Meristematic Activity Research Articles

Hydrogen Peroxide Signaling in the Maintenance of Plant Root Apical Meristem Activity.

Hydrogen peroxide (H2O2) is a prevalent reactive oxygen species (ROS) found in cells and takes a central role in plant development and stress adaptation. The root apical meristem (RAM) has evolved strong plasticity to adapt to complex and changing environmental conditions. Recent advances have made great progress in explaining the mechanism of key factors, such as auxin, WUSCHEL-RELATED HOMEOBOX 5 (WOX5), PLETHORA (PLT), SHORTROOT (SHR), and SCARECROW (SCR), in the regulation of RAM activity maintenance. H2O2 functions as an emerging signaling molecule to control the quiescent center (QC) specification and stem cell niche (SCN) activity. Auxin is a key signal for the regulation of RAM maintenance, which largely depends on the formation of auxin regional gradients. H2O2 regulates the auxin gradients by the modulation of intercellular transport. H2O2 also modulates the expression of WOX5, PLTs, SHR, and SCR to maintain RAM activity. The present review is dedicated to summarizing the key factors in the regulation of RAM activity and discussing the signaling transduction of H2O2 in the maintenance of RAM activity. H2O2 is a significant signal for plant development and environmental adaptation.

Transcription factors HB21/40/53 trigger inflorescence arrest through abscisic acid accumulation at the end of flowering.

Flowers, and hence, fruits and seeds, are produced by the activity of the inflorescence meristem after the floral transition. In plants with indeterminate inflorescences, the final number of flowers produced by the inflorescence meristem is determined by the length of the flowering period, which ends with inflorescence arrest. Inflorescence arrest depends on many different factors, such as the presence of seeds, the influence of the environment, or endogenous factors such as phytohormone levels and age, which modulate inflorescence meristem activity. The FRUITFULL-APETALA2 (FUL-AP2) pathway plays a major role in regulating the end of flowering, likely integrating both endogenous cues and those related to seed formation. Among AP2 targets, HOMEOBOX PROTEIN21 (HB21) has been identified as a putative mediator of AP2 function in the control of inflorescence arrest. HB21 is a homeodomain leucine zipper transcription factor involved in establishing axillary bud dormancy. Here, we characterized the role of HB21 in the control of the inflorescence arrest at the end of flowering in Arabidopsis (Arabidopsis thaliana). HB21, together with HB40 and HB53, are upregulated in the inflorescence apex at the end of flowering, promoting floral bud arrest. We also show that abscisic acid (ABA) accumulation occurs in the inflorescence apex in an HB-dependent manner. Our work suggests a physiological role of ABA in floral bud arrest at the end of flowering, pointing to ABA as a regulator of inflorescence arrest downstream of the HB21/40/53 genes.

The impact of drought on phellem development: Identification of novel gene regulators and evidence of photosynthetic activity in cork oak stems

Quercus suber (cork oak) is a sustainably exploited forest resource, producing a unique renewable raw material known as cork. With drought events imposing a negative impact on tree vitality, further research is needed to enhance our understanding of the genetic and environmental factors regulating cork development, to foster the resilience of cork oak ecosystems. We focused on characterizing long-term drought-induced molecular adaptations occurring in stems, and identifying key genetic pathways regulating phellem development. One-year-old cork oak plants were grown for 6 months under well-watered, or water-deficit (WD) conditions and main stems were targeted for histological characterization and transcriptomic analysis. WD treatment reduced meristem activity at both vascular cambium and phellogen, impairing secondary growth. Transcriptional analysis revealed a global downregulation of genes related to cell division, differentiation, and cell wall biogenesis in phellem, inner bark, and xylem under WD conditions. Phellem and inner bark showed upregulation of photosynthesis-related genes, highlighting a determinant role of stem photosynthesis in the adaptation to long-term drought. We show that developing phellem cells contain chloroplasts and their abundance increases under WD. Lastly, we propose new candidate regulatory genes involved in regulating phellogen activity and demonstrate the role of phellem in drought-induced bark photosynthesis in young plants.

Physiological and developmental disturbances caused by Botryosphaeria dieback in the annual stems of grapevine.

