Meristem Research Articles

Investigation of phytotoxic effects of fullerene С60 with the use of Allium-test

Aim. To investigate the cytogenetic effects of water-soluble fullerene C60 with the use of Allium test. To study the influence of various concentrations of fullerene С60 on the morpho-physiological parameters of A. cepa. Methods. The influence of different concentrations of fullerene С60 (25–100 μg/ml) on the induction of root formation and their growth, as well as its cytogenetic effects on the meristematic cells of root apices, were investigated. Results. It was found that fullerene C60 at concentrations of 50 and 75 μg/ml induced root formation and stimulated root growth, though causing minor deformations. Also, it was shown that fullerene С60 disrupts the progression of various phases of mitosis in the meristematic cells of roots, along with the occurrence of chromosomal aberrations at all investigated concentrations. Conclusions. The cytogenetic effects of water-soluble fullerene C60 were investigated for the first time using the Allium-test. Both positive effects on the morphophysiological parameters of A. cepa and its genotoxicity were demonstrated, which could indicate the disruptions in the mechanisms of spindle formation.

Peduncle Rot of Banana Caused by Fusarium petroliphilum and F. pernambucanum in Guangxi, China.

Banana is one of the main fruit crops worldwide. In October 2020, peduncles with rot were observed on bananas (Musa sp. ABB, Pisang Awak subgroup) at a about 1600 square meter commercial banana plantation in Dayu Town (23.17° N, 109.80° E), Guigang, Guangxi, China. The incidence of the disease was about 40%. The interior of the peduncle initially appeared reddish-brown and gradually turned black, and the peduncle eventually rotted. Two whole diseased bunches diseased samples were collected from banana plantations. Small pieces of tissues from the peduncle at the junction of disease and health were surface-disinfected in 75% ethanol for 10 s, 2% NaClO for 1 min, and rinsed three times in sterile water, then placed on potato dextrose agar (PDA) for incubation at 25°C. Forty-nine fungal isolates with similar morphology were recovered from diseased tissues, with 82% isolation frequency. Six isolates (GG3-1, GG3-2, GG3-3, GG4-1, GG4-2 and GG4-3) were selected for further study. Genomic DNAs of these isolates were extracted from 7-day-old mycelia. The internal transcribed spacer (ITS), translation elongation factor (TEF1), calmodulin (CAM), and partial RNA polymerase second largest subunit (RPB2) of six representative isolates were amplified and sequenced (O'Donnell et al. 2000, 2010; White et al. 1990). Sequences were deposited in GenBank. (accessions PP087392-PP087397 for ITS; PP102792-PP102797 for TEF1; PP102798-PP102803 for CAM; PP102804-P102809 for RPB2). A phylogenetic tree based on concatenated sequences of all markers using the Maximum Likelihood algorithm (Xia et al. 2019; Schroers et al. 2016). Based on phylogenetic analyses, GG3-1, -2 and -3 were identified as F. petroliphilum with 100% bootstrap support, and GG4-1, -2 and -3 were tightly clustered with F. pernambucanum with 95% bootstrap support. The two representative isolates GG3-2 and GG4-2 were selected for morphology and pathogenicity observation. Colonies of GG3-2 were light yellow and flat mycelium. They produced falciform macroconidia of 46.1 ± 5.3 × 2.6 ± 0.4 μm with 3 to 5 septates, and hyaline, ovoid microconidia of 7.6 ± 0.9 × 4.0 ± 0.6 μm with 0 septate (Brown et al. 2022). Mycelia were whitish to yellowish aerial mycelium for GG4-2. Their macroconidia were falcate of 31.6 ± 3.0 × 4.3 ± 0.3 μm with curved apical cells, foot-shaped basal cells, and 3 to 5 septates. The microconidia were fusoid of 8.9 ± 1.0 × 2.7 ± 0.3 μm with 0 to 1 septate (Santos et al. 2019). For pathogenicity tests, the ends of the banana peduncles were cut off. Needle punctures were made on the ethanol-treated peduncle pieces, followed by inoculation with 20 μL of conidial suspension (106 spores/ml) of each of the two isolates with three replications each. Sterilized water was used as a control. Peduncle pieces were placed in a humid box and incubated at 28ºC. After 7 days, reddish-brown to black lesions were observed on all inoculated peduncle pieces, while no symptoms were observed on the control pieces. The fungus was isolated from the inoculated peduncle pieces and found to match the morphological characteristics and marker sequences of the original isolates, confirming Koch's postulates. To our knowledge, this is the first report of peduncle rot on banana caused by F. petroliphilum and F. pernambucanum in China. This study will provide valuable information on causal pathogens of this disease which can contribute to improving prevention and disease management strategies for growers.

