elicitors methyl jasmonate and salicylic acid induction. Experimental method research with stages: 1) Observation, collection and identification of Solanaceae plants, 2) Preparation of experimental land, 3) Planting of plant collections, 4) Application of elicitor methyl jasmonate 60 ppm and salicylic acid 0.45 g/L, 5) Metabolomic and antioxidant analysis on fresh extracts of samples. Testing antioxidant activity using a spectrophotometer with DPPH solution. Metabolomic profile analysis using GCMS. 
 
Result: The results of the non-targeted metabolomic analysis with GCMS and antioxidant activity with IC50&nbsp;in the Solanum genus, namely Solanum betaceum, Solanum melongena, Solanum torvum, Solanum Lycopersicum and Solanum tuberosum, respectively identified 18, 16, 14, 13 and 20 compounds. The IC50&nbsp;values&nbsp;&nbsp; were 51.32 mg/L (strong category), 56.64 mg/L (strong category), 65.86 mg/L (strong category), 113.60 mg/L (medium category), 102.71 mg/L (medium category), respectively. In the Capsicum genus, namely Capsicum frutescens and Capsicum annuum, 21 and 37 compounds were identified, respectively, with IC50&nbsp;values&nbsp; of 124.76 mg/L (medium category) and 63.23 mg/L (strong category). In Physalis angulata, 19 compounds were identified, IC50&nbsp;was 44.04 mg/L (very strong category), six compounds were identified in&nbsp; Nicotiana tabacum and IC50&nbsp;was 67.55 mg/L (strong category).&nbsp; Solanaceae plants have moderate to very strong antioxidant potential. 
&nbsp;</div>

<div>Analysis of Metabolomic Compounds and Antioxidant Potential of Solanaceae Family Plants using Gcms after Elicitor Induction Methyl Jasmonate and Salicylic Acid</div>

Background: Estrus detection is crucial for reproductive performance of goats which requires accurate, economical and easy techniques. Present study was undertaken to delineate urine metabolomics profile to identify biomarkers for different phases of estrous cycle in Surti goats. 
 
Methods: Ten non-pregnant Surti goats were synchronized for estrous using intra-vaginal progesterone sponges (impregnated with 60 mg Medroxyprogesterone acetate) for 11 days followed by intramuscular injection of 125 &mu;g Cloprostenol (PGF2a analogue). Estrus was confirmed based on physiological and behavioral signs. Urine samples collected during proestrus, estrus and diestrus were analyzed using gas chromatography-mass spectrometry. Web-based interface MetaboAnalyst 5.0 was used for data preprocessing, normalization, statistical analysis and data visualization. Statistical analysis comprised data normalization, one-way ANOVA and post-hoc Fisher LSD test, hierarchical clustering, principal component analysis and partial least squares discriminant analysis. 
 
Result: Total 56 metabolites were found to vary during different phases of estrous cycle. The study was successful in generating untargeted urine metabolomics profile of Surti goats that can be used for identifying specific biomarkers for different phases of estrous. Two metabolites 4-amino-2-hydroxybenzoic acid (2-dieth-4-A-2) and 4-Methylphenol (p-Cresol) emerged as potential biomarkers for detecting estrus. These results can serve as base line reference paving way for future studies in larger group that may further validate these findings.

Untargeted Urine Metabolomics Profiling during Different Phases of Estrous Cycle in Surti Goats

Background: Varietal differences in metabolite composition play a crucial role in determining agronomic traits, nutritional quality and stress resilience in Solanum tuberosum.&nbsp; 
 
Methods: Three potatoes varieties; Longshu 7, Longshu 10 and Longshu 14 were selected and bred by the at the Potato Research Institute of Gansu Academy of Agricultural Sciences between December 2021 to December 2023. Thereafter, comparative metabolomics analysis of three new potato varieties (LS7, LS10 and LS14) was undertaken against the well-established Atlantic cultivar. We employed a bioinformatics-based experimental approach to analyze the metabolomic data obtained from the three new potato varieties (LS7, LS10 and LS14) and the Atlantic cultivar. Metabolite profiling was performed using high-throughput liquid chromatography-mass spectrometry (LC-MS) techniques. Differential metabolite analysis and pathway enrichment analysis were conducted to identify significant metabolic differences and enriched pathways among the varieties.&nbsp; 
 
