GARNet Research Roundup: April 11th 2018

This weeks GARNet research roundup begins with a microscopy-based study led by Lorenzo Frigerio from the University of Warwick that investigates the origin of Protein Storage Vacuoles. The second paper from John Doonan at Aberystwyth University looks at how differential splicing of cyclin-dependent Kinase G1 effects the thermosensory response. Reiner van de Hoorn from Oxford leads the next paper that characterises the use of activity-based protein profiling (ABPP) to identify novel α-glycosidases in model and non-model plants. Simon McQueen-Mason from York is corresponding author of the next paper that identified a new QTL from Brachypodium that is involved in cell wall formation. The fifth paper is led by Anthony Dodd from Bristol and characterises the role of the SnRK1 complex in hypocotyl elongation whilst the penultimate manuscript from Julia Davies’s lab in Cambridge performs patch clamp analysis of dorn1 mutant plants. The final paper from Brendan Davies at the University of Leeds characterises the SMG kinase, a gene that is lacking from the Arabidopsis genome, in Physcomitrella patens.


http://www.plantphysiol.org/content/early/2018/03/19/pp.18.00010.long

Feeney M, Kittelmann M, Menassa R, Hawes C, Frigerio L. Protein storage vacuoles originate from remodelled pre-existing vacuoles in Arabidopsis thaliana (2018) Plant Physiol. 2018 Mar 19. pii: pp.00010.2018. doi: 10.1104/pp.18.00010 Open Access

This collaboration between the Universities of Warwick and Oxford Brookes is led by Lorenzo Frigerio and Chris Hawes. They have investigated the origin of seed Protein Storage Vacuoles (PSV) using a two-pronged approach using confocal and immunoelectron microscopy. They looked at embryo development as well as in leaf cells that have been reprogrammed for embryonic cell fate by overexpression of the LEAFY COTYLEDON2 TF. These studies indicate that PSVs are formed following the reprogramming of pre-existing embryonic vacuole (EV) rather than from de novo assembly.


https://onlinelibrary.wiley.com/doi/abs/10.1111/tpj.13914

Cavallari N, Nibau C, Fuchs A, Dadarou D, Barta A, Doonan JH. The Cyclin Dependent Kinase G group defines a thermo-sensitive alternative splicing circuit modulating the expression of Arabidopsis ATU2AF65A (2018) Plant J. doi: 10.1111/tpj.13914 Open Access

John Doonan (Aberystwyth University) is the corresponding author on this UK-Austrian collaboration that presents the role of the cyclin-dependent Kinase G1 (CDKG1) in thermosensing in Arabidopsis. Ambient temperature change causes altered gene expression of the spliceosome component, ATU2AF65A. Interestingly the CDKG1 gene is differentially spliced and to produces two protein isoforms that are both needed to complement the expression of ATU2AF65A across a temperature range. This alternative splicing is dependent on CDKG2 and CYCLIN L1 and is a novel control mechanism in the temperature control response.


Husaini AM, Morimoto K, Chandrasekar B, Kelly S, Kaschani F, Palmero D, Jiang J, Kaiser M, Ahrazem O, Overkleeft HS, van der Hoorn RAL (2018) Multiplex fluorescent, activity-based protein profiling identifies active α-glycosidases and other hydrolases in plants. Plant Physiol. pii: pp.00250.2018. doi: 10.1104/pp.18.00250 Open Access

Renier Van de Hoorn (University of Oxford) leads this pan-european study that uses novel cyclophellitol aziridine probes that label α-glycosidase enzymes. They identified two novel α-glycosidases in Arabidopsis as well as using the technique in non-model saffron crocus. Finally they showed that this multiplex fluorescent labelling in combination with probes for serine hydrolases and cysteine proteases can be used to identify changes in hydrolase activity in response to pathogen infection.


Whitehead C, Ostos Garrido FJ, Reymond M, Simister R, Distelfeld A, Atienza SG, Piston F, Gomez LD, McQueen-Mason SJ (2018) A glycosyl transferase family 43 protein involved in xylan biosynthesis is associated with straw digestibility in Brachypodium distachyon. New Phytol. doi: 10.1111/nph.15089 Open Access

Simon McQueen-Mason (University of York) leads this study that use QTL mapping to identify a gene in Bracypodium that is involved in cell wall architecture, which might then be a target to develop plants with improved cellulose digestibility. This glycosyl transferase family (GT) 43 protein is an orthologue of Arabidopsis IRX14, which is involved in xylan biosynthesis. When RNAi was used to reduce expression of this gene the resulting plants showed increased digestibility, indicating that this BdGT43A will be a good target for future breeding plans.


Wang L, Wilkins KA, Davies JM (2018) Arabidopsis DORN1 extracellular ATP receptor; activation of plasma membrane K(+) -and Ca(2+) -permeable conductances New Phytol. 2018 Mar 25. doi: 10.1111/nph.15111. Open Access

This letter to New Phytologist from the lab of Julia Davis (University of Cambridge) outlines some experiments to determine whether the DORN1 plasma membrane receptor is responsible for transmitting a signal from extracellular ATP (eATP). They performed patch clamp analysis on isolated protoplasts and showed that DORN1 is involved in the activation of Ca+ and K+ pumps by eATP as, in contrast to wildtype, dorn1 mutant protoplast showed no voltage changes after incubation with eATP.