Modulation of regenerative responses by retinoic and ascorbic acids in human apical papilla cells

ObjectiveThis study investigated the bioactive effects of retinoic acid and ascorbic acid on hSCAPs in vitro. DesignCells were obtained from human third molars (n=4) and characterized for mesenchymal stem cell markers by flow cytometry. The experimental groups: control (α-MEM); vehicle control group (α-MEM + 0.17 % DMSO); retinoic acid 0.1, 1, and 10 µM; and ascorbic acid 3, 30, and 300 µM (n=8) were tested for cell viability (alamarBlue; 1, 3, and 7 days), total collagen synthesis (Sirius Red; 1 and 7 days), mineralized matrix formation (Alizarin red; 14 days), and the regulation of gene expression related to mineralization (ALPL and DSPP), cell migration (ITGAV and CXCL12) angiogenesis (VEGFA) and collagen synthesis (COL1A1 and COL3A1; RT-qPCR) on 1 and 7 days. ACTB and GAPDH were used as reference genes. Data were analyzed by ANOVA and complementary tests at a 5 % significance level. ResultsAscorbic acid 300 µM increased viability, and retinoic acid reduced it dose-dependently. Retinoic acid 0.1 µM and ascorbic acid 30 and 300 µM increased mineralized matrix formation and total collagen synthesis, and retinoic acid 10 µM decreased. On day 1, 0.1 µM retinoic acid upregulated the gene expression of COL1A1, COL3A1, VEGFA, CXCL12, ALPL, DSPP e ITGAV, and 300 µM ascorbic acid upregulated COL1A1, COL3A1 and DSPP. However, on day 7, retinoic acid downregulated ALPL, COL3A1, CXCL12, and VEGFA and downregulated ITGAV and VEGFA. ConclusionRetinoic acid 0.1 µM and ascorbic acid 300 µM biostimulated hSCAPs to differentiate into pro-regenerative phenotypes with potential application for REPs.

CpSPL10-CpELF4 module involves in the negative regulation of flower bud differentiation in Chinese cherry

SQUAMOSA promoter-binding protein-like (SPL) genes play a crucial role in regulating floral induction. Despite such importance, a comprehensive study of SPLs in Chinese cherry flower bud development has been absent. In this study, 32 CpSPL genes were identified. According to expression profiling, CpSPLs exhibited tissue-specific expression and distinct trends throughout flower bud differentiation. Specifically, CpSPL10 was greatly expressed at the beginning of the differentiation, and its role was further investigated. Its overexpression extended the vegetative growth of transgenic tobacco plants, delayed flowering by about 20 days. Moreover, the accumulation of NbELF4 (Early flowering 4) transcripts was enhanced due to the up-regulated levels of CpSPL10 in tobacco plants. ELF4 functions as a major element of the circadian clock; its high expression typically delays the transition from vegetative-to-reproductive growth. Further experiments revealed that CpSPL10 interacts with CpSPL9 or a transposase-derived transcription factor CpFRS5 (FAR1-RELATED SEQUENCE 5) and activates the expression of the downstream gene CpELF4. Notably, the GUS fusing reporter assay detected the activation of CpSPL10 and CpELF4 promoters in shoot apical meristems of transgenic Arabidopsis. These findings revealed the negative regulation of the CpSPL10-CpELF4 module in flower bud differentiation, providing references for supplementing the specific relationships among SPL, FRS, and ELF4.

A DNA-free and genotype-independent CRISPR/Cas9 system in soybean

Abstract Here, we report a smart genome editing system for soybean (Glycine max) using the in planta bombardment-ribonucleoprotein (iPB-RNP) method without introducing foreign DNA or requiring traditional tissue culture processes such as embryogenesis and organogenesis. Shoot apical meristem (SAM) of embryonic axes was used as the target tissue for genome editing because the SAM in soybean mature seeds has stem cells and specific cell layers that develop germ cells during the reproductive growth stage. In the iPB-RNP method, the RNP complex of the CRISPR/Cas9 system was directly delivered into SAM stem cells via particle bombardment, and genome-edited plants were generated from these SAMs. Soybean allergenic gene Gly m Bd 30K was targeted in this study. Many E0 (the first generation of genome-edited) plants in this experiment harbored mutant alleles at the targeted locus. Editing frequency of inducing mutations transmissible to the E1 generation was approximately 0.4 to 4.6 % of all E0 plants utilized in various soybean varieties. Furthermore, simultaneous mutagenesis by iPB-RNP method was also successfully performed at other loci. Our results offer a practical approach for both plant regeneration and DNA-free genome editing achieved by delivering RNP into the SAM of dicotyledonous plants.

