Plant Developmental Biology Research Articles

Editorial: Model organisms in plant developmental biology—their effectiveness and limitations

Editorial: Model organisms in plant developmental biology—their effectiveness and limitations

Full-length transcriptome sequencing of pepper fruit during development and construction of a transcript variation database.

Chili pepper is an important spice and a model plant for fruit development studies. Large-scale omics information on chili pepper plant development continues to be gathered for understanding development as well as capsaicin biosynthesis. In this study, a full-spectrum transcriptome data of eight chili pepper tissues at five growth stages using the Oxford Nanopore long-read sequencing approach was generated. Of the 485 351 transcripts, 35 336 were recorded as reference transcripts (genes), while 450 015 were novel including coding, lnc, and other non-coding RNAs. These novel transcripts belonged to unknown/intergenic (347703), those retained introns (26336), and had multi-exons with at least one junction match (20333). In terms of alternative splicing, retained intron had the highest proportion (14795). The number of tissue-specific expressed transcripts ranged from 22 925 (stem) to 40 289 (flower). The expression changes during fruit and placenta development are discussed in detail. Integration of gene expression and capsaicin content quantification throughout the placental development clarifies that capsaicin biosynthesis in pepper is mainly derived from valine, leucin, and isoleucine degradation as well as citrate cycle and/or pyrimidine metabolism pathways. Most importantly, a user-friendly Pepper Full-Length Transcriptome Variation Database (PFTVD 1.0) (http://pepper-database.cn/) has been developed. PFTVD 1.0 provides transcriptomics and genomics information and allows users to analyse the data using various tools implemented. This work highlights the potential of long-read sequencing to discover novel genes and transcripts and their diversity in plant developmental biology.

TALE gene family: identification, evolutionary and expression analysis under various exogenous hormones and waterlogging stress in Cucumis sativus L.

BackgroundThree Amino acid Loop Extension (TALE) belongs to the homeobox group of genes that are important constituents of plant systems. The TALE gene family is instrumental not only in growth and development but also plays an essential role in regulating plant response to environmental adversaries.ResultsIn the present study, we isolated 21 CsTALE genes from the cucumber (Cucumis sativus L.) genome database. Bioinformatics tools were put in place to understand the structural and functional components of the CsTALE gene family. The evolutionary analysis dissected them into seven subclades (KNOX-I, KNOX-II, and BELL-I to BELL-V). The cis-acting elements in the promoter region of CsTALE genes disclosed that they are key regulators of hormonal and stress-related processes. Additionally, the STRING database advocated the concerting role of CsTALE proteins with other key transcription factors potent in plant developmental biology. The CsmiR319 and CsmiR167a-3p targeting the CsTALE15 and CsTALE16, respectively, further assert the importance of the CsTALE gene family posttranscriptional-related processes. Tissue-specific gene expression unfolded the fundamental involvement of CsTALE genes as they were expressed throughout the developmental stages. Under waterlogging stress, the CsTALE17 expressed significantly higher values in WL, WL-NAA, and WL-ETH but not in WL-MeJA-treated samples.ConclusionsThe present study reveals the evolution and functions of the CsTALE gene family in cucumber. Our work will provide a platform that will help future researchers address the issue of waterlogging stress in the Yangtze River Delta.

Plant-specific calreticulin is localized in the nuclei of highly specialized cells in the pistil-new observations for an old hypothesis.

One of the first cellular locations of the calreticulin (CRT) chaperone in eukaryotic cells, apart from its obvious localization in the endoplasmic reticulum (ER), was the cell nucleus (Opas et al. 1991). The presence of CRT has been detected inside the nucleus and in the nuclear envelope of animal and plant cells, and a putative nuclear localization signal (NLS) in the CRT amino acid sequence has been mapped in several animal and plant species. Over the last 30years, other localization sites of this protein outside the ER and cell nucleus have also been discovered, suggesting that CRT is a multifunctional Ca2+-binding protein widely found in various cell types. In our previous studies focusing on plant developmental biology, we have demonstrated the presence of CRT inside and outside the ER in highly specialized plant cells, as well as the possibility of CRT localization in the cell nucleus. In this paper, we present a detailed analysis of immunocytochemical localization of CRT inside nuclei of the pistil transmission tract somatic cells before and after pollination. We show a similar pattern of the nuclear CRT localization in relation to exchangeable Ca2+ for two selected species of angiosperms, dicotyledonous Petunia and monocot Haemanthus, that differ in anatomical structure of the pistil and discuss the potential role of CRT in the cell nucleus.

