Alanine and glutamate catabolism collaborate to ensure the success of Bacillus subtilis sporulation.

Sporulation as a typical bacterial differentiation process has been studied for decades. However, two crucial aspects of sporulation, (i) the energy sources supporting the process, and (ii) the maintenance of spore dormancy throughout sporulation, are scarcely explored. Here, we reported the crucial role of RocG-mediated glutamate catabolism in regulating mother cell lysis, a critical step for sporulation completion of Bacillus subtilis, likely by providing energy metabolite ATP. Notably, rocG overexpression resulted in an excessive ATP accumulation in sporulating cells, leading to adverse effects on future spore properties, e.g. increased germination efficiency, reduced DPA content, and lowered heat resistance. Additionally, we revealed that Ald-mediated alanine metabolism was highly related to the inhibition of premature germination and the maintenance of spore dormancy during sporulation, which might be achieved by decreasing the typical germinant L-alanine concentration in sporulating environment. Our data inferred that sporulation of B. subtilis was a highly orchestrated biological process requiring a delicate balance in diverse metabolic pathways, hence ensuring both the completion of sporulation and production of high-quality spores.

Green-synthesized CuO and ZnO nanoparticles derived from Calotropis gigantea (Apple of Sodom): enhancing plant growth, efficient dye removal, and potent antibacterial applications.

Nanoparticles, owing to their unique physicochemical properties, have garnered significant attention in various scientific disciplines, including materials science, chemistry, biology, and environmental engineering. In recent years, the synthesis of metal oxide nanoparticles, such as NiO, Fe2O3, ZnO, SnO2, and CuO via green routes, has gained attraction due to their diverse applications in fields ranging from catalysis and electronics to medicine and environmental remediation. This study focuses on the green synthesis of copper oxide (CuO) and zinc oxide (ZnO) nanoparticles using Calotropis gigantea (Apple of Sodom) leaf extract as a reducing agent and stabilizer, with zinc nitrate (ZnNO3.6H2O) and copper nitrate (CuNO3.3H2O) as precursors. The hexagonal phase of ZnO and monoclinic plan structure of CuO with high crystallinity was confirmed by XRD and elemental composition by EDX analysis. With the help of an SEM image, particle size measured for CuO and ZnO using ImageJ software was found to be 56.08 nm and 46.49 nm, respectively. This study investigates the efficacy of nanoparticles in wastewater treatment, particularly focusing on methylene blue dye decolorization using the statistical processing of response surface methodology (RSM) using the Box-Behnken method. Additionally, it explores the impact of synthesized nanoparticles on seed growth enhancement, using Vigna radiata (green gram) seeds immersed in various doses of nanoparticles (0, 0.5, 1, 1.5, 2 mg/30 mL). Furthermore, the antibacterial activity of the nanoparticles against both gram-positive and gram-negative bacteria is evaluated. The results confirm the effectiveness of the materials for methylene blue dye removal, achieving 80.53% with CuO and 78.25% with ZnO. Significant seed growth was observed with a low nanoparticle dosage of 1.5 mg/30 mL, resulting in the highest seedling vigour index and germination percentage. This reduces the need for fertilizers and lessens environmental impact.

Exploring selection signatures in the divergence and evolution of lipid droplet (LD) associated genes in major oilseed crops.

