Leaf Development Research Articles

Evaluation of the Impact of a Fertilizer Mixture from Plant Extracts and Urea on Corn Yield in Vietnam

This study evaluates the impact of a fertilizer mixture combining plant extracts and urea on the growth and yield of NK54 maize in Vietnam. The experimental design included five treatments with different combinations of urea and plant extracts, compared against a control treatment. Growth parameters such as plant height, leaf number, leaf area, and leaf area index (LAI) were monitored throughout the growing season. The results indicated that the application of urea-coated plant extracts significantly improved growth duration, plant height, and leaf development compared to conventional urea fertilization. Specifically, treatments using the urea-plant extract mixture achieved higher plant heights and leaf area, with Treatment T3 (9 ml concentrated extract + 1 kg Urea) showing the highest theoretical and actual yields. The theoretical yield ranged from 63.62 to 68 q/ha, and the actual yield ranged from 43.7 to 48.4 q/ha, with Treatment T3 consistently outperforming others. The findings suggest that integrating plant extracts with urea can enhance maize growth and yield, highlighting the importance of optimizing fertilization strategies for improving crop productivity.

Dynamics of apex and leaf development in barley as affected by PPD-H1 alleles in two contrasting PHYC backgrounds under short or long photoperiod.

Barley development from seedling to flowering involves both external and internal changes, the latter requiring microscopic observation. Internal changes allow for the classification of preflowering development into three phases: vegetative, early reproductive, and late reproductive. Genetic and environmental factors influence the duration of these phases, impacting grain yield. Photoperiod-sensitivity genes PPD-H1 play a major role in flowering time, affecting adaptation; however, the effect might also be direct (beyond affecting phenology). In this paper, we aimed to assess how PPD-H1 alleles affect barley development, including the progression of growth phases, leaf emergence, tillering dynamics, and spikelet development. Two experiments (field and controlled conditions) were conducted with a factorial combination of (i) four near-isogenic lines (NILs) for PPD-H1 alleles (ppd-H1 or Ppd-H1) under two contrasting PHYC genetic backgrounds (PhyC-l and PhyC-e) and (ii) two photoperiod conditions (short and long days). As expected, longer photoperiods led to a shorter growth cycle. All subphases of time to flowering, final leaf number, and phyllochron were affected by photoperiod. The effects of PPD-H1 on flowering time depended on the PHYC genetic backgrounds and photoperiod conditions. PPD-H1 effects on flowering time were associated with leaf number and phyllochron; the interplay between leaf number and phyllochron affected mainly the late reproductive phase. We also found that although PPD-H1 did not affect the phyllochron of the first six leaves, the phyllochron of leaves appearing later, when grown under a short photoperiod, was consistently increased in lines carrying the ppd-H1 allele. Tillering dynamics exhibited variability, but PPD-H1 did not affect the final spike number under a 24-h photoperiod.

Phenological growth stages of Amaranthus palmeri according to the extended BBCH scale

AbstractAmaranthus palmeri S. Watson is a problematic weed that has been encountered almost all over the world because of its stronger adaptability, which causes huge economic losses and local plant diversity decline. As an invasive plant, A. palmeri has been found throughout China. We thus make a code system to describe the phenological growth process of A. palmeri according to the extended BBCH scale to better understand the phenological growth of this weed in China. The growth process of A. palmeri was divided into vegetative growth phases and reproductive growth phases. The vegetative growth phases are composed of seed germination (0), leaf development (1), formation of side shoots (2) and main stem elongation (3). Likewise, the reproductive growth phases include inflorescence emergence (5), blooming (6), fruit development (7), seed maturity (8) and senescence (9). Based on the extended BBCH scale, the first digit stands for the primary growth stages, and the third digit represents the secondary growth stages. In particular, the second digit defines the growth progress: the number 0, means the vegetative growth phases, and the number 1 stands for the reproductive growth phases. Owing to the fact that its seed can germinate multiple times within a year, a general illustration of the phenological development stages of A. palmeri germinating in different months was provided here to describe the growth dynamic characteristic at different times. These data are of great significance to develop effective strategies for weed control and management of A. palmeri.

