PROTEIN L-ISOASPARTYL METHYLTRANSFERASE protects enolase dysfunction by repairing isoaspartyl-induced damage and is positively implicated in agronomically important seed traits.

The protein-repairing enzyme (PRE) PROTEIN L-ISOASPARTYL METHYLTRANSFERASE (PIMT) influences seed vigor by repairing isoaspartyl-mediated protein damage in seeds. However, PIMTs function in other seed traits, and the mechanisms by which PIMT affects such seed traits are still poorly understood. Herein, through molecular, biochemical, and genetic studies using overexpression and RNAi lines in Oryza sativa and Arabidopsis thaliana, we demonstrate that PIMT not only affects seed vigor but also affects seed size and weight by modulating enolase (ENO) activity. We have identified ENO2, a glycolytic enzyme, as a PIMT interacting protein through Y2H cDNA library screening, and this interaction was further validated by BiFC and co-immunoprecipitation assay. We show that mutation or suppression of ENO2 expression results in reduced seed vigor, seed size, and weight. We also proved that ENO2 undergoes isoAsp modification that affects its activity in both in vivo and in vitro conditions. Further, using MS/MS analyses, amino acid residues that undergo isoAsp modification in ENO2 were identified. We also demonstrate that PIMT repairs such isoAsp modification in ENO2 protein, protecting its vital cellular functions during seed maturation and storage, and plays a vital role in regulating seed size, weight, and seed vigor. Taken together, our study identified ENO2 as a novel substrate of PIMT, and both ENO2 and PIMT in turn implicate in agronomically important seed traits.

Non-destructive detection of single corn seed vigor based on visible/near-infrared spatially resolved spectroscopy combined with chemometrics.

Seed vigor is an essential quality evaluation index for seed selection. However, accurately detecting the vigor of a single corn seed is challenging. In this study, we constructed a single-fiber spatially resolved detection device using visible/near-infrared spectroscopy to investigate the patterns and correlations between spatially resolved spectroscopy (SRS) at 500-1000 nm and seed vigor. The device collected spectral data at a light source-detector distance of 5-6.6 mm on the embryo side (S1) and endosperm side (S2) of the corn seeds. The proposed spectral ratio method based on SRS and spectral combination analysis achieved an improvement in the detection accuracy of different corn seed vigor. Modeling by SG-CARS-PLSDA using the ratio method showed further improvement in the prediction ability. The highest accuracy for both S1 and S2 in the Zhengdan 958 variety was 91.67 %, while those of S1 and S2 for the Shaandan 650 variety were 86.67 % and 88.33 %, respectively. In addition, SRS was found to be more advantageous in S2 acquisition, verifying the potential of SRS in the non-destructive testing of seed vigor. This provides a favorable reference for the comprehensive evaluation of other internal quality indices of seeds.

Revealing the seed microbiome: Navigating sequencing tools, microbial assembly, and functions to amplify plant fitness.

Microbial communities within seeds play a vital role in transmitting themselves to the next generation of plants. These microorganisms significantly impact seed vigor and early seedling growth, for successful crop establishment. Previous studies reported on seed-associated microbial communities and their influence on processes like dormancy release, germination, and disease protection. Modern sequencing and conventional methods reveal microbial community structures and environmental impacts, these information helps in microbial selection and manipulation. These studies form the foundation for using seed microbiomes to enhance crop resilience and productivity. While existing research has primarily focused on characterizing microbiota in dried mature seeds, a significant gap exists in understanding how these microbial communities assemble during seed development. The review also discusses applying seed-associated microorganisms to improve crops in the context of climate change. However, limited knowledge is available about the microbial assembly pattern on seeds, and their impact on plant growth. The review provides insight into microbial composition, functions, and significance for plant health, particularly regarding growth promotion and pest control.

OsJMJ718, a histone demethylase gene, positively regulates seed germination in rice.

