Leaf Meristem Research Articles

Diversity of phenological patterns of Handroanthus ochraceus (Bignoniaceae) in Costa Rica

The study of phenological patterns in plant communities is of vital importance for understanding the temporal cycles of ecosystems, but there is little information on the diversity of phenological patterns that can occur at the intraspecific level, as well as the genetic or environmental factors causing this variation. In Handroanthus ochraceus, a deciduous tree species of neotropical dry forests, ecophysiological studies have proposed the release of water stress as the mechanism that triggers flowering and the sprouting of new leaves. During four years, I observed the cycles of leaf fall, flowering, fruiting, and new leaf production in seven Costa Rican sites that differed in their proximity to water courses and in soil moisture. Six were located in dry forest areas, and one in premontane forest. There were two general phenological patterns: the explosive, where trees depend on the first rains for floral anthesis and the expansion of leaf meristems, and the staggered pattern, with unsynchronized flowering at the beginning and middle of the dry season, independently of rainfall. Although this phenological variation has been previously recognized, the occurrence of these two patterns is not determined by the proximity of water courses, contradicting phenological models proposed for this tree species.

Linkage between leaf development and photosynthetic response at hyperosmotic salinity in the C-4 grass Panicum antidotale

This study aimed at elucidating the impact of salinity on the differential responses of the photosynthetic apparatus at different stages of leaf development in the C4 perennial grass Panicum antidotale. Plants were grown for a period of 15 days at hyper-osmotic salinity (up to 400 mM NaCl) or at control conditions (no addition of NaCl to the culture medium). Each day plants were harvested and salt stress effect on biomass production (fresh and dry weight) was recorded. In parallel leaves were divided from leaf tip to leaf base in five segments for analysis of stress effects on the following parameters: chlorophyll fluorescence parameters (Fv/Fm, ΦPSII, ETR), MDA (an indicator of peroxidation of membrane lipids), and ion leakage.A direct correlation between membrane lipid peroxidation and (i) membrane integrity as well as (ii) inhibition of PSII function was found under salinity. In monocots the leaf meristem is located at the base and mature parenchymatic cells preferentially are found at the leaf tip. This may explain our finding that the degree of salt mediated inhibition of photosynthetic activity (ETRcontrol/ETRstressed) was 20-fold higher at leaf tip than at leaf base. Thus, we discuss observed variance in apparent salt resistance in terms of heterogeneity of the photosynthetic apparatus (due to difference in spatial and temporal development) along the leaf lamina.

One-leaf plants in the Gesneriaceae: Natural mutants of the typical shoot system.

The aerial part of seed plants is called the shoot, which is composed of stems, leaves, and axial buds. These are produced by indeterminate activity in the shoot apical meristem (SAM), whereas the morphogenesis of leaves depends on determinate activity of leaf meristems. However, one-leaf plants in the Gesneriaceae family (eudicots) do not have a typical SAM and do not produce new organs when in the vegetative phase. Instead, they have one cotyledon whose growth is indeterminate. This peculiar development is supported by the groove meristem, which corresponds to the canonical SAM, and the basal meristem, which corresponds to the typical leaf meristem. However, the former does not produce any organ and the latter is active indeterminately. Gene expression and physiological analyses have been conducted in an effort to determine the molecular nature of this peculiar organogenesis. This review summarizes the current understanding of the development of one-leaf plants to provide future perspectives in this field of research.

Positive effects of the catastrophic Hurricane Patricia on insect communities

Highly seasonal conditions of tropical dry forests determine the temporal patterns of insect abundance. However, density-independent factors such as natural disturbances can abruptly change environmental conditions, affecting insect populations. We address the effects of the Hurricane Patricia (category 5) on species density and abundance of three feeding guilds of herbivorous insects (sap-sucking, folivorous beetles and xylophagous) and predatory beetles associated to the canopy of a tropical dry forest. Hurricane Patricia has been the strongest tropical hurricane ever reported in the Western Hemisphere. Herbivorous insects (sap-sucking and xylophagous) and predatory beetles increased in species density and abundance in the following months after the hurricane, compared to samples before it. The positive response of sap-sucking insects to Hurricane Patricia was probably related to an increase in the availability of new shoots and leaf meristems after the natural coppicing by the hurricane, while xylophagous guild seems to have been positively affected by the increase in the amount and diversity of deadwood resources. The positive response of predatory beetles may be the result of a bottom-up effect due to a greater availability of arthropod preys after the hurricane. We demonstrated that catastrophic hurricane disturbances could be important events that temporarily increase the species density and abundance of insects in tropical dry forests.