Mutations in the WUSCHEL-related homeobox1 gene cause an increased leaflet number in soybean

WUSCHEL-related homeobox (WOX) transcription factors play a crucial role in lateral organ development in several plant species; however, their precise functions in soybean (Glycine max [L.] Merr.) were unclear. Here, we identified two independent multi-leaflet soybean mutants, mlw48-8 and mlw48-161, from a CRISPR/Cas9-engineered mutant library in the Williams 82 background. Both mutants exhibited irregular leaf margins, and the upper leaves were narrow and almost lanceolate at maturity. Molecular analysis revealed that these are allelic mutants with independent mutations in the WUSCHEL-related homeobox1 (GmWOX1A) gene. A transcriptome analysis demonstrated that GmWOX1A modulates the expression of auxin- and leaf development–related genes. Yeast two-hybrid and split-luciferase complementation imaging assays revealed that GmWOX1A interacts with the YABBY family protein GmYAB5, providing further evidence of its potential involvement in leaf development. Notably, the mlw48-161 mutant showed an increased seed number per plant. Consequently, our study not only provides valuable insights into the role of GmWOX1A in soybean leaf development but also offers a potential strategy for high-yield breeding.

Comprehensive analysis of hairpin small RNAs involved in resistance and pathogenesis during wheat-Puccinia triticina interactions.

Wheat leaf rust, caused by the fungus Puccinia triticina (Pt), severely affects the grain quality and quantity of bread wheat (Triticum aestivum L.). Hairpin small(s)RNAs, like micro(mi)RNAs and their variants [including isomiRNAs (isomiRs) and microRNA-like RNAs (milRNAs)], along with their corresponding target genes, bestow leaf rust disease resistance, development and progression from both interacting species. However, the regulatory networks remain inadequately understood. Thirteen differentially expressed novel miRNAs, including two isomiRs and three milRNAs were discerned from induced reads of wheat sRNA libraries, and a further 5,393 and 1,275 candidate target genes were predicted in wheat and Pt, respectively. Functional annotation divulged that wheat-originated miRNAs/isomiRs were involved in resistance, while Pt-derived milRNAs imparted pathogenesis. The identified milRNAs- Tae-Pt-milR5, Tae-Pt-milR12, and Tae-Pt-milR14b and their cleavage sites on Pt target gene MEP5 were confirmed through degradome library screening, suggesting cross-kingdom translocation of Pt virulent genes in wheat host. Co-expression analysis of miRNAs/isomiRs-target genes provided insights into combating leaf rust disease, while co-expression analysis of milRNAs-target gene pairs reflected the extent of pathogenicity exerted by Pt with varied expression levels at the analyzed time points. The analysis pinpointed leaf rust-responsive candidate hairpin sRNAs- Tae-miR8, Tae-Pt-miR12, Tae-Pt-miR14a, and Tae-Pt-miR14b in wheat and Tae-Pt-milR12 in Pt. This study provides new insights into the hairpin sRNAs involved in the resistance and pathogenesis of wheat and Pt, respectively. Furthermore, crucial hairpin sRNAs and their promising targets for future biotechnological interventions to augment stress resilience have been identified.

GmCOL4-GmZTL1 interaction co-regulates GmSBH1 to improve seed deterioration under high temperature and humidity stress and affect leaf development

GmCOL4-GmZTL1 interaction co-regulates GmSBH1 to improve seed deterioration under high temperature and humidity stress and affect leaf development

The TaWAK2-TaNAL1-TaDST pathway regulates leaf width via cytokinin signaling in wheat.