BACKGROUND: Oil bodies or lipid droplets (LDs) in the cytosol are the subcellular storage compartments of seeds and the sites of lipid metabolism providing energy to the germinating seeds. Major LD-associated proteins are lipoxygenases, phospholipaseD, oleosins, TAG-lipases, steroleosins, caleosins and SEIPINs; involved in facilitating germination and enhancing peroxidation resulting in off-flavours. However, how natural selection is balancing contradictory processes in lipid-rich seeds remains evasive. The present study was aimed at the prediction of selection signatures among orthologous clades in major oilseeds and the correlation of selection effect with gene expression. RESULTS: The LD-associated genes from the major oil-bearing crops were analyzed to predict natural selection signatures in phylogenetically close-knit ortholog clusters to understand adaptive evolution. Positive selection was the major force driving the evolution and diversification of orthologs in a lineage-specific manner. Significant positive selection effects were found in 94 genes particularly in oleosin and TAG-lipases, purifying with excess of non-synonymous substitution in 44 genes while 35 genes were neutral to selection effects. No significant selection impact was noticed in Brassicaceae as against LOX genes of oil palm. A heavy load of deleterious mutations affecting selection signatures was detected in T-lineage oleosins and LOX genes of Arachis hypogaea. The T-lineage oleosin genes were involved in mainly anther, tapetum and anther wall morphogenesis. In Ricinus communis and Sesamum indicum > 85% of PLD genes were under selection whereas selection pressures were low in Brassica juncea and Helianthus annuus. Steroleosin, caleosin and SEIPINs with large roles in lipid droplet organization expressed mostly in seeds and were under considerable positive selection pressures. Expression divergence was evident among paralogs and homeologs with one gene attaining functional superiority compared to the other. The LOX gene Glyma.13g347500 associated with off-flavor was not expressed during germination, rather its paralog Glyma.13g347600 showed expression in Glycine max. PLD-α genes were expressed on all the tissues except the seed,δ genes in seed and meristem while ß and γ genes expressed in the leaf. CONCLUSIONS: The genes involved in seed germination and lipid metabolism were under strong positive selection, although species differences were discernable. The present study identifies suitable candidate genes enhancing seed oil content and germination wherein directional selection can become more fruitful.

Impact of Germination on the Edible Quality and Nutritional Properties of Brown Rice Noodles.

Brown rice noodles are increasingly favored by consumers for their health benefits; however, their development is hindered by their poor edible qualities. The effect of germination on the cooking, textural, organoleptic and nutritional qualities of brown rice pasta was investigated. In comparison to ungerminated brown rice noodles, germination resulted in a shorter cooking time, reduced cooking losses, and decreased hardness and adhesion of noodles as well as reduced bitter taste. These changes can be attributed to germination altering the basic composition of brown rice. Meanwhile, the contents of γ-aminobutyric acid, free phenolic acid, and bound phenolic acid increased by 53.43%, 21.71%, and 7.14%, respectively, while the content of resistant starch de-creased by 21.55%. Sprouting is a promising strategy for improving the edible quality and nutritional properties of brown rice noodles.

The Discovery of Highly Efficient and Promising ABA Receptor Antagonists for Agricultural Applications Based on APAn Modification.

Abscisic acid (ABA) is one of the many naturally occurring phytohormones widely found in plants. This study focused on refining APAn, a series of previously developed agonism/antagonism switching probes. Twelve novel APAn analogues were synthesized by introducing varied branched or oxygen-containing chains at the C-6' position, and these were screened. Through germination assays conducted on A. thaliana, colza, and rice seeds, as well as investigations into stomatal movement, several highly active ABA receptor antagonists were identified. Microscale thermophoresis (MST) assays, molecular docking, and molecular dynamics simulation showed that they had stronger receptor affinity than ABA, while PP2C phosphatase assays indicated that the C-6'-tail chain extending from the 3' channel effectively prevented the ligand-receptor binary complex from binding to PP2C phosphatase, demonstrating strong antagonistic activity. These antagonists showed effective potential in promoting seed germination and stomatal opening of plants exposed to abiotic stress, particularly cold and salt stress, offering advantages for cultivating crops under adverse conditions. Moreover, their combined application with fluridone and gibberellic acid could provide more practical agricultural solutions, presenting new insights and tools for overcoming agricultural challenges.

The Impact of ZnO and Fe2O3 Nanoparticles on Sunflower Seed Germination, Phenolic Content and Antiglycation Potential.