Simon NML, Sawkins E, Dodd AN. Involvement of the SnRK1 subunit KIN10 in sucrose-induced hypocotyl elongation (2018) Plant Signal Behav. 27:1-9. doi: 10.1080/15592324.2018.1457913.

Anthony Dodd (University of Bristol) is the corresponding author of this follow-on study from one that previously featured on the GARNet YouTube channel. This study measures sucrose-induced hypocotyl elongation in two T-DNA mutants of the SnRK1 subunit KIN10 gene. These mutants had altered responses to sucrose leading to the hypothesis that the SnRK1 complex suppresses hypocotyl elongation in the presence of external sugar.


Lloyd JPB, Lang D, Zimmer AD, Causier B, Reski R, Davies B (2018) The loss of SMG1 causes defects in quality control pathways in Physcomitrella patens. Nucleic Acids Res. doi: 10.1093/nar/gky225 Open Access

Brendan Davis (University of Leeds) is the corresponding author on research that investigates the role of the SMG1 kinase during nonsense-mediated mRNA decay (NMD) in the moss Physcomitrella patens. This kinase plays a critical role in animals but as it is not present in Arabidopsis, its function is not well studied in plants. However moss smg mutants show expression changes in genes involved in a variety of processes indicating that NMD is a common control mechanism in moss. In addition these plants have increased susceptibility to DNA damage, which suggests that the SMG1 kinase is a key player in quality control mechanisms in plants.

https://academic.oup.com/nar/advance-article/doi/10.1093/nar/gky225/4955258

GARNet Research Roundup: March 29th

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Published on: March 29, 2018

This edition of the GARNet research roundup begins with an analysis of the CELLULOSE SYNTHASE COMPLEX led by Simon Turner in Manchester. Next are two papers from Ian Henderson at Cambridge who, in collaboration with Rob Martienssen in the USA, has investigated the epigenetic factors that control meiotic recombination. Next are two papers led by Hugh Nimmo (Glasgow) who is researching alternative splicing of the LATE ELONGATED HYPOCOTYL transcript. Marc Vendrell and Karl Oparka at the University of Edinburgh are co-authors in the next paper that investigates the binding specificity of the AtSUC2 protein. The seventh paper includes David Salt (Nottingham) as a co-author in an investigation in the plants response to zinc. The final three papers are methods papers on gravitropism, ChIP-Seq and calcium sensing from Nottingham University, SLCU and the John Innes Centre respectively.


Kumar M, Mishra L, Carr P, Pilling M, Gardner P, Mansfield SD, Turner SR (2018) Exploiting CELLULOSE SYNTHASE (CESA) class-specificity to probe cellulose microfibril biosynthesis. Plant Physiol. doi: 10.1104/pp.18.00263 Open Access

Simon Turner (University of Manchester) leads this study that investigates the subunit specificity of the CELLULOSE SYNTHASE COMPLEX, which is composed of many CESA components. Mutant cesa plants were used to probe the specificity of these subunits. Overall the authors found that CESA classes have similar roles in determining cellulose microfibril structure but that the rates of cellulose synthesis might be altered in a subunit-specific manner.


Choi K,, Zhao X, Tock AJ, Lambing C, Underwood CJ,, Hardcastle TJ, Serra H, Kim J, Cho HS, Kim J, Ziolkowski PA, Yelina NE, Hwang I, Martienssen RA, Henderson IR (2018) Nucleosomes and DNA methylation shape meiotic DSB frequency in Arabidopsis thaliana transposons and gene regulatory regions. Genome Res. doi: 10.1101/gr.225599.117
The research groups of Ian Henderson (University of Cambridge) and Rob Martienssen (CSHL) co-lead back-to-back papers that investigate the factors that influence meiotic recombination frequencies. The Henderson led-paper focuses on the position of the SPO11 topoisomerase and the epigenetic factors, such as H3K4me3 and DNA methylation that reside in those areas. They discovered some surprising relationships between SPO11 binding and different transposon classes.


Underwood CJ, Choi K, Lambing C, Zhao X, Serra H, Borges F, Simorowski J, Ernst E, Jacob Y, Henderson IR, Martienssen RA (2018) Epigenetic activation of meiotic recombination near Arabidopsis thaliana centromeres via loss of H3K9me2 and non-CG DNA methylation. Genome Res. doi: 10.1101/gr.227116.117

Open Access

The Martienssen–led paper focuses on epigenetic marks, such as H3K9me2 and non-CG DNA methylation that reside at pericentromeric regions. By altering the distribution of these marks, the amount of pericentrometric recombination can be changed and that the number of double stranded breaks increase in H3K9me2/non-CG mutants.


James AB, Sullivan S, Nimmo HG (2018) Global spatial analysis of Arabidopsis natural variants implicates 5’UTR splicing of LATE ELONGATED HYPOCOTYL in responses to temperature. Plant Cell Environment. doi: 10.1111/pce.13188

James AB, Calixto CPG, Tzioutziou NA, Guo W, Zhang R, Simpson CG, Jiang W, Nimmo GA, Brown JWS, Nimmo HG (2018) How does temperature affect splicing events? Isoform switching of splicing factors regulates splicing of LATE ELONGATED HYPOCOTYL (LHY). Plant Cell Environ. doi: 10.1111/pce.13193

The first of these back-to-back papers is led by Hugh Nimmo (Glasgow) in a study that characterises a set of 5’UTRs in the LATE ELONGATED HYPOCOTYL (LHY) gene and how they change in response to temperature. This is linked to a correlation of how these LHY haplotypes are global distributed.

