Mature Seeds Research Articles

Pollen fertility at different flowering times and its effect on seed setting of some bitter gourd (Momordica charantia L.) varieties in Ho Chi Minh city

In the hybridisation process, pollen grains directly participate in the process through flower pollination. Pollen grain quality determines fruit formation and seed setting. In this study, the pollen grains of the local bitter gourd variety (BD2) and F1 commercial variety were collected at 8 different flowering times (8-11h30) for fertility evaluation in the 2020 Spring-Winter crop season in Ho Chi Minh city. Self-pollinate the 2 varieties over 8-9 hours to evaluate their ability to set seeds. The obtained results indicated that the local variety (BD2) had the highest fertility pollen (61.26%) at 08:30 am and the lowest (46.22%) at 9:00 am. Meanwhile, the F1 commercial variety showed the highest fertility pollen (94.05%) at 11:30 am and the lowest (79.58%) at 8:00 am. The fertility of variety BD2 is 60.21%, resulting in 33.50 seeds per fruit in which the percentage of mature seed is 71.72%. Similarly, the commercial variety found the pollen fertility is 85.55%, resulting in 11.25 seeds per fruit in which the percentage of mature seeds is 84.90%. This information is useful for bitter gourd hybridisation.

Cone allometry and seed protection from fire are similar in serotinous and nonserotinous conifers.

Serotiny is an adaptive trait that allows certain woody plants to persist in stand-replacing fire regimes. However, the mechanisms by which serotinous cones avoid seed necrosis and nonserotinous species persist in landscapes with short fire cycles and serotinous competitors remain poorly understood. To investigate whether ovulate cone traits that enhance seed survival differ between serotinous and nonserotinous species, we examined cone traits in 24 species within Pinaceae and Cupressaceae based on physical measurements and cone heating simulations using a computational fluid dynamics model. Fire-relevant cone traits were largely similar between cone types; those that differed (e.g. density and moisture) conferred little seed survival advantage under simulated fire. The most important traits influencing seed survival were cone size and seed depth within the cone, which was found to be an allometric function of cone mass for both cone types. Thus, nonserotinous cones should not suffer significantly greater seed necrosis than serotinous cones of equal size. Closed nonserotinous cones containing mature seeds may achieve substantial regeneration after fire if they are sufficiently large relative to fire duration and temperature. To our knowledge, this is the most comprehensive study of the effects of fire-relevant cone traits on conifer regeneration supported by physics-based fire simulation.

Maturity detection of single maize seeds based on hyperspectral imaging and transfer learning

Seed maturity is closely related to seed germination, seed storage and the yield of the next generation of crops and it is an important indicator of seed quality. Maize is an important food, feed, and industrial raw material crop in China. Hyperspectral imaging technology has the advantages of spectral technology and machine vision technology and it can obtain information inside and outside the seed without damaging it. On the basis of hyperspectral imaging technology, transfer learning algorithms were used in this study to explore the transfer feasibility of maize seed maturity models amongst different varieties. Three transfer tasks were designed between different varieties on the basis of the model established for the Xianyu 335 variety. Cross-variety detection models were constructed using the classical machine-learning algorithm partial least square-linear discriminant analysis (PLS-LDA), the transfer learning algorithm transfer component analysis (TCA), and manifold embedded distribution alignment (MEDA). Results showed that the PLS-DA direct transfer model had varying degrees of model failure in all three transfer tasks. However, the average recognition accuracies of the TCA migration model and the MEDA migration model were 89.3% and 91.2%, respectively. Both of the two Transfer learning models can effectively solve the problem of different data feature distributions amongst different varieties, making the model more universal. It solves the problem of transferring the single-grain maize seed maturity detection model amongst different varieties, providing effective, stable and universal technical support for maize seed detection, which is conducive to the promotion of single-grain sowing technology and the improvement of agricultural automation level.

