Vegetative Meristem Research Articles

Initial proplastid-to-chloroplast differentiation in the developing vegetative shoot apical meristem of Arabidopsis

In dicot plants, the process by which undifferentiated plastids, termed proplastids, differentiate into mature functional chloroplasts begins in the shoot apical meristem (SAM) and young leaf primordia, and continues along leaf development. In this work, we followed initial chloroplast biogenesis in cells of the SAM in Arabidopsis, during the early stages of germination, using chlorophyll fluorescence as a marker. We found that cells bound to form the SAM in the mature seed embryo lack chlorophyll, while plastids in the rest of the embryo cells are somewhat developed. The initial appearance of chlorophyll in the SAM occurred two days after the onset of germination, was not expedited by higher light intensities, and required a light period of between five to ten hours within these two days. In addition, we found that biogenesis of chloroplasts occurred only in the upper layer of the SAM, as opposed to the two central subtending cell layers. This pattern was maintained, mirroring the developmental status of plastids in the mature vegetative SAM. The work also presents another model for studying proplastid-to-chloroplast development, in which differentiation can be followed in SAM cells in a defined time-wise fashion.

New Symptoms Identified in Phytoplasma-Infected Plants Reveal Extra Stages of Pathogen-Induced Meristem Fate-Derailment.

In flowering plants, the transition of a shoot apical meristem from vegetative to reproductive destiny is a graduated, multistage process that involves sequential conversion of the vegetative meristem to an inflorescence meristem, initiation of floral meristems, emergence of flower organ primordia, and formation of floral organs. This orderly process can be derailed by phytoplasma, a bacterium that parasitizes phloem sieve cells. In a previous study, we showed that phytoplasma-induced malformation of flowers reflects stage-specific derailment of shoot apical meristems from their genetically preprogrammed reproductive destiny. Our current study unveiled new symptoms of abnormal morphogenesis, pointing to derailment of meristem transition at additional stages previously unidentified. We also found that the fate of developing meristems may be derailed even after normal termination of the floral meristem and onset of seed production. Although previous reports by others have indicated that different symptoms may be induced by different phytoplasmal effectors, the phenomenon observed in our experiment raises interesting questions as to (i) whether effectors can act at specific stages of meristem transition and (ii) whether specific floral abnormalities are attributable to meristem fate-derailment events triggered by different effectors that each act at a specific stage in meristem transition. Research addressing such questions may lead to discoveries of an array of phytoplasmal effectors.

Fruit-dependent epigenetic regulation of flowering in Citrus.

In many perennial plants, seasonal flowering is primarily controlled by environmental conditions, but in certain polycarpic plants, environmental signals are locally gated by the presence of developing fruits initiated in the previous season through an unknown mechanism. Polycarpy is defined as the ability of plants to undergo several rounds of reproduction during their lifetime, alternating vegetative and reproductive meristems in the same individual. To understand how fruits regulate flowering in polycarpic plants, we focused on alternate bearing in Citrus trees that had been experimentally established as fully flowering or nonflowering. We found that the presence of the fruit causes epigenetic changes correlating with the induction of the CcMADS19 floral repressor, which prevents the activation of the floral promoter CiFT2 even in the presence of the floral inductive signals. By contrast, newly emerging shoots display an opposite epigenetic scenario associated with CcMADS19 repression, thereby allowing the activation of CiFT2 the following cold season.

Mechanisms of ROS Regulation of Plant Development and Stress Responses.

Plants are subjected to various environmental stresses throughout their life cycle. Reactive oxygen species (ROS) play important roles in maintaining normal plant growth, and improving their tolerance to stress. This review describes the production and removal of ROS in plants, summarizes recent progress in understanding the role of ROS during plant vegetative apical meristem development, organogenesis, and abiotic stress responses, and some novel findings in recent years are discussed. More importantly, interplay between ROS and epigenetic modifications in regulating gene expression is specifically discussed. To summarize, plants integrate ROS with genetic, epigenetic, hormones and external signals to promote development and environmental adaptation.

Confocal Live Imaging of Shoot Apical Meristems from Different Plant Species.