The enhancement of seed germination by using nanoparticles (NPs) holds the potential to elicit the synthesis of more desired compounds with important biomedical applications, such as preventing protein glycation, which occurs in diabetes. Here, we used 7 nm and 100 nm ZnO and 4.5 nm and 16.7 nm Fe2O3 NPs to treat sunflower seeds. We evaluated the effects on germination, total phenolic content, and the anti-glycation potential of extracted polyphenols. Sunflower seeds were allowed to germinate in vitro after soaking in NP solutions of different concentrations. Polyphenols were extracted, dosed, and used in serum albumin glycation experiments. The germination speed of seeds was significantly increased by the 100 nm ZnO NPs and significantly decreased by the 4.5 nm Fe2O3 NPs. The total phenolic content (TPC) of seeds was influenced by the type of NP, as ZnO NPs enhanced TPC, and the size of the NPs, as smaller NPs led to improved parameters. The polyphenols extracted from seeds inhibited protein glycation, especially those extracted from seeds treated with 7 nm ZnO. The usage of NPs impacted the germination speed and total polyphenol content of sunflower seeds, highlighting the importance of NP type and size in the germination process.

Variation in Seed Morphological Traits Affects the Dispersal Strategies of Chromolaena odorata Following Invasion.

Seed germination and dispersal have an important impact on the establishment and spread of invasive plants. Understanding the extent of intraspecific seed trait variations can enhance our understanding of how invasive plants respond to environmental change after introduction and help predict the dynamic of invasive species under future environmental conditions. However, less attention has been given to the variation in seed traits within species as opposed to among species. We compared seed production, seed morphological traits, dispersal ability, and seedling performance of Chromolaena odorata from 10 introduced populations in Asia and 12 native populations in America in a common garden. The results showed that range (introduced vs. native) and climate affected these traits. Compared with the native population, the introduced populations had higher seed numbers per capitula, lighter seeds, and higher potential dispersal ability seeds (lower terminal velocity) but lower germination rates and seedling lengths. Climatic clines in seed numbers per capitula and pappus length were observed; however, the clines in pappus length differed between the introduced and native populations. Trait covariation patterns were also different between both ranges. In the native populations, there was a trade-off between seed numbers per capitula and seed mass, while this relationship was not found for the introduced populations. These results indicate that C. odorata alters the ecological strategy of seed following invasion, which facilitates its establishment and fast dispersal and contributes to successful invasion in the introduced ranges.

Effect of Light Quality on the seed Germination and Development of Coffee Seedlings (Coffea arabica).

Coffee (Coffea arabica) cultivation is vital to the global economic, social and cultural life of farmers. However, senescent and disease-susceptible plantations affect coffee productivity. Therefore, it is crucial to improve biotechnological strategies such as micropropagation to increase the number of plants for replanting. In this study, the dark condition (T1) and different light qualities (T0-white light 400-700 nm; T2-red light 660 nm and T3-blue light 460 nm) were evaluated to optimize the in vitro propagation of 4 and 9 month-old coffee seeds. The results showed that red light had the highest percentage, an outstanding germination rate index, which may suggest that in the case of coffee seeds could be involved phytochromes that promote germination in a red light quality. In summary, the ideal conditions for in vitro micropropagation of coffee are under white and red light condition.

Effects of Photoperiod and Light Quality on Germination and Growth of Camellia sinensis 'HuangKui'.