Leaves play a crucial role in photosynthesis and respiration, ultimately affecting the final grain yield of crops, including wheat (Triticum aestivum L.); however, the molecular mechanisms underlying wheat leaf development remain largely unknown. Here, we isolated a narrow-leaf gene, TaWAK2-A, through a map-based cloning strategy. TaWAK2-A encodes a wall-associated kinase (WAK), for which a single Ala-to-Val amino acid substitution reduces the protein stability, leading to a narrow-leaf phenotype in wheat. Further investigation suggests that TaWAK2 directly interacts with and phosphorylates TaNAL1, a trypsin-like serine/cysteine protease. The phosphorylated TaNAL1 is then involved in the degradation of the zinc finger transcription factor TaDST, which acts as a repressor of leaf expansion by activating the expression of the cytokinin oxidase gene TaCKX9 and triggering in vivo cytokinin degradation. Therefore, our findings elucidate a signaling cascade involving TaWAK2-TaNAL1-TaDST that sheds light on the regulation of wheat leaf development.

Analysis and profiling of the purple acid phosphatase gene family in wheat (Triticum aestivum L.).

Purple acid phosphatases (PAPs) play a vital role in plant phosphorus nutrition, serving as a crucial family of metallo-phosphoesterase enzymes. This research aimed to identify the PAP genes from the A/B/D genomes of Triticum aestivum to elucidate evolutionary mechanisms of the gene family in plants and provide genomic information for subsequent research on phosphorous-use efficiency in wheat crops. In total, 105 PAP genes (TaPAPs) were identified from the A/B/D genomes by using the Arabidopsis thaliana and Oryza sativa PAP protein sequences as queries for BLASTP against the wheat protein database. The TaPAPs were grouped into six subfamilies, Ia (17), Ib (26), IIa (11), IIb (30), IIIa (12), and IIIb (9), based on their similarities in the structure of genes and the presence of conserved protein motifs. A majority of TaPAPs were derived from tandemly (20) or segmentally (87) duplicated, with the homoeologous chromosomes 5A/B/D harboring the most duplicated PAP genes. Further analysis indicated that TaPAPs were responsible for the modulation of seed, root, and leaf development and hormone synthesis and signaling, as well as plant responses to abiotic stresses, including low temperatures, drought, and anaerobic conditions. Nine TaPAPs (TaPAP9-4A/4B/4D, TaPAP24-6A/6B/6D, and TaPAP28-7A/7B/7D) were constitutively expressed in diverse tissues such as root, shoot, leaf, spike, and seed, while the remaining genes exhibited tissue-specific expression patterns. Concerning the response to phosphate (Pi) deprivation, 57 TaPAPs were highly expressed in roots under Pi stress, including TaPAP31-4A, 4B, and 4D homeologs from the subfamily IIIb. A TaPAP31-4A transgene in A. thaliana promoted plant growth and development while increasing plant resistance to Pi-deficiency stress by enhancing the secretion of phosphatase. These discoveries provide a scientific foundation for comprehending the role of TaPAPs, offering valuable insights for identifying additional candidate genes and fostering the development of new wheat varieties with enhanced tolerance to low phosphorus conditions.

Physiological and Molecular Mechanisms of Rice Tolerance to Salt and Drought Stress: Advances and Future Directions.

Rice, a globally important food crop, faces significant challenges due to salt and drought stress. These abiotic stresses severely impact rice growth and yield, manifesting as reduced plant height, decreased tillering, reduced biomass, and poor leaf development. Recent advances in molecular biology and genomics have uncovered key physiological and molecular mechanisms that rice employs to cope with these stresses, including osmotic regulation, ion balance, antioxidant responses, signal transduction, and gene expression regulation. Transcription factors such as DREB, NAC, and bZIP, as well as plant hormones like ABA and GA, have been identified as crucial regulators. Utilizing CRISPR/Cas9 technology for gene editing holds promise for significantly enhancing rice stress tolerance. Future research should integrate multi-omics approaches and smart agriculture technologies to develop rice varieties with enhanced stress resistance, ensuring food security and sustainable agriculture in the face of global environmental changes.