The shoot apical meristem (SAM) functions as a conserved stem cell reservoir and it generates almost all aboveground tissues during the postembryonic development. The activity and morphology of SAMs determine important agronomic traits, such as shoot architecture, size and number of reproductive organs, and most importantly, grain yield. Here, we provide a detailed protocol for analyzing both the surface morphology and the internal cellular structure of the living SAMs from different species through laser scanning confocal microscope. The whole procedure from the sample preparation to the acquisition of high resolution three-dimensional (3D) images can be accomplished within as short as 20 minutes. We demonstrate that this protocol is highly efficient for studying not only the inflorescence SAMs of the model species but also the vegetative meristems from different crops, providing a simple but powerful tool to study the organization and development of meristems across different plant species.

Overexpression of both flowering time genes AtSOC1 and SaFUL revealed huge influence onto plant habitus in poplar

The MADS box genes SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1) and FRUITFULL (FUL) are widely known to be involved in the determination of flowering time and the transition from vegetative to reproductive meristem identity in A. thaliana and many other plant species. Analog to the induction of SOC1 and FUL, double mutants in Arabidopsis revealed both secondary cell wall and wood formation. We tested a putative influence of the overexpression of both genes onto growth and plant habitus in the perennial genus Populus. Two different poplar hybrid clones were transformed with SOC1 from A. thaliana (AtSOC1) and FUL from Sinapis alba (SaFUL). Both genes were under the control of the CaMV-35S promoter to induce constitutive overexpression. Regenerated single and double transgenic lines were transferred to the glasshouse and cultivated for up to two vegetation periods. Parallel overexpression of both genes led to plants with a significantly restricted plant growth compared to both untransformed controls and single transgenic plants. Extreme phenotypes also exhibit a different overall habitus of plants and leaf morphology. Thus, both genes seem to be involved in plant habitus and biomass formation in poplar. This study confirms the hypothesis that the two MADS box genes SOC1 and FUL are not only part of reproductive meristem regulation but also involved in vegetative meristem growth. In the next step, downregulation of all SOC1 and FUL genes in poplar by genome editing could lead to increased plant growth and biomass formation.

RNA-Seq reveals new DELLA targets and regulation in transgenic GA-insensitive grapevines

BackgroundGibberellins (GAs) and their regulator DELLA are involved in many aspects of plant growth and development and most of our current knowledge in the DELLA-facilitated GA signaling was obtained from the studies of annual species. To understand GA-DELLA signaling in perennial species, we created ten GA-insensitive transgenic grapevines carrying a DELLA mutant allele (Vvgai1) in the background of Vitis vinifera ‘Thompson Seedless’ and conducted comprehensive analysis of their RNA expression profiles in the shoot, leaf and root tissues.ResultsThe transgenic lines showed varying degrees of dwarf stature and other typical DELLA mutant phenotypes tightly correlated with the levels of Vvgai1 expression. A large number of differentially expressed genes (DEGs) were identified in the shoot, leaf and root tissues of the transgenic lines and these DEGs were involved in diverse biological processes; many of the DEGs showed strong tissue specificity and about 30% them carried a DELLA motif. We further discovered unexpected expression patterns of several key flowering induction genes VvCO, VvCOL1 and VvTFL1.ConclusionsOur results not only confirmed many previous DELLA study findings in annual species, but also revealed new DELLA targets and responses in grapevine, including the roles of homeodomain transcription factors as potential co-regulators with DELLA in controlling the development of grapevine which uniquely possess both vegetative and reproductive meristems at the same time. The contrasting responses of some key flowering induction pathway genes provides new insights into the divergence of GA-DELLA regulations between annual and perennial species in GA-DELLA signaling.

FLOWERING LOCUS T2 regulates spike development and fertility in temperate cereals.

FLOWERING LOCUS T2 (FT2) is the closest paralog of the FT1 flowering gene in the temperate grasses. Here we show that overexpression of FT2 in Brachypodium distachyon and barley results in precocious flowering and reduced spikelet number, while down-regulation by RNA interference results in delayed flowering and a reduced percentage of filled florets. Similarly, truncation mutations of FT2 homeologs in tetraploid wheat delayed flowering (2-4 d) and reduced fertility. The wheat ft2 mutants also showed a significant increase in the number of spikelets per spike, with a longer spike development period potentially contributing to the delayed heading time. In the wheat leaves, FT2 was expressed later than FT1, suggesting a relatively smaller role for FT2 in the initiation of the reproductive phase. FT2 transcripts were detected in the shoot apical meristem and increased during early spike development. Transversal sections of the developing spike showed the highest FT2 transcript levels in the distal part, where new spikelets are formed. Our results suggest that, in wheat, FT2 plays an important role in spike development and fertility and a limited role in the timing of the transition between the vegetative and reproductive shoot apical meristem.