Systemic research of plant growth and stress in a specially designed climatic hydroponics system

AbstractModern plant physiology deals with a plant at different hierarchical levels, from the ecosystem to molecular ones, so there is a demand to integrate the knowledge obtained at these levels. Examining plant growth and cold resistance, namely the intermediate levels (organism, organs, tissues, multicellular) often reveals the peculiarities of the functional–structural organization of the entire plant. The paper presents an integrated climatic hydroponic system, designed to implement a physical‐holistic experiment, accompanied by a particular methodology of systemic research. It covers the following main steps, which are the verification of the experimental system, production of high‐quality experimental material, including direct measurements and mathematical data analysis. The latter involves the decomposition of the phenomena under study, synthesis of data and the extraction of non‐linear components. The studies on the growth and stress of Poaceae plants (Lolium multiflorum, Festuca pratensis, Festulolium braunii) have already provided some significant results. A new boundary of the leaf meristem was proposed, adaptive growth bifurcations were discovered in the meristem, and a cells rounding phenomenon was detected. Using this top‐down approach, further research directions are delineated towards the lower hierarchical levels. Finally, the presented approach allows for the integration of data obtained at different plant investigation levels.

Seagrass rhizosphere microenvironment alters plant-associated microbial community composition.

The seagrass rhizosphere harbors dynamic microenvironments, where plant-driven gradients of O2 and dissolved organic carbon form microhabitats that select for distinct microbial communities. To examine how seagrass-mediated alterations of rhizosphere geochemistry affect microbial communities at the microscale level, we applied 16S rRNA amplicon sequencing of artificial sediments surrounding the meristematic tissues of the seagrass Zostera muelleri together with microsensor measurements of the chemical conditions at the basal leaf meristem (BLM).Radial O2 loss (ROL) from the BLM led to ∼300µm thick oxic microzones, wherein pronounced decreases in H2 S and pH occurred. Significantly higher relative abundances of sulphate-reducing bacteria were observed around the meristematic tissues compared to the bulk sediment, especially around the root apical meristems (RAM; ∼57% of sequences). Within oxic microniches, elevated abundances of sulphide-oxidizing bacteria were observed compared to the bulk sediment and around the RAM. However, sulphide oxidisers within the oxic microzone did not enhance sediment detoxification, as rates of H2 S re-oxidation here were similar to those observed in a pre-sterilized root/rhizome environment. Our results provide novel insights into how chemical and microbiological processes in the seagrass rhizosphere modulate plant-microbe interactions potentially affecting seagrass health.

Comparative systemic analysis of the cellular growth of leaves and roots in controlled conditions

The comparative cytological analysis of the leaf/root growth of Lolium multiflorum has been performed. It revealed differences of the mentioned above/under-ground organs that express the whole plant’s polarity. To perform accurate and simultaneous growth comparison a climatic-hydroponics system has been implemented. A sharp increase in the epidermis cell length of the leaf meristem has been detected for the first time. It allows the proposal of a new way to demarcate the boundary of the meristem and suggests a lengthed leaf meristem, that is 4 times longer than the root meristem. As the cell cycle duration in leaves and roots is similar, the prolonged leaf meristem and a higher leaf growth rate could be determined by the longer life span of cells in meristem, resulting in more cell cycles. The prolonged meristem provides a significantly higher leaf growth rate, ensuring a functional balance with roots. The elongation zone of the roots is significantly shorter than in leaves, which is caused by the larger relative root elongation rate and the slower meristemic root growth rate. The novelty formulated, i.e., the prolonged leaf meristem, opens theoretical perspectives in longitudinal zonation, in finding molecular markers and provides practical significance for the biology of productivity.

Leaf ontogeny of Schinus molle L. plants under cadmium contamination: the meristematic origin of leaf structural changes.

