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1.
Mol Biol Rep ; 51(1): 605, 2024 May 03.
Article in English | MEDLINE | ID: mdl-38700570

ABSTRACT

BACKGROUND: Cultivation of Crocus sativus (saffron) faces challenges due to inconsistent flowering patterns and variations in yield. Flowering takes place in a graded way with smaller corms unable to produce flowers. Enhancing the productivity requires a comprehensive understanding of the underlying genetic mechanisms that govern this size-based flowering initiation and commitment. Therefore, samples enriched with non-flowering and flowering apical buds from small (< 6 g) and large (> 14 g) corms were sequenced. METHODS AND RESULTS: Apical bud enriched samples from small and large corms were collected immediately after dormancy break in July. RNA sequencing was performed using Illumina Novaseq 6000 to access the gene expression profiles associated with size dependent flowering. De novo transcriptome assembly and analysis using flowering committed buds from large corms at post-dormancy and their comparison with vegetative shoot primordia from small corms pointed out the major role of starch and sucrose metabolism, Auxin and ABA hormonal regulation. Many genes with known dual responses in flowering development and circadian rhythm like Flowering locus T and Cryptochrome 1 along with a transcript showing homology with small auxin upregulated RNA (SAUR) exhibited induced expression in flowering buds. Thorough prediction of Crocus sativus non-coding RNA repertoire has been carried out for the first time. Enolase was found to be acting as a major hub with protein-protein interaction analysis using Arabidopsis counterparts. CONCLUSION: Transcripts belong to key pathways including phenylpropanoid biosynthesis, hormone signaling and carbon metabolism were found significantly modulated. KEGG assessment and protein-protein interaction analysis confirm the expression data. Findings unravel the genetic determinants driving the size dependent flowering in Crocus sativus.


Subject(s)
Crocus , Flowers , Gene Expression Profiling , Gene Expression Regulation, Plant , Indoleacetic Acids , Meristem , Signal Transduction , Flowers/genetics , Flowers/growth & development , Flowers/metabolism , Crocus/genetics , Crocus/growth & development , Crocus/metabolism , Gene Expression Regulation, Plant/genetics , Indoleacetic Acids/metabolism , Gene Expression Profiling/methods , Meristem/genetics , Meristem/growth & development , Meristem/metabolism , Signal Transduction/genetics , Plant Proteins/genetics , Plant Proteins/metabolism , Transcriptome/genetics , Sugars/metabolism , Plant Growth Regulators/metabolism
2.
Sci Rep ; 14(1): 11148, 2024 05 15.
Article in English | MEDLINE | ID: mdl-38750143

ABSTRACT

The one-leaf plant Monophyllaea glabra exhibits a unique developmental manner in which only one cotyledon continues growing without producing new vegetative organs. This morphology is formed by specific meristems, the groove meristem (GM) and the basal meristem (BM), which are thought to be modified shoot apical meristem (SAM) and leaf meristem. In this study, we analysed the expression of the organ boundary gene CUP-SHAPED COTYLEDON (CUC) and the SAM maintenance gene SHOOT MERISTEMLESS (STM) orthologs by whole-mount in situ hybridisation. We found that CUCs did not show clear border patterns around GM and BM during the vegetative phase. Furthermore, double-colour detection analysis at the cellular level revealed that CUC and STM expression overlapped in the GM region during the vegetative phase. We also found that this overlap is dissolved in the reproductive phase when normal shoot organogenesis is observed. Since co-expression of these genes occurs during SAM initiation under embryogenesis in Arabidopsis, our results demonstrate that GM is a prolonged stage of pre-mature SAM. Therefore, we propose that neotenic meristems could be a novel plant trait acquired by one-leaf plants.


Subject(s)
Cotyledon , Gene Expression Regulation, Plant , Meristem , Meristem/genetics , Meristem/growth & development , Meristem/metabolism , Cotyledon/genetics , Cotyledon/growth & development , Plant Shoots/genetics , Plant Shoots/growth & development , Plant Shoots/metabolism , Plant Leaves/genetics , Plant Leaves/metabolism , Plant Leaves/growth & development , Plant Proteins/genetics , Plant Proteins/metabolism , Arabidopsis/genetics , Arabidopsis/growth & development
3.
Nat Plants ; 10(5): 815-827, 2024 May.
Article in English | MEDLINE | ID: mdl-38745100