The signaling peptide-encoding genes CLE16, CLE17 and CLE27 are dispensable for Arabidopsis shoot apical meristem activity.

The shoot apical meristem produces all of the leaves, stems and flowers of a flowering plant from a reservoir of stem cells at its growing tip. In Arabidopsis, the small polypeptide signaling molecule CLAVATA3 (CLV3), a member of the CLV3/EMBRYO SURROUNDING REGION-RELATED (CLE) gene family, is a key component of a negative feedback loop that maintains stem cell activity in shoot and floral meristems throughout development. Because in some plant species multiple CLE genes are involved in regulating shoot apical meristem activity, we tested the hypothesis that CLE genes other than CLV3 might function in stem cell homeostasis in Arabidopsis. We identified three Arabidopsis CLE genes expressed in the post-embryonic shoot apical meristem, generated loss-of-function alleles using genome editing, and analyzed the meristem phenotypes of the resulting mutant plants. We found that null mutations in CLE16, CLE17 or CLE27 affected neither vegetative nor reproductive shoot meristem activity under normal growth conditions, although CLE27 appears to slightly prolong vegetative growth. Our results indicate that the CLE16, CLE17 and CLE27 genes have largely redundant roles in the Arabidopsis shoot apical meristem and/or regulate meristem activity only under specific environmental conditions.

Developmental transcriptome analysis of floral transition in Rosa odorata var. gigantea.

Expression analyses revealed that floral transition of Rosa odorata var. gigantea is mainly regulated by VRN1, COLs, DELLA and KSN, with contributions by the effects of phytohormone and starch metabolism. Seasonal plants utilize changing environmental and developmental cues to control the transition from vegetative growth to flowering at the correct time of year. This study investigated global gene expression profiles at different developmental stages of Rosa odorata var. gigantea by RNA-sequencing, combined with phenotypic characterization and physiological changes. Gene ontology enrichment analysis of the differentially expressed genes (DEGs) between four different developmental stages (vegetative meristem, pre-floral meristem, floral meristem and secondary axillary buds) indicated that DNA methylation and the light reaction played a large role in inducing the rose floral transition. The expression of SUF and FLC, which are known to play a role in delaying flowering until vernalization, was down-regulated from the vegetative to the pre-floral meristem stage. In contrast, the expression of VRN1, which promotes flowering by repressing FLC expression, increased. The expression of DELLA proteins, which function as central nodes in hormone signaling pathways, and probably involve interactions between GA, auxin, and ABA to promote the floral transition, was well correlated with the expression of floral integrators, such as AGL24, COL4. We also identified DEGs associated with starch metabolism correlated with SOC1, AGL15, SPL3, AGL24, respectively. Taken together, our results suggest that vernalization and photoperiod are prominent cues to induce the rose floral transition, and that DELLA proteins also act as key regulators. The results summarized in the study on the floral transition of the seasonal rose lay a foundation for further functional demonstration, and have profound economic and ornamental values.

Plant architecture of 'Albion' strawberry (<i>Fragaria</i> × <i>ananassa</i> Duch.) is not influenced by light source during conditioning