Previous works show the development of thicker leaves on tolerant plants growing under cadmium (Cd2+) contamination. The aim of this study was to evaluate the Cd2+ effects on the leaf meristems of the tolerant species Schinus molle. Plants were grown in nutrient solution containing 0, 10, and 50μM of Cd2+. Anatomical analysis was performed on leaf primordia sampled at regular time intervals. Under the lowest Cd2+ level (10μM), increased ground meristem thickness, diameter of the cells, cell elongation rate, and leaf dry mass were found. However, 50μM of Cd2+ reduced all these variables. In addition, the ground meristem cells became larger when exposed to any Cd2+ level. The epidermis, palisade parenchyma, and vascular tissues developed earlier in Cd2+-exposed leaves. The modifications found on the ground meristem may be related to the development of thicker leaves on S. molle plants exposed to low Cd2+ levels. Furthermore, older leaves showed higher Cd2+ content when compared to the younger ones, preventing the Cd2+ toxicity to these leaves. Thus, low Cd2+ concentrations change the ground meristem structure and function reflecting on the development of thicker and enhanced leaves.

High Antioxidant Activity Facilitates Maintenance of Cell Division in Leaves of Drought Tolerant Maize Hybrids.

We studied the impact of drought on growth regulation in leaves of 13 maize varieties with different drought sensitivity and geographic origins (Western Europe, Egypt, South Africa) and the inbred line B73. Combining kinematic analysis of the maize leaf growth zone with biochemical measurements at a high spatial resolution allowed us to examine the correlation between the regulation of the cellular processes cell division and elongation, and the molecular redox-regulation in response to drought. Moreover, we demonstrated differences in the response of the maize lines to mild and severe levels of water deficit. Kinematic analysis indicated that drought tolerant lines experienced less impact on leaf elongation rate due to a smaller reduction of cell production, which, in turn, was due to a smaller decrease of meristem size and number of cells in the leaf meristem. Clear differences in growth responses between the groups of lines with different geographic origin were observed in response to drought. The difference in drought tolerance between the Egyptian hybrids was significantly larger than between the European and South-African hybrids. Through biochemical analyses, we investigated whether antioxidant activity in the growth zone, contributes to the drought sensitivity differences. We used a hierarchical clustering to visualize the patterns of lipid peroxidation, H2O2 and antioxidant concentrations, and enzyme activities throughout the growth zone, in response to stress. The results showed that the lines with different geographic region used different molecular strategies to cope with the stress, with the Egyptian hybrids responding more at the metabolite level and African and the European hybrids at the enzyme level. However, drought tolerance correlated with both, higher antioxidant levels throughout the growth zone and higher activities of the redox-regulating enzymes CAT, POX, APX, and GR specifically in leaf meristems. These findings provide evidence for a link between antioxidant regulation in the leaf meristem, cell division, and drought tolerance.

Active suppression of a leaf meristem orchestrates determinate leaf growth.

Leaves are flat determinate organs derived from indeterminate shoot apical meristems. The presence of a specific leaf meristem is debated, as anatomical features typical of meristems are not present in leaves. Here we demonstrate that multiple NGATHA (NGA) and CINCINNATA-class-TCP (CIN-TCP) transcription factors act redundantly, shortly after leaf initiation, to gradually restrict the activity of a leaf meristem in Arabidopsis thaliana to marginal and basal domains, and that their absence confers persistent marginal growth to leaves, cotyledons and floral organs. Following primordia initiation, the restriction of the broadly acting leaf meristem to the margins is mediated by the juxtaposition of adaxial and abaxial domains and maintained by WOX homeobox transcription factors, whereas other marginal elaboration genes are dispensable for its maintenance. This genetic framework parallels the morphogenetic program of shoot apical meristems and may represent a relic of an ancestral shoot system from which seed plant leaves evolved.

Molecular characterization and evolution of carnivorous sundew (Drosera rotundifolia L.) class V β-1,3-glucanase.