The second paper is an extension of this study and includes Hugh Nimmo (Glasgow) and John Brown (JHI, Dundee) as co-corresponding authors. They that show RNA-binding splicing factors (SFs) are necessary for temperature-induced changes in the LHY transcript. LHY might be considered a molecular thermostat whose splicing can response to changes as little as 2°C.


De Moliner F, Knox K, Reinders A, Ward J, McLaughlin P, Oparka K, Vendrell M (2018) Probing binding specificity of the sucrose transporter AtSUC2 with fluorescent coumarin glucosides. J Exp Bot. doi: 10.1093/jxb/ery075 Open Access

Marc Vendrell and Karl Oparka (University of Edinburgh) are the corresponding authors on this bioimaging study that probes the specificity of the AtSUC2 phloem sucrose transporter. They use structural varieties in coumarin glucosides to precisely define the binding characteristics of AtSUC2.


Chen ZR, Kuang L, Gao YQ, Wang YL, Salt DE, Chao DY (2018) AtHMA4 Drives Natural Variation in Leaf Zn Concentration of Arabidopsis thaliana. Front Plant Sci. doi: 10.3389/fpls.2018.00270

Open Access

David Salt (University of Nottingham) is a co-author on this Chinese study that characterises the role of the Heavy Metal-ATPase 4 (HMA4) in the respond to zinc.


Muller L, Bennett MJ, French A, Wells DM, Swarup R (2018) Root Gravitropism: Quantification, Challenges, and Solutions. Methods Mol Biol. doi: 10.1007/978-1-4939-7747-5_8

Ranjan Swarup (University of Nottingham) leads this methods paper that describes techniques for the automated measurement of root gravitropic responses.


Cortijo S, Charoensawan V, Roudier F, Wigge PA (2018) Chromatin Immunoprecipitation Sequencing (ChIP-Seq) for Transcription Factors and Chromatin Factors in Arabidopsis thaliana Roots: From Material Collection to Data Analysis. Methods Mol Biol. doi: 10.1007/978-1-4939-7747-5_18

Phillip Wigge (SLCU) leads this methods paper that outlines the technical details for the now common and important technique of ChIP-Seq from Arabidopsis roots.


Kelner A, Leitão N, Chabaud M, Charpentier M, de Carvalho-Niebel F (2018) Dual Color Sensors for Simultaneous Analysis of Calcium Signal Dynamics in the Nuclear and Cytoplasmic Compartments of Plant Cells. Front Plant Sci. doi: 10.3389/fpls.2018.00245 Open Access

Miriam Charpentier (John Innes Centre) is a co-author on this work that uses fluorescent protein-based Ca2+ sensors, the GECOs, to successfully monitor the calcium response to a range of biotic and abiotic elicitors. These GECO-based sensors represent an exciting new tool for the study of calcium dynamics.

Arabidopsis Research Roundup: March 9th

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Published on: March 9, 2018

This Arabidopsis Research Roundup has five papers that includes two from the John Innes Centre and two from the University of Edinburgh. Firstly Kristen Bomblies’s group at the JIC have investigated the relationship between temperature and meiotic recombination rates. Secondly Veronica Grieneisen and Stan Maree have developed a mathematical model to characterise cell morphologies taken from a 2D image. Andrew Miller from Edinburgh is a co-corresponding author on a study that shows how the Arabidopsis proteome changes in different photoperiods. In the fourth paper Peter Doerner is a co-author on work that looks at the phosphate starvation response. Finally researchers from Bristol and Nottingham contribute to an investigation into a novel genetic component that controls auxin-induced root hair development.


Lloyd A, Morgan C, Franklin C, Bomblies K (2018) Plasticity of Meiotic Recombination Rates in Response to Temperature in Arabidopsis. Genetics. doi: 10.1534/genetics.117.300588

Open Access

Kristen Bomblies (John Innes Centre) leads this study that investigates the influence of temperature on meiotic recombination rate. They show that in Arabidopsis the number of crossovers positively correlates with increasing temperature. However the mechanistic explanation for the increase at higher temperatures remains opaque as, in contrast to findings from other plants, synaptonemal complex length negatively correlates with temperature.


Sánchez-Corrales YE, Hartley M, van Rooij J, Marée AFM, Grieneisen VA (2018) Morphometrics of complex cell shapes: Lobe Contribution Elliptic Fourier Analysis (LOCO-EFA). Development. doi: 10.1242/dev.15677

http://dev.biologists.org/content/early/2018/02/08/dev.156778.long

Open Access

Veronica Grieneisen and Stan Maree (John Innes Centre) lead this study that has developed the Lobe Contribution Elliptical Fourier Analysis (LOCO-EFA) method. This generates meaningful descriptors from a 2D image of cells that can then be linked to morphological features. This tool allows for the efficient phenotyping of cell morphologies that they demonstrate by analysing images of Arabidopsis leaf pavement cells. They extend this analysis to larger populations where they used LOCO-EFA to predict how cell shapes change when they move into a more crowded space.