Light, as a critical environmental factor, plays a pivotal role in photosynthesis, ultimately influencing the timing of bud flush in tea plants. However, the synergistic effects of different photoperiods and light qualities on the timing of bud flush in the albino tea cultivar 'HuangKui' (later germination variety) remain unknown. Thus, the objective of this study was to investigate the effects of different photoperiods (12L/12D, 14L/10D, 16L/8D, and 18L/6D, where L = the number of daylight hours and D = the number of hours of darkness) and ratios of red (R) to blue (B) light (R/B 1:1, R/B 1:2, R/B 1:3, and R/B 2:1) on the germination and growth of the albino tea variety 'HuangKui'. In our study, we examined how different photoperiods and red light and blue light affected tea germination and growth by investigating the timing of bud flush, photosynthesis, chlorophyll content, and growth indicators. First, our study showed that 'HuangKui' germinated 4 days, 2 days, and 1 day earlier under the 16L/8D photoperiod at the one bud and one leaf period compared with plants cultivated under the 12L/12D, 14L/10D, and 18L/6D photoperiods under light simulating the solar spectrum. Also, the growth of 'HuangKui' was maximumly promoted under the 16L/8D photoperiod treatment. Additionally, the earliest germination of 'HuangKui' was observed for the 16L/8D photoperiod under the R/B 2:1 (red/blue) treatment compared with the other treatments. Moreover, the greatest plant height, length of the new shoots, and new leaf areas were detected in the albino tea variety 'HuangKui' under R/B 2:1. Moreover, the contents of auxin (indole acetic acid, IAA) and trans-zeatin (tZ) under R/B 2:1 were significantly higher than those under the R/B 1:1 and control treatments with the 16L/8D photoperiod. Additionally, the auxin-related expression levels of CsIAA13, CsGH3.1, CsAUX1, and CsARF2 under the R/B 2:1 treatment were significantly higher than those in the control. The expression of CsARR-B, a positive regulator of cytokinin-related genes, was significantly higher under the R/B 2:1 treatment than under the control treatment, while the opposite result was found for the expression of the negative regulator CsARR-A. Therefore, the R/B 2:1 treatment with the 16L/8D photoperiod was an appropriate means of timing the bud flush for the albino tea variety 'HuangKui', which may be related to IAA or tZ signal transduction. In conclusion, our research offers a novel lighting strategy that promotes the germination and growth of albino tea cultivars.

When Size Matters: New Insights on How Seed Size Can Contribute to the Early Stages of Plant Development.

The seed habit is the most complex and successful method of sexual reproduction in vascular plants. It represents a remarkable moment in the evolution of plants that afterward spread on land. In particular, seed size had a pivotal role in evolutionary success and agronomic traits, especially in the field of crop domestication. Given that crop seeds constitute one of the primary products for consumption, it follows that seed size represents a fundamental determinant of crop yield. This adaptative feature is strictly controlled by genetic traits from both maternal and zygotic tissues, although seed development and growth are also affected by environmental cues. Despite being a highly exploited topic for both basic and applied research, there are still many issues to be elucidated for developmental biology as well as for agronomic science. This review addresses a number of open questions related to cues that influence seed growth and size and how they influence seed germination. Moreover, new insights on the genetic-molecular control of this adaptive trait are presented.

Rupture Test: A New Method for Evaluating Maize (Zea mays) Seed Vigour.

To explore the application of seed germination biomechanical event(s) in seed vigour tests, a new procedure for the evaluation of maize seed vigour tests based on pericarp-testa rupture (PR) and coleorhiza rupture (CR) during seed germination was developed. Twenty-four lots of hybrid maize were used to determine the feasibility of the rupture test (RT) as a seed vigour test in Zea mays. The results showed that the physiological quality pattern of 24 maize seed lots assessed through RT was similar to that obtained through analysis with other seed test methods. Correlation and regression analyses revealed that the percentage of CR and percentage of PR + CR at "15 ± 0.5 °C for 120 h ± 1 h" and "20 ± 0.5 °C for 72 h ± 15 min" exhibited positive correlations with the field seedling emergence data (p < 0.01). Hence, the proposed method (the rupture test) is cogent and effective, thus providing an important reference for more crops to select for seed germination event(s) and establishing corresponding new methods for seed vigour tests in the future.

Comparative Cytological and Gene Expression Analysis Reveals That a Common Wild Rice Inbred Line Showed Stronger Drought Tolerance Compared with the Cultivar Rice.