Botryosphaeria dieback is a grapevine trunk disease caused by fungi of the Botryosphaeriaceae family, which attacks more specifically the woody tissues. The infection leads to different symptoms including a severe form with a leaf drop as well as premature plant death. Botryosphaeria dieback causes major economic losses, since no effective treatment is yet available. A better understanding is necessary to find solutions to fight this disease. In this study, our objective was to characterize the "leaf drop" form by (1) looking for the presence of pathogens in the basal internodes of stems, (2) quantifying blocked vessels by tylosis and/or gummosis, and (3) describing the impact of the disease on vine physiology (gene expression and metabolome) and development (establishment and functioning of the cambium and phellogen) at the level of xylem and phloem of basal stem internodes. Our study has shown that Botryosphaeriaceae were present in both phloem and xylem of the basal internodes of the annual stem, with xylem vessels obturated. We have also clearly demonstrated that gene expression and metabolite profiles were strongly modified in both xylem and phloem of diseased plants. Differences in stems between healthy (control, C) and diseased (D) plants were low at flowering (vines not yet symptomatic), higher at the onset of symptom expression and still present, although less marked, at full disease expression. qRT-PCR analysis showed in both phloem and xylem an overexpression of genes involved in plant defense, and a repression of genes related to meristematic activity (i.e. vascular cambium and phellogen). Metabolomic analysis showed specific fingerprints in stems of healthy and diseased plants from the onset of symptom expression, with an increase of the level of phytoalexins and mannitol, and a decrease of 1-kestose one. At the structural level, many alterations were observed in internodes, even before the onset of symptoms: a classical obstruction of xylem vessels and, for the first time, a disorganization of the secondary phloem with an obstruction of the sieve plates by callose. The disease modifies the development of both secondary phloem (liber) and phellogen. Altogether, this study combining different approaches allowed to highlight deep vine dysfunction in the internodes at the base of stems, that may explain vine decline due to Botryosphaeria dieback.

Concurrent improvement of rice grain yield and abiotic stress tolerance by overexpression of cytokinin activating enzyme LONELY GUY (OsLOG)

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.

LE AF DEVELOPMENT IN CURIO MUIRII (L. BOLUS) VAN JAARSV. (ASTERACEAE - SENECIONEAE)

Curio muirii is the only species in the genus with bifacial leaves, making it crucial for understanding leaf evolution in this group. Leaf development can be divided into three stages: initiation, bizonal primordium, and intercalary growth. The developing leaves of C. muirii are bifacial and resemble those of other angiosperms from inception to the end of the bizonal primordium stage. However, as in monocots, the primary vascular bundles differentiate at the end of the bizonal primordium stage. The unifacial forerunner tip, which is recognized for Curio species but unknown for other eudicots, is the rst discernible part of the de nitive leaf. The leaf blade and petiole in C. muirii are distinctive in their biphasic development during the intercalary stage. The petiole grows faster than the leaf blade during the rst phase. During the second phase, the lamina overcomes the petiole in terms of length. In the developing leaf blade, the marginal meristem was not found. Adaxial meristem activity causes the leaf base to thicken unevenly during intercalary growth and envelopes the younger leaf primordium in the resultant depression.

“Hurdley technologies” utilized to improve postharvest life of asparagus spears (Asparagus officinalis L.)

Asparagus (Asparagus officinalis L.) has short shelf-life due to the high metabolic activity of the apical meristems. Storage at low temperature and high relative humidity is used commercially to keep fresh asparagus spears. Techniques denominated “Hurdley technologies” (UV-C or gamma irradiation) have been tested in fruits and vegetables to extend postharvest life. These technologies were used to extend postharvest shelf life of asparagus spears by inhibition of meristematic activity. Spears were irradiated with UV-C at dosages of 2.46, and 4.93 kJ m-2 and gamma irradiation at 1 and 1.5 kGy, before storage at 2°C and 90% relative humidity (RH) for 20 days. Metabolic heat (Rq) was measured in apical meristems, as well as whole spear respiration, sugars content, water potential components and color descriptors. Metabolic heat and whole spears respiration rate did not show differences due to effect of UV-C treatments, while spears treated with gamma radiation showed a metabolic activity inhibition of 10 and 15% for 1 and 1.5 kGy, respectively, while whole spear respiration rate was not affected. Changes in color variables showed a slight reduction in gloss. Sugars content in UV-C remained unchanged, while gamma radiation induced a reduction in glucose. An increase in fresh weight loss was noticeable on those treated with gamma irradiation. No changes in water potential components were observed. It was concluded the treatments used did not reported positive benefits in extending asparagus spears shelf life.