Architectural analysis describes the position and fate (vegetative or floral) of plant meristems to account for differences in meristem sensitivity to stimuli depending on developmental stage and position on the plant. To provide further insight into the flowering responses of long-day strawberries to nitrogen (N), photoperiod and light source, ‘Albion’ strawberry plants were conditioned with 100 or 800 ppm N under ND (natural daylength) or LD (long days, natural days plus 24-hr supplementary illumination provided by either 60- or 7-watt incandescent bulbs) and greenhouse growth was evaluated for a total of 10 weeks following conditioning. After greenhouse forcing, plants were dissected and their floral architecture evaluated. Additional plants were established in early July in off-season plasticulture production where fruit, crown and stolon production were evaluated. Both light sources were equally effective in eliciting long-day photoperiod responses. No photoperiod effect on floral precocity, leaf, crown, or runner production was observed during greenhouse forcing. Plants under ND tended to produce more inflorescences during the first 5 weeks while LD enhanced inflorescence and flower production during the last 3 weeks of forcing. In dissected plants, maximum floral initiation was observed in plants receiving elevated N under LD. LD inhibited branch crown formation, but had no effect on the number of vegetative, floral or stolon producing axillary meristems regardless of N treatment. LD conditioning enhanced early yield (through 4 September). Field stolon and branch crown formation was supressed in plants receiving low N with LD conditioning. Stolon and branch crown inhibition by LD conditioning was not observed with elevated N. Growth data combined with architectural mapping of meristems allows more conclusive statements regarding treatment effects on specific stages of floral physiology (i.e. induction, initiation, differentiation and development) compared to more generalized conclusions obtained with growth data alone. The separation of direct and indirect effects on floral physiology is possible with floral architectural analysis.

Plasticity in Meristem Allocation as an Adaptive Strategy of a Desert Shrub under Contrasting Environments.

The pattern of resource allocation to reproduction vs. vegetative growth is a core component of a plant’s life-history strategy. Plants can modify their biomass allocation patterns to adapt to contrasting environments. Meristems can have alternative fates to commit to vegetative growth, reproduction, or remaining inactive (dormant or senescent/dead). However, knowledge about whether meristem fates can interpret adaptive changes in biomass allocation remains largely unknown. We measured aboveground plant biomass (a proxy of plant size) and meristem number of a dominant shrub Artemisia ordosica in three populations occupying different habitats in the Mu Us Desert of northern China. Size-dependent biomass allocation and meristem allocation among habitats were compared. The size-dependent biomass allocation and meristem allocation of A. ordosica strongly varied across habitats. There were significant positive linear relationships between meristem allocation and biomass allocation in all habitats, indicating that meristem allocation is an indicator of the estimated resource allocation to reproductive and vegetative organs in this species. Plasticity in meristem allocation was more likely caused by larger individuals having less active meristems due to environmental stress. Vegetative meristems (VM) were likely more vulnerable to environmental limitation than reproductive ones, resulting in the ratio of resource investment between vegetative and reproductive functions exhibiting plasticity in different habitats. A. ordosica invested a higher fraction of its resource to reproduction in the adverse habitat, while more resource to vegetative growth in the favorable habitat. A. ordosica adopts different resource allocation patterns to adapt to contrasting habitat conditions through altering its meristem fates. Our results suggest that the arid-adapted shrub A. ordosica deactivates more VM than reproductive ones to hedge against environmental stress, representing an important adaptive strategy. This information contributes to understand the life-history strategies of long-lived plants under stressful environments.

Model for the role of auxin polar transport in patterning of the leaf adaxial-abaxial axis.

Leaf adaxial-abaxial polarity refers to the two leaf faces, which have different types of cells performing distinct biological functions. In 1951, Ian Sussex reported that when an incipient leaf primordium was surgically isolated by an incision across the vegetative shoot apical meristem (SAM), a radialized structure without an adaxial domain would form. This led to the proposal that a signal, now called the Sussex signal, is transported from the SAM to emerging primordia to direct leaf adaxial-abaxial patterning. It was recently proposed that instead of the Sussex signal, polar transport of the plant hormone auxin is critical in leaf polarity formation. However, how auxin polar transport functions in the process is unknown. Through live imaging, we established a profile of auxin polar transport in and around young leaf primordia. Here we show that auxin polar transport in lateral regions of an incipient primordium forms auxin convergence points. We demonstrated that blocking auxin polar transport in the lateral regions of the incipient primordium by incisions abolished the auxin convergence points and caused abaxialized leaves to form. The lateral incisions also blocked the formation of leaf middle domain and margins and disrupted expression of the middle domain/margin-associated marker gene WUSCHEL-RELATED HOMEOBOX 1 (SlWOX1). Based on these results we propose that the auxin convergence points are required for the formation of leaf middle domain and margins, and the functional middle domain and margins ensure leaf adaxial-abaxial polarity. How middle domain and margins function in the process is discussed.