Plasma membrane aquaporins function in moisture regulation during seed germination and leaf hydration in eelgrass

Eelgrass (Zostera marina) is a marine angiosperm distributed in shallow seas that has a unique mechanism for regulating water content. Water transfer across the plasma membrane is facilitated by aquaporins, which are membrane proteins. In this study, we investigated the physiological functions of plasma membrane intrinsic proteins (PIPs) in eelgrass. ZoPIP1;1 and ZoPIP2;1 in eelgrass leaves were amplified by RT-PCR. Both ZoPIPs localized to the plasma membrane, and only ZoPIP2;1 showed water permeability. ZoPIP1;1 and ZoPIP2;1 were expressed in all tissues except male flowers. ZoPIPs were expressed during seed and fruit development, and their expression levels were low in mature seeds and elevated in germinating seeds. Moisture content decreased in the later phases of seed development and increased during seed germination. These findings imply that ZoPIPs participate in water uptake during seed germination. In leaves, ZoPIP expression was upregulated by dehydration but downregulated by rehydration. Water reabsorption of leaves was inhibited by HgCl2, an inhibitor of aquaporins. These findings indicate that ZoPIPs are upregulated by dehydration during extremely low tides and promote water uptake when the tide turns. Therefore, eelgrass PIPs function in moisture regulation during growth, seed germination, and the flood-ebb tidal cycle in seawater.

Endosperm cell death: roles and regulation in angiosperms.

Double fertilization in angiosperms results in the formation of a second zygote, the fertilized endosperm. Unlike its embryo sibling, the endosperm is a transient structure that eventually undergoes developmentally controlled programmed cell death (PCD) at specific time points of seed development or germination. The nature of endosperm PCD exhibits a considerable diversity, both across different angiosperm taxa and within distinct endosperm tissues. In endosperm-less species, PCD might cause central cell degeneration as a mechanism preventing the formation of a fertilized endosperm. In most other angiosperms, embryo growth necessitates the elimination of surrounding endosperm cells. Nevertheless, complete elimination of the endosperm is rare and, in most cases, specific endosperm tissues persist. In mature seeds, these persisting cells may be dead, such as the starchy endosperm in cereals, or remain alive to die only during germination, like the cereal aleurone or the endosperm of castor beans. In this review, we explore current knowledge surrounding the cellular, molecular, and genetic aspects of endosperm PCD, and the influence environmental stresses have on PCD processes. Overall, this review provides an exhaustive overview of endosperm PCD processes in angiosperms, shedding light on its diverse mechanisms and its significance in seed development and seedling establishment.

The Impact of Magnetic Field and Gibberellin Treatment on the Release of Dormancy and Internal Nutrient Transformation in Tilia miqueliana Maxim. Seeds

The seeds of Tilia miqueliana Maxim. exhibit deep dormancy, which is categorized as combinational dormancy. This study utilized a comprehensive treatment involving magnetic fields, gibberellin (GA3), and cold stratification to promote the release of seed physiological dormancy and enhance germination rates. After being soaked in 98% H2SO4 for 15 min, mature seeds of Tilia were exposed to magnetic field treatments (150 MT, 250 MT) for different durations (25 min, 45 min, 65 min, and 85 min), as well as GA3 solution soaking (concentration: 0 μmol·L−1, 1443 μmol·L−1). Subsequently, cold stratification (0–5 °C) was applied to investigate the effects of these treatments on seed dormancy release and nutrient transformation. The results indicated that the comprehensive treatment involving magnetic field, GA3 solution soaking, and cold stratification effectively released the physiological dormancy of Tilia seeds and improved germination rates. Among the treatments, M150T85G1443 (magnetic field intensity: 150 MT, magnetic field treatment time: 85 min, GA3 soaking concentration: 1433 μmol·L−1) exhibited the most favorable outcome. After 75 days of cold stratification following the comprehensive treatments, the germination rate of M150T85G1443 seeds reached 89%. Additionally, the levels of storage substances such as starches and crude fats within the seeds decreased, while the utilization of soluble sugars and soluble proteins increased. The M150T85G1443 treatment exhibited the highest degree of variation, leading to gradual increases in metabolic activities of the seeds and a transition from dormancy to germination.

Analysis of genes specific to the early maturation stage of Sesamum indicum seeds by subtraction method*,*.

