Germination test, seedling growth, and physiochemical traits are used to screen okra varieties for salt tolerance.

Excess soil salinity is a major stress factor that inhibits plant growth, development, and production. Among the growth stages, seed germination is particularly susceptible to salt stress. Okra, a nutraceutical vegetable, has a low germination percentage. Literature has revealed genetic diversity in okra, which can be studied to develop salt-tolerant varieties. This study examined the salt tolerance of 13 okra varieties using germination tests and then tested five varieties in pot experiments with different NaCl levels (75, 100, and 125 mM NaCl). Results showed that salt levels affected all varieties, with differential variations in stress response. Salt stress reduced agronomic, and physiochemical traits in the studied varieties. In variety "MALAV-27", the highest salt concentration significantly reduced the shoot length (68.12 %), root length (65.11 %), shoot fresh weight (78.73 %), root fresh weight (68.32 %), shoot dry weight (75.60 %), and root dry weight (75.81 %), along with different physiochemical traits. Variety "NAYAB-F1" performed the best, and maintained the highest shoot length (57.12 %), root length (58.72 %), shoot fresh weight (68.26 %), and root fresh weight (58.34 %), shoot dry weight (69.23 %), root dry weight (62.50 %), and numerous physiochemical traits such as sugar (0.74 µg/g), proline (0.51 µmol/g), and chlorophyll 'a' (7.97 mg/g), chlorophyll 'b' (9.56 mg/g). The study recommended 'NAYAB-F1', 'Arka anamika', and 'Shehzadi' as salt-tolerant varieties suitable for selection in salt-tolerant okra breeding programs.

Pea Seed Priming with Pluronic P85-Grafted Single-Walled Carbon Nanotubes Affects Photosynthetic Gas Exchange but Not Photosynthetic Light Reactions.

Nanotechnology is rapidly advancing towards the development of applications for sustainable plant growth and photosynthesis optimization. The nanomaterial/plant interaction has been intensively investigated; however, there is still a gap in knowledge regarding their effect on crop seed development and photosynthetic performance. In the present work, we apply a priming procedure with 10 and 50 mg/L Pluronic-P85-grafted single-walled carbon nanotubes (P85-SWCNT) on garden pea seeds and examine the germination, development, and photosynthetic activity of young seedlings grown on soil substrate. The applied treatments result in a distorted topology of the seed surface and suppressed (by 10-19%) shoot emergence. No priming-induced alterations in the structural and functional features of the photosynthetic apparatus in 14-day-old plants are found. However, photosynthetic gas exchange measurements reveal reduced stomatal conductance (by up to 15%) and increased intrinsic water use efficiency (by 12-15%), as compared to hydro-primed variants, suggesting the better ability of plants to cope with drought stress-an assumption that needs further verification. Our study prompts further research on the stomatal behavior and dark reactions of photosynthesis in order to gain new insights into the effect of carbon nanotubes on plant performance.

Biochar Amendments and Phytoremediation: A Combined Approach for Effective Lead Removal in Shooting Range Soils.

The increasing contamination of soil with heavy metals poses a problem to environmental sustainability. Among these pollutants, lead is particularly concerning due to its persistence in the environment, with harmful effects on human health and ecosystems. Various strategies that combine phytoremediation techniques with soil amendments have emerged to mitigate lead contamination. In this context, biochar has gained significant attention for its potential to enhance soil quality and remediate metal-contaminated environments. This study aims to investigate the combined effect of biochar amendments on the phytoremediation of lead-contaminated shooting range soils. A series of experiments were conducted to determine the impact of the amount and distribution of biochar on lead removal from soil. Soil samples were incubated with biochar for one week, after which two types of seeds (Brassica rapa and Lolium perenne) were planted. Plant and root lengths, as well as the number of germinated seeds, were measured, and a statistical analysis was conducted to determine the influence of the amendments. After one month, the Pb concentration decreased by more than 70%. Our results demonstrate that seed germination and plant growth were significantly better in soil samples where biochar was mixed rather than applied superficially, with the optimal performance observed at a 10% wt. biochar amendment. Additionally, the combined use of biochar and phytoremediation proved highly effective in immobilizing lead and reducing its bioavailability. These findings suggest that the combination of biochar, particularly when mixed at appropriate concentrations, and Brassica rapa significantly improved lead removal efficiency.