Optimization of peapod peel biochar amendment for sustainable agriculture by surface response methodology towards water-food-environment nexus

Valorization of food waste to biochar offers economic and environmental opportunities for sustainable agriculture production. Peapod (Pisum sativum) peel is a high-content lignocellulose vegetable waste and was used in this study to produce biochar through pyrolysis temperatures of 350, 450, and 550 °C, with a heating rate of 5 °C/min for 30 min and thereafter characterized for its various properties. The biochar produced at 350 °C, exhibiting the best soil amendment properties was used for pot testing with mung bean (Vigna radiata). The optimization of plant growth, soil water retention capacity, and microbial activity were determined using response surface methodology (RSM) with three design factors: (a) biochar loading rates (0 %, 0.5 %, and 1 %), (b) temperature (30–40 °C), and (c) quantity of water supplied (50–60 mL/day). The results showed that applying 0.5 % biochar with a water supply of 55 mL/day at 35 °C resulted in optimal responses for seed germination, plant height, and leaf development. Applying 1 % biochar with a water supply of 55 mL/day at 35 °C showed optimal responses for leaf area and leaf chlorophyll content. Stomatal conductance and catalase activity showed optimal responses for 1 % biochar applied with a water supply of 60 mL/day at 40 °C. Urease activity and water retention capacity showed optimal responses when 1 % biochar was applied with a water supply of 60 mL/day at 30 °C, respectively. This study demonstrates the potential use of peapod peel biochar in water-food-environment nexus, with a lower dose and lesser volume of water demand.

Genetic Evidence of SpGH9A3 in Leaf Morphology Variation of Spathiphyllum 'Mojo'.

Leaves play a crucial role as ornamental organs in Spathiphyllum, exhibiting distinct differences across various Spathiphyllum varieties. Leaf development is intricately linked to processes of cell proliferation and expansion, with cell morphology often regulated by plant cell walls, primarily composed of cellulose. Alterations in cellulose content can impact cell morphology, subsequently influencing the overall shape of plant organs. Although cellulases have been shown to affect cellulose levels in plant cells, genetic evidence linking them to the regulation of leaf shape remains limited. This study took the leaves of Spathiphyllum 'Mojo' and its somatic variants as the research objects. We screened four cellulase gene family members from the transcriptome and then measured the leaf cellulose content, cellulase activity, and expression levels of cellulase-related genes. Correlation analysis pinpointed the gene SpGH9A3 as closely associated with leaf shape variations in the mutant. Green fluorescent fusion protein assays revealed that the SpGH9A3 protein was localized to the cell membrane. Notably, the expression of the SpGH9A3 gene in mutant leaves peaked during the early spread stage, resulting in smaller overall leaf size and reduced cellulose content upon overexpression in Arabidopsis.

Hypodermis or multiple epidermis: Leaf ontogenesis in nine species of subfamily Myrtoideae (Myrtaceae)

The ninth largest family of angiosperms is Myrtaceae, which comprises 127 genera and over 6000 species. The leaf anatomy of Myrtaceae has been widely studied and can be used in phylogenetic analyses and species delimitations in complexes within the family. A multiple epidermis and hypodermis have been described for the leaves of representatives of the family but have not been confirmed using ontogenetic studies, which could lead to incorrect interpretations about these tissues. Thus, we studied the leaf ontogenesis of the following 9 species in subfamily Myrtoideae: tribe Syzigieae - Syzigium cumini; tribe Myrteae - Myrceugenia alpigena and M. euosma (subtribe Luminae), Psidium sartorianum, P. guajava, Campomanesia adamantium and Pimenta pseudocaryophyllus (subtribe Pimentinae), Myrcianthes pungens and M. gigantea (subtribe Eugeniinae). Shoot apices and leaves up to the fourth node were transversally and longitudinally sectioned following techniques used for plant anatomy. Leaf development in all species is from apical, intercalary, dorsal, marginal and plate meristematic activity. Protodermal periclinal divisions only occur to give rise to secretory cavities. The results show that in all the species the subepidermal layer is a hypodermis, which originates from periclinal divisions of the ground meristem. The occurrence of this layer could have phylogenetic implications according to reports in the literature, reinforcing the relationships among the subtribes in current topologies of Myrteae. However, the occurrence of a hypodermis in representatives of other subtribes of Myrteae and Syzigieae need to be better studied and discussed in relation to the phylogeny of Myrtaceae.