RNA-seq analysis of apical meristem reveals integrative regulatory network of ROS and chilling potentially related to flowering in Litchi chinensis

Litchi is an important woody fruit tree. Floral initiation in litchi is triggered by low temperatures. However, defective flowering is a major challenge for litchi production in times of climate change and global warming. Previous studies have shown that the reactive oxygen species (ROS) generated by methyl viologen dichloride hydrate (MV) promotes flowering. In this study, potted trees were transferred to growth chambers for low-temperature (LT), medium-temperature (MT), and high-temperature (HT) treatments. Trees at MT were subjected to ROS treatment to promote flowering, and those at LT were induced to flower. RNA-sequencing was applied to obtain a global transcriptome of the apical meristem and reveal potential gene networks controlling the transformation from vegetative meristems (VM) into inflorescence meristems (IM). We assembled 73,117 unigenes with a mean size of 790 bp and 11741 unigenes were identified as both chilling and ROS responsive genes (CRRGs), of which 48 were identified as flowering-related CRRGs, 59 were plant hormone signal transduction CRRGs, and 146 were plant hormone biosynthesis-related CRRGs. Genes co-expression network analysis indicated inner relationships, suggesting that ROS and chilling promotes the VM to IM transition through a regulatory gene network of transcription factors, hormones, and flowering regulators.

Maize YABBY Genes drooping leaf1 and drooping leaf2 Regulate Plant Architecture.

Leaf architecture directly influences canopy structure, consequentially affecting yield. We discovered a maize (Zea mays) mutant with aberrant leaf architecture, which we named drooping leaf1 (drl1). Pleiotropic mutations in drl1 affect leaf length and width, leaf angle, and internode length and diameter. These phenotypes are enhanced by natural variation at the drl2 enhancer locus, including reduced expression of the drl2-Mo17 allele in the Mo17 inbred. A second drl2 allele, produced by transposon mutagenesis, interacted synergistically with drl1 mutants and reduced drl2 transcript levels. The drl genes are required for proper leaf patterning, development and cell proliferation of leaf support tissues, and for restricting auricle expansion at the midrib. The paralogous loci encode maize CRABS CLAW co-orthologs in the YABBY family of transcriptional regulators. The drl genes are coexpressed in incipient and emergent leaf primordia at the shoot apex, but not in the vegetative meristem or stem. Genome-wide association studies using maize NAM-RIL (nested association mapping-recombinant inbred line) populations indicated that the drl loci reside within quantitative trait locus regions for leaf angle, leaf width, and internode length and identified rare single nucleotide polymorphisms with large phenotypic effects for the latter two traits. This study demonstrates that drl genes control the development of key agronomic traits in maize.

Transcriptome of the floral transition in Rosa chinensis \u2018Old Blush\u2019

BackgroundThe floral transition plays a vital role in the life of ornamental plants. Despite progress in model plants, the molecular mechanisms of flowering regulation remain unknown in perennial plants. Rosa chinensis ‘Old Blush’ is a unique plant that can flower continuously year-round. In this study, gene expression profiles associated with the flowering transition were comprehensively analyzed during floral transition in the rose.ResultsAccording to the transcriptomic profiles, 85,663 unigenes and 1,637 differentially expressed genes (DEGs) were identified, among which 32 unigenes were involved in the circadian clock, sugar metabolism, hormone, and autonomous pathways. A hypothetical model for the regulation of floral transition was proposed in which the candidate genes function synergistically the floral transition process. Hormone contents and biosynthesis and metabolism genes fluctuated during the rose floral transition process. Gibberellins (GAs) inhibited rose floral transition, the content of GAs gradually decreased and GA2ox and SCL13 were upregulated from vegetative (VM) meristem to floral meristem (FM). Auxin plays an affirmative part in mediating floral transition, auxin content and auxin-related gene expression levels were gradually upregulated during the floral transition of the rose. However, ABA content and ABA signal genes were gradually downregulated, suggesting that ABA passively regulates the rose floral transition by participating in sugar signaling. Furthermore, sugar content and sugar metabolism genes increased during floral transition in the rose, which may be a further florigenic signal that activates floral transition. Additionally, FRI, FY, DRM1, ELIP, COP1, CO, and COL16 are involved in the circadian clock and autonomous pathway, respectively, and they play a positively activating role in regulating floral transition. Overall, physiological changes associated with genes involved in the circadian clock or autonomous pathway collectively regulated the rose floral transition.ConclusionsOur results summarize a valuable collective of gene expression profiles characterizing the rose floral transition. The DEGs are candidates for functional analyses of genes affecting the floral transition in the rose, which is a precious resource that reveals the molecular mechanism of mediating floral transition in other perennial plants.