Seed vigor has major impact on the rate and uniformity of seedling growth, crop yield, and quality. However, the epigenetic regulatory mechanism of crop seed vigor remains unclear. In this study, a (jumonji C) JmjC gene of the histone lysine demethylase OsJMJ718 was cloned in rice, and its roles in seed germination and its epigenetic regulation mechanism were investigated. OsJMJ718 was located in the nucleus and was engaged in H3K9 methylation. Histochemical GUS staining analysis revealed OsJMJ718 was highly expressed in seed embryos. Abiotic stress strongly induced the OsJMJ718 transcriptional accumulation level. Germination percentage and seedling vigor index of OsJMJ718 knockout lines (OsJMJ718-CR) were lower than those of the wild type (WT). Chromatin immunoprecipitation followed by sequencing (ChIP-seq) of seeds imbibed for 24 h showed an increase in H3K9me3 deposition of thousands of genes in OsJMJ718-CR. ChIP-seq results and transcriptome analysis showed that differentially expressed genes were enriched in ABA and ethylene signal transduction pathways. The content of ABA in OsJMJ718-CR was higher than that in WT seeds. OsJMJ718 overexpression enhanced sensitivity to ABA during germination and early seedling growth. In the seed imbibition stage, ABA and ethylene content diminished and augmented, separately, suggesting that OsJMJ718 may adjust rice seed germination through the ABA and ethylene signal transduction pathways. This study displayed the important function of OsJMJ718 in adjusting rice seed germination and vigor, which will provide an essential reference for practical issues, such as improving rice vigor and promoting direct rice sowing production.

Genomic identification and expression profiling of WRKY genes in alfalfa (Medicago sativa) elucidate their responsiveness to seed vigor.

BACKGROUND: Seed aging is a critical factor contributing to vigor loss, leading to delayed forage seed germination and seedling growth. Numerous studies have revealed the regulatory role of WRKY transcription factors in seed development, germination, and seed vigor. However, a comprehensive genome-wide analysis of WRKY genes in Zhongmu No.1 alfalfa has not yet been conducted. RESULTS: In this study, a total of 91 MsWRKY genes were identified from the genome of alfalfa. Phylogenetic analysis revealed that these MsWRKY genes could be categorized into seven distinct subgroups. Furthermore, 88 MsWRKY genes were unevenly mapped on eight chromosomes in alfalfa. Gene duplication analysis revealed segmental duplication as the principal driving force for the expansion of this gene family during the course of evolution. Expression analysis of the 91 MsWRKY genes across various tissues and during seed germination exhibited differential expression patterns. Subsequent RT-qPCR analysis highlighted significant induction of nine selected MsWRKY genes in response to seed aging treatment, suggesting their potential roles in regulating seed vigor. CONCLUSION: This study investigated WRKY genes in alfalfa and identified nine candidate WRKY transcription factors involved in the regulation of seed vigor. While this finding provides valuable insights into understanding the molecular mechanisms underlying vigor loss and developing new strategies to enhance alfalfa seed germinability, further research is required to comprehensively elucidate the precise pathways through which the MsWRKY genes modulate seed vigor.

Rice seed vigor detection based on near-infrared hyperspectral imaging and deep transfer learning.

Vigor is one of the important factors that affects rice yield and quality. Rapid and accurate detection of rice seed vigor is of great importance for rice production. In this study, near-infrared hyperspectral imaging technique and transfer learning were combined to detect rice seed vigor. Four varieties of artificial-aged rice seeds (Yongyou12, Yongyou1540, Suxiangjing100, and Longjingyou1212) were studied. Different convolutional neural network (CNN) models were built to detect the vigor of the rice seeds. Two transfer strategies, fine-tuning and MixStyle, were used to transfer knowledge among different rice varieties for vigor detection. The experimental results showed that the convolutional neural network model of Yongyou12 classified the vigor of Yongyou1540, Suxiangjing100, and Longjingyou1212 through MixStyle transfer knowledge, and the accuracy reached 90.00%, 80.33%, and 85.00% in validation sets, respectively, which was better or close to the initial modeling performances of each variety. MixStyle statistics are based on probabilistic mixed instance-level features of cross-source domain training samples. When training instances, new domains can be synthesized, which increases the domain diversity of the source domain, thereby improving the generalization ability of the trained model. This study would help rapid and accurate detection of a large varieties of crop seeds.