The mechanisms regulating the content ratio of unsaturated fatty acid in sesame oil need to be clarified in order to breed novel varieties with high contents of unsaturated fatty acids. Full-length cDNA libraries prepared from sesame seeds 1 to 3 weeks after flowering were subtracted with cDNAs from plantlets of 4 weeks after germination. A total of 1545 cDNA clones was sequenced. The functions of novel genes expressed specifically during the early maturation of sesame seeds were investigated by the transformation of Arabidopsis thaliana. Thirteen genes for a transcription factor were identified, four of which were involved in ethylene signaling. Fifty-nine genes, including those for the aquaporin-like protein and ethylene response factor, were analyzed by overexpression in A. thaliana. The overexpression of novel genes and the aquaporin-like protein gene in A. thaliana increased the content of unsaturated fatty acids. The localization of these products was investigated by the induction of the expression vectors for the GFP fusion protein into onion epidermal cells and sesame root cells with a particle gun. As a result, two cDNA clones were identified as good candidate genes to clarify the regulation in the yield and the ratio of unsaturated fatty acids in sesame seeds. Sein60414 (Accession No. LC603128), an intrinsic membrane protein, may be involved in the increase of unsaturated fatty acids, and Sein61074 (Accession No. LC709278) MAP3K δ-1 protein kinase in the regulation of the total ratio of unsaturated fatty acids in sesame seeds.

Quantitation of Lupinus spp. Quinolizidine Alkaloids by qNMR and Accelerated Debittering with a Resin-Based Protocol.

Quinolizidine alkaloids (QAs) are toxic secondary metabolites of the Lupinus species, the presence of which limits the expansion of lupin beans consumption, despite their high protein content. Evaluation of the level of alkaloids in edible Lupinus species is crucial from a food safety point of view. However, quantitation of QAs is complicated by the fact that not all important alkaloids used for quantitation are commercially available. In this context, we developed a method for the simultaneous quantitation of eight major lupin alkaloids using quantitative NMR spectroscopy (qNMR). Quantitation and analysis were performed in 15 different seed extracts of 11 Lupinus spp. some of which belonged to the same species, with different geographical origins and time of harvest, as well as in all aerial parts of L. pilosus. The mature seeds of L. pilosus were found to be a uniquely rich source of multiflorine. Additionally, we developed a protocol using adsorption or ionic resins for easy, fast, and efficient debittering of the lupine seeds. The protocol was applied to L. albus, leading to a decrease of the time required for alkaloids removal as well as water consumption and to a method for QA isolation from the debittering wastewater.

Studies on genetic diversity, correlation and path coefficient analysis for yield and yield contributing characters in pigeonpea [Cajanus cajan (L). Millips

The experiment was conducted during Kharif 2023 for study of genetic diversity of 42 genotypes of pigeonpea using D2 statistics method of Mahalanobis. Genetic diversity of the forty two genotypes of pigeonpea was accessed for nine characters in a Randomized Block Design with two replications at Agriculture Research Station, Badnapur. Forty two genotypes of pigeonpea were grouped into nine clusters which indicated diversity. Cluster I had the 25 maximum number of genotypes, Cluster II had 10, while, Cluster III, IV, V, VI, VII, VIII, IX had 01 genotype each, respectively. Correlation between yield and yield attribute component traits in forty two genotypes of pigeonpea revealed that seed yield per plant had positive and highly significant association with number of pods per plant, number of primary branches per plant, plant height, number of secondary branches per plant, days to maturity and 100 seed weight . Selection criterion can be done based on these characters for seed yield will give successful results for yield improvement in pigeonpea. Path coefficient analysis of different traits on seed yield per plant recorded that traits viz., 100 seed weight, number of primary branches per plant, days to maturity, plant height and number of seeds per pod showed highest positive direct effect.