Seaton DD, Graf A, Baerenfaller K, Stitt M, Millar AJ, Gruissem W (2018) Photoperiodic control of the Arabidopsis proteome reveals a translational coincidence mechanism. Mol Syst Biol. doi: 10.15252/msb.20177962 Open Access

http://onlinelibrary.wiley.com/doi/10.15252/msb.20177962/abstract

Andrew Miller (University of Edinburgh) is the corresponding author on this collaboration with German and Swiss colleagues that compares the Arabidopsis proteome across four photoperiods. They shows coordinated changes across the proteome, most notably at longer photoperiods in the abundance of proteins involved in photosynthesis and metabolism. They show higher translation rates during the day that correspond with the increased RNA abundance that is a characteristic of circadian rhythms. This ‘translational coincidence’ describes the alignment of higher translation rates with high transcript levels and they assigned a mathematical model in an attempt to explain this phenomenon.


Hanchi M, Thibaud MC, Légeret B, Kuwata K, Pochon N, Beisson F, Cao A, Cuyas L, David P, Doerner P, Ferjani A, Lai F, Li-Beisson Y, Mutterer J, Philibert M, Raghothama KG, Rivasseau C, Secco D, Whelan J, Nussaume L, Javot H (2018) The phosphate fast-responsive genes PECP1 and PPsPase1 affect phosphocholine and phosphoethanolamine content. Plant Physiol. doi: 10.1104/pp.17.01246 Open Access

Peter Doerner (University of Edinburgh) is a co-author on this global study that characterises the phosphate starvation-mediated induction of the HAD-type phosphatases PPsPase1 (AT1G73010) and PECP1 (AT1G17710). They show that expression of these genes closely follows phosphate status but that their activity does not alter phospate content. The role of these proteins is to control phosphocholine and phosphoethanolamine content, which is a output of changing phosphate conditions. The authors conclude that expression of these genes can be an excellent molecular marker for the phosphate starvation response.


www.cell.com/current-biology/fulltext/S0960-9822(18)30083-6

Schoenaers S, Balcerowicz D, Breen G, Hill K, Zdanio M, Mouille G, Holman TJ, Oh J, Wilson MH, Nikonorova N, Vu LD, De Smet I, Swarup R, De Vos WH, Pintelon I, Adriaensen D, Grierson C, Bennett MJ, Vissenberg K (2018) The Auxin-Regulated CrRLK1L Kinase ERULUS Controls Cell Wall Composition during Root Hair Tip Growth. Current Biology doi: 10.1016/j.cub.2018.01.050

This Belgian-led study includes contributions from Claire Greirson’s and Malcolm Bennett’s labs in Bristol and Nottingham respectively. They investigate the role of the ERULUS (ERU) protein, an auxin-induced receptor-like kinase, during the development of root hairs. ERU localises to the apical root hair plasma membrane and regulates cell wall composition by altering pectin dynamic. The authors conclude that ERU is a key regulator of auxin-mediated control of root hair development.

Arabidopsis Research Roundup: February 22nd 2018

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Published on: February 22, 2018

This edition of the Arabidopsis Research roundup beings with a study from SLCU that provides a molecular context to the changes that occur at graft junctions. Second is a study from Edinburgh that reports on the findings of a citizen science plant phenotyping project. Third are two studies from the John Innes Centre that follow-on from previous studies. These characterise the molecular response to seasonal transitions and the factors that control floral development.

The fifth paper is led by Chris Hawes at Oxford Brookes and characterises a novel sub-group of ER localized reticulon proteins. The next paper from the University of Sheffield looks at the whole plant response to changing global carbon dioxide concentrations. The seventh paper from Bristol and York also broadly looks at CO2 but this time at the molecular factors that control stomatal closure in response to both ABA and CO2 signals. Christine Foyer (Leeds) is a co-author on the penultimate paper that characterises the role of ascorbic acid in hormone signaling whilst the final paper from Julian Hibberd at the University of Cambridge analyses a regulatory element that contributes to the evolutionary transition to C4 photosynthesis.


Melnyk CW, Gabel A, Hardcastle TJ, Robinson S, Miyashima S, Grosse I, Meyerowitz EM (2018) Transcriptome dynamics at Arabidopsis graft junctions reveal an intertissue recognition mechanism that activates vascular regeneration. Proc Natl Acad Sci U S A. doi: 10.1073/pnas.1718263115 Open Access
This research was led by Charles Melynk during this time working with Elliot Meyerowitz at SLCU. Grafting is an important classic and contemporary technique in plant biology and this study investigates the gene expression changes that occur on either side of a graft junction. They show asymmetry changes in gene expression on either side of a graft that include an increase in vascular initiation but only in grafted tissues and not in those that are cut and then separated. This study provides an exciting insight into the molecular changes that occur during tissue grafting.


Giuffrida MV, Chen F, Scharr H, Tsaftaris SA (2018) Citizen crowds and experts: observer variability in image-based plant phenotyping. Plant Methods. doi: 10.1186/s13007-018-0278-7 Open Access

This UK, German and Italian study is led by Sotirios Tsaftaris at the University of Edinburgh and reports on a fascinating citizen science study that evaluated the ability of experts and non-experts to use plant phenotyping software. They demonstrate that non-experts can be effectively involved in plant phenotyping annotation tasks given enough statistical power and if the study is suitably designed.