ABSTRACT

A comprehensive understanding of inflorescence development is crucial for crop genetic improvement, as inflorescence meristems give rise to reproductive organs and determine grain yield. However, dissecting inflorescence development at the cellular level has been challenging owing to a lack of specific marker genes to distinguish among cell types, particularly in different types of meristems that are vital for organ formation. In this study, we used spatial enhanced resolution omics-sequencing (Stereo-seq) to construct a precise spatial transcriptome map of the developing maize ear primordium, identifying 12 cell types, including 4 newly defined cell types found mainly in the inflorescence meristem. By extracting the meristem components for detailed clustering, we identified three subtypes of meristem and validated two MADS-box genes that were specifically expressed at the apex of determinate meristems and involved in stem cell determinacy. Furthermore, by integrating single-cell RNA transcriptomes, we identified a series of spatially specific networks and hub genes that may provide new insights into the formation of different tissues. In summary, this study provides a valuable resource for research on cereal inflorescence development, offering new clues for yield improvement.


Subject(s)
Inflorescence , Meristem , Transcriptome , Zea mays , Zea mays/genetics , Zea mays/growth & development , Zea mays/metabolism , Inflorescence/genetics , Inflorescence/growth & development , Meristem/genetics , Meristem/growth & development , Meristem/metabolism , Gene Expression Regulation, Plant , Gene Expression Profiling
4.
Planta ; 259(6): 148, 2024 May 08.
Article in English | MEDLINE | ID: mdl-38717679

ABSTRACT

MAIN CONCLUSION: Mutation of OsSHR2 adversely impacted root and shoot growth and impaired plant response to N conditions, further reducing the yield per plant. Nitrogen (N) is a crucial factor that regulates the plant architecture. There is still a lack of research on it. In our study, it was observed that the knockout of the SHORTROOT 2 (OsSHR2) which was induced by N deficiency, can significantly affect the regulation of plant architecture response to N in rice. Under N deficiency, the mutation of OsSHR2 significantly reduced root growth, and impaired the sensitivity of the root meristem length to N deficiency. The mutants were found to have approximately a 15% reduction in plant height compared to wild type. But mutants showed a significant increase in tillering at post-heading stage, approximately 26% more than the wild type, particularly in high N conditions. In addition, due to reduced seed setting rate and 1000-grain weight, mutant yield was significantly decreased by approximately 33% under low N fertilizer supply. The mutation also changed the distribution of N between the vegetative and reproductive organs. Our findings suggest that the transcription factor OsSHR2 plays a regulatory role in the response of plant architecture and yield per plant to N in rice.


Subject(s)
Gene Expression Regulation, Plant , Nitrogen , Oryza , Transcription Factors , Gene Expression Regulation, Plant/drug effects , Meristem/genetics , Meristem/growth & development , Meristem/drug effects , Mutation , Nitrogen/metabolism , Nitrogen/pharmacology , Oryza/genetics , Oryza/growth & development , Oryza/metabolism , Oryza/drug effects , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Roots/growth & development , Plant Roots/genetics , Plant Roots/drug effects , Plant Roots/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism
5.
Curr Biol ; 34(11): 2344-2358.e5, 2024 Jun 03.
Article in English | MEDLINE | ID: mdl-38781954

ABSTRACT

Inflorescence architecture and crop productivity are often tightly coupled in our major cereal crops. However, the underlying genetic mechanisms controlling cereal inflorescence development remain poorly understood. Here, we identified recessive alleles of barley (Hordeum vulgare L.) HvALOG1 (Arabidopsis thaliana LSH1 and Oryza G1) that produce non-canonical extra spikelets and fused glumes abaxially to the central spikelet from the upper-mid portion until the tip of the inflorescence. Notably, we found that HvALOG1 exhibits a boundary-specific expression pattern that specifically excludes reproductive meristems, implying the involvement of previously proposed localized signaling centers for branch regulation. Importantly, during early spikelet formation, non-cell-autonomous signals associated with HvALOG1 expression may specify spikelet meristem determinacy, while boundary formation of floret organs appears to be coordinated in a cell-autonomous manner. Moreover, barley ALOG family members synergistically modulate inflorescence morphology, with HvALOG1 predominantly governing meristem maintenance and floral organ development. We further propose that spatiotemporal redundancies of expressed HvALOG members specifically in the basal inflorescence may be accountable for proper patterning of spikelet formation in mutant plants. Our research offers new perspectives on regulatory signaling roles of ALOG transcription factors during the development of reproductive meristems in cereal inflorescences.