Phyto-toxicological effect of fipronil to plant seedlings: Assessing germination attributes, root-tip morphology, oxidative stress, and cellular respiration indices

Pesticides including insecticides are applied in agricultural practices to control insect pests. However, their excessive usage often poses a severe threat to the growth, physiology, and biochemistry of plants. Here, responses of chickpea and greengram seedlings exposed to three fipronil (FIP) concentrations i. e. 100 (1×), 200 (2×) and 300 (3×) μg mL−1 was evaluated under in vitro. Among doses, 3× had a greater negative impact on germination attributes, root-shoot elongation, vigor indices, length ratios, and survival of seedlings. Besides, the morphological distortion in root tips, oxidative stress generation, and cellular death in fipronil-supplemented root seedlings were observed under scanning electron (SEM) and confocal laser scanning (CLSM), respectively. A significant (p ≤ 0.05) and pronounced upsurge in plant stressor metabolites such as proline, malondialdehyde (MDA), electrolyte leakage (EL), hydrogen peroxide (H2O2) content, and antioxidants enzymes in plant seedlings further confirmed the fipronil toxicity. In addition, a concentration-dependent decrease in respiration efficiency (RE) and ATP content in FIP-treated seedlings was observed. Reduced mitotic index (MI) and numerous chromosomal anomalies (CAs) in root meristematic cells of seedlings are a clear indication of insecticide-induced cytotoxicity. Furthermore, a dose-dependent increase in DNA damage in root meristematic cells of greengram revealed the genotoxic potential of fipronil. Conclusively, fipronil suggested phyto and cyto-genotoxic effects that emphasize their careful monitoring in soils before application and their optimum addition in soil-plant systems. It is high time to prepare both target-specific and slow-released agrochemical formulations for crop protection with concurrent safeguarding of the soil.

Light at the end of the tunnel: FRAP assays combined with super resolution microscopy confirm the presence of a tubular vacuole network in meristematic plant cells.

Plant vacuoles play key roles in cellular homeostasis, performing catabolic and storage functions, and regulating pH and ion balance. Despite their essential role, there is still no consensus on how vacuoles are established. A model proposing that the endoplasmic reticulum is the main contributor of membrane for growing vacuoles in meristematic cells has been challenged by a study proposing that plant vacuoles are formed de novo by homotypic fusion of multivesicular bodies (MVBs). Here, we use the Arabidopsis thaliana root as a model system to provide a systematic overview of successive vacuole biogenesis stages, starting from the youngest cells proximate to the quiescent center. We combine in vivo high- and super-resolution (STED) microscopy to demonstrate the presence of tubular and connected vacuolar structures in all meristematic cells. Using customized fluorescence recovery after photobleaching (FRAP) assays, we establish different modes of connectivity and demonstrate that thin, tubular vacuoles, as observed in cells near the quiescent center, form an interconnected network. Finally, we argue that a growing body of evidence indicates that vacuolar structures cannot originate from MVBs alone but receive membrane material from different sources simultaneously.

Saltbush seedlings (Atriplex spp.) shed border-like cells from closed-type root apical meristems

Australian saltbush (Atriplex spp.) survive in exceptionally saline environments and are often used for pasture in semi-arid areas. To investigate the impact of salinity on saltbush root morphology and root exudates, three Australian native saltbush species (Atriplex nummularia, Atriplex amnicola, and Atriplex vesicaria) were grown in vitro in optimised sterile, semi-hydroponic systems in media supplemented with different concentrations of salt (NaCl). Histological stains and chromatographic techniques were used to characterise the root apical meristem (RAM) type and root exudate composition of the saltbush seedlings. We report that saltbush species have closed-type RAMs, which release border-like cells (BLCs). Monosaccharide content, including glucose and fructose, in the root mucilage of saltbush was found to be uniquely low, suggesting that saltbush may minimise carbon release in polysaccharides of root exudates. Root mucilage also contained notable levels of salt, plus increasing levels of unidentified compounds at peak salinity. Un-esterified homogalacturonan, xyloglucan, and arabinogalactan proteins between and on the surface of BLCs may aid intercellular adhesion. At the highest salinity levels, root cap morphology was altered but root:shoot ratio remained consistent. While questions remain about the identity of some components in saltbush root mucilage other than the key monosaccharides, this new information about root cap morphology and cell surface polysaccharides provides avenues for future research.