Hepworth J, Antoniou-Kourounioti RL, Bloomer RH, Selga C, Berggren K, Cox D, Collier Harris BR, Irwin JA, Holm S, Säll T,Howard M, Dean C (2018) Absence of warmth permits epigenetic memory of winter in Arabidopsis. Nat Commun. doi: 10.1038/s41467-018-03065-7 Open Access

Caroline Dean and Martin Howard (John Innes Centre) lead this study that further characterise the relationship between the VERNALIZATION INSENSITIVE3 (VIN3) an FLOWERING LOCUS C (FLC) genes in two separate thermosensory processes that monitor long term temperature changes. They suggest that the regulatory strategies currently employed by plants might become less effective as the climate becomes more variable and will have a knock-on effect on plant growth and productivity.


Simonini S, Stephenson P, Østergaard L (2018) A molecular framework controlling style morphology in Brassicaceae. Development. doi: 10.1242/dev.158105 Open Access
Lars Ostergaard (John Innes Centre) leads this study that characterises how the activity of five transcription factors (TF) integrate with auxin signaling in the control of gynoecium development. The auxin response factor ETTIN is a central controller of this relationship across members of the Brassicaceae and that variation in an ETTIN sub-domain effects TF affinities, interaction strength and gynoecium morphology


Kriechbaumer V, Maneta-Peyret L, Fouillen L, Botchway SW, Upson J, Hughes L, Richardson J, Kittelmann M, Moreau P, Hawes C

The odd one out: Arabidopsis reticulon 20 does not bend ER membranes but has a role in lipid regulation. Sci Rep. doi: 10.1038/s41598-018-20840-0

This study is led by Chris Hawes (Oxford Brookes) and continues his labs work on the plant ER. They are working on a subgroup of reticulons, which are ER membrane proteins, that have an extended N-terminal domain. Three members of this subgroup show different localisation patterns that indicates that along their sequences are similar they might play different cellular roles.


Williams A, Pétriacq P, Schwarzenbacher RE, Beerling DJ, Ton J (2018) Mechanisms of glacial-to-future atmospheric CO2 effects on plant immunity. New Phytol. doi: 10.1111/nph.15018 Open Access
This article from the University of Sheffield uses Arabidopsis to investigate the impact that changing climatic CO2 concentrations might have on plant immunity. The authors performed a global analysis on the response to sub-ambient and elevated CO2 and found that both changes causes alterations to salicyclic acid or jasmonic acid response pathways. However these responses are not always opposite, revealing new insights in the response to changing CO2 concentrations.


 

Isner JC, Begum A, Nuehse T, Hetherington AM, Maathuis FJM (2018) KIN7 Kinase Regulates the Vacuolar TPK1 K+ Channel during Stomatal Closure. Curr Biol. doi: 10.1016/j.cub.2017.12.046.

This is collaborative work between the Universities of York and Bristol and analyses factors that control stomatal closure. They show the TPK1 vacuolar K+ channel is important for ABA and CO2 mediated closure and that the function of this protein is regulated by the KIN7 receptor-like kinase. These activities result in potassium release from the vacuole leading to osmotic changes that contribute to stomatal closure.


Caviglia M, Mazorra Morales LM, Concellón A, Gergoff Grozeff GE, Wilson M, Foyer CH, Bartoli CG (2018) Ethylene signaling triggered by low concentrations of ascorbic acid regulates biomass accumulation in Arabidopsis thaliana. Free Radic Biol Med. doi: 10.1016/j.freeradbiomed.2018.01.032

Christine Foyer (University of Leeds) is a co-author on this research showing that a defect in ascorbic acid production leads to elevated levels of the hormone ethylene as well as having a wider impact on the control of growth-mediating hormone signalling. This result indicates that the cellular redox buffer AA is a significant contributor to hormone signalling pathways.


Reyna-Llorens I, Burgess SJ, Reeves G, Singh P, Stevenson SR, Williams BP, Stanley S, Hibberd JM (2018) Ancient duons may underpin spatial patterning of gene expression in C4 leaves.Proc Natl Acad Sci U S A. doi: 10.1073/pnas.1720576115

Julian Hibberd (University of Cambridge) is the corresponding author on this study that is part of his labs overarching aim of discovering what is necessary to transfer C4 photosynthesis into C3 plants. As part of this work they are searching for master regulator sequences that have allowed for the multiple independent evolution of C4 photosynthesis. They have identified a regulatory duon that is a pair of cis-elements located in coding sequences of genes preferentially expressed in bundle sheath cells of C4 leaves and are also present in C3 plants and algae. Therefore they discuss how C4 plants have co-opted these regulatory elements and how it might be exploited in future molecular engineering projects.

Arabidopsis Research Roundup: February 12th

This weeks Arabidopsis Research Roundup begins with a study from SLCU that investigates the interaction between nitrate and cytokinin signaling in the shoot meristem. Next is research from Sheffield that studies changes to the macromolecular composition of the photosynthetic apparatus following the transition from dark to light. Third are three papers that include University of Edinburgh faculty members as co-authors; Gary Loake is involved in a global study on NO signaling, Karen Halliday is included on a study into the relationship between clock components and the PIF-mediated hypocotyl elongation and Naomi Nakayama contributes to the development of a model that explains PIN protein localisation. Cyril Zipfel (TSL) is a co-author on the fifth paper, which introduces a new signaling component in the defence response and whilst the penultimate paper includes Denis Murphy (University of South Wales) and investigates the effect of dioxins on seed development. The final paper documents research from Manchester and Nottingham that uses a cress endosperm as a model to test the elastic properties of thin biological membranes.