Subject(s)
Hordeum , Inflorescence , Meristem , Plant Proteins , Signal Transduction , Hordeum/genetics , Hordeum/growth & development , Hordeum/metabolism , Meristem/growth & development , Meristem/genetics , Meristem/metabolism , Inflorescence/growth & development , Inflorescence/genetics , Inflorescence/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Gene Expression Regulation, Plant
6.
Plant Mol Biol ; 114(3): 65, 2024 May 30.
Article in English | MEDLINE | ID: mdl-38816532

ABSTRACT

Telomeres are conserved chromosomal structures necessary for continued cell division and proliferation. In addition to the classical telomerase pathway, multiple other genes including those involved in ribosome metabolism and chromatin modification contribute to telomere length maintenance. We previously reported that Arabidopsis thaliana ribosome biogenesis genes OLI2/NOP2A, OLI5/RPL5A and OLI7/RPL5B have critical roles in telomere length regulation. These three OLIGOCELLULA genes were also shown to function in cell proliferation and expansion control and to genetically interact with the transcriptional co-activator ANGUSTIFOLIA3 (AN3). Here we show that AN3-deficient plants progressively lose telomeric DNA in early homozygous mutant generations, but ultimately establish a new shorter telomere length setpoint by the fifth mutant generation with a telomere length similar to oli2/nop2a -deficient plants. Analysis of double an3 oli2 mutants indicates that the two genes are epistatic for telomere length control. Telomere shortening in an3 and oli mutants is not caused by telomerase inhibition; wild type levels of telomerase activity are detected in all analyzed mutants in vitro. Late generations of an3 and oli mutants are prone to stem cell damage in the root apical meristem, implying that genes regulating telomere length may have conserved functional roles in stem cell maintenance mechanisms. Multiple instances of anaphase fusions in late generations of oli5 and oli7 mutants were observed, highlighting an unexpected effect of ribosome biogenesis factors on chromosome integrity. Overall, our data implicate AN3 transcription coactivator and OLIGOCELLULA proteins in the establishment of telomere length set point in plants and further suggest that multiple regulators with pleiotropic functions can connect telomere biology with cell proliferation and cell expansion pathways.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Cell Division , Telomerase , Telomere , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Telomere/genetics , Telomere/metabolism , Cell Division/genetics , Telomerase/genetics , Telomerase/metabolism , Telomere Homeostasis/genetics , Gene Expression Regulation, Plant , Mutation , Transcription Factors/metabolism , Transcription Factors/genetics , Cell Proliferation/genetics , Meristem/genetics , Meristem/metabolism
7.
Methods Mol Biol ; 2787: 95-103, 2024.
Article in English | MEDLINE | ID: mdl-38656484

ABSTRACT

Our method describes how to collect forest tree root tips in the field, to store them for transfer to the lab, to pretreat root tips in order to arrest cells in metaphase, fix root tips to preserve specific morphological organizations, to stain fixed root tips by Feulgen's Reaction in order to increase contrast, and to prepare the root meristem for analyzing mitotic stages and chromosomal aberrations via light microscopy. We further describe how to classify chromosomal abnormalities and quantify them via aberration indices.


Subject(s)
Meristem , Trees , Meristem/genetics , Trees/genetics , Chromosome Aberrations , Plant Roots/genetics , Plant Roots/growth & development , Cytogenetic Analysis/methods
8.
Methods Mol Biol ; 2787: 81-94, 2024.
Article in English | MEDLINE | ID: mdl-38656483

ABSTRACT

Plant genetics plays a key role in determining root hair initiation and development. A complex network of genetic interactions therefore closely monitors and influences root hair phenotype and morphology. The significance of these genes can be studied by employing, for instance, loss-of-function mutants, overexpression plant lines, and fluorescently labeled constructs. Confocal laser scanning microscopy is a great tool to visually observe and document these morphological features. This chapter elaborates the techniques involved in handling of microscopic setup to acquire images displaying root hair distribution along the fully elongated zone of Arabidopsis thaliana roots. Additionally, we illustrate an approach to visualize early fate determination of epidermal cells in the root apical meristem, by describing a method for imaging YFP tagged transgenic plant lines.


Subject(s)
Arabidopsis , Microscopy, Confocal , Plant Roots , Microscopy, Confocal/methods , Plant Roots/growth & development , Plant Roots/genetics , Plant Roots/cytology , Arabidopsis/genetics , Plants, Genetically Modified/genetics , Meristem/growth & development , Meristem/genetics
9.
Physiol Plant ; 176(3): e14320, 2024.
Article in English | MEDLINE | ID: mdl-38686642