Identification of a Seed Vigor-Related QTL Cluster Associated with Weed Competitive Ability in Direct-Seeded Rice (Oryza Sativa L.).

Direct seeding of rice (Oryza sativa L.) is a low-labor and sustainable cultivation method that is used worldwide. Seed vigor and early vigor are important traits associated with seedling stand density (SSD) and weed competitive ability (WCA), which are key factors in direct-seeded rice (DSR) cultivation systems. Here, we developed a set of chromosome segment substitution lines with Xiushui134 as receptor parent and Yangdao6 as donor parent and used these lines as a mapping population to identify quantitative trait loci (QTLs) for seed vigor, which we evaluated based on germinability-related indicators (germination percentage (GP), germination energy (GE), and germination index (GI)) and seedling vigor-related indicators (root number (RN), root length (RL), and shoot length (SL) at 14 days after imbibition) under controlled conditions in an incubator. Ten QTLs were detected across four chromosomes, of which a cluster of QTLs (qGP11, qGE11, qGI11, and qRL11) co-localized on Chr. 11 with high LOD values (12.03, 8.13, 7.14, and 8.75, respectively). Fine mapping narrowed down the QTL cluster to a 0.7-Mb interval between RM26797 and RM6680. Further analysis showed that the QTL cluster has a significant effect (p < 0.01) on early vigor under hydroponic culture (root length, total dry weight) and direct seeding conditions (tiller number, aboveground dry weight). Thus, our combined analysis revealed that the QTL cluster influenced both seed vigor and early vigor. Identifying favorable alleles at this QTL cluster could facilitate the improvement of SSD and WCA, thereby addressing both major factors in DSR cultivation systems.

Genomic Identification and Expression Profiling of Lesion Simulating Disease Genes in Alfalfa (Medicago sativa) Elucidate Their Responsiveness to Seed Vigor.

Seed aging, a common physiological phenomenon during forage seed storage, is a crucial factor contributing to a loss of vigor, resulting in delayed seed germination and seedling growth, as well as limiting the production of hay. Extensive bodies of research are dedicated to the study of seed aging, with a particular focus on the role of the production and accumulation of reactive oxygen species (ROS) and the ensuing oxidative damage during storage as a primary cause of decreases in seed vigor. To preserve optimal seed vigor, ROS levels must be regulated. The excessive accumulation of ROS can trigger programmed cell death (PCD), which causes the seed to lose vigor permanently. LESION SIMULATING DISEASE (LSD) is one of the proteins that regulate PCD, encodes a small C2C2 zinc finger protein, and plays a molecular function as a transcriptional regulator and scaffold protein. However, genome-wide analysis of LSD genes has not been performed for alfalfa (Medicago sativa), as one of the most important crop species, and, presently, the molecular regulation mechanism of seed aging is not clear enough. Numerous studies have also been unable to explain the essence of seed aging for LSD gene regulating PCD and affecting seed vigor. In this study, we obtained six MsLSD genes in total from the alfalfa (cultivar Zhongmu No. 1) genome. Phylogenetic analysis demonstrated that the MsLSD genes could be classified into three subgroups. In addition, six MsLSD genes were unevenly mapped on three chromosomes in alfalfa. Gene duplication analysis demonstrated that segmental duplication was the key driving force for the expansion of this gene family during evolution. Expression analysis of six MsLSD genes in various tissues and germinating seeds presented their different expressions. RT-qPCR analysis revealed that the expression of three MsLSD genes, including MsLSD2, MsLSD5, and MsLSD6, was significantly induced by seed aging treatment, suggesting that they might play an important role in maintaining seed vigor. Although this finding will provide valuable insights into unveiling the molecular mechanism involved in losing vigor and new strategies to improve alfalfa seed germinability, additional research must comprehensively elucidate the precise pathways through which the MsLSD genes regulate seed vigor.

