Active Control of Cell Size Generates Spatial Detail during Plant Organogenesis

SummaryHow cells regulate their dimensions is a long-standing question [1, 2]. In fission and budding yeast, cell-cycle progression depends on cell size, although it is still unclear how size is assessed [3–5]. In animals, it has been suggested that cell size is modulated primarily by the balance of external signals controlling growth and the cell cycle [1], although there is evidence of cell-autonomous control in cell cultures [6–9]. Regardless of whether regulation is external or cell autonomous, the role of cell-size control in the development of multicellular organisms remains unclear. Plants are a convenient system to study this question: the shoot meristem, which continuously provides new cells to form new organs, maintains a population of actively dividing and characteristically small cells for extended periods [10]. Here, we used live imaging and quantitative, 4D image analysis to measure the sources of cell-size variability in the meristem and then used these measurements in computer simulations to show that the uniform cell sizes seen in the meristem likely require coordinated control of cell growth and cell cycle in individual cells. A genetically induced transient increase in cell size was quickly corrected by more frequent cell division, showing that the cell cycle was adjusted to maintain cell-size homeostasis. Genetically altered cell sizes had little effect on tissue growth but perturbed the establishment of organ boundaries and the emergence of organ primordia. We conclude that meristem cells actively control their sizes to achieve the resolution required to pattern small-scale structures

The Same Regulatory Point Mutation Changed Seed-Dispersal Structures in Evolution and Domestication

SummaryIt is unclear whether gene regulatory changes that drive evolution at the population and species levels [1–3] can be extrapolated to higher taxonomic levels [4, 5]. Here, we investigated the role of cis-regulatory changes in fruit evolution within the Brassicaceae family. REPLUMLESS (RPL, At5g02030) controls development of the replum, a structure with an important role in fruit opening and seed dispersal [6]. We show that reduced repla resembling the Arabidopsis rpl mutant correlated across the Brassicaceae with a point mutation in a conserved cis-element of RPL. When introduced in Arabidopsis, this nucleotide change specifically reduced RPL expression and function in the fruit. Conversely, Brassica RPL containing the Arabidopsis version of the cis-element was sufficient to convert the Brassica replum to an Arabidopsis-like morphology. A mutation in the same nucleotide position of the same cis-element in a RPL ortholog has been independently selected to reduce seed dispersal during domestication of rice [7], in spite of its very different fruit anatomy. Thus, single-nucleotide regulatory mutations at the same position explain developmental variation in seed-dispersal structures at the population and family levels and suggest that the same genetic toolkit is relevant to domestication and natural evolution in widely diverged species

A flower-specific Myb protein activates transcription of phenylpropanoid biosynthetic genes

10 pages, 9 figures.-- PMID: 8306956 [PubMed].-- PMCID: PMC394786.Synthesis of flavonoid pigments in flowers requires the co-ordinated expression of genes encoding enzymes in th phenylpropanoid biosynthetic pathway. Some cis-elements involved in the transcriptional control of these genes have been defined. We report binding of petal-specific activities from tobacco and Antirrhinum majus (snapdragon) to an element conserved in promoters of phenylpropanoid biosynthetic genes and implicated in expression in flowers. These binding activities were inhibited by antibodies raised against Myb305, a flower-specific Myb protein previously cloned from Antirrhinum by sequence homology. Myb305 bound to the same element and formed a DNA-protein complex with the same mobility as the Antirrhinum petal protein in electrophoretic mobility shift experiments. Myb305 activated expression from its binding site in yeast and in tobacco protoplasts. In protoplasts, activation also required a G-box-like element, suggesting co-operation with other elements and factors. The results strongly suggest a role for Myb305-related proteins in the activation of phenylpropanoid biosynthetic genes in flowers. This is consistent with the genetically demonstrated role of plant Myb proteins in the regulation of genes involved in flavonoid synthesis.We are grateful to Dr V.Chandler (University of Oregon) for anti-Cl serum, to Dr J.Lipsick (Stanford University) for anti-Cl and anti-v-Myb sera, to C.Smith for tobacco transformations, to Dr J.Munoz-Blanco and Dr M.Holdsworth for advice and vectors for yeast transformation, to Dr D.Hatton for advice on tobacco protoplast transformation, to Dr V.Hocher and Y.Kishima for access to unpublished data. R.W.M.S. received an EC pre-doctoral fellowship (ECLAIR programme, contract AGRE913013), E.M. was supported by a FEBS Fellowship and F.C-M. was supported by the EC Bridge Programme (contract BIOT0164CEDB).Peer reviewe