Indukce organogeneze a multiplikace korkovníku amurského (Phellodendron amurense Rupr.) v in vitro podmínkách

Amurk cork tree (Phellodendron amurense Rupr.) is known for its anti-inflammatory, antipyretic, cholagogue, and antibacterial properties, and it is also a potential source of industrial cork. Shoot organogenesis and subsequent plant multiplication of Amur cork tree were established from mature seeds of trees growing on research plots in the Czech Republic. Germination percentage was increased by mechanical disruption of the seed coat, the highest number of germinated plants was achieved within 8–10 weeks of culture on MS medium containing 2 mg.l-1 BAP and 0,2 mg.l-1 NAA. Within 4 weeks of transfer to the multiplication medium containing 2 mg.l-1 BAP and 0,2 mg.l-1 NAA, chlorosis and reduced growth were observed in all cultured microcuttings. Lowering the BAP concentration to 0,4 mg.l-1 led to a resumption of growth, the newly emerging shoots and leaves were vital and the multiplication rate was around 90%. Our developed plant regeneration method may be useful in future studies on ex vitro establishment of in vitro derived plants.

Métodos para estimar el nivel de poliembrionía en semillas de lima mexicana y limón italiano

Polyembryony is a common trait in most citrus varieties. The level of polyembryony has the potential to be used as a descriptor in the characterization of varieties or to select parents in conventional hybridization work. The aim of this research was to find an efficient methodology to detect, count and estimate the potential number of embryos per seed, the percentage of polyembryonic seeds and the level of polyembryony in the varieties ‘Colimex’ and ‘Lise’ of Mexican lime, and the variety ‘Rosenberger’ of true lemon. Four methodologies were evaluated: 1) direct counting of embryos in mature seeds (DCEMS), 2) direct counting of embryos in immature seeds (DCEIS), 3) counting of seedlings derived from mature seeds germinated in vitro (CSMSGV) and 4) counting of seedlings derived from mature seeds germinated in substrate (CSMSGS). The highest averages of embryos per seed and percentage of polyembryonic seeds were recorded with the DCEMS and CSMSGV methods (P ≤ 0.01), so it was concluded that they are adequate to estimate these variables in the genotypes studied. The varieties ‘Colimex’ and ‘Lise’ had 80.65 and 78.27 %, respectively, of seeds with two or more embryos, resulting in a high level of polyembryony, while ‘Rosenberger’, with 18.37 %, was slightly polyembryonic (P ≤ 0.01).

Особенности развития видов Verbascum в Центральной Якутии

The possibility of cultivating mullein species in Central Yakutia, which are relatively undemanding plants in terms of moisture, is of scientific and practical interest in order to determine their biological potential and the possibility of using them as ornamental and medicinal plants. The aim of the work is to identify the peculiarities of the developmental rhythms of some species of the genus Verbascum in the permafrost conditions of Central Yakutia. The objects of the study were 11 samples of the species V. chaixii subsp. austriacum, V. densiflorum, V. nigrum, V. nigrum subsp. abietinum, V. phoeniceum and V. thapsus. The material was obtained in the form of seeds in 2018–2021 from institutions in Russia, Denmark, Romania and Poland. The seeds of the first V. phoeniceum sample were sown in a greenhouse and the resulting seedlings were transplanted into the field collection. The remaining seeds were sown outdoors in the second half of September. The plants overwintered under the snow without any additional protection. Phenological phases in the conditions of Central Yakutia were 3–4 weeks later in the calendar than in Central Russia; the end of the growing season was limited by the onset of autumn frosts. The results of the introduction in Central Yakutia showed high resistance in culture of the species V. phoeniceum, V. nigrum, V. nigrum subsp. abietinum, V. chaixii subsp. austriacum and V. thapsus. When sown in the fall, plants of these species flower profusely in the second year and produce mature seeds, except for V. chaixii subsp. austriacum, which flowers profusely in the first year. These species are very decorative and can be recommended for inclusion in the range of ornamental herbaceous plants for Central Yakutia. V. thapsus is also promising as a medicinal plant. V. densiflorum plants developed as annuals without having time to form mature seeds. To use V. densiflorum as an ornamental or medicinal plant, a seedling period is required.

Exploring genetic diversity of potential legume, Vigna angularis (Willd.) Ohwi and Ohashi through agro-morphological traits and SSR markers analysis.