In vitro exploration of Acinetobacter strain (SG-5) for antioxidative potential and phytohormone biosynthesis in maize (Zea mays L.) cultivars differing in cadmium tolerance.

Cadmium (Cd) poses serious threats to plant growth and development, whereas the use of plant growth-promoting rhizobacteria (PGPR) has emerged a promising approach to diminish Cd retention in crops. A pot experiment was conducted to evaluate the effect of Cd tolerant strain Acinetobacter sp. SG-5 on growth, phytohormonal response, and Cd uptake of two maize cultivars (3062 and 31P41) under various Cd stress levels (0, 5, 12, 18, 26, and 30 µM CdCl2). The results revealed that CdCl2 treatment significantly suppressed the seed germination and growth together with higher Cd retention in maize cultivars in a dose-dependent and cultivar-specific manner with pronounced negative effect in 31P41. However, SG-5 strain exerted positive impact by up-regulating seed germination traits, plant biomass, photosynthetic pigments, enzymatic and non-enzymatic antioxidants, endogenous hormone level indole-3-acetic acid (IAA), abscisic acid (ABA), and sustained optimal nutrient's levels in both cultivars but predominantly in Cd-sensitive one (31P41). Further, Cd-resistant PGPR decreased the formation of reactive oxygen species in terms of malondialdehyde (MDA) and hydrogen peroxide (H2O2) verified through 3, 3'-diaminobenzidine (DAB) and nitroblue tetrazolium (NBT) analysis in conjunction with reduced Cd uptake and translocation in maize root and shoots in comparison to controls, advocating its sufficiency for bacterial-assisted Cd bioremediation. In conclusion, both SG-5 inoculated cultivars exhibited maximum Cd tolerance but substantial Cd tolerance was acquired by Cd susceptible cultivar-31P41 than Cd-tolerant one (3062). Current work recommended SG-5 strain as a promising candidate for plant growth promotion and bacterial-assisted phytomanagement of metal-polluted agricultural soils.

Arabidopsis DNA glycosylase MBD4L improves recovery of aged seeds.

DNA glycosylases initiate the base excision repair (BER) pathway by catalyzing the removal of damaged or mismatched bases from DNA. The Arabidopsis DNA glycosylase methyl-CpG-binding domain protein 4 like (MBD4L) is a nuclear enzyme triggering BER in response to the genotoxic agents 5-fluorouracil and 5-bromouracil. To date, the involvement of MBD4L in plant physiological processes has not been analyzed. To address this, we studied the enzyme functions in seeds. We found that imbibition induced the MBD4L gene expression by generating two alternative transcripts, MBD4L.3 and MBD4L.4. Gene activation was stronger in aged than in non-aged seeds. Seeds from mbd4l-1 mutants displayed germination failures when maintained under control or ageing conditions, while 35S:MBD4L.3/mbd4l-1 and 35S:MBD4L.4/mbd4l-1 seeds reversed these phenotypes. Seed nuclear DNA repair, assessed by comet assays, was exacerbated in an MBD4L-dependent manner at 24 h post-imbibition. Under this condition, the BER genes ARP, APE1L, and LIG1 showed higher expression in 35S:MBD4L.3/mbd4l-1 and 35S:MBD4L.4/mbd4l-1 than in mbd4l-1 seeds, suggesting that these components could coordinate with MBD4L to repair damaged DNA bases in seeds. Interestingly, the ATM, ATR, BRCA1, RAD51, and WEE1 genes associated with the DNA damage response (DDR) pathway were activated in mbd4l-1, but not in 35S:MBD4L.3/mbd4l-1 or 35S:MBD4L.4/mbd4l-1 seeds. These results indicate that MBD4L is a key enzyme of a BER cascade that operates during seed imbibition, whose deficiency would cause genomic damage detected by DDR, generating a delay or reduction in germination.