A dynamic regulome of shoot-apical-meristem-related homeobox transcription factors modulates plant architecture in maize

BackgroundThe shoot apical meristem (SAM), from which all above-ground tissues of plants are derived, is critical to plant morphology and development. In maize (Zea mays), loss-of-function mutant studies have identified several SAM-related genes, most encoding homeobox transcription factors (TFs), located upstream of hierarchical networks of hundreds of genes.ResultsHere, we collect 46 transcriptome and 16 translatome datasets across 62 different tissues or stages from the maize inbred line B73. We construct a dynamic regulome for 27 members of three SAM-related homeobox subfamilies (KNOX, WOX, and ZF-HD) through machine-learning models for the detection of TF targets across different tissues and stages by combining tsCUT&Tag, ATAC-seq, and expression profiling. This dynamic regulome demonstrates the distinct binding specificity and co-factors for these homeobox subfamilies, indicative of functional divergence between and within them. Furthermore, we assemble a SAM dynamic regulome, illustrating potential functional mechanisms associated with plant architecture. Lastly, we generate a wox13a mutant that provides evidence that WOX13A directly regulates Gn1 expression to modulate plant height, validating the regulome of SAM-related homeobox genes.ConclusionsThe SAM-related homeobox transcription-factor regulome presents an unprecedented opportunity to dissect the molecular mechanisms governing SAM maintenance and development, thereby advancing our understanding of maize growth and shoot architecture.

Effects of Plant Meristem-Cell-Based Cosmetics on Menopausal Skin: Clinical Data and Mechanisms.

A randomised open clinical/laboratory study was performed to evaluate the safety and cosmetic efficacy of facial cosmetics for females during the menopausal period. The cosmetics contain active ingredients of meristem cells derived from the medicinal plants Leontopodium alpinum, Buddeleja davidii, Centella asiatica, and Echinacea angustifolia. Recently, the major bioactive molecules of these medicinal plants (leontopodic acid, verbascoside, asiaticoside, and echinacoside, respectively) have been thoroughly evaluated in vitro for molecular pathways and cellular mechanisms and their preventive/curative effects on human skin cells exposed to factors promoting premature skin ageing and cellular senescence. Nevertheless, clinical data on their safety/efficacy to ageing human skin are scarce. This clinical study enrolled 104 Caucasian females in pre-menopause, menopause, or post-menopause periods. They applied cosmetic serums daily for 1 month. Questionnaires and instrumental and biochemical methods were used to assess dermatological/ophthalmological safety and cosmetic efficacy through changes of the skin physiology markers characteristic of ageing/menopause (elasticity, barrier functions, moisture, sebum, ultrasonic properties, and collagen content and structure). Quantitative microbiological tests were carried out for skin microbiota fluctuations. Data showed that the cosmetics were safe, and they shifted the skin physiology parameters to a younger biological age, enhanced collagen synthesis, inhibited lipid peroxidation, and favoured normal microbiota.

Algorithm for Locating Apical Meristematic Tissue of Weeds Based on YOLO Instance Segmentation