Result: Results revealed substantial differences in metabolite composition and pathway enrichment between the new potato varieties and the Atlantic cultivar. Metabolites involved in diverse metabolic pathways, including glucoside and hydroxyproline in LS7, kaempferol and ascorbic acid in LS10 and phenyl-butyryl-glutamine and ascorbic acid in LS14, exhibited significant upregulation or downregulation compared to the Atlantic cultivar. Pathway enrichment analysis highlighted the prominence of ABC transporters across all comparisons, suggesting their universal role in mediating cellular processes and stress responses in potato varieties. In conclusion, these findings contribute to our understanding of potato metabolism and pave the way for the development of novel breeding approaches to meet the evolving demands of global agriculture and food security.

Comparative Metabolomics Analysis of 3 New Potato Varieties against the Atlantic Cultivar; Bioinformatics based Experimental Study

Background: Low-temperature stress is a major abiotic factor that significantly impacts plant growth and productivity, especially in cold climatic conditions. Trifolium ambiguum M. Bieb. (Caucasian clover), a perennial legume known for its cold tolerance and it is an ideal model for studying cold adaptation mechanisms. However, the metabolic pathways that underpin its cold tolerance are not well understood. This study used widely targeted metabolomics to examine the metabolic reprogramming of T. ambiguum under low-temperature stress. 
 
Methods: Trifolium ambiguum seedlings were grown under controlled conditions and 14-day-old plants were exposed to two treatments: low-temperature stress (4&deg;C) and control conditions (25&deg;C). Samples were collected two hours after treatment. Widely targeted metabolomics profiling was performed to analyze metabolic changes induced by low-temperature stress. Key pathways and metabolites involved in the low-temperature stress response were identified through kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment and metabolic network analysis. 
 
Result: Low-temperature stress induced significant metabolic reprogramming in Trifolium ambiguum, with notable alterations observed in fatty acid metabolism. This enhancement improved cell membrane fluidity and stability, thereby strengthening the plant&rsquo;s towards cold tolerance. Furthermore, the significant accumulation of proline and glutamate underscores their critical roles in osmotic regulation and antioxidative defense mechanisms. Additionally, the upregulation of secondary metabolites like (+)-Forbesione and flavonoids plays a key role in scavenging reactive oxygen species (ROS) and regulating stress-related signaling. KEGG pathway enrichment analysis revealed that C5-Branched dibasic acid metabolism and alpha-Linolenic acid metabolism play central roles in cold adaptation. C5-Branched dibasic acid metabolism provided essential carbon skeletons and energy substrates, while alpha-linolenic acid metabolism contributed to maintaining membrane stability and supporting antioxidative responses. The enrichment of the TCA cycle and glycerophospholipid metabolism further ensured energy supply and membrane integrity under stress conditions.

Metabolic Response of Trifolium ambiguum to Low-temperature Stress based on Widely Targeted Metabolomics

Background: There is a current lack of research on the molecular pathways by which lysine regulates feed intake in chicks. Regulation is controlled by the hypothalamus and lysine metabolism links the availability of amino acids and nutritional perception with food intake. The mechanism underlying this phenomenon remains unclear. To investigate the regulatory mechanism of lysine on feed intake in chicks, this study used Yao chickens as a research model. The levels of lysine in the feed of chicks were adjusted to determine the effect on feed intake. The hypothalamus tissue of the chicks was analyzed using RNA-seq and metabolomics to identify the relevant genes. We employed Gene Ontology and identification of signaling pathways to characterize the molecular regulatory biological pathways of lysine in poultry feeding. 
 
Methods: This study used RNA-seq technology to investigate the genes and marker metabolites involved in lysine regulation of feed intake in chickens. The study explored the molecular mechanism by which dietary lysine regulates feed intake in chickens via the hypothalamus. Four hundred healthy 1-day-old chicks (Yao chickens) were randomly divided&nbsp; into four groups, with five replicates in each group (n = 20). 
 