An AGO10:miR165/6 module regulates meristem activity and xylem development in the Arabidopsis root.

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.

Stochastic modelling of development and biomass allocation: Computation applied to architecture of young mahogany trees (Khaya senegalensis Desr. A. Juss), a native African savannah emblematic agroforestry species

The architectural plasticity forms observed in trees is a result of meristem functioning, which generates new organs and branches, and adjusts growth processes in response to heterogeneous climatic adaptations that affect biomass allocation. Analyzing this plasticity should enable the selection of adapted individuals for optimizing successful cropping systems. Mahogany tree (Khaya senegalensis) is a rhythmically growing indigenous agroforestry tree that is heavily exploited for its multiple uses. Understanding its growth characteristics, as well as the complexity of its structure (randomness, rhythmicity, etc. of Mahogany tree), could facilitate its conservation and sustainable management. This study aims to model the architecture and physiology of young mahogany trees based on field data using an organ-level structural–functional model called 'GreenLab', which is founded on source-sink relationships. Ninety trees aged 6, 12, and 24 months were measured in the field. Development was calculated using a dual-scale automaton based on the Monte Carlo process, while biomass production and its distribution to different plant organs (source and sink) were calibrated using Pressler's law using Markov chains. Meristematic activity laws, combined with organ sinks (D: Demand), photosynthesis (Q: Supply), and organic series (Q/D: Trophic pressure), were employed to simulate individual architecture. The results demonstrate that the model realistically and flexibly describes topological development and replicates biomass production and allocation processes for rhythmically growing trees. This model will enable the identification of mahogany ideotypes suited for enhancing agroforestry cropping systems based on this species and several other threatened species. These findings introduce and thus lay the groundwork for a computational plant model tailored to the needs of agroforestry from a novel perspective, offering new avenues for agronomic and forestry applications in West Africa and Côte d'Ivoire.

Decoding the leaf apical meristem of Guarea glabra Vahl (Meliaceae): insight into the evolution of indeterminate pinnate leaves

In seed plants, growth of shoots and roots is indeterminate, while leaves are typically determinate organs that cease to grow after a certain developmental stage. This is due to the characteristics of the leaf meristem, where cell proliferation activity is retained only for a limited period. However, several plants exhibit indeterminacy in their leaves, exemplified by the pinnate compound leaves of Guarea and Chisocheton genera in the Meliaceae family. In these plants, the leaf meristem at the tip of the leaf retains meristematic activity and produces leaflets over years, resulting in a single leaf that resembles a twig. The molecular mechanism underlying the indeterminate leaf meristem of these plants has not been examined. In this research, we used Guarea glabra as a model to investigate the development of indeterminate pinnate leaves. Transcriptome analyses revealed that the gene expression profile in leaf apex tissue differed from that in the shoot apex. However, a class 1 KNOTTED-LIKE HOMEOBOX (KNOX1) gene which is lost in Brassicaceae was highly expressed in both tissues. We established an in situ hybridisation system for this species using Technovit 9100 to analyse the spatial expression patterns of genes. We revealed that the leaf meristematic region of G. glabra expresses KNOX1, LEAFY and ANGUSTIFORIA3 simultaneously, suggesting the involvement of these genes in the indeterminacy of the leaf meristem.