A gene for β-1,3-glucanase was isolated from carnivorous sundew. It is active in leaves and roots, but not in digestive glands. Analyses in transgenic tobacco suggest its function in germination. Ancestral plant β-1,3-glucanases (EC 3.2.1.39) played a role in cell division and cell wall remodelling, but divergent evolution has extended their roles in plant defense against stresses to decomposition of prey in carnivorous plants. As available gene sequences from carnivorous plants are rare, we isolated a glucanase gene from roundleaf sundew (Drosera rotundifolia L.) by a genome walking approach. Computational predictions recognized typical gene features and protein motifs described for other plant β-1,3-glucanases. Phylogenetic reconstructions suggest strong support for evolutionary relatedness to class V β-1,3-glucanases, including homologs that are active in the traps of related carnivorous species. The gene is expressed in sundew vegetative tissues but not in flowers and digestive glands, and encodes for a functional enzyme when expressed in transgenic tobacco. Detailed analyses of the supposed promoter both in silico and in transgenic tobacco suggest that this glucanase plays a role in development. Specific spatiotemporal activity was observed during transgenic seed germination. Later during growth, the sundew promoter was active in marginal and sub-marginal areas of apical true leaf meristems of young tobacco plants. These results suggest that the isolated glucanase gene is regulated endogenously, possibly by auxin. This is the first report on a nuclear gene study from sundew.

Leaf meristems: an easily ignored component of the response to human disturbance in alpine grasslands.

Grazing and fencing are two important factors that influence productivity and biomass allocation in alpine grasslands. The relationship between root (R) and shoot (S) biomass and the root:shoot ratio (R/S) are critical parameters for estimating the terrestrial carbon stocks and biomass allocation mechanism responses to human activities. Previous studies have often used the belowground:aboveground biomass ratio (M b/M a) to replace the R/S in alpine ecosystems. However, these studies may have neglected the leaf meristem biomass, which belongs to the shoot but occurs below the soil surface, leading to a significant overestimation of the R/S ratio. We conducted a comparative study to explore the differences between the R/S and M b/M a at both the species (Stipa purpurea, Carex moorcroftii, and Artemisia nanschanica) and community levels on a Tibetan alpine grassland with grazing and fencing management blocks. The results revealed that the use of the M b/M a to express the R/S appeared to overestimate the actual value of the R/S, both at species and community levels. For S. purpurea, the M b/M a was three times higher than the R/S. The M b/M a was approximately two times higher than the R/S for the species of C. moorcroftii and A. nanschanica and at the community level. The relationships between the R‐S and M b‐M a exhibited different slopes for the alpine plants across all the management practices. Compared to the fenced grasslands, the plants in the grazing blocks not only allocated more biomass to the roots but also to the leaf meristems. The present study highlights the contribution of leaf meristems to the accurate assessment of shoot and belowground biomasses. The R/S and M b/M a should be cautiously used in combination in the future research. The understanding of the distinction between the R‐S and M b‐M a may help to improve the biomass allocation mechanism response to human disturbances in an alpine area.

Cloning and Functional Analysis of a WRKY28-like Gene in Soybean

WRKY transcription factors play important roles in regulating plant growth and development, and participate in plant response to abiotic and biotic stresses. Arabidopsis WRKY28 is an essential transcription factor for plant response to necrotrophic pathogen and abiotic stress. To explore the function of the homologues gene of AtWRKY28 in soybean, we cloned GmWRKY28- like (Glyma.14G028900) and performed bioinformatics analysis. Its sub-cellular localization and tissue expression patterns were further investigated. We also analyzed the expression levels of GmWRKY28-like under ABA, PEG, and NaCl stress treatments. Our results showed that the coding DNA sequence (CDS) of GmWRKY28-like gene is 1008 bp in length, encoding 335 amino acids. GmWRKY28-like contains a conserved WRKY domain with 22 serine, one threonine and two tyrosine, without any trans- membrane domain or signal peptide. Phylogenetic analysis showed that the WRKY28 from Phaseolus vulgaris was most similar to GmWRKY28-like. GmWRKY28-like was verified to be located in the nucleus. This gene expressed at low levels in root and seed while high levels in true leaf, flower and shoot apical meristem. The 1500 bp upstream region of GmWRKY28-like contains a

Behavior of Leaf Meristems and Their Modification.

A major source of diversity in flowering plant form is the extensive variability of leaf shape and size. Leaf formation is initiated by recruitment of a handful of cells flanking the shoot apical meristem (SAM) to develop into a complex three-dimensional structure. Leaf organogenesis depends on activities of several distinct meristems that are established and spatiotemporally differentiated after the initiation of leaf primordia. Here, we review recent findings in the gene regulatory networks that orchestrate leaf meristem activities in a model plant Arabidopsis thaliana. We then discuss recent key studies investigating the natural variation in leaf morphology to understand how the gene regulatory networks modulate leaf meristems to yield a substantial diversity of leaf forms during the course of evolution.