Common wild rice (Oryza rufipogon Griff.) is an important germplasm resource containing valuable genes. Our previous analysis reported a stable wild rice inbred line, Huaye3, which derives from the common wild rice of Guangdong Province. However, there was no information about its drought tolerance ability. Here, we assessed the germination characteristics and seedling growth between the Dawennuo and Huaye3 under five concentrations of PEG6000 treatment (0, 5%, 10%, 15%, and 20%). Huaye3 showed a stronger drought tolerance ability, and its seed germination rate still reached more than 52.50% compared with Dawennuo, which was only 25.83% under the 20% PEG6000 treatment. Cytological observations between the Dawennuo and Huaye3 indicated the root tip elongation zone and buds of Huaye3 were less affected by the PEG6000 treatment, resulting in a lower percentage of abnormalities of cortical cells, stele, and shrinkage of epidermal cells. Using the re-sequencing analysis, we detected 13,909 genes that existed in the genetic variation compared with Dawennuo. Of these genes, 39 were annotated as drought stress-related genes and their variance existed in the CDS region. Our study proved the strong drought stress tolerance ability of Huaye3, which provides the theoretical basis for the drought resistance germplasm selection in rice.

Comparative Phenotypic and Transcriptomic Analyses Provide Novel Insights into the Molecular Mechanism of Seed Germination in Response to Low Temperature Stress in Alfalfa.

Low temperature is the most common abiotic factor that usually occurs during the seed germination of alfalfa (Medicago sativa L.). However, the potential regulatory mechanisms involved in alfalfa seed germination under low temperature stress are still ambiguous. Therefore, to determine the relevant key genes and pathways, the phenotypic and transcriptomic analyses of low-temperature sensitive (Instict) and low-temperature tolerant (Sardi10) alfalfa were conducted at 6 and 15 h of seed germination under normal (20 °C) and low (10 °C) temperature conditions. Germination phenotypic results showed that Sardi10 had the strongest germination ability under low temperatures, which was manifested by the higher germination-related indicators. Further transcriptome analysis indicated that differentially expressed genes were mainly enriched in galactose metabolism and carbon metabolism pathways, which were the most commonly enriched in two alfalfa genotypes. Additionally, fatty acid metabolism and glutathione metabolism pathways were preferably enriched in Sardi10 alfalfa. The Weighted Gene Co-Expression Network Analysis (WGCNA) suggested that genes were closely related to galactose metabolism, fatty acid metabolism, and glutathione metabolism in Sardi10 alfalfa at the module with the highest correlation (6 h of germination under low temperature). Finally, qRT-PCR analysis further validated the related genes involved in the above pathways, which might play crucial roles in regulating seed germination of alfalfa under low temperature conditions. These findings provide new insights into the molecular mechanisms of seed germination underlying the low temperature stress in alfalfa.

Automated Imaging to Evaluate the Exogenous Gibberellin (Ga3) Impact on Seedlings from Salt-Stressed Lettuce Seeds.

Salinity stress is a common challenge in plant growth, impacting seed quality, germination, and general plant health. Sodium chloride (NaCl) ions disrupt membranes, causing ion leakage and reducing seed viability. Gibberellic acid (GA3) treatments have been found to promote germination and mitigate salinity stress on germination and plant growth. 'Bauer' and 'Muir' lettuce (Lactuca sativa) seeds were soaked in distilled water (control), 100 mM NaCl, 100 mM NaCl + 50 mg/L GA3, and 100 mM NaCl + 150 mg/L GA3 in Petri dishes and kept in a dark growth chamber at 25 °C for 24 h. After germination, seedlings were monitored using embedded cameras, capturing red, green, and blue (RGB) images from seeding to final harvest. Despite consistent germination rates, 'Bauer' seeds treated with NaCl showed reduced germination. Surprisingly, the 'Muir' cultivar's final dry weight differed across treatments, with the NaCl and high GA3 concentration combination yielding the poorest results (p < 0.05). This study highlights the efficacy of GA3 applications in improving germination rates. However, at elevated concentrations, it induced excessive hypocotyl elongation and pale seedlings, posing challenges for two-dimensional imaging. Nonetheless, a sigmoidal regression model using projected canopy size accurately predicted dry weight across growth stages and cultivars, emphasizing its reliability despite treatment variations (R2 = 0.96, RMSE = 0.11, p < 0.001).

Seed dormancy types and germination response of 15 plant species in temperate montane peatlands.