Laser technology can be used to control weeds by irradiating the apical meristematic tissue (AMT) of weeds when they are still seedlings. Two factors are necessary for the successful large-scale implementation of this technique: the ability to accurately identify the apical meristematic tissue and the effectiveness of the localization algorithm used in the process. Based on this, this study proposes a lightweight weed AMT localization algorithm based on YOLO (look only once) instance segmentation. The YOLOv8n-seg network undergoes a lightweight design enhancement by integrating the FasterNet lightweight network as its backbone, resulting in the F-YOLOv8n-seg model. This modification effectively reduces the number of parameters and computational demands during the convolution process, thereby achieving a more efficient model. Subsequently, F-YOLOv8n-seg is combined with the connected domain analysis algorithm (CDA), yielding the F-YOLOv8n-seg-CDA model. This integration enables the precise localization of the AMT of weeds by calculating the center-of-mass coordinates of the connected domains. The experimental results indicate that the optimized model significantly outperforms the original model; the optimized model reduces floating-point computations by 26.7% and the model size by 38.2%. In particular, the floating-point calculation is decreased to 8.9 GFLOPs, and the model size is lowered to 4.2 MB. Comparing this improved model against YOLOv5s-seg and YOLOv10n-seg, it is lighter. Furthermore, it exhibits exceptional segmentation accuracy, with a 97.2% accuracy rate. Experimental tests conducted on five different weed species demonstrated that F-YOLOv8n-seg-CDA exhibits strong generalization capabilities. The combined accuracy of the algorithm for detecting these weeds was 81%. Notably, dicotyledonous weeds were detected with up to 94%. Additionally, the algorithm achieved an average inference speed of 82.9 frames per second. These results indicate that the algorithm is suitable for the real-time detection of apical meristematic tissues across multiple weed species. Furthermore, the experimental results demonstrated the impact of distinctive variations in weed morphology on identifying the location of the AMT of weeds. It was discovered that dicotyledonous and monocotyledonous weeds differed significantly in terms of the detection effect, with dicotyledonous weeds having significantly higher detection accuracy than monocotyledonous weeds. This discovery can offer novel insights and avenues for future investigation into the identification and location of the AMT of weeds.

Tyrosylprotein Sulfotransferase Positively Regulates Symbiotic Nodulation and Root Growth.

Posttranslational tyrosine sulfation of peptides and proteins is catalysed by tyrosylprotein sulfotransferases (TPSTs). In Arabidopsis, tyrosine sulfation is essential for the activities of peptide hormones, such as phytosulfokine (PSK) and root meristem growth factor (RGF). Here, we identified a TPST-encoding gene, MtTPST, from model legume Medicago truncatula. MtTPST expression was detected in all organs, with the highest level in root nodules. A promoter:GUS assay revealed that MtTPST was highly expressed in the root apical meristem, nodule primordium and nodule apical meristem. The loss-of-function mutant mttpst exhibited a stunted phenotype with short roots and reduced nodule number and size. Application of both of the sulfated peptides PSK and RGF3 partially restored the defective root length of mttpst. The reduction in symbiotic nodulation in mttpst was partially recovered by treatment with sulfated PSK peptide. MtTPST-PSK module functions downstream of the Nod factor signalling to promote nodule initiation via regulating accumulation and/or signalling of cytokinin and auxin. Additionally, the small-nodule phenotype of mttpst, which resulted from decreased apical meristematic activity, was partially complemented by sulfated RGF3 treatment. Together, these results demonstrate that MtTPST, through its substrates PSK, RGF3 and other sulfated peptide(s), positively regulates nodule development and root growth.

Connecting high-resolution 3D chromatin maps with cell division and cell differentiation at the root apical meristem

Key messageWe used marker-free technologies to study chromatin at cellular resolution. Our results show asymmetric chromatin distribution, explore chromatin dynamics during mitosis, and reveal structural differences between trichoblast and atrichoblast cell.The shapes, sizes, and structural organizations of plant nuclei vary considerably among cell types, tissues, and species. This diversity is dependent on various factors, including cellular function, developmental stage, and environmental or physiological conditions. The differences in nuclear structure reflect the state of chromatin, which, in turn, controls gene expression and regulates cell fate. To examine the interrelationship between nuclear structure, cell morphology, and tissue-specific cell proliferation and differentiation processes, we conducted multiple visualizations of H3K4me1, H3K9me2, 4′,6-diamidino-2-phenylindole, 5-ethynyl 2′-deoxyuridine, and SCRI Renaissance 2200, followed by subsequent quantitative analysis of individual cells and nuclei. By assigning cylindrical coordinates to the nuclei in the iRoCS toolbox, we were able to construct in situ digital three-dimensional chromatin maps for all the tissue layers of individual roots. A detailed analysis of the nuclei features of H3K4me1 and H3K9me2 in the mitotic and the elongation zones in trichoblast and atrichoblast cells at the root apical meristem revealed cell type-specific chromatin dynamics with asymmetric distribution of euchromatin and heterochromatin marks that may be associated with cell cycle and cell differentiation characteristics of specific cells. Furthermore, the spatial distribution of nuclei stained with 5-ethynyl 2′-deoxyuridine in the epidermis and cortex tissues suggests short-range coordination of cell division and nuclear migration in a linear sequence through an unknown regulatory mechanism.