CDK8 of the mediator kinase module connects leaf development to the establishment of correct stomata patterning by regulating the levels of the transcription factor SPEECHLESS (SPCH).

The components of the mediator kinase module are highly conserved across all eukaryotic lineages, and cyclin-dependent kinase 8 (CDK8) is essential for correct cell proliferation and differentiation in diverse eukaryotic systems. We show that CDK8 couples leaf development with the establishment of correct stomata patterning for prevailing CO2 conditions. In Arabidopsis, the basic helix-loop-helix(bHLH) transcription factor SPEECHLESS(SPCH) controls cellular entry into the stomatal cell lineage, and CDK8 interacts with and phosphorylates SPCH, controlling SPCH protein levels and thereby also expression of the SPCH target genes encoding key regulators of cell fate and asymmetric cell divisions. The lack of the CDK8-mediated control of SPCH results in an increased number of meristemoid and guard mother cells, and increased stomata index in the cdk8 mutants. Increasing atmospheric CO2 concentrations trigger a developmental programme controlling cell entry into stomatal lineage by limiting the asymmetric divisions. In cdk8, the number of meristemoids and guard mother cells remains the same under ambient and high CO2 concentrations, as the accumulated levels of SPCH caused by the lack of CDK8 appear to override the negative regulation of increased CO2. Thus, our work provides novel mechanistic understanding of how plants alter critical leaf properties in response to increasing atmospheric CO2.

Coding and description of phenological stages of blue honeysuckle (Lonicera caerulea L.) according to the BBCH scale

Blue honeysuckle (Lonicera caerulea L.) is an emerging fruit crop from the temperate to circumpolar climate regions with outstanding agronomic traits, substantial nutritional value, and versatile processing applications. However, the limited understanding of its phenology poses substantial challenges to its cultivation, breeding, and research. In this study, we characterized the progressive phenological growth stages of blue honeysuckle according to the BBCH (Biologische Bundesanstalt, Bundessortenamt und Chemische Industrie) scale by investigating three major cultivars (L. caerulea) and a wild accession (L. caerulea var. edulis) in Harbin, China (hot-summer humid continental climate) during 2022–2023. The annual growth of blue honeysuckle was characterized by eight primary stages, including bud development (0), leaf development (1), shoot development (3), inflorescence emergence (5), flowering (6), fruit development (7), fruit ripening (8), and senescence and dormancy (9). Additionally, 35 secondary growth stages were documented, among which eight stages are related to fruit development and ripening. The annual growth period of blue honeysuckle spans 6.5–7 months, starting from late March and ending in early October. The flowering season lasts for 2–3 weeks, ranging from late April to mid-May. The fruit development and ripening last for 7–9 weeks, beginning in early May and ending in early July. This study provides valuable insights for the cultivation and research of blue honeysuckle.

Effects of Phosphorus and Potassium Supply on Photosynthetic Nitrogen Metabolism, Nitrogen Absorption, and Nitrogen Utilization of Hydroponic Rice

Phosphorus (P) and potassium (K) stress significantly affect the growth, physiological characteristics, and nutrient uptake of rice plants. This study investigated the photosynthetic nitrogen (N) metabolism, N uptake, and N utilization of plants under varied P and K supplies. Two local conventional high-yield rice varieties (Shennong 265 and Liaojing 294) were used. These varieties were subjected to the following hydroponic experimental treatments: HPHK (normal P and K concentrations), HPLK (normal P and 1/20 normal K concentration), LPHK (normal K and 1/20 normal P concentration), and LPLK (1/20 normal P and K concentrations). The results showed that the mesophyll cells had a relatively complete nuclear and chloroplast structures, and the antioxidant enzymes of the plants were significantly reduced under the HPHK treatment. Compared to the LP treatments (LPHK and LPLK), the HPHK treatment was found to have the following potential effects: effectively optimize plant configuration; promote leaf development (Pn, E, Ci, and Tr, chlorophyll, and leaf area index); significantly increase the N-metabolism-related enzyme activity of leaves and roots and the accumulation of N in the plant in the main growth stages; and significantly increase the rice yield and N-related efficiency. In conclusion, the HPHK treatment was found to be beneficial in improving the plant configuration, promoting photosynthetic N metabolism, and increasing grain yield and N-related utilization efficiency.