ABSTRACT

Many nucleoside triphosphate-diphosphohydrolases (NTPDases/APYRASEs, APYs) play a key role in modulating extracellular nucleotide levels. However, the Golgi-localized APYs, which help control glycosylation, have rarely been studied. Here, we identified AtAPY1, a gene encoding an NTPDase in the Golgi apparatus, which is required for cell wall integrity and plant growth under boron (B) limited availability. Loss of function in AtAPY1 hindered cell elongation and division in root tips while increasing the number of cortical cell layers, leading to swelling of the root tip and abundant root hairs under low B stress. Further, expression pattern analysis revealed that B deficiency significantly induced AtAPY1, especially in the root meristem and stele. Fluorescent-labeled AtAPY1-GFP localized to the Golgi stack. Biochemical analysis showed that AtAPY1 exhibited a preference of UDP and GDP hydrolysis activities. Consequently, the loss of function in AtAPY1 might disturb the homoeostasis of NMP-driven NDP-sugar transport, which was closely related to the synthesis of cell wall polysaccharides. Further, cell wall-composition analysis showed that pectin content increased and borate-dimerized RG-II decreased in apy1 mutants, along with a decrease in cellulose content. Eventually, altered polysaccharide characteristics presumably cause growth defects in apy1 mutants under B deficiency. Altogether, these data strongly support a novel role for AtAPY1 in mediating responses to low B availability by regulating cell wall integrity.


Subject(s)
Apyrase , Arabidopsis Proteins , Arabidopsis , Boron , Cell Wall , Golgi Apparatus , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis/enzymology , Arabidopsis/metabolism , Cell Wall/metabolism , Boron/metabolism , Boron/deficiency , Golgi Apparatus/metabolism , Arabidopsis Proteins/metabolism , Arabidopsis Proteins/genetics , Apyrase/metabolism , Apyrase/genetics , Gene Expression Regulation, Plant , Plant Roots/growth & development , Plant Roots/genetics , Plant Roots/metabolism , Meristem/genetics , Meristem/growth & development , Meristem/metabolism , Pectins/metabolism
10.
Plant Physiol Biochem ; 211: 108635, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38688114

ABSTRACT

Meristem activity is important for normal plant growth as well as adaptive plastic development under abiotic stresses. Cytokinin has been recognized to have a major role in regulating meristem function which is controlled by cytokinin activating enzymes by fine-tuning the concentrations and spatial distribution of its bioactive forms. It was previously reported that LONELY GUY (LOG) acts in the direct activation pathway of cytokinin in rice shoot meristems. LOG has a cytokinin specific phosphoribohydrolase activity, which transforms inactive cytokinin nucleotides into active free bases. Here, we explored the role of OsLOG in controlling meristem activity mediated by cytokinin and its effects on growth, development, and stress resilience of rice plants. Overexpression of OsLOG in rice led to significant alterations in cytokinin levels in the inflorescence meristem, leading to enhanced plant growth, biomass and grain yield under both non-stress as well as stress conditions such as drought and salinity. Moreover, our study provides insight into how overexpression of OsLOG improves the ability of plants to withstand stress. The OsLOG-overexpressing lines exhibit reduced accumulation of H2O2 along with elevated antioxidant enzyme activities, thereby maintaining better redox homeostasis under stress conditions. This ultimately reduces the negative impact of stresses on grain yield and improves harvest index, as evidenced by observations in the OsLOG-overexpressing lines. In summary, our study emphasizes the diverse role of OsLOG, not only in regulating plant growth and yield via cytokinin but also in enhancing adaptability to abiotic stresses. This highlights its potential to improve crop yield and promote sustainable agriculture.


Subject(s)
Cytokinins , Oryza , Plant Proteins , Stress, Physiological , Oryza/genetics , Oryza/enzymology , Oryza/growth & development , Oryza/metabolism , Cytokinins/metabolism , Plant Proteins/metabolism , Plant Proteins/genetics , Plants, Genetically Modified , Edible Grain/growth & development , Edible Grain/genetics , Gene Expression Regulation, Plant , Meristem/growth & development , Meristem/genetics , Meristem/metabolism , Droughts
11.
EMBO J ; 43(9): 1843-1869, 2024 May.
Article in English | MEDLINE | ID: mdl-38565948