Heritable gene editing in tomato through viral delivery of isopentenyl transferase and single-guide RNAs to latent axillary meristematic cells

Realizing the full potential of genome editing for crop improvement has been slow due to inefficient methods for reagent delivery and the reliance on tissue culture for creating gene-edited plants. RNA viral vectors offer an alternative approach for delivering gene engineering reagents and bypassing the tissue culture requirement. Viruses, however, are often excluded from the shoot apical meristem, making virus-mediated gene editing inefficient in some species. Here, we developed effective approaches for generating gene-edited shoots in Cas9-expressing transgenic tomato plants using RNA virus-mediated delivery of single-guide RNAs (sgRNAs). RNA viral vectors expressing sgRNAs were either delivered to leaves or sites near axillary meristems. Trimming of the apical and axillary meristems induced new shoots to form from edited somatic cells. To further encourage the induction of shoots, we used RNA viral vectors to deliver sgRNAs along with the cytokinin biosynthesis gene, isopentenyl transferase. Abundant, phenotypically normal, gene-edited shoots were induced per infected plant with single and multiplexed gene edits fixed in the germline. The use of viruses to deliver both gene editing reagents and developmental regulators overcomes the bottleneck in applying virus-induced gene editing to dicotyledonous crops such as tomato and reduces the dependency on tissue culture.

Preventive effects of dill oil on potassium bromated-induced oxidative DNA damage in garlic root tips

In this study, the potential antimutagenic/cytoprotective effect of the oil extract (2.5µg/ml) from Anethum graveolens L. (Dill) on oxidative DNA damage was tested. The experiments were carried out to inhibit mutagenicity induced by potassium bromated (KBrO3-9g/l) in Allium assay using mitotic activities and nuclear DNA contents. Result showed a clear reduction in radical length in all KBrO3-treated groups compared to control plants. Mitotic activity, measured by mitotic index (MI) and mitotic stages, were significantly decreased in meristematic cells after KBrO3 application, The results showed that mitotic index (MI) was decreased to reach about 0.8 at 24 hrs compared with 23.9% in untreated plants. The analysis of potential cyto and genoprotective of dill extract on nuclear DNA contents showed a statistically significant increase in the nuclear DNA content compared to KBrO3-treated roots was observed. Clearly, dill oil showed highly performance in protecting the mitotic cells against mutagenic KBrO3 effects by increasing the MI and decreasing the abnormal chromosomes. Also, the percentage of damage reduction was observed in the pre- (28.9, 57.9 and 46.4 %), simultaneous (53.0, 70.6 and 35.7 %) and post- (37.3, 61.8 and 50 %) treatment. The results of the qualitative study showed that this dill extracts were rich in chemical compounds belonging to several phytochemical classes. GC-MS analysis detected 29 chemical compounds with different concentrations. The predominant ones were Octadecanoic acid (31.75%) and 2-pentanone, 4-hydroxy-4-methyl (16.77). So that these experiments will help to understand and uncover the mechanism of action dill oil against the mutagenic effects of KBrO3 (i.e whether they have a prophylactic or treatment activity).

Large-volume fully automated cell reconstruction generates a cell atlas of plant tissues.

The geometric shape and arrangement of individual cells play a role in shaping organ functions. However, analyzing multicellular features and exploring their connectomes in centimeter-scale plant organs remain challenging. Here, we established a set of frameworks named Large-Volume Fully Automated Cell Reconstruction (LVACR), enabling the exploration of three-dimensional (3D) cytological features and cellular connectivity in plant tissues. Through benchmark testing, our framework demonstrated superior efficiency in cell segmentation and aggregation, successfully addressing the inherent challenges posed by light sheet fluorescence microscopy (LSFM) imaging. Using LVACR, we successfully established a cell atlas of different plant tissues. Cellular morphology analysis revealed differences of cell clusters and shapes in between different poplar (P. simonii Carr. and P. canadensis Moench.) seeds, whereas topological analysis revealed that they maintained conserved cellular connectivity. Furthermore, LVACR spatiotemporally demonstrated an initial burst of cell proliferation, accompanied by morphological transformations at an early stage in developing the shoot apical meristem. During subsequent development, cell differentiation produced anisotropic features, thereby resulting in various cell shapes. Overall, our findings provided valuable insights into the precise spatial arrangement and cellular behavior of multicellular organisms, thus enhancing our understanding of the complex processes underlying plant growth and differentiation.