Adzuki bean, an underutilized grain legume, has a significant potential for enhancing food and nutritional security. The main obstacles to developing new cultivars and promoting the adzuki bean as a mainstream pulse crop are a lack of awareness about its potential and insufficient information on crop its genetic diversity. Here, we aimed to explore the untapped potential of adzuki bean germplasm by evaluating its agro-morphological traits and diversity at the molecular level and also to identify trait-specific germplasm by utilizing 100 adzuki bean accessions conserved in the Indian National Genebank. Significant variations was recorded for the morphological traits and identified promising accessions exhibiting desirable traits, such as early flowering (IC341945, EC340257 and EC340283), number of primary branches (IC341945 and IC469175), number of clusters per plant (EC000264, IC167611 and IC341939), number of pods per plant (IC469175, EC34264, EC000264), early maturity (EC340283; EC120460; IC341941) and number of seeds per pod (EC340240, IC455396 and IC341955). Molecular characterization of diverse accessions using 22 polymorphic SSR markers identified a total of 50 alleles, with a mean of 2.27 alleles per loci. The polymorphic information content (PIC) ranged from 0.03 to 0.46, indicating informativeness of markers in distinguishing diverse accessions. Further, the gene diversity among the accessions ranged from 0.03 to 0.57 with a mean of 0.19. Population structure analysis grouped the accessions into three genetic groups, supported by Principal Coordinate Analysis (PCoA) and a phylogenetic tree. Additionally, Analysis of Molecular Variance (AMOVA) confirmed a substantial genetic diversity among the adzuki bean accessions. Thus, the combined assessment of agro-morphological traits and molecular markers effectively distinguished adzuki bean accessions and provided valuable insights in understanding untapped variation at both morphological and molecular levels. The promising accessions identified in the study hold potential for integration into legume improvement programs through introgression breeding, contributing to the development of adzuki bean varieties with target trait.

Optimising Juniperus excelsa (Cupressaceae) germination for sustainable ecosystem restoration in the Prespa area (Western Macedonia, Greece)

In the Mediterranean Region, species of the genus Juniperus face conservation challenges due to human activities and climate change threats. Juniperus species are reported to present low regeneration rate, while their seeds often exhibit physical and physiological dormancy. Viability and germination of mature Juniperus excelsa seeds from population originating from the Prespa area (Florina, Greece), alongside with seedling survival were investigated to study the reproduction dynamic of species population known to represent a EU priority habitat type of limited distribution and with low regeneration rate. The seed cut test of an uncleaned seed lot revealed a high percentage of empty seeds (90.03%) and a low embryo viability (2%). Cleaned seeds were subjected to a total of 44 pretreatments, categorised into eight groups targeting physical and physiological dormancy, along with combinations of these techniques. Despite most treatments showing limited success in enhancing germination, three pretreatments exhibited promising results (germination between 22.22% and 39.40%). Subsequent germination tests involving these treatments in controlled chambers, as well as in field conditions, along with hydration-dehydration cycles, were conducted using newly collected seeds. Seed germination rates in controlled chambers remained low, suggesting a deep primary dormancy. Pretreated seeds, under controlled field conditions, passed two winter periods fully saturated, with 13 hydrating-dehydrating cycles in-between. Germination was maximised during the second year of the field experiment, reaching almost 100% after mechanical scarification for 6 sec., followed by immersion in 3% H2O2 for 2 h. However, seedling survival was low, reaching approximately 38.26%. The findings emphasise the challenges in germinating Juniperus excelsa seeds and highlight the importance of the optimised protocols for reforestation in order to conserve this habitat type.

Embryological observations on seed abortion in Hibiscus syriacus L. and physiological studies on nutrients, enzyme activity and endogenous hormones