Identification of novel candidate loci and genes for seed vigor-related traits in upland cotton (Gossypium hirsutum L.) via GWAS.

Seed vigor (SV) is a crucial trait determining the quality of crop seeds. Currently, over 80% of China's cotton-planting area is in Xinjiang Province, where a fully mechanized planting model is adopted, accounting for more than 90% of the total fiber production. Therefore, identifying SV-related loci and genes is crucial for improving cotton yield in Xinjiang. In this study, three seed vigor-related traits, including germination potential, germination rate, and germination index, were investigated across three environments in a panel of 355 diverse accessions based on 2,261,854 high-quality single-nucleotide polymorphisms (SNPs). A total of 26 significant SNPs were detected and divided into six quantitative trait locus regions, including 121 predicted candidate genes. By combining gene expression, gene annotation, and haplotype analysis, two novel candidate genes (Ghir_A09G002730 and Ghir_D03G009280) within qGR-A09-1 and qGI/GP/GR-D03-3 were associated with vigor-related traits, and Ghir_A09G002730 was found to be involved in artificial selection during cotton breeding by population genetic analysis. Thus, understanding the genetic mechanisms underlying seed vigor-related traits in cotton could help increase the efficiency of direct seeding by molecular marker-assisted selection breeding.

Progress in the study of functional genes related to direct seeding of rice.

Rice is a major food crop in the world. Owing to the shortage of rural labor and the development of agricultural mechanization, direct seeding has become the main method of rice cultivation. At present, the main problems faced by direct seeding of rice are low whole seedling rate, serious weeds, and easy lodging of rice in the middle and late stages of growth. Along with the rapid development of functional genomics, the functions of a large number of genes have been confirmed, including seed vigor, low-temperature tolerance germination, low oxygen tolerance growth, early seedling vigor, early root vigor, resistance to lodging, and other functional genes related to the direct seeding of rice. A review of the related functional genes has not yet been reported. In this study, the genes related to direct seeding of rice are summarized to comprehensively understand the genetic basis and mechanism of action in direct seeding of rice and to lay the foundation for further basic theoretical research and breeding application research in direct seeding of rice.

Hydrotime Model Parameters Estimate Seed Vigor and Predict Seedling Emergence Performance of Astragalus sinicus under Various Environmental Conditions.

Seed vigor is an important aspect of seed quality. High-vigor seeds show rapid and uniform germination and emerge well, especially under adverse environmental conditions. Here, we determined hydrotime model parameters by incubating seeds at different water potentials (0.0, -0.2, -0.4, -0.6, and -0.8 MPa) in the laboratory, for 12 seed lots of Chinese milk vetch (Astragalus sinicus) (CMV), a globally important legume used as forage, green manure, and a rotation crop. Pot experiments were conducted to investigate the seedling emergence performance of 12 CMV seed lots under control, water stress, salinity stress, deep sowing, and cold stress conditions. Meanwhile, the field emergence performance was evaluated on two sowing dates in June and October 2022. Correlation and regression analyses were implemented to explore the relationships between hydrotime model parameters and seedling emergence performance under various environmental conditions. The seed germination percentage did not differ significantly between seed lots when seeds were incubated at 0.0 MPa, whereas it did differ significantly between seed lots at water potentials of -0.2, -0.4, and -0.6 MPa. The emergence percentage, seedling dry weight, and simplified vigor index also differed significantly between the 12 seed lots under various environmental conditions. Ψb(50) showed a significant correlation with germination and emergence performance under various environmental conditions; however, little correlation was observed between θH or σφb and germination and emergence. These results indicate that Ψb(50) can be used to estimate seed vigor and predict seedling emergence performance under diverse environmental conditions for CMV and similar forage legumes. This study will enable seed researchers, plant breeders, and government program directors to target higher seed vigor more effectively for forage legumes.

Integrated Analysis of miRNAome and Transcriptome Identify Regulators of Elm Seed Aging.