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.

Decoding histone 3 lysine methylation: Insights into seed germination and flowering.

Histone lysine methylation is a highly conserved epigenetic modification across eukaryotes that contributes to creating different dynamic chromatin states, which may result in transcriptional changes. Over the years, an accumulated set of evidence has shown that histone methylation allows plants to align their development with their surroundings, enabling them to respond and memorize past events due to changes in the environment. In this review, we discuss the molecular mechanisms of histone methylation in plants. Writers, readers, and erasers of Arabidopsis histone methylation marks are described with an emphasis on their role in two of the most important developmental transition phases in plants, seed germination and flowering. Further, the crosstalk between different methylation marks is also discussed. An overview of the mechanisms of histone methylation modifications and their biological outcomes will shed light on existing research gaps and may provide novel perspectives to increase crop yield and resistance in the era of global climate change.

Spatial specificity of metabolism regulation of abscisic acid-imposed seed germination inhibition in Korean pine (Pinus koraiensis sieb et zucc).

Introduction: Abscisic acid (ABA) can negatively regulate seed germination, but the mechanisms of ABA-mediated metabolism modulation are not well understood. Moreover, it remains unclear whether metabolic pathways vary with the different tissue parts of the embryo, such as the radicle, hypocotyl and cotyledon. Methods: In this report, we performed the first comprehensive metabolome analysis of the radicle and hypocotyl + cotyledon in Pinus koraiensis seeds in response to ABA treatment during germination. Results and discussion: Metabolome profiling showed that following ABA treatment, 67 significantly differentially accumulated metabolites in the embryo were closely associated with pyrimidine metabolism, phenylalanine metabolism, cysteine and methionine metabolism, galactose metabolism, terpenoid backbone biosynthesis, and glutathione metabolism. Meanwhile, 62 metabolites in the hypocotyl + cotyledon were primarily involved in glycerophospholipid metabolism and glycolysis/gluconeogenesis. We can conclude that ABA may inhibit Korean pine seed germination primarily by disrupting the biosynthesis of certain plant hormones mediated by cysteine and methionine metabolism and terpenoid backbone biosynthesis, as well as reducing the reactive oxygen species scavenging ability regulated by glutathione metabolism and shikimate pathway in radicle. ABA may strongly disrupt the structure and function of cellular membranes due to alterations in glycerophospholipid metabolism, and weaken glycolysis/gluconeogenesis in the hypocotyl + cotyledon, both of which are major contributors to ABA-mediated inhibition of seed germination. These results highlight that the spatial modulation of metabolic pathways in Pinus koraiensis seeds underlies the germination response to ABA.

Optimisation of indole acetic acid production by Neopestalotiopsis aotearoa endophyte isolated from Thymus vulgaris and its impact on seed germination of Ocimum basilicum.

BACKGROUND: Microbial growth during plant tissue culture is a common problem that causes significant losses in the plant micro-propagation system. Most of these endophytic microbes have the ability to propagate through horizontal and vertical transmission. On the one hand, these microbes provide a rich source of several beneficial metabolites. RESULTS: The present study reports on the isolation of fungal species from different in vitro medicinal plants (i.e., Breynia disticha major, Breynia disticha, Duranta plumieri, Thymus vulgaris, Salvia officinalis, Rosmarinus officinalis, and Ocimum basilicum l) cultures. These species were tested for their indole acetic acid (IAA) production capability. The most effective species for IAA production was that isolated from Thymus vulgaris plant (11.16 µg/mL) followed by that isolated from sweet basil plant (8.78 µg/mL). On screening for maximum IAA productivity, medium, "MOS + tryptophan" was chosen that gave 18.02 µg/mL. The macroscopic, microscopic examination and the 18S rRNA sequence analysis indicated that the isolate that given code T4 was identified as Neopestalotiopsis aotearoa (T4). The production of IAA by N. aotearoa was statistically modeled using the Box-Behnken design and optimized for maximum level, reaching 63.13 µg/mL. Also, IAA extract was administered to sweet basil seeds in vitro to determine its effect on plant growth traits. All concentrations of IAA extract boosted germination parameters as compared to controls, and 100 ppm of IAA extract exhibited a significant growth promotion effect for all seed germination measurements. CONCLUSIONS: The IAA produced from N. aotearoa (T4) demonstrated an essential role in the enhancement of sweet basil (Ocimum basilicum) growth, suggesting that it can be employed to promote the plant development while lowering the deleterious effect of using synthetic compounds in the environment.