Spatiotemporal coordination of cell division and growth during organ morphogenesis

A developing plant organ exhibits complex spatiotemporal patterns of growth, cell division, cell size, cell shape, and organ shape. Explaining these patterns presents a challenge because of their dynamics and cross-correlations, which can make it difficult to disentangle causes from effects. To address these problems, we used live imaging to determine the spatiotemporal patterns of leaf growth and division in different genetic and tissue contexts. In the simplifying background of the speechless (spch) mutant, which lacks stomatal lineages, the epidermal cell layer exhibits defined patterns of division, cell size, cell shape, and growth along the proximodistal and mediolateral axes. The patterns and correlations are distinctive from those observed in the connected subepidermal layer and also different from the epidermal layer of wild type. Through computational modelling we show that the results can be accounted for by a dual control model in which spatiotemporal control operates on both growth and cell division, with cross-connections between them. The interactions between resulting growth and division patterns lead to a dynamic distributions of cell sizes and shapes within a deforming leaf. By modulating parameters of the model, we illustrate how phenotypes with correlated changes in cell size, cell number, and organ size may be generated. The model thus provides an integrated view of growth and division that can act as a framework for further experimental study

The Tarantula Massive Binary Monitoring

We present the first SB2 orbital solution and disentanglement of the massive Wolf-Rayet binary R145 (P = 159d) located in the Large Magellanic Cloud. The primary was claimed to have a stellar mass greater than 300Msun, making it a candidate for the most massive star known. While the primary is a known late type, H-rich Wolf-Rayet star (WN6h), the secondary could not be so far unambiguously detected. Using moderate resolution spectra, we are able to derive accurate radial velocities for both components. By performing simultaneous orbital and polarimetric analyses, we derive the complete set of orbital parameters, including the inclination. The spectra are disentangled and spectroscopically analyzed, and an analysis of the wind-wind collision zone is conducted. The disentangled spectra and our models are consistent with a WN6h type for the primary, and suggest that the secondary is an O3.5 If*/WN7 type star. We derive a high eccentricity of e = 0.78 and minimum masses of M1 sin^3 i ~ M2 sin^3 i ~ 13 +- 2 Msun, with q = M2 / M1 = 1.01 +- 0.07. An analysis of emission excess stemming from a wind-wind collision yields a similar inclination to that obtained from polarimetry (i = 39 +- 6deg). Our analysis thus implies M1 = 53^{+40}_{-20} and M2 = 54^{+40}_{-20} Msun, excluding M1 > 300Msun. A detailed comparison with evolution tracks calculated for single and binary stars, as well as the high eccentricity, suggest that the components of the system underwent quasi-homogeneous evolution and avoided mass-transfer. This scenario would suggest current masses of ~ 80 Msun and initial masses of Mi,1 ~ 105 and Mi,2 ~ 90Msun, consistent with the upper limits of our derived orbital masses, and would imply an age of ~2.2 Myr.Comment: Accepted for Publication in A&A, 16 pages, 17 figures and 4 table

Cycling in a crowd: coordination of plant cell division, growth, and cell fate

The reiterative organogenesis that drives plant growth relies on the constant production of new cells, which remain encased by interconnected cell walls. For these reasons, plant morphogenesis strictly depends on the rate and orientation of both cell division and cell growth. Important progress has been made in recent years in understanding how cell cycle progression and the orientation of cell divisions are coordinated with cell and organ growth and with the acquisition of specialized cell fates. We review basic concepts and players in plant cell cycle and division, and then focus on their links to growth-related cues, such as metabolic state, cell size, cell geometry, and cell mechanics, and on how cell cycle progression and cell division are linked to specific cell fates. The retinoblastoma pathway has emerged as a major player in the coordination of the cell cycle with both growth and cell identity, while microtubule dynamics are central in the coordination of oriented cell divisions. Future challenges include clarifying feedbacks between growth and cell cycle progression, revealing the molecular basis of cell division orientation in response to mechanical and chemical signals, and probing the links between cell fate changes and chromatin dynamics during the cell cycleEuropean Commission (ERC-2018-AdG_833617), the Ministry of Science and Innovation (RTI2018-094793-B-I00) and institutional grants from Fundación Ramón Areces and Banco de Santander to the Centro de Biologia Molecular Severo OchoaPeer reviewe