Under natural conditions, most Hibiscus syriacus L. individuals form very few mature seeds or the mature seeds that do form are of poor quality. As a result, seed yield is poor and seeds have low natural germinability. These phenomena strongly hinder utilization of the excellent germplasm resources of H. syriacus. The study has shown that pollen activity and stigma receptivity were high on the day of anthesis, and the pistils and stamens were fertile. Pollen release and stigma receptivity were synchronous. But in styles following self and cross-pollination, pollen tube abnormalities (distortion and twisting of the pollen tubes) and callose deposition were observed. Cross-pollinated pollen tubes elongated faster and fewer pollen tube abnormalities were observed compared with self-pollinated pollen tubes. And during embryo development, abnormalities during the heart-shaped embryo stage led to embryo abortion. Imbalance in antioxidant enzyme activities and low contents of auxin and cytokinin during early stages of embryo development may affect embryo development. Therefore, a low frequency of outcrossing and mid-development embryo abortion may be important developmental causes of H. syriacus seed abortion. Nutrient deficiencies, imbalance in antioxidant enzyme activities, and a high content of abscisic acid at advanced stages of seed development may be physiological causes of seed abortion.

Characterization of the chemical composition of different parts of Dolichos lablab L. and revelation of its anti-ulcerative colitis effects by modulating the gut microbiota and host metabolism

Ethnopharmacological relevanceUlcerative colitis (UC) is a non-specific inflammatory disease characterized by long duration and easy relapse. Dolichos lablab L. (DLL) belongs to the family Fabaceae, was listed in a famous Chinese medical classic, Compendium of Materia Medic, and described as possessing features that invigorate the spleen, alleviate dampness, provide diarrhea relief, and other effects. The DLL-dried white mature seeds (DS) and dried flower (DF), which hold significant medicinal value in China, were used in clinical prescriptions to prevent and treat UC. DS and DF have appeared in different editions of the Pharmacopoeia of the People's Republic of China from 1977 to 2020. However, their chemical composition, pharmacological effects, and mechanism of treating UC are unclear. Aim of the studyThis study aimed to characterize the chemical composition of different parts of DLL (seeds and flowers), further explore their pharmacological effects, and elaborate its underlying mechanism of treating UC. MethodsThe chemical composition of DS and DF crude polysaccharides (DSP and DFP) and ethanolic extracts (DSE and DFE) were characterized by high-performance anion-exchange chromatography (HPAEC), ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS), and gas chromatography-mass spectrometry (GC-MS). Then, based on the acute UC mice model, the pharmacodynamic effects were investigated by Western blotting, ELISA, and other methods. Finally, the 16S rRNA gene sequencing and metabonomic analysis were used to explore the regulatory effects of DS and DF on intestinal microbiota and host metabolism. ResultsDSE and DFE inhibited the oxidative stress response, reducing proinflammatory factor production and maintaining intestinal barrier integrity in UC mice. The 16S rRNA gene sequencing and metabonomic analysis revealed that DS and DF treated UC by regulating the intestinal microbiota structure and reversing the abnormal metabolism of the host. ConclusionThis study suggested that different parts of DLL (flowers and seeds) may be potential medicines for treating UC, which exert their therapeutic effects through various active ingredients and might contribute significantly to reducing the economic pressures and challenges of UC treatment worldwide.

Assessment of inter-relationships between yield and yieldattributes in safflower (Carthamus tinctorius L.)

Character association studies in 29 genotypes of safflower for seven characters conducted at Agricultural Research Station, Tandur revealed significant positive association of single plant yield with test weight, number of effective capitula per plant, plant height and number of seeds per capitulum. Estimates of direct and indirect effects of component characters on single plant yield at genotypic level revealed that test weight contributed for maximum positive direct effect on single plant yield followed by days to maturity, plant height and number of seeds per capitulum. Also, these traits had positive and significant association with grain yield. Critical analysis of character association and path analysis showed that these characters should be given importance to isolate superior lines with genetic potentiality for high grain yield.

Somatic embryogenesis from mature sorghum seeds: An underutilized genome editing recipient system