Dynamic interplay of reactive oxygen and nitrogen species (ROS and RNS) in plant resilience: unveiling the signaling pathways and metabolic responses to biotic and abiotic stresses.

KEY MESSAGE: Plants respond to environmental challenges by producing reactive species such as ROS and RNS, which play critical roles in signaling pathways that lead to adaptation and survival strategies. Understanding these pathways, as well as their detection methods and effects on plant development and metabolism, provides insight into increasing crop tolerance to combined stresses. Plants encounter various environmental stresses (abiotic and biotic) that affect plant growth and development. Plants sense biotic and abiotic stresses by producing different molecules, including reactive species, that act as signaling molecules and stimulate secondary messengers and subsequent gene transcription. Reactive oxygen and nitrogen species (ROS and RNS) are produced in both physiological and pathological conditions in the plasma membranes, chloroplasts, mitochondria, and endoplasmic reticulum. Various techniques, including spectroscopy, chromatography, and fluorescence methods, are used to detect highly reactive, short-half-life ROS and RNS either directly or indirectly. In this review, we highlight the roles of ROS and RNS in seed germination, root development, senescence, mineral nutrition, and post-harvest control. In addition, we provide information on the specialized metabolism involved in plant growth and development. Secondary metabolites, including alkaloids, flavonoids, and terpenoids, are produced in low concentrations in plants for signaling and metabolism. Strategies for improving crop performance under combined drought and pathogen stress conditions are discussed in this review.

Diet and seed dispersal of bearded capuchin monkeys (Sapajus libidinosus) in Brasilia National Park.

The dietary ecology of a species can provide information on habitat requirements, food resources, and trophic interactions, important to guide conservation efforts of wildlife populations in endangered habitats. In this study, we investigated the dietary ecology of bearded capuchin monkeys (Sapajus libidinosus) in Brasilia National Park, in the endangered Cerrado biome of central Brazil. To obtain diet composition and evaluate the role of these primates as seed dispersers of local tree species, fecal sample collections and feeding observations were performed for a 7-month period. To determine whether seeds germinated better after passing through a primate gut, we conducted germination trials with (i) pulped seeds from trees, (ii) depulped seeds from trees, (iii) seeds from feces planted with feces, and (iv) seeds from feces planted without feces. During experimental procedures, 7308 seeds from 8 families and 10 species were planted. We found that S. libidinosus spent more time feeding on fruits than on any other food item and the diet consisted of 33 plant species from 21 families. However, 20% of their diet consisted of anthropic food. Most seeds planted with feces germinated faster compared to seeds in other experimental treatments, suggesting that passing through the gut and being deposited with fecal material is advantageous. The bearded capuchins also defecated many medium- (5 species) and large-sized (2 species) seeds that may be inaccessible to smaller arboreal frugivores. The results obtained emphasize the important role of bearded capuchins as seed dispersers for the maintenance and conservation of the endangered Cerrado biome.

Elucidating the regulatory role of long non-coding RNAs in drought stress response during seed germination in leaf mustard.