Defoliation, Shoot Plasticity, Sward Structure and Herbage Utilization in Pasture: Review of the Underlying Ecophysiological Processes

Sward structure affects herbage growth, pasture species dynamics, and herbage utilization. Defoliation management has a major impact on sward structure. In particular, tiller size-tiller density compensations allow for the maintenance of herbage growth. Tiller size and tiller density are determined by several major morphogenetical components. Defoliation affects these morphogenetical components, depending on its frequency and its intensity, through several direct and indirect physiological and environmental processes. Due to the implications of leaf area removal, defoliation has a direct effect on the mobilization of C and N reserves and their supply to growing leaves. In addition, defoliation has an indirect effect on leaf and tiller morphogenesis, due to its impact on the light environment within the canopy as well as plant responses to light signals (blue light, red far red ratio). Defoliation may also in some cases have a direct negative effect on leaf growth by damaging leaf meristems. Understanding the respective role of these various physiological and environmental processes requires studies where defoliation, photosynthetic active radiation and light signals are manipulated independently. Past and recent knowledge on these direct and indirect effects of defoliation on plant morphogenesis are discussed, leading to an overall integrated view of physiological and environmental processes that lead to adaptations of sward structure in response to defoliation management. Major consequences for herbage utilization efficiency are presented.

An Efficient Regeneration System for Pigeonpea (Cajanus cajan L.)

Pigeonpea is an important protein rich legume food crop grown in marginal environments of the world. Environmental stress, pests and disease cause massive yield losses due to limited genetic diversity of pigeonpea in the primary gene pool. Genetic manipulation is becoming an important method for pigeonpea improvement. In the present study, a reproducible and highly efficient plant-regeneration protocol in pigeonpea varieties, viz., ‘Phule Rajeshwari’ and ‘Vipula’ was developed. Multiple shoot induction was achieved using leaf and apical meristem with attached leaf as explant by use of various concentrations and combinations of growth regulators. Optimum embryogenic callus induction from leaf explant was observed on Murashige and Skoogs medium supplemented with 2.0 mg/l thidiazuron. Apical meristem with attached leaf explants showed early shoot initiation, high frequency of callusing (100%), and shoot regeneration (100%) as compared with leaf explant in both varieties. In vitro rooting (93.33%) was obtained within 10 days on MS medium supplemented with 2.0 mg/l indole butyric acid. The in vitro rooted plantlets were successfully established in polycarbonated polyhouse with 80% survival rate. This optimized regeneration protocol can be efficiently used for genetic transformation in pigeonpea.

Daily changes in temperature, not the circadian clock, regulate growth rate in Brachypodium distachyon.

Plant growth is commonly regulated by external cues such as light, temperature, water availability, and internal cues generated by the circadian clock. Changes in the rate of growth within the course of a day have been observed in the leaves, stems, and roots of numerous species. However, the relative impact of the circadian clock on the growth of grasses has not been thoroughly characterized. We examined the influence of diurnal temperature and light changes, and that of the circadian clock on leaf length growth patterns in Brachypodium distachyon using high-resolution time-lapse imaging. Pronounced changes in growth rate were observed under combined photocyles and thermocycles or with thermocycles alone. A considerably more rapid growth rate was observed at 28°C than 12°C, irrespective of the presence or absence of light. In spite of clear circadian clock regulated gene expression, plants exhibited no change in growth rate under conditions of constant light and temperature, and little or no effect under photocycles alone. Therefore, temperature appears to be the primary cue influencing observed oscillations in growth rate and not the circadian clock or photoreceptor activity. Furthermore, the size of the leaf meristem and final cell length did not change in response to changes in temperature. Therefore, the nearly five-fold difference in growth rate observed across thermocycles can be attributed to proportionate changes in the rate of cell division and expansion. A better understanding of the growth cues in B. distachyon will further our ability to model metabolism and biomass accumulation in grasses.