ABSTRACT

The RNA-silencing effector ARGONAUTE10 influences cell fate in plant shoot and floral meristems. ARGONAUTE10 also accumulates in the root apical meristem (RAM), yet its function(s) therein remain elusive. Here, we show that ARGONAUTE10 is expressed in the root cell initials where it controls overall RAM activity and length. ARGONAUTE10 is also expressed in the stele, where post-transcriptional regulation confines it to the root tip's pro-vascular region. There, variations in ARGONAUTE10 levels modulate metaxylem-vs-protoxylem specification. Both ARGONAUTE10 functions entail its selective, high-affinity binding to mobile miR165/166 transcribed in the neighboring endodermis. ARGONAUTE10-bound miR165/166 is degraded, likely via SMALL-RNA-DEGRADING-NUCLEASES1/2, thus reducing miR165/166 ability to silence, via ARGONAUTE1, the transcripts of cell fate-influencing transcription factors. These include PHABULOSA (PHB), which controls meristem activity in the initials and xylem differentiation in the pro-vasculature. During early germination, PHB transcription increases while dynamic, spatially-restricted transcriptional and post-transcriptional mechanisms reduce and confine ARGONAUTE10 accumulation to the provascular cells surrounding the newly-forming xylem axis. Adequate miR165/166 concentrations are thereby channeled along the ARGONAUTE10-deficient yet ARGONAUTE1-proficient axis. Consequently, inversely-correlated miR165/166 and PHB gradients form preferentially along the axis despite ubiquitous PHB transcription and widespread miR165/166 delivery inside the whole vascular cylinder.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Argonaute Proteins , Gene Expression Regulation, Plant , Meristem , MicroRNAs , Plant Roots , Xylem , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis/growth & development , MicroRNAs/metabolism , MicroRNAs/genetics , Meristem/metabolism , Meristem/growth & development , Meristem/genetics , Arabidopsis Proteins/metabolism , Arabidopsis Proteins/genetics , Argonaute Proteins/metabolism , Argonaute Proteins/genetics , Xylem/metabolism , Xylem/growth & development , Xylem/genetics , Plant Roots/metabolism , Plant Roots/growth & development , Plant Roots/genetics
12.
Plant Cell Rep ; 43(4): 87, 2024 Mar 09.
Article in English | MEDLINE | ID: mdl-38460026

ABSTRACT

KEY MESSAGE: Low concentrations of hydroxyurea, an inhibitor of DNA replication, induced oxidative and replicative stress in root apical meristem (RAM) cells of Vicia faba. Plant cells are constantly exposed to low-level endogenous stress factors that can affect DNA replication and lead to DNA damage. Long-term treatments of Vicia faba root apical meristems (RAMs) with HU leads to the appearance of atypical cells with intranuclear asynchrony. This rare form of abnormality was manifested by a gradual condensation of chromatin, from interphase to mitosis (so-called IM cells). Moreover, HU-treated root cells revealed abnormal chromosome structure, persisting DNA replication, and elevated levels of intracellular hydrogen peroxide (H2O2) and superoxide anion (O2∙-). Immunocytochemical studies have shown an increased number of fluorescent foci of H3 histones acetylated at lysine 56 (H3K56Ac; canonically connected with the DNA replication process). We show that continuous 3-day exposure to low concentrations (0.75 mM) of hydroxyurea (HU; an inhibitor of DNA replication) induces cellular response to reactive oxygen species and to DNA replication stress conditions.


Subject(s)
Hydroxyurea , Vicia faba , Hydroxyurea/pharmacology , Meristem/genetics , Vicia faba/genetics , Hydrogen Peroxide , Oxidative Stress
13.
J Exp Bot ; 75(11): 3220-3232, 2024 Jun 07.
Article in English | MEDLINE | ID: mdl-38527334

ABSTRACT

The first TALE homeodomain transcription factor gene to be described in plants was maize knotted1 (kn1). Dominant mutations in kn1 disrupt leaf development, with abnormal knots of tissue forming in the leaf blade. kn1 was found to be expressed in the shoot meristem but not in a peripheral region that gives rise to leaves. Furthermore, KN1 and closely related proteins were excluded from initiating and developing leaves. These findings were a prelude to a large body of work wherein TALE homeodomain proteins have been identified as vital regulators of meristem homeostasis and organ development in plants. KN1 homologues are widely represented across land plant taxa. Thus, studying the regulation and mechanistic action of this gene class has allowed investigations into the evolution of diverse plant morphologies. This review will focus on the function of TALE homeodomain transcription factors in leaf development in eudicots. Here, we discuss how TALE homeodomain proteins contribute to a spectrum of leaf forms, from the simple leaves of Arabidopsis thaliana to the compound leaves of Cardamine hirsuta and species beyond the Brassicaceae.