Leaf mustard (Brassica juncea L. Czern & Coss), an important vegetable crop, experiences pronounced adversity due to seasonal drought stress, particularly at the seed germination stage. Although there is partial comprehension of drought-responsive genes, the role of long non-coding RNAs (lncRNAs) in adjusting mustard's drought stress response is largely unexplored. In this study, we showed that the drought-tolerant cultivar 'Weiliang' manifested a markedly lower base water potential (-1.073 MPa vs -0.437 MPa) and higher germination percentage (41.2% vs 0%) than the drought-susceptible cultivar 'Shuidong' under drought conditions. High throughput RNA sequencing techniques revealed a significant repertoire of lncRNAs from both cultivars during germination under drought stress, resulting in the identification of 2,087 differentially expressed lncRNAs (DELs) and their correspondingly linked 12,433 target genes. It was noted that 84 genes targeted by DEL exhibited enrichment in the photosynthesis pathway. Gene network construction showed that MSTRG.150397, a regulatory lncRNA, was inferred to potentially modulate key photosynthetic genes (Psb27, PetC, PetH, and PsbW), whilst MSTRG.107159 was indicated as an inhibitory regulator of six drought-responsive PIP genes. Further, weighted gene co-expression network analysis (WGCNA) corroborated the involvement of light intensity and stress response genes targeted by the identified DELs. The precision and regulatory impact of lncRNA were verified through qPCR. This study extends our knowledge of the regulatory mechanisms governing drought stress responses in mustard, which will help strategies to augment drought tolerance in this crop.

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.

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.

Seed germination, morphology and fruit phenology insight of Cylicomorpha solmsii (Urb.) Urb: a step towards sustainable restoration planning.

Cylicomorpha solmsii (Urb.) Urb (Caricaceae) is a wild relative of domesticated Carica papaya native to the humid tropical forest of Cameroon. C. solmsii is becoming extinct due to rapid urbanization of its habitat. There is currently no restoration planning, no available data on seed germination, details on morphological description and fruit phenology. We investigated the effects of light and soil on seed germination, updated its morphological description and provided cues of its fruit phenology. In two series of experiments, a germination test was first conducted under light and dark conditions with three seed pre-treatments (scarification, drying and cold). Secondly, pre-treated seeds were sown in native soils of C. solmsii habitat collected at Eloumden I and II, two ex-situ and mixtures soil with sand. Qualitative and quantitative data were collected on different part of the plant and analyzed using R package version 4.3.2. Our findings showed that C. solmsii seeds can germinate only under light. The seeds manifested a physiological embryonic dormancy. The native soils showed the highest germination percentage and seedling establishment. The dioicy of C. solmsii was clearly described with incomplete staminate and pistillate unisexual flower whorls. C. solmsii was observed to produce fruits throughout the year at varying intensity. This information is a vital cue to species restoration and policy makers towards C. solmsii conservation.

Promotion of seedling germination in Arabidopsis by B-box zinc-finger protein BBX32.

Seed germination represents a determinant for plants to enter ecosystems and is thus regarded as a key ecological and agronomic trait. It is tightly regulated by a variety of environmental cues to ensure that seeds germinate under favorable conditions. Here, we characterize BBX32, a B-box zinc-finger protein, as an imbibition-stimulated positive regulator of seed germination. Belonging to subgroup V of the BBX family, BBX32 exhibits distinct characteristics compared with its close counterparts within the same subgroup. BBX32 is transiently induced at both the transcriptional and post-transcriptional levels in the embryo upon water absorption. Genetic evidence indicates that BBX32 acts upstream of the master transcription factor PHYTOCHROME-INTERACTING FACTOR 1 (PIF1) to facilitate light-induced seed germination. BBX32 directly interacts with PIF1, suppressing its protein-interacting and DNA-binding capabilities, thereby relieving PIF1's repression on seed germination. Furthermore, the imbibition-stimulated BBX32 functions in parallel with the light-induced transcription regulator HFR1 to collectively attenuate the transcriptional activities of PIF1. The BBX32-PIF1 de-repression module serves as a molecular connection that enables plants to integrate signals of water availability and light exposure, effectively coordinating the initiation of seed germination.

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.

Topping and NPK fertilization alter seed germination, plant growth and active components of Valeriana amurensis.