Hormonal and epigenetic regulation of root responses to salinity stress

Salinity stress is a major environmental stress affecting crop productivity, and its negative impact on global food security is only going to increase, due to current climate trends. Salinity tolerance was present in wild crop relatives but significantly weakened during domestication. Regaining it back requires a good understanding of molecular mechanisms and traits involved in control of plant ionic and ROS homeostasis. This review summarizes our current knowledge on the role of major plant hormones (auxin, cytokinins, abscisic acid, salicylic acid, and jasmonate) in plants adaptation to soil salinity. We firstly discuss the role of hormones in controlling root tropisms, root growth and architecture (primary root elongation, meristematic activity, lateral root development, and root hairs formation). Hormone-mediated control of uptake and sequestration of key inorganic ions (sodium, potassium, and calcium) is then discussed followed by regulation of cell redox balance and ROS signaling in salt-stressed roots. Finally, the role of epigenetic alterations such as DNA methylation and histone modifications in control of plant ion and ROS homeostasis and signaling is discussed. This data may help develop novel strategies for breeding and cultivating salt-tolerant crops and improving agricultural productivity in saline regions.

Intercellular Communication in Shoot Meristems.

The shoot meristem of land plants maintains the capacity for organ generation throughout its lifespan due to a group of undifferentiated stem cells. Most meristems are shaped like a dome with a precise spatial arrangement of functional domains, and, within and between these domains, cells interact through a network of interconnected signaling pathways. Intercellular communication in meristems is mediated by mobile transcription factors, small RNAs, hormones, and secreted peptides that are perceived by membrane-localized receptors. In recent years, we have gained deeper insight into the underlying molecular processes of the shoot meristem, and we discuss here how plants integrate internal and external inputs to control shoot meristem activities.

Transcriptomic analyses to summarize gene expression patterns that occur during leaf initiation of Chinese cabbage.

In Chinese cabbage, rosette leaves expose their adaxial side to the light converting light energy into chemical energy, acting as a source for the growth of the leafy head. In the leafy head, the outer heading leaves expose their abaxial side to the light while the inner leaves are shielded from the light and have become a sink organ of the growing Chinese cabbage plant. Interestingly, variation in several ad/abaxial polarity genes is associated with the typical leafy head morphotype. The initiation of leaf primordia and the establishment of leaf ad/abaxial polarity are essential steps in the initiation of marginal meristem activity leading to leaf formation. Understanding the molecular genetic mechanisms of leaf primordia formation, polar differentiation, and leaf expansion is thus relevant to understand leafy head formation. As Brassica's are mesa-hexaploids, many genes have multiple paralogues, complicating analysis of the genetic regulation of leaf development. In this study, we used laser dissection of Chinese cabbage leaf primordia and the shoot apical meristem (SAM) to compare gene expression profiles between both adaxial and abaxial sides and the SAM aiming to capture transcriptome changes underlying leaf primordia development. We highlight genes with roles in hormone pathways and transcription factors. We also assessed gene expression gradients along expanded leaf blades from the same plants to analyze regulatory links between SAM, leaf primordia and the expanding rosette leaf. The catalogue of differentially expressed genes provides insights in gene expression patterns involved in leaf development and form a starting point to unravel leafy head formation.

Cotton Meristem Transcriptomes: Constructing an RNA-Seq Pipeline to Explore Crop Architecture Regulation.

Plants stem cells, known as meristems, specify all patterns of growth and organ size. Differences in meristem activities contribute to diverse shoot architectures. As many architectural traits, such as branching patterns, flowering time, and fruit size, are yield determinants, meristem regulation is of fundamental importance to crop productivity. Cotton (Gossypium spp.) produces our most prevalent natural fiber that finds its way into products ranging from industrial cellulose, medical supplies, and paper currency, to a broad diversity of textiles, not least of which is our clothing. However, the cotton plant has growth habits that challenge management practices and limit harvest yield and quality. Unraveling and leveraging the genetic networks regulating meristem activities offers the potential to overcome these limitations. We use virus-based technologies in cotton to perturb signals regulating meristem fate and size. In this chapter, we describe our pipeline for altering cotton meristem dynamics and preparing, analyzing, and exploring the transcriptomes from isolated meristems.

Control of lateral root formation by rapamycin-induced dimerization of engineered RGI/BAK1 and by BIR3 chimera.