Subject(s)
Homeodomain Proteins , Plant Leaves , Plant Proteins , Transcription Factors , Plant Leaves/metabolism , Plant Leaves/growth & development , Plant Leaves/genetics , Homeodomain Proteins/metabolism , Homeodomain Proteins/genetics , Transcription Factors/metabolism , Transcription Factors/genetics , Plant Proteins/metabolism , Plant Proteins/genetics , Gene Expression Regulation, Plant , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis/growth & development , Meristem/genetics , Meristem/growth & development , Meristem/metabolism
14.
Plant Cell Environ ; 47(6): 2027-2043, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38391415

ABSTRACT

Understanding the regulation of flowering time is crucial for adaptation of crops to new environment. In this study, we examined the timing of floral transition and analysed transcriptomes in leaf and shoot apical meristems of photoperiod-sensitive and -insensitive quinoa accessions. Histological analysis showed that floral transition in quinoa initiates 2-3 weeks after sowing. We found four groups of differentially expressed genes in quinoa genome that responded to plant development and floral transition: (i) 222 genes responsive to photoperiod in leaves, (ii) 1812 genes differentially expressed between accessions under long-day conditions in leaves, (iii) 57 genes responding to developmental changes under short-day conditions in leaves and (iv) 911 genes responding to floral transition within the shoot apical meristem. Interestingly, among numerous candidate genes, two putative FT orthologs together with other genes (e.g. SOC1, COL, AP1) were previously reported as key regulators of flowering time in other species. Additionally, we used coexpression networks to associate novel transcripts to a putative biological process based on the annotated genes within the same coexpression cluster. The candidate genes in this study would benefit quinoa breeding by identifying and integrating their beneficial haplotypes in crossing programs to develop adapted cultivars to diverse environmental conditions.


Subject(s)
Chenopodium quinoa , Gene Expression Regulation, Plant , Meristem , Photoperiod , Plant Leaves , Transcriptome , Chenopodium quinoa/genetics , Chenopodium quinoa/growth & development , Chenopodium quinoa/physiology , Meristem/genetics , Meristem/growth & development , Plant Leaves/genetics , Plant Leaves/growth & development , Transcriptome/genetics , Flowers/genetics , Flowers/growth & development , Plant Shoots/genetics , Plant Shoots/growth & development , Plant Proteins/genetics , Plant Proteins/metabolism , Gene Expression Profiling
15.
Plant J ; 118(3): 607-625, 2024 May.
Article in English | MEDLINE | ID: mdl-38361340

ABSTRACT

The conservation of GOLVEN (GLV)/ROOT MERISTEM GROWTH FACTOR (RGF) peptide encoding genes across plant genomes capable of forming roots or root-like structures underscores their potential significance in the terrestrial adaptation of plants. This study investigates the function and role of GOLVEN peptide-coding genes in Medicago truncatula. Five out of fifteen GLV/RGF genes were notably upregulated during nodule organogenesis and were differentially responsive to nitrogen deficiency and auxin treatment. Specifically, the expression of MtGLV9 and MtGLV10 at nodule initiation sites was contingent upon the NODULE INCEPTION transcription factor. Overexpression of these five nodule-induced GLV genes in hairy roots of M. truncatula and application of their synthetic peptide analogues led to a decrease in nodule count by 25-50%. Uniquely, the GOLVEN10 peptide altered the positioning of the first formed lateral root and nodule on the primary root axis, an observation we term 'noduletaxis'; this decreased the length of the lateral organ formation zone on roots. Histological section of roots treated with synthetic GOLVEN10 peptide revealed an increased cell number within the root cortical cell layers without a corresponding increase in cell length, leading to an elongation of the root likely introducing a spatiotemporal delay in organ formation. At the transcription level, the GOLVEN10 peptide suppressed expression of microtubule-related genes and exerted its effects by changing expression of a large subset of Auxin responsive genes. These findings advance our understanding of the molecular mechanisms by which GOLVEN peptides modulate root morphology, nodule ontogeny, and interactions with key transcriptional pathways.


Subject(s)
Gene Expression Regulation, Plant , Medicago truncatula , Plant Proteins , Plant Roots , Root Nodules, Plant , Medicago truncatula/genetics , Medicago truncatula/growth & development , Medicago truncatula/metabolism , Medicago truncatula/drug effects , Medicago truncatula/physiology , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Roots/growth & development , Plant Roots/genetics , Plant Roots/drug effects , Plant Roots/metabolism , Root Nodules, Plant/genetics , Root Nodules, Plant/growth & development , Root Nodules, Plant/metabolism , Root Nodules, Plant/drug effects , Indoleacetic Acids/metabolism , Indoleacetic Acids/pharmacology , Plant Root Nodulation/genetics , Meristem/genetics , Meristem/growth & development , Meristem/drug effects , Peptides/metabolism , Peptides/genetics
16.
Mutat Res ; 828: 111851, 2024.
Article in English | MEDLINE | ID: mdl-38382175