Landrein B, Formosa-Jordan P, Malivert A,, Schuster C, Melnyk CW,, Yang W, Turnbull C, Meyerowitz EM, Locke JCW,, Jönsson H (2018) Nitrate modulates stem cell dynamics in Arabidopsis shoot meristems through cytokinins. PNAS doi: 10.1073/pnas.1718670115.

Open Access

http://www.pnas.org/content/early/2018/01/22/1718670115

Henrik Jonsson and James Locke (SLCU) are corresponding authors on this investigation into the relationship between nitrate and cytokinin signalling in the Arabidopsis shoot meristem (SAM). They show that nitrate availability determines the size of the SAM, which is controlled by the transport of cytokinin precursors from the root to the shoot. A discussion about this paper with lead author Benoit Landrien and Professor Jonsson is available on the GARNet YouTube and iTunes channels.


Wood WHJ, MacGregor-Chatwin C, Barnett SFH, Mayneord GE, Huang X, Hobbs JK, Hunter CN, Johnson MP (2018) Dynamic thylakoid stacking regulates the balance between linear and cyclic photosynthetic electron transfer. Nature Plants. doi: 10.1038/s41477-017-0092-7

Open with this link

This research in this manuscript has come from the University of Sheffield with Matthew Johnson as the corresponding author. They have used atomic force microscopy (AFM) to investigate how the transition from dark to light affects the macromolecular architecture of the photosynthetic apparatus within the thylakoid membrane. This transition does not alter the antenna size of either photosystem yet increases the number of thylakoid grana. Overall these changes serve to regulate the balance between light harvesting, CO2 fixation and enabling the protection of PSII activity from the destructive effects of non-photochemical quenching.


Imran QM, Hussain A, Lee SU, Mun BG, Falak N, Loake GJ, Yun BW (2018) Transcriptome profile of NO-induced Arabidopsis transcription factor genes suggests their putative regulatory role in multiple biological processes. Sci Rep. doi: 10.1038/s41598-017-18850-5.

https://www.nature.com/articles/s41598-017-18850-5

Open Access

Gary Loake (University of Edinburgh) is a contributor to this Korean-led manuscript that has performed expression analysis on plants treated with S-nitrosocysteine (CySNO). They have identified many novel NO-responsive transcription factors and were able to confirm the role of three random TFs in this response following analysis of loss of function mutants. This paper provides new insights into the molecular components that contribute to NO signalling during plant defence and immunity.


Martín G, Rovira A, Veciana N, Soy J, Toledo-Ortiz G, Gommers CMM, Boix M, Henriques R, Minguet EG, Alabadí D, Halliday KJ, Leivar P, Monte E Circadian Waves of Transcriptional Repression Shape PIF-Regulated Photoperiod-Responsive Growth in Arabidopsis. Curr Biol. doi: 10.1016/j.cub.2017.12.021

Karen Halliday (University of Ediburgh) is a co-author on this Spanish-led study that investigates how the expression of PHYTOCHROME-INTERACTING FACTORS (PIFs) genes is controlled. The activity of PIFs are responsible for determining the rate of hypocotyl elongation in different light conditions and this paper demonstrates that PSEUDO-RESPONSE REGULATORS PRR9/7/5 proteins act antagonistically to the PIFs by interacting at the promotor of the CDF5 transcription factor. This provides a mechanism to explain the circadian-controlled regulation of hypocotyl cell elongation.


Hernandez V, Barrio RA, Benítez M, Nakayama N, Romero-Arias JR, Villarreal Lujan C (2018) A physico-genetic module for the polarisation of auxin efflux carriers PIN-FORMED (PIN). Phys Biol. doi: 10.1088/1478-3975/aaac99

Naomi Nakayama (University of Edinburgh) is a co-author on this Mexican-led study that proposes a physico-genetic model that explains the localization of PIN auxin transporter proteins to the Arabidopsis plasma membrane. This model confirms experimental observations and allows the prediction that mechanical forces can predominate over molecular components.


www.cell.com/molecular-cell/fulltext/S1097-2765(17)30983-8

Wang J, Grubb LE, Wang J, Liang X, Li L, Gao C, Ma M, Feng F, Li M, Li L, Zhang X, Yu F, Xie Q, Chen S, Zipfel C, Monaghan J, Zhou JM (2018) A Regulatory Module Controlling Homeostasis of a Plant Immune Kinase. Mol Cell. doi: 10.1016/j.molcel.2017.12.026

This Chinese-led paper includes Cyril Zipfel (TSL) as a co-author and identifies the U-box proteins PUB25 and PUB26 as E3 ligases for the cytoplasmic kinase BIK1, which is a key rate limiting component of the plant defence response. This multi-protein regulatory module provides another level of complexity to our understanding of the molecular factors involved in plant immunity.