Leucine-rich repeat receptor kinases (LRR-RKs) play a pivotal role in diverse aspects of growth, development, and immunity in plants by sensing extracellular signals. Typically, LRR-RKs are activated through the ligand-induced interaction with a SOMATIC EMBRYOGENESIS RECEPTOR KINASE (SERK) coreceptor, triggering downstream signaling. ROOT MERISTEM GROWTH FACTOR1 (RGF1) INSENSITIVEs (RGIs) LRR-RLK receptors promote primary root meristem activity while inhibiting lateral root (LR) development in response to RGF peptide. In this study, we employed rapamycin-induced dimerization (RiD) and BAK1-INTERACTING RECEPTOR-LIKE KINASE3 (BIR3) chimera approaches to explore the gain-of-function of RGI1, RGI4, and RGI5. Rapamycin induced the association of cytosolic kinase domains (CKDs) of RGI1 and the BAK1 coreceptor, activating both mitogen-activated protein kinase 3 (MPK3) and MPK6. Rapamycin significantly inhibited LR formation in RiD-RGI1/RGI4/RGI5-BAK1 plants. Using transgenic Arabidopsis expressing RGI1CKD fused to the BIR3-LRR chimera under estradiol control, we observed a substantial reduction in LR density upon β-estradiol treatment. Additionally, we identified a decrease in root gravitropism in BIR3 chimera plants. In contrast, RiD-RGI/BAK1 plants did not exhibit defects in root gravitropism, implying the importance of combinatorial interactions between RGIs and SERK coreceptors in the inhibition of root gravitropism. Constitutive activation of RGIs with BAK1 in RiD-RGI/BAK1 plants by rapamycin treatment resulted in the inhibition of primary root growth, resembling the inhibitory effects observed with high concentrations of phytohormones on primary root elongation. Our findings highlight that the interactions between CKDs of RGIs and BAK1, constitutively induced by rapamycin or BIR3 chimera, efficiently control LR development.

Heat stress impairs floral meristem termination and fruit development by affecting the BR-SlCRCa cascade in tomato

Floral meristem termination is a key step leading to carpel initiation and fruit development. The frequent occurrence of heat stress due to global warming often disrupts floral determinacy, resulting in defective fruit formation. However, the detailed mechanism behind this phenomenon is largely unknown. Here, we identify CRABS CLAW a (SlCRCa) as a key regulator of floral meristem termination in tomato. SlCRCa functions as an indispensable floral meristem terminator by suppressing SlWUS activity through the TOMATO AGAMOUS 1 (TAG1)–KNUCKLES (SlKNU)–INHIBITOR OF MERISTEM ACTIVITY (SlIMA) network. A direct binding assay revealed that SlCRCa specifically binds to the promoter and second intron of WUSCHEL (SlWUS). We also demonstrate that SlCRCa expression depends on brassinosteroid homeostasis in the floral meristem, which is repressed by heat stress via the circadian factor EARLY FLOWERING 3 (SlELF3). These results provide new insights into floral meristem termination and the heat stress response in flowers and fruits of tomato and suggest that SlCRCa provides a platform for multiple protein interactions that may epigenetically abrogate stem cell activity at the transition from floral meristem to carpel initiation.

Establishing surface sterilization protocol for clonal apple rootstock MM106

Surface sterilization is crucial in preparing viable and uncontaminated explants for tissue culture. Utilizing the right sterilizing agent in this procedure can efficiently eliminate the majority of surface contaminants. The present study aimed to establish the best sterilization procedure for clonal apple rootstock MM106 using shoot tips and nodal segments. Two important sterilizing agents, mercuric chloride (ranging from 0.05% to 0.3%) for various durations (2 to 6 minutes) and sodium hypochlorite (ranging from 3.0% to 12%) for different durations (10 to 30 minutes), were employed. The results indicated that the maximum aseptic conditions (80.00% and 80.67%) and survival rates of the cultures (60.66% and 67.33%) for shoot tips and nodal segments, respectively, were achieved through surface sterilization with 0.1% mercuric chloride for 4 minutes, specifically during April, in comparison to March and May of 2021 due to low phenol content and high meristematic activity. The results yielded valuable insights for efficiently propagating clonal apple rootstocks on a local scale, and they have the potential to offer guidance for establishing commercial facilities dedicated to producing these specific clonal apple rootstocks and varieties.