Transgressive and subgenome expression level dominance and co-expression network analyses at the early fiber development in allopolyploid Gossypium

Polyploidization can lead to the emergence of new species. Allotetraploid cotton arose through hybridization and whole genome duplication of its diploid A and D genome progenitors. As a prominent global fiber crop, cotton serves as an excellent model organism for exploring plant evolutionary biology and fiber development, particularly in polyploidization. This study utilized transcriptomics to analyze expression difference between diploid and allotetraploid cotton, aiming to further the understanding of transcriptomic dynamics associated at different fiber developmental stages during polyploidization process. In this study, we identified additive and non-additive genes and clarified the contributions of non-additive genes during the tetraploid formation process. Furthermore, we constructed a gene co-expression network, providing new insights into the transcriptional regulation of polyploid cotton fiber development. Among non-additive genes, more A genome-biased expression level dominance (ELD-A) than D genome (ELD-D) occurred in the tetraploids. In Gossypium hirsutum, ELD-A genes played major roles during initial elongation, exerting greater dominance in fiber development. Transgressive genes impacted early fiber development in Gossypium barbadense. While the At subgenome profoundly influences tetraploid fiber development, Dt sub-genome activation, inheritance and sub/neo-functionalization promote superior high-yield traits. By constructing co-expression networks, we identified key fiber developmental candidate genes, providing valuable resources for functional research and breeding perspectives to develop superior cotton varieties.

Best practices for the execution, analysis, and data storage of plant single-cell/nucleus transcriptomics.

Single-cell and single-nucleus RNA-sequencing technologies capture the expression of plant genes at an unprecedented resolution. Therefore, these technologies are gaining traction in plant molecular and developmental biology for elucidating the transcriptional changes across cell types in a specific tissue or organ, upon treatments, in response to biotic and abiotic stresses, or between genotypes. Despite the rapidly accelerating use of these technologies, collective and standardized experimental and analytical procedures to support the acquisition of high-quality data sets are still missing. In this commentary, we discuss common challenges associated with the use of single-cell transcriptomics in plants and propose general guidelines to improve reproducibility, quality, comparability, and interpretation and to make the data readily available to the community in this fast-developing field of research.

Beyond stomatal development: SMF transcription factors as versatile toolkits for land plant evolution.

As master transcription factors of stomatal development, SPEECHLESS, MUTE, and FAMA, collectively termed SMFs, are primary targets of molecular genetic analyses in the model plant Arabidopsis thaliana. Studies in other model systems identified SMF orthologs as key players in evolutionary developmental biology studies on stomata. However, recent studies on the astomatous liverwort Marchantia polymorpha revealed that the functions of these genes are not limited to the stomatal development, but extend to other types of tissues, namely sporophytic setal and gametophytic epidermal tissues. These studies provide insightful examples of gene-regulatory network co-opting, and highlight SMFs and related transcription factors as general toolkits for novel trait evolution in land plant lineages. Here, we critically review recent literature on the SMF-like gene in M. polymorpha and discuss their implications for plant evolutionary biology.

Rediscovering Chara as a model organism for molecular and evo-devo studies.

Chara has been used as a model for decades in the field of plant physiology, enabling the investigation of fundamental physiological processes. In electrophysiological studies, Chara has been utilized thanks to its large internodal cells that can be easily manipulated. Additionally, Chara played a pioneering role in elucidating the presence and function of the cytoskeleton in cytoplasmic streaming, predating similar findings in terrestrial plants. Its representation considerably declined following the establishment and routine application of genetic transformation techniques in Arabidopsis. Nevertheless, the recent surge in evo-devo studies can be attributed to the whole genome sequencing of the Chara braunii, which has shed light on ancestral traits prevalent in land plants. Surprisingly, the Chara braunii genome encompasses numerous genes that were previously regarded as exclusive to land plants, suggesting their acquisition prior to the colonization of terrestrial habitats. This review summarizes the established methods used to study Chara, while incorporating recent molecular data, to showcase its renewed importance as a model organism in advancing plant evolutionary developmental biology.