A gain-of-function mutation at the C-terminus of FT-D1 promotes heading by interacting with 14-3-3A and FDL6 in wheat.

Vernalization and photoperiod pathways converging at FT1 control the transition to flowering in wheat. Here, we identified a gain-of-function mutation in FT-D1 that results in earlier heading date (HD), and shorter plant height and spike length in the gamma ray-induced eh1 wheat mutant. Knockout of the wild-type and overexpression of the mutated FT-D1 indicate that both alleles are functional to affect HD and plant height. Protein interaction assays demonstrated that the frameshift mutation in FT-D1eh1 exon 3 led to gain-of-function interactions with 14-3-3A and FDL6, thereby enabling the formation of florigen activation complex (FAC) and consequently activating a flowering-related transcriptomic programme. This mutation did not affect FT-D1eh1 interactions with TaNaKR5 or TaFTIP7, both of which could modulate HD, potentially via mediating FT-D1 translocation to the shoot apical meristem. Furthermore, the 'Segment B' external loop is essential for FT-D1 interaction with FDL6, while residue Y85 is required for interactions with TaNaKR5 and TaFTIP7. Finally, the flowering regulatory hub gene, ELF5, was identified as the FT-D1 regulatory target. This study illustrates FT-D1 function in determining wheat HD with a suite of interaction partners and provides genetic resources for tuning HD in elite wheat lines.

Magnetic Resonance Imaging-Based Monitoring of the Accumulation of Polyethylene Terephthalate Nanoplastics.

Polyethylene terephthalate (PET) is one of the most produced plastic materials in the world. The emergence of microplastics and nanoplastics (MPs/NPs) as a significant environmental contaminant has become a matter of increasing concern. While the toxicological effects of PET NPs have been widely researched, there is a lack of methodologies for studying their accumulation. The present study introduces a novel method to monitor the distribution of PET NPs in germinating wheat (Triticum aestivum L.) seeds. This involves the functionalization of superparamagnetic iron oxide nanoparticles (SPIONs) with PET NPs (PET-fSPIONs) coupled with magnetic resonance microimaging (µMRI) to provide insight into their distribution within the seed. The present study has demonstrated that PET-fSPIONs accumulate in specific regions of germinating wheat seeds, including the shoot apical meristem, the radicle, the coleoptile, the plumule, and the scutellum. Furthermore, the accumulation of PET-fSPIONs has been shown to exert a discernible effect on spin-spin relaxation (T2), as observed via MRI and quantitative T2 relaxation time analysis. The accumulation of PET NPs in embryo regions was also confirmed by SEM. Diffusion-weighted magnetic resonance imaging (DW-MRI) and non-invasive chemical shift imaging analyses demonstrated that PET NPs resulted in restricted diffusion within the highlighted areas, as well as an impact on lipid content. Our study reveals that using µMRI with fSPIONs provides a non-invasive method to monitor the biodistribution of PET nanoparticles in wheat seeds. Additionally, it offers valuable insights into the microstructural interactions of PET.

The WOX9-WUS modules are indispensable for the maintenance of stem cell homeostasis in Arabidopsis thaliana.

The dynamic balance between the self-renewal and differentiation of stem cells in plants is precisely regulated by a series of developmental regulated genes that exhibit spatiotemporal-specific expression patterns. Several studies have demonstrated that the WOX family transcription factors play critical roles in maintaining the identity of stem cells in Arabidopsis thaliana. In this study, we obtained amiR-WOX9 transgenic plants, which displayed terminating prematurely of shoot apical meristem (SAM) development, along with alterations in inflorescence meristem and flower development. The phenotype of amiR-WOX9 plants exhibited similarities to that of wus-101 mutant, characterized by a stop-and-go growth pattern. It was also found that the expression of WUS in amiR-WOX9 lines was decreased significantly, while in UBQ10::WOX9-GFP transgenic plants, the WUS expression was increased significantly despite no substantial alteration in meristem size compared to Col. Therefore, these data substantiated the indispensable role of WOX9 in maintaining the proper expression of WUS. Further investigations unveiled the direct binding of WOX9 to the WUS promoter via the TAAT motif, thereby activating its expression. It was also found that WUS recognized identical the same TAAT motif cis-elements in its own promoter, thereby repress self-expression. Next, we successfully identified a physical interaction between WOX9 and WUS, and verified that it was harmful to the expression of WUS. Finally, our experimental findings demonstrate that WOX9 was responsible for the direct activating of WUS, which however was interfered by the ways of WUS binding its own promoter and the interaction of WUS and WOX9, thereby ensuring the appropriate expression pattern of WUS and then the stem cell stability. This study contributes to an enhanced comprehension of the regulatory network of the WOX9-WUS module in maintaining the equilibrium of the SAM.