Dual roles of the MPK3 and MPK6 mitogen-activated protein kinases in regulating Arabidopsis stomatal development.

An Arabidopsis (Arabidopsis thaliana) mitogen-activated protein kinase (MAPK) cascade composed of YODA (YDA)-MKK4/MKK5-MPK3/MPK6 plays an essential role downstream of the ERECTA (ER)/ER-LIKE (ERL) receptor complex in regulating stomatal development in the leaf epidermis. STOMAGEN (STO), a peptide ligand produced in mesophyll cells, competes with EPIDERMAL PATTERNING FACTOR2 (EPF2) for binding ER/ERL receptors to promote stomatal formation. In this study, we found that activation of MPK3/MPK6 suppresses STO expression. Using MUTE and STO promoters that confer epidermis- and mesophyll-specific expression, respectively, we generated lines with cell-specific activation and suppression of MPK3/MPK6. The activation or suppression of MPK3/MPK6 in either epidermis or mesophyll cells is sufficient to alter stomatal differentiation. Epistatic analyses demonstrated that STO overexpression can rescue the suppression of stomatal formation conferred by the mesophyll-specific expression of the constitutively active MKK4DD or MKK5DD, but not by the epidermis-specific expression of these constitutively active MKKs. These data suggest that STO is downstream of MPK3/MPK6 in mesophyll cells, but upstream of MPK3/MPK6 in epidermal cells in stomatal development signaling. This function of the MPK3/MPK6 cascade allows it to coordinate plant epidermis development based on its activity in mesophyll cells during leaf development.

Water wisteria genome reveals environmental adaptation and heterophylly regulation in amphibious plants.

Heterophylly is a phenomenon whereby an individual plant dramatically changes leaf shape in response to the surroundings. Hygrophila difformis (Acanthaceae; water wisteria), has recently emerged as a model plant to study heterophylly because of its striking leaf shape variation in response to various environmental factors. When submerged, H. difformis often develops complex leaves, but on land it develops simple leaves. Leaf complexity is also influenced by other factors, such as light density, humidity, and temperature. Here, we sequenced and assembled the H. difformis chromosome-level genome (scaffold N50: 60.43 Mb, genome size: 871.92 Mb), which revealed 36 099 predicted protein-coding genes distributed over 15 pseudochromosomes. H. difformis diverged from its relatives during the Oligocene climate-change period and expanded gene families related to its amphibious habit. Genes related to environmental stimuli, leaf development, and other pathways were differentially expressed in submerged and terrestrial conditions, possibly modulating morphological and physiological acclimation to changing environments. We also found that auxin plays a role in H. difformis heterophylly. Finally, we discovered candidate genes that respond to different environmental conditions and elucidated the role of LATE MERISTEM IDENTITY 1 (LMI1) in heterophylly. We established H. difformis as a model for studying interconnections between environmental adaptation and morphogenesis.