Nitric oxide controls shoot meristem activity via regulation of DNA methylation

Despite the importance of Nitric Oxide (NO) as signaling molecule in both plant and animal development, the regulatory mechanisms downstream of NO remain largely unclear. Here, we show that NO is involved in Arabidopsis shoot stem cell control via modifying expression and activity of ARGONAUTE 4 (AGO4), a core component of the RNA-directed DNA Methylation (RdDM) pathway. Mutations in components of the RdDM pathway cause meristematic defects, and reduce responses of the stem cell system to NO signaling. Importantly, we find that the stem cell inducing WUSCHEL transcription factor directly interacts with AGO4 in a NO dependent manner, explaining how these two signaling systems may converge to modify DNA methylation patterns. Taken together, our results reveal that NO signaling plays an important role in controlling plant stem cell homeostasis via the regulation of de novo DNA methylation.

Control of stem cell behavior by CLE-JINGASA signaling in the shoot apical meristem in Marchantia polymorpha

Land plants undergo indeterminate growth by the activity of meristems in both gametophyte (haploid) and sporophyte (diploid) generations. In the sporophyte of the flowering plant Arabidopsis thaliana, the apical meristems are located at the shoot and root tips in which a number of regulatory gene homologs are shared for their development, implying deep evolutionary origins. However, little is known about their functional conservation with gametophytic meristems in distantly related land plants such as bryophytes, even thoughgenomic studies have revealed that the subfamily-level diversity of regulatory genes is mostly conserved throughout land plants. Here, we show that a NAM/ATAF/CUC (NAC) domain transcription factor, JINGASA (MpJIN), acts downstream of CLAVATA3 (CLV3)/ESR-related (CLE) peptide signaling and controls stem cell behavior in the gametophytic shoot apical meristem of the liverwort Marchantia polymorpha. In the meristem, strong MpJIN expression was associated with the periclinal cell division at the periphery of the stem cell zone (SCZ), whereas faint MpJIN expression was found at the center of the SCZ. Time course observation indicates that the MpJIN-negative cells are lost from the SCZ and respecified de novo at two separate positions during the dichotomous branching event. Consistently, the induction of MpJIN results in ectopic periclinal cell division in the SCZ and meristem termination. Based on the comparative expression data, we speculate that the function of JIN/FEZ subfamily genes was shared among the shoot apical meristems in the gametophyte and sporophyte generations in early land plants but was lost in certain lineages, including the flowering plant A.thaliana.

Genome-Wide Classification and Evolutionary Analysis of the KNOX Gene Family in Plants

The Knotted1-like homeobox (KNOX) gene family plays a pivotal role in regulating meristem activity, organ differentiation, and cell meristematic identity. However, there has been a lack of large-scale, systematic, and comprehensive comparative analyses to explore their expression patterns and evolutionary mechanisms. In this study, a total of 1425 KNOX genes were identified across 118 plant species. The result showed that higher plants exhibited a significantly higher abundance of KNOX genes compared to lower plants. Phylogenetic analysis revealed that all KNOX genes can be divided into two classes (class I and II) and evolved independently after species differentiation. An analysis of gene duplication or loss showed that gene loss was more common than gene duplication in lower plants within the KNOX gene family. These findings suggest that gene loss in the KNOX gene family occurs after events such as whole-genome duplication (WGD) or whole-genome triplication (WGT). In addition, conserved motif analysis was also conducted to uncover the evolutionary trajectories of KNOX genes. We found that three motifs (M1, M2, and M4) were present in nearly all KNOX genes, while four novel motifs (M7–M10) were lost in lower plants but present in higher plants. Moreover, the loss of certain motifs in the KNOX genes was also observed in higher plants, indicating sequence divergence in KNOX genes throughout evolution. To understand the expression patterns of KNOX genes, a gene expression pattern analysis was performed in A. thaliana and O. sativa. The results showed that class I KNOX genes exhibit conserved high expression in stems, suggesting their potential similar biological roles across different plant species and the conservation of their functions during evolution. Additionally, we analyzed the KNOX genes in the Citrus genus and closely related species, and we found that the number of KNOX genes evolved at a slower rate in these species, indicating a relatively conservative nature. In conclusion, this study provides valuable resources for the investigation of KNOX gene family evolution and function in plants.