First report of Neopestalotiopsis clavispora Causing guava scab in China.

Psidium guajava L. is widely cultivated in southern China. In May 2021, guava scab on cv. Zhenzhu was observed in Zhanjiang (21.18° N, 110.21° E), Guangdong province, China. Guavascab was corky with ovoid or round lesions on the surfaces of green fruits. Gradually the lesions sunk. Disease incidence was estimated as 85% in 500 investigated plants in about 50 ha. Twenty diseased fruits were collected from twenty trees in the field. From each fruit the margin of the diseased tissues was cut into 2 mm × 2 mm pieces; surface disinfected with 75% ethanol and 2% sodium hypochlorite for 30 and 60 s, successively; and rinsed thrice with sterile water. The tissues were plated onto potato dextrose agar (PDA) medium and incubated at 28 ℃. Thirty-four isolates were obtained. Single-spore isolation method (Liu et al. 2021) was used to recover pure cultures of three isolates (PGNC-1, PGNC-2, and PGNC-3) . The colonies were initially white with cottony aerial mycelium at 7 days on PDA. Then, these colonies form black acervular conidiomata at 10 days. Conidia were clavate to fusiform, four-septate, straight or slightly curved, and measured 15.8 to 21.2 µm × 4.5 to 6.5 µm (n = 40). The three median cells were versicolored, whereas the basal and apical cells were hyaline. Conidia had a single basal appendage (4.5 to 5.5 µm long; n = 40) and three apical appendages (19.2 to 24.5 µm long; n = 40). The morphological characteristics of the isolates were consistent with the description of Neopestalotiopsisclavispora (Maharachchikumbura et al. 2012). Molecular identification was performed using PCR method with MightyAmp DNA Polymerase (Takara-Bio, Dalian, China) (Lu et al. 2012). Sequences were generated from the isolates using primers for the rDNA ITS (ITS1/ITS4), TEF1-α (EF1-728F/EF1-986R), and β-tubulin (T1/βt2b) loci (Maharachchikumbura et al. 2012). The sequences of the isolates were submitted to GenBank (ITS, OQ996557 to OQ996559; TEF, OR101037 to OR101039; β-tubulin, OR100971 to OR100973). The sequences of the isolates were 100% identical to the type strain MFLUCC12-0281 (accession nos. JX398979, JX399014, and JX399045) through BLAST analysis. The isolates clustered with N. clavispora (MFLUCC12-0280 and MFLUCC12-0281). N. clavispora and Pestalotiopsis clavispora are synonyms. The pathogenicity was tested in vivo. Plants (cv. Zhenzhu) were grown ( 3 years old) in a quarantine orchard at 25℃ to 32℃ with 60 to 80% relative humidity in May 2022. Disease-free green fruits were inoculated. Sterile cotton balls were immersed in the spore suspension (1 × 105 per mL) and sterile distilled water (control) for about 15 s before they were fixed on the wounded fruits with transparent tape. Five fruits on one plant per isolate were inoculated. Five fruits on one plant severed as control. The test was performed thrice. Disease symptoms were found on the inoculated fruits after 20 days, whereas the controls remained healthy. The pathogen was re-isolated from infected fruits and was phenotypically identical to the original isolates thus fulfilling Koch's postulates. Neopestalotiopsis or Pestalotiopsis spp. were reported to be the causal agents of guava scab in Colombia and in Hawaii (Keith et al. 2006; Solarte et al. 2018). N. clavispora has been reported to cause disease in a broad range of hosts (Ge et al. 2009; Chen et al. 2018), but not in guava. This is the first report of N. clavispora causing guava scab in China. There would be no harvest if this disease is left unmanaged.