Hanano A, Almousally I, Shaban M, Murphy DJ (2018) Exposure of Arabidopsis Plants to Dioxin Results in a Wrinkled Seed Phenotype that is likely due to 20S Proteasomal Degradation of WRI1. J Exp Bot. doi: 10.1093/jxb/ery027

Denis Murphy (University of South Wales) is a co-author on this Syrian-led study that uses Arabidopsis seeds to test the negative effects of dioxins. Seeds treated with dioxins have a wrinked phenotype that corresponds to changes in the expression of genes related to lipid and carbohydrate metabolism. Overall this study reveals a novel set of genetic changes effects caused by dioxins that explain the profound effects on seed development.


S. P. Pearce, J. R. King, T. Steinbrecher, G. Leubner-Metzger, N. M. Everitt, M. J. Holdsworth (2018) Finite indentation of highly curved elastic shells Proceedings of the Royal Society A doi: 10.1098/rspa.2017.0482

Open Access

Plant scientist Mike Holdsworth (University of Nottingham) is a co-author on this paper that has used the endosperm from garden cress (Lepidium sativum) as the experimental model to define the elastic properties of a thin biological surface. Indentation experiments have been classically used to measure these properties and then develop mathematically models that explain their characteristics. These models rely on an assumed flat surface whereas in reality any surface will often be curved. By obtaining measurements from identations studies on the cress endosperm they are able to better refine the models that explain the properties of the membrane in this context.

Arabidopsis Research Roundup: December 8th.

This weeks Research Roundup begins with two papers from the University of Edinburgh on very different topics of Arabidopsis research. Firstly Alistair McCormick and Sofirtios Tsaftaris introduce a new low-cost phenotyping platform whilst Gerben Ooijen’s group has analysed the role of SUMOylation in the control of the circadian clock. The next three papers each involve wide UK collaborations and either look at plant nutrient composition (Nottingham, Dundee, York), the role of N-end rule pathway in the control of seed storage mobilisation (Rothamsted, Nottingham, Oxford, Birmingham, Cambridge) or the development of a new tool for the study of phloem sieve elements (Leeds, Rothamsted, Cambridge, Newcastle). The penultimate paper from Daniel Zilbermann (JIC) highlights the global mechanisms of methyltransferase function in Arabidopsis and mice whilst the final paper from Alexandre Ruban (QMUL) and co-authors continues his groups work to unpick the specifics of NPQ.


Dobrescu A, Scorza LCT, Tsaftaris SA, McCormick AJ (2017) A “Do-It-Yourself” phenotyping system: measuring growth and morphology throughout the diel cycle in rosette shaped plants. Plant Methods. doi: 10.1186/s13007-017-0247-6

Open Access

University of Edinburgh colleagues Alistair McCormick and Sofirtios Tsaftaris lead this work that presents a low cost phenotyping system for the analysis of the growth rate and phenotypic characteristics of Arabidopsis thaliana rosettes. The software that they have developed allows the accurate segmentation of multiple rosettes within a single image and overall offers a straightforward solution for automated phenotyping across a range of growth environments.


Hansen LL, van den Burg HA, van Ooijen G (2017) Sumoylation Contributes to Timekeeping and Temperature Compensation of the Plant Circadian Clock. J Biol Rhythms. doi: 10.1177/0748730417737633

Gerben van Ooijen (University of Edinburgh) is the corresponding author of this work that has identified SUMOylation as a novel mechanism of regulating circadian clock genes in Arabidopsis. Plants with defects in sumoylation have altered circadian periods that exhibit incorrect temperature compensation. Overall these results indicate that sumoylation importantly buffers clock function in response to changing temperatures.


Alcock TD, Havlickova L, He Z, Bancroft I, White PJ, Broadley MR, Graham NS (2017) Identification of Candidate Genes for Calcium and Magnesium Accumulation in Brassica napus L. by Association Genetics. Front Plant Sci. doi: 10.3389/fpls.2017.01968

Open Access

Neil Graham and Martin Broadley (University of Nottingham) are the corresponding authors of this study that has taken advantage of the Brassica napus Associative Transcriptomes RIPR diversity panel developed by Ian Bancroft’s lab in York. Novel loci involved with an altered response to calcium and magnesium were identified in B.napus before mineral composition was analysed in Arabidopsis mutants defective in orthologous genes. The analysed plants exhibited alteration in mineral composition, meaning that the associated Brassica loci might be targets for future breeding strategies aimed at improving plant nutrient compositions.


Zhang H, Gannon L, Hassall KL, Deery MJ, Gibbs DJ, Holdsworth MJ, van der Hoorn RAL, Lilley KS, Theodoulou FL (2017) N-terminomics reveals control of Arabidopsis seed storage proteins and proteases by the Arg/N-end rule pathway. New Phytol. doi: 10.1111/nph.14909

Freddie Theodoulou (Rothamsted Research) is the corresponding author of this research that involved a collaboration with colleagues in Cambridge, Birmingham, Nottingham and Oxford. They have performed a proteomic analysis on etiolated seedlings to identify those proteins designated for degradation by the N-end rule pathway. They analysed prt6 mutant plants that lack the function of the E3 ligase PROTEOLYSIS6 (PRT6) and discovered that N-terminal peptides from 45 protein groups were upregulated in this mutant, corresponding to the equivalent downregulation of several known N-end rule proteases. Overall the authors show that PRT6 plays an important role in the regulation of seed storage mobilisation in young seedlings and is therefore a possible future target to manipulate the plant responses to adverse environmental conditions. Dr Kirsty Hassall, a statistician at Rothamsted, is an author on this paper and in the latest edition of the GARNish newsletter explains how she interacts with plant scientists during her work.