In-Depth Quantification ofCell Division andElongation Dynamics at the Tip ofGrowing Arabidopsis Roots Using 4D Microscopy, AI-Assisted Image Processing andData Sonification.

One of the fundamental questions in plant developmental biology is how cell proliferation and cell expansion coordinately determine organ growth and morphology. An amenable system to address this question is the Arabidopsis root tip, where cell proliferation and elongation occur in spatially separated domains, and cell morphologies can easily be observed using a confocal microscope. While past studies revealed numerous elements of root growth regulation including gene regulatory networks, hormone transport and signaling, cell mechanics and environmental perception, how cells divide and elongate under possible constraints from cell lineages and neighboring cell files has not been analyzed quantitatively. This is mainly due to the technical difficulties in capturing cell division and elongation dynamics at the tip of growing roots, as well as an extremely labor-intensive task of tracing the lineages of frequently dividing cells. Here, we developed a motion-tracking confocal microscope and an Artificial Intelligence (AI)-assisted image-processing pipeline that enables semi-automated quantification of cell division and elongation dynamics at the tip of vertically growing Arabidopsis roots. We also implemented a data sonification tool that facilitates human recognition of cell division synchrony. Using these tools, we revealed previously unnoted lineage-constrained dynamics of cell division and elongation, and their contribution to the root zonation boundaries.

Phenotypic Characterization and Gene Mapping of a Spiral Leaf and Dwarf (sld) Mutant from Tetraploid Common Tobacco (Nicotiana tabacum L.)

Leaf morphology and plant height are two agronomic traits closely related to tobacco (Nicotiana tabacum L.) yield and quality. The study of leaf morphology and plant stature mutants will greatly contribute to the fields of plant architecture breeding and developmental biology. Here, we report the characterization of a spiral leaf and dwarf (sld) mutant identified from an ethylmethane sulfonate (EMS)-induced common tobacco population. The sld mutant displayed the phenotype of wrinkled, spiral, and miniature leaves, with the growth point as the central axis and plant dwarfing with shortened internodes. The inheritance pattern of the sld mutant phenotype was manipulated by a recessive nuclear monogene, which was linked to six tobacco simple sequence repeat (SSR) markers from linkage group 5 via gene mapping. Utilizing an F2 population, the sld mutant gene the sld mutant gene was located between the co-segregated markers PT51778, PT54913, and the marker PT61414, with an equal genetic distance of 0.16 cM. Taking advantage of a BC1F1 population, the markers PT51778, PT54913, the sld gene, and the marker PT61414 demonstrated co-segregation, located between the markers PT40040 and PT60933, respectively, with a genetic distance of 1.37 cM and 6.32 cM, respectively. These findings will be helpful in cloning the sld gene and in the further characterization of the regulatory genes controlling the spiral and dwarfing phenotypes in tobacco.

Plants use molecular mechanisms mediated by biomolecular condensates to integrate environmental cues with development.

This review highlights recent literature on biomolecular condensates in plant development and discusses challenges for fully dissecting their functional roles. Plant developmental biology has been inundated with descriptive examples of biomolecular condensate formation, but it is only recently that mechanistic understanding has been forthcoming. Here, we discuss recent examples based on current findings in plant and cell biology at different stages of the plant life cycle. We group these examples based on putative molecular functions, including: sequestering interacting components, enhancing dwell time, and interacting with cytoplasmic biophysical properties in response to environmental change. We explore how these mechanisms modulate plant development in response to environmental inputs and discuss challenges and opportunities for further research into deciphering molecular mechanisms to better understand the diverse roles that biomolecular condensates exert on life.

Molecular mechanisms of vascular tissue patterning in Arabidopsis thaliana L. roots.