ABSTRACT

Bleomycin, commonly employed in treating Hodgkin's lymphoma and testicular cancer, is associated with significant pulmonary toxicity. While various studies have assessed the toxic impact of chemotherapeutic agents on aquatic and terrestrial environments, limited data exist on bleomycin's effects, especially concerning higher plants. To address this gap, we utilized the Allium cepa assays, renowned for evaluating chemical and biochemical agents' toxic effects, to investigate bleomycin's impact on the terrestrial ecosystem. Our study aimed to assess bleomycin's cyto-genotoxic effects on A. cepa root tip cells at minimal concentrations (10-40 µg mL-1) and varied exposure durations (2, 4, 6, and 24 h). Analysis of nuclear and mitotic abnormalities in bleomycin-treated A. cepa root tip cells, alongside an acridine orange-ethidium bromide double staining assay, illuminated its influence on cell viability. Additionally, agarose gel electrophoresis determined the drug's potential for DNA degradation, unveiling the underlying mechanisms of cyto-genotoxicity. Results also demonstrated a decline in the mitotic index with increased bleomycin concentrations and exposure time, elevated frequencies of various cyto-genotoxic abnormalities, including sticky chromosomes, chromatid breaks, laggards, bridges, polar deviations, nuclear lesions, and hyperchromasia. The study indicated the potential risks of bleomycin even at low concentrations and brief exposures, highlighting its severe adverse effects on genetic material of plant, potentially contributing to cell death. Consequently, this investigation unveils bleomycin's cyto-genotoxic effects on higher plant system, underscoring its threat to terrestrial ecosystems, particularly upon chronic and unmonitored exposure.


Subject(s)
Bleomycin , Meristem , Onions , Bleomycin/toxicity , Onions/drug effects , Onions/genetics , Meristem/drug effects , Meristem/genetics , Cell Cycle/drug effects , DNA Damage/drug effects , Cell Survival/drug effects , Mutagenicity Tests/methods , Antibiotics, Antineoplastic/toxicity , Mutagens/toxicity , Chromosome Aberrations/chemically induced , Mitotic Index
17.
Plant J ; 118(3): 802-822, 2024 May.
Article in English | MEDLINE | ID: mdl-38305492

ABSTRACT

Floral patterns are unique to rice and contribute significantly to its reproductive success. SL1 encodes a C2H2 transcription factor that plays a critical role in flower development in rice, but the molecular mechanism regulated by it remains poorly understood. Here, we describe interactions of the SL1 with floral homeotic genes, SPW1, and DL in specifying floral organ identities and floral meristem fate. First, the sl1 spw1 double mutant exhibited a stamen-to-pistil transition similar to that of sl1, spw1, suggesting that SL1 and SPW1 may located in the same pathway regulating stamen development. Expression analysis revealed that SL1 is located upstream of SPW1 to maintain its high level of expression and that SPW1, in turn, activates the B-class genes OsMADS2 and OsMADS4 to suppress DL expression indirectly. Secondly, sl1 dl displayed a severe loss of floral meristem determinacy and produced amorphous tissues in the third/fourth whorl. Expression analysis revealed that the meristem identity gene OSH1 was ectopically expressed in sl1 dl in the fourth whorl, suggesting that SL1 and DL synergistically terminate the floral meristem fate. Another meristem identity gene, FON1, was significantly decreased in expression in sl1 background mutants, suggesting that SL1 may directly activate its expression to regulate floral meristem fate. Finally, molecular evidence supported the direct genomic binding of SL1 to SPW1 and FON1 and the subsequent activation of their expression. In conclusion, we present a model to illustrate the roles of SL1, SPW1, and DL in floral organ specification and regulation of floral meristem fate in rice.


Subject(s)
Flowers , Gene Expression Regulation, Plant , Meristem , Oryza , Plant Proteins , Oryza/genetics , Oryza/growth & development , Oryza/metabolism , Meristem/genetics , Meristem/growth & development , Meristem/metabolism , Flowers/genetics , Flowers/growth & development , Flowers/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Transcription Factors/metabolism , Transcription Factors/genetics , Plants, Genetically Modified , Mutation
18.
Plant Cell ; 36(6): 2140-2159, 2024 May 29.
Article in English | MEDLINE | ID: mdl-38391349