Result: There was a significant difference in feed intake among chicks fed with diets containing 0.65% (abbreviated as MGO), 0.85% (FIH), 1.0% (MGS) and 1.2% (FIL) lysine levels. The group with the highest feed intake was FIH, while the group with the lowest feed intake was FIL. By conducting RNA-seq of the hypothalamic tissue of chickens in both the highest and lowest feed intake groups, we obtained 2006 differentially expressed genes, comprising 1275 upregulated genes and 731 downregulated genes. Fourteen genes were related to food intake, including growth hormone, recombinant glutamate receptor, metabotropic 1&nbsp;(GRM1) and recombinant glutamate receptor, metabotropic 3&nbsp;(GRM3). Using liquid chromatograph mass spectrometer (LC-MS) metabolomics, we identified 25 differential metabolites. Nine metabolites, including C3H5NO4, C10H15NO2&nbsp;and C13H14N203, were downregulated, while 16 metabolites, including C2H8NO3P, C3H7NO3&nbsp;and C7H7NO2, were upregulated. Marker metabolites included 5-hydroxytryptophan and L-valine. A KEGG enrichment analysis revealed that the differential metabolites between the FIH and FIL groups were enriched in 26 pathways, with the significantly enriched pathway being the ABC transporter protein pathway (ID: map02010). RNA-seq and LC-MS analyses revealed that in chicks, the expression of the neuropeptides cocaine- and Amphetamine-Regulated Transcript Protein (CARTPT) and neuropeptide Y (NPY)/agouti-related protein (AGRP), along with genes such as sucrose synthase (SS 2), &nbsp;recombinant cholecystokinin A receptor (CCKAR) and recombinant cholecystokinin B receptor (CCKBR), was regulated by the hypothalamic feeding regulation center that senses the level of lysine in the feed. These molecules bind to specific receptors and initiate a signaling cascade in specific hypothalamic neuronal groups, thereby regulating chick feed intake and in turn affecting growth, development and production performance. 
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Investigating the Molecular Mechanisms of Lysine&rsquo;s Effect on Feed Intake in Chicks Through the Hypothalamus using RNA-seq and Metabolomics

Background: The aim of this experiment was to investigate the differences in subcutaneous fat metabolism expression in yaks under natural grazing and housed feeding conditions in the cold season. Eighteen 18-month-old yaks with similar body weights and non-significant differences in dorsal subcutaneous fat were randomly divided into three groups.&nbsp; 
 
Methods: One group (G18_SF) of yaks was selected for slaughter at the beginning of the experiment (October), one group continued to graze for 6 months and then slaughtered (G24_SF) and one group was housed for 6 months and then slaughtered (F24_SF) and subcutaneous dorsal fat of the three groups was collected for metabolomic analysis, screening of differential metabolites and enrichment of KEGG pathway. 
 
Result: In this experiment, 110 differential metabolites were detected in G24_SF vs F24_SF and 83 metabolites were screened for down-regulation and 27 metabolites were up-regulated and the differential metabolites were mainly enriched in Alanine, aspartate and glutamate metabolism, Arachidonic acid metabolism and D-Glutamine and D-glutamate metabolism pathways. Ninety-seven differential metabolites were detected in G18_SF vs G24_SF and 41 metabolites were screened for down-regulation and 56 metabolites were screened for up-regulation, with the two groups of differential metabolites mainly enriched in Alanine, aspartate and glutamate metabolism, Caffeine metabolism and Starch and sucrose metabolism pathways. The results of this experiment revealed the information of differential metabolites and enrichment pathways of yak fat in different feeding methods and different months of age and laid the foundation for further in-depth study of subcutaneous fat deposition in yaks.

Lipid Metabolomics Reveals Metabolic Responses of Subcutaneous Fat in Yaks under Different Feeding Conditions during the Cold Season

<div>Background:&nbsp;The pulse beetle, Callosobruchus chinensis L. is one of the main biotic threats to chickpea in both field and storage in the world. The evaluation of chickpea genotypes for pulse beetle resistance has increased motivation for the identification and application of host plant resistance as a critical component of pest management.&nbsp; 
 
Methods:&nbsp;In the present study, different chickpea genotypes were evaluated under laboratory conditions to identify the antixenosis, antibiosis and GC-MS based untargeted metabolomics of promising chickpea genotypes to the C. chinensis. 
 