After maturity, seed vigor irreversibly decreases. Understanding the underlying mechanism is important to germplasm preservation. MicroRNAs (miRNAs) play vital regulatory roles in plants. However, little is known about how miRNAs regulate seed aging. Here, elm (Ulmus pumila L.) seeds of three aging stages were subjected to a multi-omics analysis including transcriptome, small RNAome and degradome, to find regulators of seed aging. In the small RNAome, 119 miRNAs were identified, including 111 conservative miRNAs and eight novel miRNAs specific to elm seeds, named upu-miRn1-8. A total of 4900 differentially expressed genes, 22 differentially expressed miRNAs, and 528 miRNA-target pairs were identified during seed ageing. The target genes were mainly involved in the processing of proteins in the endoplasmic reticulum, metabolism, plant hormone signal transduction, and spliceosome. The expression of several DEGs and miRNAs were verified by qRT-PCR. The degradome data showed the exact degradation sites of upu-miR399a on ABCG25, and upu-miR414a on GIF1, etc. The dual-luciferase assay verified the negative regulation of upu-miR399a on ABCG25 and upu-miR414a on GIF1 in tobacco leaves. This study outlined the regulation network of mRNA, miRNA and miRNA-target genes during seed aging, which is helpful in integrating the regulation mechanisms of seed vigor at the transcriptional and post-transcriptional levels.

Evaluation and Heritability Analysis of the Seed Vigor of Soybean Strains Tested in the Huanghuaihai Regional Test of China.

High seed vigor is a prerequisite for high and stable yield. At present, seed vigor is not considered a goal of soybean breeding in China. Therefore, the seed vigor status of soybean strains is unclear. In this study, the seed vigor of 131 soybean strains included in the Huanghuaihai regional test in 2019 was measured using the artificial accelerated aging method. Significant (p < 0.01) differences in seed vigor were found, with a coefficient of variation of four vigor indexes being more than 48%. The strains with high vigor only accounted for 28.24%, indicating the seed quality of the tested strains was generally not high. Furthermore, the seed vigor of four representative strains selected from each of three vigor types was evaluated using multiyear and multilocation experiments. The seed vigor indexes of high-vigor-type strains were the most stable, while medium- and low-vigor-type strains varied under different environmental conditions. The generalized heritability of the four vigor indexes of the three vigor types was more than 51% and in the order of high vigor type > low vigor type > medium vigor type. Our results suggested that the genotypes of high-vigor-type strains have a greater influence on seed vigor, so to ensure varieties with high seed vigor, this criterion should be prioritized in soybean breeding programs in China.

Effect of Low Water Availability on Seed Yield and Seed Quality of Basil (Ocimum basilicum).

Basil (Ocimum basilicum L.) is an aromatic and medicinal plant with important properties and is used as an alternative crop in many countries of the world because of its medicinal, economic, industrial, and nutritional importance. The objective of the present study was to determine the effect of low water availability on seed production and seed quality of five cultivars of basil (Mrs Burns, Cinnamon, Sweet, Red Rubin, and Thai). Irrigation levels and cultivars affected seed yield and thousand seed weight. In addition, plants that were exposed to low water availability produced seeds that germinated in a greater percentage. Additionally, root length was increased as the PEG concentration was increased in the germination solution and was affected by the low water availability of the mother plants. The length of the shoot, the length of the root and the seed vigor could not be used as indicators of low water availability on the mother plants, but these characteristics and especially the seed vigor could be used as indicators of low water availability of the seed. Furthermore, the root length and the seed vigor indicated that there is a possibility of an epigenetic effect of water availability on the seed produced under low water availability, though more work is needed.

Maize PIMT2 repairs damaged 3-METHYLCROTONYL COA CARBOXYLASE in mitochondria, affecting seed vigor.