A vascular system in plants is a product of aromorphosis that enabled them to colonize land because it delivers water, mineral and organic compounds to plant organs and provides effective communications between organs and mechanical support. Vascular system development is a common object of fundamental research in plant development biology. In the model plant Arabidopsis thaliana, early stages of vascular tissue formation in the root are a bright example of the self-organization of a bisymmetric (having two planes of symmetry) pattern of hormone distribution, which determines vascular cell fates. In the root, vascular tissue development comprises four stages: (1) specification of progenitor cells for the provascular meristem in early embryonic stages, (2) the growth and patterning of the embryo provascular meristem, (3) postembryonic maintenance of the cell identity in the vascular tissue initials within the root apical meristem, and (4) differentiation of their descendants. Although the anatomical details of A. thaliana root vasculature development have long been known and described in detail, our knowledge of the underlying molecular and genetic mechanisms remains limited. In recent years, several important advances have been made, shedding light on the regulation of the earliest events in provascular cells specification. In this review, we summarize the latest data on the molecular and genetic mechanisms of vascular tissue patterning in A. thaliana root. The first part of the review describes the root vasculature ontogeny, and the second reconstructs the sequence of regulatory events that underlie this histogenesis and determine the development of the progenitors of the vascular initials in the embryo and organization of vascular initials in the seedling root.

The origin and evolution of carpels and fruits from an evo-devo perspective.

The flower is an evolutionary innovation in angiosperms that drives the evolution of biodiversity. The carpel is integral to a flower and develops into fruits after fertilization, while the perianth, consisting of the calyx and corolla, is decorative to facilitate pollination and protect the internal organs, including the carpels and stamens. Therefore, the nature of flower origin is carpel and stamen origin, which represents one of the greatest and fundamental unresolved issues in plant evolutionary biology. Here, we briefly summarize the main progress and key genes identified for understanding floral development, focusing on the origin and development of the carpels. Floral ABC models have played pioneering roles in elucidating flower development, but remain insufficient for resolving flower and carpel origin. The genetic basis for carpel origin and subsequent diversification leading to fruit diversity also remains elusive. Based on current research progress and technological advances, simplified floral models and integrative evolutionary-developmental (evo-devo) strategies are proposed for elucidating the genetics of carpel origin and fruit evolution. Stepwise birth of a few master regulatory genes and subsequent functional diversification might play a pivotal role in these evolutionary processes. Among the identified transcription factors, AGAMOUS (AG) and CRABS CLAW (CRC) may be the two core regulatory genes for carpel origin as they determine carpel organ identity, determinacy, and functionality. Therefore, a comparative identification of their protein-protein interactions and downstream target genes between flowering and non-flowering plants from an evo-devo perspective may be primary projects for elucidating carpel origin and development.

A blast from the past: Understanding stem cell specification in plant roots using laser ablation.

In the Arabidopsis root, growth is sustained by the meristem. Signalling from organiser cells, also termed the quiescent centre (QC), is essential for the maintenance and replenishment of the stem cells. Here, we highlight three publications from the founder of the concept of the stem cell niche in Arabidopsis and a pioneer in unravelling regulatory modules governing stem cell specification and maintenance, as well as tissue patterning in the root meristem: Ben Scheres. His research has tremendously impacted the plant field. We have selected three publications from the Scheres legacy, which can be considered a breakthrough in the field of plant developmental biology. van den Berg et al. (1995) and van den Berg et al. (1997) uncovered that positional information-directed patterning. Sabatini et al. (1999), discovered that auxin maxima determine tissue patterning and polarity. We describe how simple but elegant experimental designs have provided the foundation of our current understanding of the functioning of the root meristem.

Live Fluorescence Visualization of Cellulose and Pectin in Plant Cell Walls.

The plant cell wall comprises various types of macromolecules whose abundance and spatial distribution change dynamically and are crucial for plant architecture. High-resolution live cell imaging of plant cell wall components is, therefore, a powerful tool for plant cell biology and plant developmental biology. To acquire suitable data, the experimental setup for staining and imaging of non-fixed samples must be straightforward and avoid creating stress-induced artifacts. We present a detailed sample preparation and live image acquisition protocol for fluorescence visualization of cell wall components using commercially available probes and stains.