ABSTRACT

Transcription factors (TFs) are essential for the regulation of gene expression and cell fate determination. Characterizing the transcriptional activity of TF genes in space and time is a critical step toward understanding complex biological systems. The vegetative gametophyte meristems of bryophytes share some characteristics with the shoot apical meristems of flowering plants. However, the identity and expression profiles of TFs associated with gametophyte organization are largely unknown. With only ∼450 putative TF genes, Marchantia (Marchantia polymorpha) is an outstanding model system for plant systems biology. We have generated a near-complete collection of promoter elements derived from Marchantia TF genes. We experimentally tested reporter fusions for all the TF promoters in the collection and systematically analyzed expression patterns in Marchantia gemmae. This allowed us to build a map of expression domains in early vegetative development and identify a set of TF-derived promoters that are active in the stem-cell zone. The cell markers provide additional tools and insight into the dynamic regulation of the gametophytic meristem and its evolution. In addition, we provide an online database of expression patterns for all promoters in the collection. We expect that these promoter elements will be useful for cell-type-specific expression, synthetic biology applications, and functional genomics.


Subject(s)
Gene Expression Regulation, Plant , Marchantia , Meristem , Plant Proteins , Promoter Regions, Genetic , Transcription Factors , Marchantia/genetics , Marchantia/growth & development , Promoter Regions, Genetic/genetics , Transcription Factors/genetics , Transcription Factors/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Meristem/genetics , Meristem/growth & development
19.
Int J Mol Sci ; 25(4)2024 Feb 06.
Article in English | MEDLINE | ID: mdl-38396652

ABSTRACT

The architecture of the root system is fundamental to plant productivity. The rate of root growth, the density of lateral roots, and the spatial structure of lateral and adventitious roots determine the developmental plasticity of the root system in response to changes in environmental conditions. One of the genes involved in the regulation of the slope angle of lateral roots is DEEPER ROOTING 1 (DRO1). Its orthologs and paralogs have been identified in rice, Arabidopsis, and several other species. However, nothing is known about the formation of the slope angle of lateral roots in species with the initiation of lateral root primordia within the parental root meristem. To address this knowledge gap, we identified orthologs and paralogs of the DRO1 gene in cucumber (Cucumis sativus) using a phylogenetic analysis of IGT protein family members. Differences in the transcriptional response of CsDRO1, CsDRO1-LIKE1 (CsDRO1L1), and CsDRO1-LIKE2 (CsDRO1L2) to exogenous auxin were analyzed. The results showed that only CsDRO1L1 is auxin-responsive. An analysis of promoter-reporter fusions demonstrated that the CsDRO1, CsDRO1L1, and CsDRO1L2 genes were expressed in the meristem in cell files of the central cylinder, endodermis, and cortex; the three genes displayed different expression patterns in cucumber roots with only partial overlap. A knockout of individual CsDRO1, CsDRO1L1, and CsDRO1L2 genes was performed via CRISPR/Cas9 gene editing. Our study suggests that the knockout of individual genes does not affect the slope angle formation during lateral root primordia development in the cucumber parental root.


Subject(s)
Arabidopsis , Cucumis sativus , Cucumis sativus/metabolism , Plant Roots/metabolism , Phylogeny , Indoleacetic Acids/metabolism , Meristem/genetics , Meristem/metabolism , Arabidopsis/metabolism , Gene Expression Regulation, Plant
20.
Plant Cell Environ ; 47(5): 1513-1525, 2024 May.
Article in English | MEDLINE | ID: mdl-38251425

ABSTRACT

The DNA damage response avoids mutations into dividing cells. Here, we analysed the role of photoreceptors on the restriction of root growth imposed by genotoxic agents and its relationship with cell viability and performance of meristems. Comparison of root growth of Arabidopsis WT, phyA-211, phyB-9, and phyA-211phyB-9 double mutants unveiled a critical role for phytochrome A (PhyA) in protecting roots from genotoxic stress, regeneration and cell replenishment in the meristematic zone. PhyA was located on primary root tips, where it influences genes related to the repair of DNA, including ERF115 and RAD51. Interestingly, phyA-211 mutants treated with zeocin failed to induce the expression of the repressor of cell cycle MYB3R3, which correlated with expression of the mitotic cyclin CycB1, suggesting that PhyA is required for safeguarding the DNA integrity during cell division. Moreover, the growth of the primary roots of PhyA downstream component HY5 and root growth analyses in darkness suggest that cell viability and DNA damage responses within root meristems may act independently from light and photomorphogenesis. These data support novel roles for PhyA as a key player for stem cell niche maintenance and DNA damage responses, which are critical for proper root growth.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Phytochrome , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Cell Death , DNA/metabolism , DNA Repair/genetics , Light , Meristem/genetics , Meristem/metabolism , Mutation , Phytochrome/metabolism , Phytochrome A/genetics , Phytochrome A/metabolism , Phytochrome B/metabolism
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