PROTEIN l-ISOASPARTYL O-METHYLTRANSFERASE (PIMT) affects seed vigor by repairing damaged proteins. While PIMT is capable of isoaspartyl (isoAsp) repair in all proteins, those proteins most susceptible to isoAsp formation have not been well characterized, and the mechanisms by which PIMT affects seed vigor remain largely unknown. Using co-immunoprecipitation and LC-MS/MS, we found that maize (Zea mays) PIMT2 (ZmPIMT2) interacted predominantly with both subunits of maize 3-METHYLCROTONYL COA CARBOXYLASE (ZmMCC). ZmPIMT2 is specifically expressed in the maize embryo. Both mRNA and protein levels of ZmPIMT2 increased during seed maturation and declined during imbibition. Maize seed vigor was decreased in the zmpimt2 mutant line, while overexpression of ZmPIMT2 in maize and Arabidopsis thaliana increased seed vigor upon artificial aging. ZmPIMT2 was localized in the mitochondria, as determined by subcellular localization assays using maize protoplasts. ZmPIMT2 binding to ZmMCCα was confirmed by luciferase complementation tests in both tobacco (Nicotiana benthamiana) leaves and maize protoplasts. Knockdown of ZmMCCα decreased maize seed aging tolerance. Furthermore, overexpression of ZmPIMT2 decreased the accumulation of isoAsp of ZmMCCα protein in seed embryos that underwent accelerated aging treatment. Taken together, our results demonstrate that ZmPIMT2 binds ZmMCCα in mitochondria, repairs isoAsp damage, and positively affects maize seed vigor.

Development and Optimization of Label-Free Quantitative Proteomics under Different Crossing Periods of Bottle Gourd.

Bottle gourd, a common vegetable in the human diet, has been valued for its medicinal and energetic properties. In this experiment, the time-resolved analysis of the changes in the proteins' electrophoretic patterning of the seed development at different crossing periods was studied in bottle gourd using label-free quantitative proteomics. Hybrid HBGH-35 had the highest observed protein levels at the 4th week of the crossing period (F4) compared to the parental lines, viz. G-2 (M) and Pusa Naveen (F). The crossing period is significantly correlated with grain filling and reserve accumulation. The observed protein expression profile after storage was related to seed maturation and grain filling in bottle gourds. A total of 2517 proteins were identified in differentially treated bottle gourd fruits, and 372 proteins were differentially expressed between different crossing periods. Proteins related to carbohydrate and energy metabolism, anthocyanin biosynthesis, cell stress response, and fruit firmness were characterized and quantified. Some proteins were involved in the development, while others were engaged in desiccation and the early grain-filling stage. F4 was distinguished by an increase in the accumulation of low molecular weight proteins and enzymes such as amylase, a serine protease, and trypsin inhibitors. The seed vigor also followed similar patterns of differential expression of seed storage proteins. Our findings defined a new window during seed production, which showed that at F4, maximum photosynthetic assimilates accumulated, resulting in an enhanced source-sink relationship and improved seed production. Our study attempts to observe the protein expression profiling pattern under different crossing periods using label-free quantitative proteomics in bottle gourd. It will facilitate future detailed investigation of the protein associated with quality traits and the agronomic importance of bottle gourd through selective breeding programs.

Protein l-isoAspartyl Methyltransferase (PIMT) and antioxidants in plants.

All life forms, including plants, accumulate reactive oxygen species (ROS) as a byproduct of metabolism; however, environmental stresses, including abiotic stresses and pathogen attacks, cause enhanced accumulation of ROS in plants. The increased accumulation of ROS often causes oxidative damage to cells. Organisms are able to maintain levels of ROS below permissible limits by several mechanisms, including efficient antioxidant systems. In addition to antioxidant systems, recent studies suggest that protein l-isoaspartyl methyltransferase (PIMT), a highly conserved protein repair enzyme across evolutionary diverse organisms, plays a critical role in maintaining ROS homeostasis by repairing isoaspartyl-mediated damage to antioxidants in plants. Under stress conditions, antioxidant proteins undergo spontaneous isoaspartyl (isoAsp) modification which is often detrimental to protein structure and function. This reduces the catalytic action of antioxidants and disturbs the ROS homeostasis of cells. This chapter focuses on PIMT and its interaction with antioxidants in plants, where PIMT constitutes a secondary level of protection by shielding a primary level of antioxidants from dysfunction and permitting them to guard during unfavorable situations.