Introduction: Valeriana amurensis, a tall herbaceous plant, has been traditionally utilized as a herbal remedy both in China and Russia. Methods: This study was set to explore how to cultivate high quality seedlings, considering factors such as seedling time, seeding density, shading, and plastic film mulching. In addition, we investigated the impact of topping and fertilizer on the growth and quality of V. amurensis. Results: According to the percentage of seed germination, the seeds of V. amurensis were sowed with 0.88 g m-2 density using plastic film mulching from late April to mid-May (germination percentage was more than 30%). The optimal Transplanting time was identified as late September, ensuring a high survival rate of 98%. Generally, topping showed the most improved growth indices in late fruit period (21.87 for number of radical leaves) and withering period (6.50 for number of buds and 234.81 for number of roots). Topping increased the yields of valtrate (10.91 mg per plant), valepotriates (809.51 mg per plant) and essential oil (395.64 mg per plant) in withering period. Nitrogen fertilizer promoted maximum root growth and increased the biomass of V. amurensis. Meanwhile, N fertilizer significantly increased the yields of valtrate to 10.46 mg per plant and valepotriates to 772.32 mg per plant among three types of fertilizers. Seedlings are obtained according to rational sowing factors and transplanting time. Topping and nitrogen fertilization emerge as superior strategies to enhance the growth and medicinal quality of this valuable plant. Discussion: This study provides actionable insights for the cultivation V. amurensis.

Physical seed damage, not rodent's saliva, accelerates seed germination of trees in a subtropical forest.

Many tree species adopt fast seed germination to escape the predation risk by rodents. Physical seed damage and the saliva of rodents on partially consumed seeds may act as cues for seeds to accelerate germination process. However, the impacts of these factors on seed germination rate and speed remain unclear. In this study, we investigated such impacts on the germination rate and speed (reversal of germination time) of four tree species (Quercus variabilis, Q. serrata, Q. acutissima, Q. glauca) after partial consumption by four rodent species (Leopoldamys edwards, Niviventer fulvescens, N. confucianus, Apodemus draco), through a series of experiments. We also examined how seed traits may affect the seed damage degree by rodents by analyzing the relationship between the germination rate and time of rodent-damaged seed and the traits. We found that, artificially- and rodent-damaged seeds exhibited a significantly higher seed germination rate and speed, compared to intact seeds. Also, the rodent saliva on seeds showed no significant effect on seed germination rate and speed. Furthermore, we observed significant positive correlations between several seed traits (including the seed mass, coat thickness, and protein content) and the seed germination rate and speed. These correlations are likely due to their beneficial traits countering seed damage by rodents. Overall, our results highlight the significant role of physical seed damage by rodents (rather than their saliva) in facilitating seed germination of tree species, and potential mutualism between rodents and trees. Additionally, our results may have some implications in forest restoration, such that intentionally sowing or dispersing slightly damaged seeds by humans or drones may increase the likelihood of successful seed regeneration.

Temporal dynamics of chloroplast biogenesis revealed initiation of photosynthesis-related gene expression and protein complexes during alfalfa seed germination.

The chloroplast biogenesis occurs in cotyledon during alfalfa seed germination before true leaf formation, and is extremely important for the followed plant development and growth. In this study, we conducted a simulation of alfalfa seed germination in the soil by using tin foil and focused on 10 pivotal time points of chloroplast biogenesis in cotyledons before and after light exposure, which showed significant differences in multispectral images, and covered the whole process of chloroplast biogenesis from proplastid, etioplast to mature chloroplast. We revealed three phases that referred to the programmed involvements of photosynthesis promotion, ultrastructure maturity, transcriptomic expression, and protein complex construction, and observed distinct transcriptional expressions of genes from nuclear and chloroplast genomes. In phase I at dark germination before light exposure, chloroplast-encoded genes showed up-regulated expressions together with the importation of chloroplast proteins. In phase II for the first day after light exposure, nuclear-encoded genes' expressions were initiated at 2 h after light exposure (E2h), followed by swift assembly of chloroplast thylakoid membrane protein complexes, and roaring Fv/Fm and contents of chlorophyll a, chlorophyll b and carotenoid. The initiation at E2h was pronounced by the observation of gradual accumulation of single lamella, and facilitated the formation of granum stacks (thylakoid) at E8h in phase II. In phase III from the second day after light exposure, chloroplast became gradually complete with the fully established photosynthetic capacity. Altogether, our results layed a theoretical foundation for enhancing potential photosynthetic efficiency in alfalfa and related species.