Identification of candidate genes for drought tolerance by whole-genome resequencing in maize

BackgroundDrought stress is one of the major limiting factors for maize production. With the availability of maize B73 reference genome and whole-genome resequencing of 15 maize inbreds, common variants (CV) and clustering analyses were applied to identify non-synonymous SNPs (nsSNPs) and corresponding candidate genes for drought tolerance.ResultsA total of 524 nsSNPs that were associated with 271 candidate genes involved in plant hormone regulation, carbohydrate and sugar metabolism, signaling molecules regulation, redox reaction and acclimation of photosynthesis to environment were detected by CV and cluster analyses. Most of the nsSNPs identified were clustered in bin 1.07 region that harbored six previously reported QTL with relatively high phenotypic variation explained for drought tolerance. Genes Ontology (GO) analysis of candidate genes revealed that there were 35 GO terms related to biotic stimulus and membrane-bounded organelle, showing significant differences between the candidate genes and the reference B73 background. Changes of expression level in these candidate genes for drought tolerance were detected using RNA sequencing for fertilized ovary, basal leaf meristem tissue and roots collected under drought stressed and well-watered conditions. The results indicated that 70% of candidate genes showed significantly expression changes under two water treatments and our strategies for mining candidate genes are feasible and relatively efficient.ConclusionsOur results successfully revealed candidate nsSNPs and associated genes for drought tolerance by comparative sequence analysis of 16 maize inbred lines. Both methods we applied were proved to be efficient for identifying candidate genes for complex traits through the next-generation sequencing technologies (NGS). These selected genes will not only facilitate understanding of genetic basis of drought stress response, but also accelerate genetic improvement through marker-assisted selection in maize.

The nitrate reductase inhibitor, tungsten, disrupts actin microfilaments in Zea mays L.

Tungsten is a widely used inhibitor of nitrate reductase, applied to diminish the nitric oxide levels in plants. It was recently shown that tungsten also has heavy metal attributes. Since information about the toxic effects of tungsten on actin is limited, and considering that actin microfilaments are involved in the entry of tungsten inside plant cells, the effects of tungsten on them were studied in Zea mays seedlings. Treatments with sodium tungstate for 3, 6, 12 or 24h were performed on intact seedlings and seedlings with truncated roots. Afterwards, actin microfilaments in meristematic root and leaf tissues were stained with fluorescent phalloidin, and the specimens were examined by confocal laser scanning microscopy. While the actin microfilament network was well organized in untreated seedlings, in tungstate-treated ones it was disrupted in a time-dependent manner. In protodermal root cells, the effects of tungsten were stronger as cortical microfilaments were almost completely depolymerized and the intracellular ones appeared highly bundled. Fluorescence intensity measurements confirmed the above results. In the meristematic leaf tissue of intact seedlings, no depolymerization of actin microfilaments was noticed. However, when root tips were severed prior to tungstate application, both cortical and endoplasmic actin networks of leaf cells were disrupted and bundled after 24h of treatment. The differential response of root and leaf tissues to tungsten toxicity may be due to differential penetration and absorption, while the effects on actin microfilaments could not be attributed to the nitric oxide depletion by tungsten.

Asymmetric patterns of recovery in two habitat forming seagrass species following simulated overgrazing by urchins

The persistence of seagrass meadows reflects variation in factors that influence their productivity and consumption. Sea urchins (Amblypneustes pallidus) can over-graze seagrass (Amphibolis antarctica) to create sparse meadows in South Australia, but this effect is not observed in adjacent Posidonia sinuosa meadows despite greater densities of inhabiting urchins. To test the effect of urchin grazing on seagrass biomass, we elevated the density of urchins in meadows of A. antarctica and P. sinuosa and quantified seagrass decline. Urchins removed similar amounts of biomass from both seagrass species, but the loss of leaf meristems was 11-times greater in A. antarctica than in P. sinuosa. In a second experiment to assess the recovery of seagrass, we simulated urchin grazing by clipping seagrass to mimic impacts measured in the first experiment, as well as completely removing all above ground biomass in one treatment. Following simulated grazing, P. sinuosa showed a rapid trajectory toward recovery, while A. antarctica meadows continued to decline relative to control treatments. While both A. antarctica and P. sinuosa were susceptible to heavy grazing loss, consumption of the exposed meristems of A. antarctica appears to reduce its capacity to recover, which may increase its vulnerability to long-term habitat phase-shifts and associated cascading ecosystem changes.