An in-frame deletion mutation in the degron tail of auxin coreceptor IAA2 confers resistance to the herbicide 2,4-D in Sisymbrium orientale.

The natural auxin indole-3-acetic acid (IAA) is a key regulator of many aspects of plant growth and development. Synthetic auxin herbicides such as 2,4-D mimic the effects of IAA by inducing strong auxinic-signaling responses in plants. To determine the mechanism of 2,4-D resistance in a Sisymbrium orientale (Indian hedge mustard) weed population, we performed a transcriptome analysis of 2,4-D-resistant (R) and -susceptible (S) genotypes that revealed an in-frame 27-nucleotide deletion removing nine amino acids in the degron tail (DT) of the auxin coreceptor Aux/IAA2 (SoIAA2). The deletion allele cosegregated with 2,4-D resistance in recombinant inbred lines. Further, this deletion was also detected in several 2,4-D-resistant field populations of this species. Arabidopsis transgenic lines expressing the SoIAA2 mutant allele were resistant to 2,4-D and dicamba. The IAA2-DT deletion reduced binding to TIR1 invitro with both natural and synthetic auxins, causing reduced association and increased dissociation rates. This mechanism of synthetic auxin herbicide resistance assigns an in planta function to the DT region of this Aux/IAA coreceptor for its role in synthetic auxin binding kinetics and reveals a potential biotechnological approach to produce synthetic auxin-resistant crops using gene-editing.

AtCube: Performing pathogen-root infection tests on Arabidopsis thaliana in a completely controlled way

Arabidopsis thaliana represents the main model plant for the experimental analysis of plant genetics and developmental biology of photoautotrophic organisms, due to characteristics such as size, life cycle, fecundity, genetics and its easy experimental manipulation. In turn, A. thaliana is used as a model plant in plant-microorganism interaction studies, being of great importance in the knowledge of the form of infection of numerous plant pathogens. The present work is based on the development of an A. thaliana plant growth system in Phytatray II boxes and culture substrate, trying to establish a system that is free of contamination, completely controlled, self-sufficient, reproducible and standardized, called AtCube. Through the use of different necrotrophic and biotrophic/hemibiotrophic root-pathogens, bacteria, fungi and oomycetes, and the comparison with a conventional system of growth in plots, various results are analyzed on the effects of pathogens on plants and the advantages and differences of the AtCube system. This work makes it possible to highlight that the AtCube system represents a methodology that allows obtaining results similar to other systems but with important advantages regarding its standardization, rigor and reproducibility with respect to studies with A. thaliana and root pathogens. Furthermore, it could represent an equally efficient system in studies with other plant species and the application of foliar pathogens and/or beneficial microorganisms.

The role of genus and life span in predicting seed and vegetative trait variation and correlation in Lathyrus, Phaseolus, and Vicia.

PremiseAnnual and perennial life history transitions are abundant among angiosperms, and understanding the phenotypic variation underlying life span shifts is a key endeavor of plant evolutionary biology. Comparative analyses of trait variation and correlation networks among annual and perennial plants is increasingly important as new herbaceous perennial crops are being developed for edible seed. However, it remains unclear how seed to vegetative growth trait relationships correlate with life span.MethodsTo assess the relative roles of genus and life span in predicting phenotypic variation and trait correlations, we measured seed size and shape, germination proportion, and early‐life‐stage plant height and leaf growth over 3 mo in 29 annual and perennial, herbaceous congeneric species from three legume genera (Lathyrus, Phaseolus, and Vicia).ResultsGenus was the strongest predictor of seed size and shape variation, and life span consistently predicted plant height and leaf number at single time points. Correlation networks revealed that annual species had significant associations between seed traits and vegetative traits, whereas perennials had no significant seed‐vegetative associations. Each genus also differed in the extent of integration between seed and vegetative traits, as well as within‐vegetative‐trait correlation patterns.ConclusionsGenus and life span were important for predicting aspects of early‐life‐stage phenotypic variation and trait relationships. Differences in phenotypic correlation may indicate that selection on seed size traits will impact vegetative growth differently depending on life span, which has important implications for nascent perennial breeding programs.

An in situ sequencing approach maps PLASTOCHRON1 at the boundary between indeterminate and determinate cells.