Result:&nbsp;Genotype NBeG1004 (36.61 eggs/100 seeds) was less prone to egg laying, while ICCV14872 (180.45 eggs/100 seeds) was the most preferred. However, maximal developmental periods, minimum per cent adult emergence and growth index was observed in ICCV92944 Genotype HC1 showed substantial resistance with maximum phenolic, flavonoid and protease inhibitors content. The results revealed that presence of 9-Octadecenoic acid (Z) in HC1 conferred resistance to C. chinensis. These results show assurance for incorporation into an IPM program against C. chinensis.</div>

Gas Chromatography-Mass Spectrometry (GC-MS) based Metabolomics of Promising Chickpea Genotypes against Callosobruchus chinensis (L.)

Background: Production of female offspring is vital for maximizing milk yield in the dairy industry, making embryo sexing essential. Metabolic differences between male and female embryos may be reflected in the culture medium (CM). This study explored non-invasive sex detection of in vitro produced buffalo embryos by analyzing metabolites in spent CM. 
 
Methods: Buffalo ovaries were collected from a slaughterhouse and oocytes were matured and fertilized in vitro. Presumptive zygotes were cultured in either a sequential medium, Synthetic oviduct fluid (SOF) or a single-step medium, Potassium Simplex Optimized Medium (KSOM). After 5 days, morulae were placed in 25 &micro;l of CM. On day 7, embryos and spent medium were collected and stored at -20oC. Embryo sex was determined using PCR. Spent media were analyzed using Attenuated Total Reflection - Fourier Transform Infrared Spectroscopy (ATR-FTIR) spectroscopy to detect sex-specific metabolites. 
 
Result: This approach enabled non-invasive identification of embryo sex and revealed the impact of different culture media on sex ratio. The findings highlight the potential of spent CM analysis for predicting embryo sex, offering significant benefits for the dairy industry.

Comparative Analysis of Culture Media and ATR-FTIR Metabolomics for Non-invasive Sex Determination in In vitro Produced Buffalo Embryos

Background: Harvesting period is an important feature that not only affects the yield quality of Astragalus herbs but also the content of astragaloside IV and calycosin 7-O-&beta;-D-glucopyranoside which&nbsp; are considered as the core indicators of the quality of Astragalus herbs. Herb yield and quality undergo rapid and substantial changes during the harvesting process. Part of enzymes and their encoding genes have been identified as potential candidates for the biosynthesis of herb quality-related triterpenoid saponins and isoflavonoids. However, wide gap still exists in the understanding of the pathways involving growth, development and biosynthesis of these two important active components in the roots of Astragalus. 
&nbsp; 
Methods: In this study, we used Astragalus roots as material to analyze its metabolome and transcriptome of herbs at harvesting stage. 
&nbsp; 
Result: The results indicated that changes in the harvesting period had a significant effect on the yield and quality of the herbs. Correlation of metabolome and RNA-seq results constructed using weighted gene co-expression network analysis (WGCNA) with herb growth and development indicated that the expression patterns of 19 hub metabolites (Oxamniquine, 4-ene-Valproic acid, Traumatic Acid, L-Histidinol, Salidroside, Temsavir, Retaspimycin, Isosakuranetin, etc.) and 16 structural genes with annotations (CNX2, talA, ABCF2, HIBCH, crtZ, etc.) had the highest connectivity with root growth and secondary metabolism of Astragalus. To search for potential regulatory genes related to root growth and development, we screened 54 relevant genes using 16 hub genes with annotations. The results advance the molecular basis of biosynthesis of secondary metabolites related to the harvesting period of Astragalus herbs, which can provide a theoretical basis for determining the regulatory mechanism of yield and quality.

Weighted Correlation Network Analysis Mining Key Metabolites and Genes for Regulation of Root Growth and Secondary Metabolism of Astragalus membranaceus mongholicus