Despite their crucial role in determining the fate of seeds, the type and breaking mode of seed dormancy in peatland plants in temperate Asia with a continental monsoon climate are rarely known. Fifteen common peatland plant species were used to test their seed germination response to various dormancy-breaking treatments, including dry storage (D), gibberellin acid soaking (GA), cold stratification (CS), warm followed cold stratification (WCS), GA soaking + cold stratification (GA + CS) and GA soaking + warm followed cold stratification (GA + WCS). Germination experiment, viability and imbibition test, and morphological observation of embryos were conducted. Of the 15 species, nine showed physiological dormancy (PD), with non-deep PD being the dominant type. Four species, Angelica pubescens, Cicuta virosa, Iris laevigata, and Iris setosa exhibited morphophysiological dormancy. Two species, Lycopus uniflorus and Spiraea salicifolia, demonstrated nondormancy. Overall, the effect hierarchy of dormancy-breaking is: CS > GA > WCS > GA + CS > D > GA + WCS. Principal component analysis demonstrated that seed traits, including embryo length: seed length ratio, seed size, and monocot/eudicot divergence, are more likely to influence seed dormancy than environmental factors. Our study suggests that nearly 90% of the tested peatland plant species in the Changbai Mountains demonstrated seed dormancy, and seed traits (e.g. embryo-to-seed ratio and seed size) and abiotic environmental factors (e.g. pH and temperature seasonality) are related to germination behavior, suggesting seed dormancy being a common adaptation strategy for the peatland plants in the temperate montane environment.

Assessing Seed Germination Response of Parasitic Plant Striga hermonthica with Small-Molecule Probes.

Seed germination of a parasitic plant Striga hermonthica is elicited by strigolactones which are exuded from roots of host plants. Here, we describe a high-throughput germination assay and a method for visualizing in vivo strigolactone receptor functions with a fluorogenic probe.

Assessment of Dormancy Level and Germination Ability of Seeds with Physiological Dormancy.

Germination test is fundamental and commonly used technique for seed dormancy and germination studies, and proper assessment of dormancy level and germination ability of a given set of seeds is prerequisite for most of the studies. However, germination is very sensitive to imbibition conditions, and dormancy development is also sensitive to growth conditions of the mother plants. In this chapter, we describe tips for plant growth and germination test mainly for physiological and molecular genetic studies with Arabidopsis. This protocol can be applied for other plant species with relatively small seeds and for various studies to analyze the effect of light, phytohormones, and other chemicals in seed germination.

Functional Analysis of Seed Dormancy Genes by Biolistic Transient Gene Expression in Immature Embryos of Wheat.

Seed dormancy genes typically suppress germination and cell division. Therefore, overexpressing these genes can negatively affect tissue culture, interfering with the generation of transgenic plants and thus hampering the analysis of gene function. Transient expression in target cells is a useful approach for studying the function of seed dormancy genes. Here, we describe a protocol for transiently expressing genes related to seed dormancy in the scutellum of immature wheat (Triticum aestivum) embryos to analyze their effects on germination.

A Seed Storage Protocol to Determine Longevity.

The longevity of seeds, also known as storability, is the period of time for which a seed lot maintains its viability during storage. The method aims to determine longevity of a seed lot during storage in a controlled environment. Seeds are first rehydrated to a preset water content (or relative humidity, RH) and then incubated under controlled conditions for various periods of time to allow for deterioration to occur. At increasing intervals during storage, seeds are retrieved and viability is tested by scoring germination of the seed lot (i.e., radicle protrusion). From these data, a survival curve can be drawn depicting loss of germination during time of storage from which different parameters estimating longevity can be inferred. These parameters can be used to compare longevity between different seed lots, genotypes, or species at similar storage conditions. This test can also be used as a proxy to measure seed vigor or physiological seed quality.

Evaluation Methods for Preharvest Sprouting Resistance in Japanese Wheat Breeding Programs.

Different methodologies have been applied for the selection of preharvest sprouting resistance in cereal breeding programs. We describe here a series of methods used in practical wheat breeding programs in Japan, including phenotyping based on germination score after artificial rain treatments and genotyping using DNA markers. These methods can be modified and applied to breeding programs in which preharvest sprouting is a problem during cereal cultivation.