The plant shoot apex houses the shoot apical meristem, a highly organized and active stem-cell tissue where molecular signaling in discrete cells determines when and where leaves are initiated. We optimized a spatial transcriptomics approach, in situ sequencing (ISS), to colocalize the transcripts of 90 genes simultaneously on the same section of tissue from the maize (Zea mays) shoot apex. The RNA ISS technology reported expression profiles that were highly comparable with those obtained by in situ hybridizations (ISHs) and allowed the discrimination between tissue domains. Furthermore, the application of spatial transcriptomics to the shoot apex, which inherently comprised phytomers that are in gradual developmental stages, provided a spatiotemporal sequence of transcriptional events. We illustrate the power of the technology through PLASTOCHRON1 (PLA1), which was specifically expressed at the boundary between indeterminate and determinate cells and partially overlapped with ROUGH SHEATH1 and OUTER CELL LAYER4 transcripts. Also, in the inflorescence, PLA1 transcripts localized in cells subtending the lateral primordia or bordering the newly established meristematic region, suggesting a more general role of PLA1 in signaling between indeterminate and determinate cells during the formation of lateral organs. Spatial transcriptomics builds on RNA ISH, which assays relatively few transcripts at a time and provides a powerful complement to single-cell transcriptomics that inherently removes cells from their native spatial context. Further improvements in resolution and sensitivity will greatly advance research in plant developmental biology.

International Association for Plant Taxonomy 2021 Stebbins Medal

Luke T. Dunning, Jill K. Olofsson, Christian Parisod, Rimjhim R. Choudhury, Jose J. Moreno-Villena, Yang Yang, Jacqueline Dionora, W. Paul Quick, Minkyu Park, Jeffrey L. Bennetzen, Guillaume Besnard, Patrik Nosil, Colin P. Osborne & Pascal-Antoine Christin. 2019. Lateral transfers of large DNA fragments spread functional genes among grasses. Proc. Natl. Acad. Sci. U.S.A. 116(10): 4416–4425. https://www.pnas.org/content/116/10/4416.short The Stebbins Medal was first awarded in 2003 in honour of the American botanist and geneticist George Ledyard Stebbins (1906–2000), one of the world's leading plant evolutionary biologists of the 20th century. The medal is awarded biennially by IAPT for an outstanding paper in phylogenetic systematics and evolution of plants. The 2021 award considered papers published in 2018 and 2019. In their paper, Dunning & al. presented the most thorough and far-reaching study to date that convincingly demonstrates extensive lateral gene transfer (LGT) in plants. By comparing genomes of many grasses, they showed that large blocks of DNA containing functional genes are laterally passed among species that are distantly related across the grass phylogeny. Through stringent phylogenomic analyses, they discovered 59 LGTs clustered on 23 laterally acquired genomic fragments that are up to 170 kb long in the genome of the grass Alloteropsis semialata (R.Br.) Hitchc., involving at least nine different donor species. They also showed that some of these laterally transferred genes have added functions to the recipient species genomes. This suggests that LGT represents a potent evolutionary force capable of spreading functional genes among distantly related lineages of grasses. Luke Dunning (Fig. 1), the first author of the 2021 Stebbins Medal paper, studied for his Bachelor's Degree at Cardiff University, Wales, United Kingdom and his Ph.D. at the University of Auckland in New Zealand. Since October 2020, he holds a Natural Environment Research Council (NERC) Independent Research Fellowship in the Department of Animal and Plant Sciences at the University of Sheffield, U.K. Previously, he worked as a postdoctoral researcher with Pascal-Antoine Christin, the senior author of the winning paper, also at the University of Sheffield. Luke's work focuses on the genomics of rapid adaptation in grasses and combines cutting-edge genomic techniques, comparative analyses and experimental approaches to understand how organisms adapt to their environment (https://dunning-lab.group.shef.ac.uk). Many congratulations to Luke Dunning and colleagues! The Stebbins Award committee comprises Paula Rudall (U.K.), Loren Rieseberg (Canada) and Colin Hughes, Chair (Switzerland). The IAPT Council is grateful to the committee for their hard work in making this outstanding selection.