An end-to-end seed vigor prediction model for imbalanced samples using hyperspectral image.

Hyperspectral imaging is a key technology for non-destructive detection of seed vigor presently due to its capability to capture variations of optical properties in seeds. As the seed vigor data depends on the actual germination rate, it inevitably results in an imbalance between positive and negative samples. Additionally, hyperspectral image (HSI) suffers from feature redundancy and collinearity due to its inclusion of hundreds of wavelengths. It also creates a challenge to extract effective wavelength information in feature selection, however, which limits the ability of deep learning to extract features from HSI and accurately predict seed vigor. Accordingly, in this paper, we proposed a Focal-WAResNet network to predict seed vigor end-to-end, which improves the network performance and feature representation capability, and improves the accuracy of seed vigor prediction. Firstly, the focal loss function is utilized to adjust the loss weights of different sample categories to solve the problem of sample imbalance. Secondly, a WAResNet network is proposed to select characteristic wavelengths and predict seed vigor end-to-end, focusing on wavelengths with higher network weights, which enhance the ability of seed vigor prediction. To validate the effectiveness of this method, this study collected HSI of maize seeds for experimental verification, providing a reference for plant breeding. The experimental results demonstrate a significant improvement in classification performance compared to other state-of-the-art methods, with an accuracy up to 98.48% and an F1 score of 95.9%.

Integrated Transcriptomic and Metabolomic Analyses Identify Critical Genes and Metabolites Associated with Seed Vigor of Common Wheat.

Seed aging is a common physiological phenomenon during storage which has a great impact on seed quality. An in-depth analysis of the physiological and molecular mechanisms of wheat seed aging is of great significance for cultivating high-vigor wheat varieties. This study reveals the physiological mechanisms of wheat seed aging in two cultivars differing in seed vigor, combining metabolome and transcriptome analyses. Differences between cultivars were examined based on metabolomic differential analysis. Artificial aging had a significant impact on the metabolism of wheat seeds. A total of 7470 (3641 upregulated and 3829 downregulated) DEGs were detected between non-aging HT and LT seeds; however, 10,648 (4506 up and 6142 down) were detected between the two cultivars after aging treatment. Eleven, eight, and four key metabolic-related gene families were identified in the glycolysis/gluconeogenesis and TCA cycle pathways, starch and sucrose metabolism pathways, and galactose metabolism pathways, respectively. In addition, 111 up-regulated transcription factor genes and 85 down-regulated transcription factor genes were identified in the LT 48h group. A total of 548 metabolites were detected across all samples. Cultivar comparisons between the non-aged groups and aged groups revealed 46 (30 upregulated and 16 downregulated) and 62 (38 upregulated and 24 downregulated) DIMs, respectively. Network analysis of the metabolites indicated that glucarate O-phosphoric acid, L-methionine sulfoxide, isocitric acid, and Gln-Gly might be the most crucial DIMs between HT and LT. The main related metabolites were enriched in pathways such as glyoxylate and dicarboxylate metabolism, biosynthesis of secondary metabolites, fatty acid degradation, etc. However, metabolites that exhibited differences between cultivars were mainly enriched in carbon metabolism, the TCA cycle, etc. Through combined metabolome and transcriptome analyses, it was found that artificial aging significantly affected glycolysis/gluconeogenesis, pyruvate metabolism, and glyoxylate and dicarboxylate metabolism, which involved key genes such as ACS, F16P2, and PPDK1. We thus speculate that these genes may be crucial in regulating physiological changes in seeds during artificial aging. In addition, an analysis of cultivar differences identified pathways related to amino acid and polypeptide metabolism, such as cysteine and methionine metabolism, glutathione metabolism, and amino sugar and nucleotide sugar metabolism, involving key genes such as BCAT3, CHI1, GAUT1, and GAUT4, which may play pivotal roles in vigor differences between cultivars.