Differential Gene Expression in the Meristem and during Early Fruit Growth of Pisum sativum L. Identifies Potential Targets for Breeding.

For successful molecular breeding it is important to identify targets to the gene family level, and in the specific species of interest, in this case Pisum sativum L. The cytokinins have been identified as a key breeding target due to their influence on plant architecture, and on seed size and sink activity. We focused on the cytokinin biosynthetic gene family (the IPTs) and the gene family key to the destruction of cytokinins (the CKXs), as well as other gene families potentially affected by changing cytokinin levels. These included key meristem genes (WUS and BAM1) and the transporter gene families, sucrose transporters (SUTs) and amino acid permeases (AAPs). We used reverse transcription quantitative PCR (RT-qPCR) to monitor gene expression in the vegetative meristem and in pre- and post-fertilisation young pea fruits. PsWUS expression was specific to the shoot apical meristem while PsBAM1 was highly expressed in the shoot apical meristem (SAM) but was also expressed at a low level in the young fruit. Differential expression was shown between genes and within gene families for IPT, CKX, SUT, and AAP. PsCKX7 showed strong gene family member-specific expression in the SAM, and was also expressed in young pea fruits. We suggest that PsCKX7 is a potential target for downregulation via molecular breeding or gene editing.

SQUAMOSA Promoter Binding Protein-like Transcription Factors: Targets for Improving Cereal Grain Yield

Grain size and shape are prime targets for crop breeders, as they affect both yield and quality of rice. In addition, breeders target plant vegetative and reproductive shoot architecture, because these traits influence grain number, and thus yield potential. Accordingly, genetic control of grain size and shape as well as shoot architecture has been extensively investigated in recent years. In a recent study, Bin Han and colleagues have used genome-wide association analysis of grain size in a diverse collection of rice varieties of worldwide origin and identified a major quantitative trait locus GLW7, which encodes a SQUAMOSA promoter binding protein-like 13 (OsSPL13), a member of the plant-specific SBP domain family of transcription factors (Si et al., 2016).

Separate elements of the TERMINAL FLOWER 1 cis-regulatory region integrate pathways to control flowering time and shoot meristem identity.

TERMINAL FLOWER 1 (TFL1) is a key regulator of Arabidopsis plant architecture that responds to developmental and environmental signals to control flowering time and the fate of shoot meristems. TFL1 expression is dynamic, being found in all shoot meristems, but not in floral meristems, with the level and distribution changing throughout development. Using a variety of experimental approaches we have analysed the TFL1 promoter to elucidate its functional structure. TFL1 expression is based on distinct cis-regulatory regions, the most important being located 3' of the coding sequence. Our results indicate that TFL1 expression in the shoot apical versus lateral inflorescence meristems is controlled through distinct cis-regulatory elements, suggesting that different signals control expression in these meristem types. Moreover, we identified a cis-regulatory region necessary for TFL1 expression in the vegetative shoot and required for a wild-type flowering time, supporting that TFL1 expression in the vegetative meristem controls flowering time. Our study provides a model for the functional organisation of TFL1 cis-regulatory regions, contributing to our understanding of how developmental pathways are integrated at the genomic level of a key regulator to control plant architecture.

The shoot concept of the flower: Still up to date?

The shoot concept of the flower suggests that flowers correspond to vegetative short-shoots except the fact that their lateral appendages are floral and not vegetative leaves. However, in view of the different properties of vegetative and flower meristems, this concept should be questioned. Differential meristem activity resulting in tubes, hypanthia and inferior ovaries, continuous meristem expansion providing space for stamen fascicles and additional structures and the process of (repeated) fractionation using a given space completely, are characteristics of flower meristems hardly explainable with the shoot concept. Linking instead flower development with recent findings in molecular biology and computational modeling widens the view to the fundamental relation between growth and form. Given that the same general principles characterize plant growth at all life stages, the loss of apical growth appears to play the major role in changing geometry, space availability and genetic regulation in flower meristems. The flower, thus, turns out to be the sporangia bearing tip of a shoot.