Somatic embryogenesis is a process of cell totipotency in vitro, whereby an embryogenic cell develops from vegetative tissues rather than from zygotes after double fertilization. Sorghum is a recalcitrant crop in genetic transformation; previous recipient systems have usually been from immature zygotic embryos, which needed more time and labors to prepare. Here, an efficient 2,4-dichlorophenoxyacetic acid (2,4-D)-induced somatic embryogenesis system from mature sorghum seeds was introduced. 2,4-D can induce two types of calli from a plumular axis section. Low-concentration 2,4-D (e.g., 2 mg/L) induces white and loose non-embryogenic calli (type 1), while high-concentration 2,4-D (e.g., 8 mg/L) induces yellow and compact embryogenic calli (type 2), which can be clearly distinguished by Sudan red staining. Germinating seeds have a long 2-day window for SE induction. Somatic embryogenesis can be enhanced by HDAC inhibitor, trichostatin A (TSA), a histone deacetylase treatment, which shows more SE productivity and a bigger size. Importantly, this easily prepared protocol does not show obvious genotype dependency in sorghum hybrids. In this study, a high-concentration 2,4-D-induced SE system was established from mature sorghum seeds. This finding provides a technical option for the genome editing recipient in sorghum.

Down-Regulation of Rice Glutelin by CRISPR-Cas9 Gene Editing Decreases Carbohydrate Content and Grain Weight and Modulates Synthesis of Seed Storage Proteins during Seed Maturation

The glutelins are a family of abundant plant proteins comprised of four glutelin subfamilies (GluA, GluB, GluC, and GluD) encoded by 15 genes. In this study, expression of subsets of rice glutelins were suppressed using CRISPR-Cas9 gene-editing technology to generate three transgenic rice variant lines, GluA1, GluB2, and GluC1. Suppression of the targeted glutelin genes was confirmed by SDS-PAGE, Western blot, and q-RT-PCR. Transgenic rice variants GluA1, GluB2, and GluC1 showed reduced amylose and starch content, increased prolamine content, reduced grain weight, and irregularly shaped protein aggregates/protein bodies in mature seeds. Targeted transcriptional profiling of immature seeds was performed with a focus on genes associated with grain quality, starch content, and grain weight, and the results were analyzed using the Pearson correlation test (requiring correlation coefficient absolute value ≥ 0.7 for significance). Significantly up- or down-regulated genes were associated with gene ontology (GO) and KEGG pathway functional annotations related to RNA processing (spliceosomal RNAs, group II catalytic introns, small nucleolar RNAs, microRNAs), as well as protein translation (transfer RNA, ribosomal RNA and other ribosome and translation factors). These results suggest that rice glutelin genes may interact during seed development with genes that regulate synthesis of starch and seed storage proteins and modulate their expression via post-transcriptional and translational mechanisms.

Integration of pharmacodynamics, metabolomics and network pharmacology to elucidate the effect of Prunella vulgaris seed oil in the treatment of hyperlipidemia

To investigate the lipid-lowering activity of PVSO (a natural oil derived from the mature seeds of Prunella vulgaris L.), supercritical CO2 extraction technology was used to extract, and the components were characterized using GC–MS/MS and LC-MS/MS techniques. The lipid-lowering activity of PVSO was evaluated in vitro and in vivo. The results indicated that the supercritical CO2 extraction rate of PVSO was 16.63 %, and the ratio of linolenic acid to linoleic acid was 3:1. In addition, PVSO also contains bioactive components such as phenols, terpenes, flavonoids, and squalene. In vitro experiments showed that PVSO has a strong inhibitory effect on porcine pancreatic lipase, cholesterol esterase, and cholesterol micellization in a significant dose-dependent manner. In vivo experiments showed that PVSO (500 and 800 mg/kg) intervention markedly attenuated body-weight gain, liver weights, hepatic steatosis, oxidative stress biomarkers, and dyslipidemia induced by HFD (p < 0.05). Serum metabolomics studies revealed that 29 endogenous metabolites were significantly altered by HFD and recovered following intervention with PVSO (p < 0.05). PVSO suppressed hyperlipidemia mainly by regulating oxidative stress, choline metabolism, glycine, serine and threonine metabolism, and cholesterol metabolism. Combining metabolomics and network pharmacology analysis, 17 intersection targets, 21 GO and 6 KEGG pathways were obtained. Molecular docking showed that ACHE, CPT1A, HMGCR, LYPLA1, and PLD2 were the key targets of PVSO against hyperlipidemia. In summary, PVSO might be a potential active resource to prevent oxidative stress and lipid metabolism disorders in HFD-induced hyperlipidemia.