The Role of BrKS in Leafy Head Formation Was Confirmed by Two Allelic Mutants of Chinese Cabbage (Brassica rapa L. ssp. pekinensis)

Considering that the presence of a leafy head is a key morphological characteristic that determines the yield and quality of Chinese cabbage, identifying the major genes regulating the formation of a leafy head is crucial for variety improvement. A heading-related gene, BrKS, was previously predicted from a non-heading mutant, nhm1, derived from a heading variety, which encodes a key enzyme involved in gibberellin biosynthesis. Here, another mutant, nhm2, was identified from the same EMS-mutagenized population, and the phenotype of nhm2 was consistent with that of nhm1. We crossed mutants nhm1 and nhm2, and their F1 plants exhibited the mutant phenotype, which indicated that their mutant genes were allelic. A single non-synonymous mutation in the fourth exon of BrKS in mutant nhm1 and another single non-synonymous mutation in the tenth exon of BrKS in mutant nhm2. The same gene mutation in mutants nhm1 and nhm2 produced a similar non-heading phenotype, which confirmed the role of BrKS in the leafy head formation of Chinese cabbage. RNA-Seq analysis indicated that a transcription factor gene, BrERF1A, which is associated with leaf development, significantly down-regulated expression in mutant nhm1, and after the mutant was treated with GA3, the expression level of BrERF1A was recovered, which indicated that BrKS might be involved in leafy head formation through regulating the expression level of BrERF1A. Our findings provide important clues for revealing the molecular mechanism of leafy head formation in Chinese cabbage.

Using a thermal gradient table to study plant temperature signalling and response across a temperature spectrum

Plants must cope with ever-changing temperature conditions in their environment. In many plant species, suboptimal high and low temperatures can induce adaptive mechanisms that allow optimal performance. Thermomorphogenesis is the acclimation to high ambient temperature, whereas cold acclimation refers to the acquisition of cold tolerance following a period of low temperatures. The molecular mechanisms underlying thermomorphogenesis and cold acclimation are increasingly well understood but neither signalling components that have an apparent role in acclimation to both cold and warmth, nor factors determining dose-responsiveness, are currently well defined. This can be explained in part by practical limitations, as applying temperature gradients requires the use of multiple growth conditions simultaneously, usually unavailable in research laboratories. Here we demonstrate that commercially available thermal gradient tables can be used to grow and assess plants over a defined and adjustable steep temperature gradient within one experiment. We describe technical and thermodynamic aspects and provide considerations for plant growth and treatment. We show that plants display the expected morphological, physiological, developmental and molecular responses that are typically associated with high temperature and cold acclimation. This includes temperature dose-response effects on seed germination, hypocotyl elongation, leaf development, hyponasty, rosette growth, temperature marker gene expression, stomatal conductance, chlorophyll content, ion leakage and hydrogen peroxide levels. In conclusion, thermal gradient table systems enable standardized and predictable environments to study plant responses to varying temperature regimes and can be swiftly implemented in research on temperature signalling and response.

Integration of High-Resolution LC-Q-TOF Mass Spectrometry and Multidimensional Chemical-Biological Analysis to Detect Nanomolar-Level Acetylcholinesterase Inhibitors from Different Parts of Zanthoxylum nitidum.

Zanthoxyli radix is a popular tea among the elderly, and it is believed to have a positive effect on Alzheimer's disease. In this study, a highly effective three-step strategy was proposed for comprehensive analysis of the active components and biological functions of Zanthoxylum nitidum (ZN), including high-resolution LC-Q-TOF mass spectrometry (HRMS), multivariate statistical analysis for heterogeneity (MSAH), and experimental and virtual screening for bioactivity analysis (EVBA). A total of 117 compounds were identified from the root, stem, and leaf of ZN through HRMS. Bioactivity assays showed that the order of acetylcholinesterase (AChE) inhibitory activity from strong to weak was root > stem > leaf. Nitidine, chelerythrine, and sanguinarine were found to be the main differential components of root, stem, and leaf by OPLS-DA. The IC50 values of the three compounds are 0.81 ± 0.02, 0.14 ± 0.01, and 0.48 ± 0.01 μM respectively, indicating that they are potent and high-quality AChE inhibitors. Molecular docking showed that pi-pi T-shaped interactions and pi-lone pairs played important roles in AChE inhibition. This study not only explains the biological function of Zanthoxyli radix in alleviating Alzheimer's disease to some extent, but also lays the foundation for the development of stem and leaf of ZN.