Torode TA, O’Neill RE, Marcus SE, Cornuault V, Pose-Albacete S, Lauder RP, Kracun SK, Gro Rydahl M, Andersen MCF, Willats WGT, Braybrook SA, Townsend BJ, Clausen MH, Knox JP (2017) Branched pectic galactan in phloem-sieve-element cell walls: implications for cell mechanics. Plant Physiol. doi: 10.1104/pp.17.01568 Open Access

Paul Knox (University of Leeds) is the corresponding author of this study that includes contributions from researchers at SLCU, Newcastle and Rothamsted. This work is based around the development of a monoclonal antibody, LM26 that is able to recognize a β-1,6-galactosyl substitution of β-1,4-galactan. LM26 has allowed the identification of this unusual branched galactan that is specific to phloem elements and the authors hope that it can be a useful tool in future studies on the biology of phloem elements


Lyons DB, Zilberman D (2017) DDM1 and Lsh remodelers allow methylation of DNA wrapped in nucleosomes. Elife. doi: 10.7554/eLife.30674 Open Access

Daniel Zilberman has recently moved to the John Innes Centre and is the lead author of this work that was conducted when he was working in US. This research is a cross-kingdom analysis showing that nucleosome-free DNA is the preferred target for methyltransferases in both Arabidopsis and mice, and that nucleosomes appear to be a barrier to the function of these enzymes. Furthermore they demonstrate that linker-specific methylation that is usually absent in Arabidopsis can be introduced by removal of histone H1. This shows that flowering plants still possess this ability despite its loss, during the evolution of H1, over a billion years ago.


Tutkus M, Chmeliov J, Rutkauskas D, Ruban AV, Valkunas L (2017) Influence of the Carotenoid Composition on the Conformational Dynamics of Photosynthetic Light-Harvesting Complexes. J Phys Chem Lett. doi: 10.1021/acs.jpclett.7b02634

Alexandre Ruban (QMUL) is a co-author on this study that investigates the role that carotenoid composition plays in the control of Non-photochemical quenching (NPQ), a mechanism that protects the photosynthetic apparatus from light-damage. Arabidopsis mutants with differing carotenoid compositions were analysed for the dynamics of the conformation switches that occur during NPQ. Interestingly they show that LHCII has robust function  that is resistant to different carotenoid concentrations.

Arabidopsis Research Roundup: August 14th

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Published on: August 14, 2017

This weeks UK Arabidopsis Research Roundup includes a study from the lab of GARNet Chairman Steven Spoel that describes their work on the control of ROS signaling. The other two papers looks at different aspects of photosynthetic control. Firstly Giles Johnson (University of Manchester) investigates the proteomic changes that occur during dynamic acclimation. Finally Alex Webb’s lab (University of Cambridge) investigate how the products of photosynthesis feed into ethylene-dependent control of the circadian clock.


Kneeshaw S, Keyani R, Delorme-Hinoux V, Imrie L, Loake GJ, Le Bihan T, Reichheld JP, Spoel SH (2017) Nucleoredoxin guards against oxidative stress by protecting antioxidant enzymes.

PNAS https://doi.org/10.1073/pnas.1703344114 Open Access

GARNet Chairman Steven Spoel (University of Edinburgh) leads this Franco-UK collaboration that investigated the role of the pathogen-induced oxidoreductase Nucleoredoxin 1 (NRX1) during the response to oxidative stress. They show that NRX1 targets a range of enzymes that are responsible for scavenging damaging cellular H2O2 and subsequently protect these enzymes from damage. This unexpected regulatory mechanism demonstrates a key role for NRX1 in protecting plants from the oxidative stress, which is an important component of the plants defence response.

Steven discussed this paper below. This audio is also on GARNet YouTube and iTunes channels.


Miller MAE, O’Cualain R, Selley J, Knight D, Karim MF, Hubbard SJ, Johnson GN (2017) Dynamic Acclimation to High Light in Arabidopsis thaliana Involves Widespread Reengineering of the Leaf Proteome Front Plant Sci. https://doi.org/10.3389/fpls.2017.01239 Open Access

Giles Johnson leads this collaborations between two departments from the University of Manchester that has looked at changes to the leaf proteome that occur under differing light conditions. Transferring leaves from low to high light conditions causes a process called dynamic acclimation, which is perturbed in gpt2 mutants. Proteomic analysis of wildtype and gpt2 leaves reveals that a similar number of proteins are changed during dynamic acclimation but the identify of these is altered, with a reduced abundance of proteins involved in photosynthesis. Interestingly gpt2 showed an increase in the number of proteins involved in the stress response.

In 2016 Giles talked to GARNet about a different paper that looks at cold acclimation in Arabidopsis.


Haydon MJ, Mielczarek O, Frank A, Román Á, Webb AA (2017) Sucrose and ethylene signaling interact to modulate the circadian clock. Plant Physiol. https://doi.org/10.1104/pp.17.00592 Open Access

Alex Webb (University of Cambridge) collaborates with Mike Laydon (previously at York, now at the University of Melbourne) on this study that links photosynthesis with circadian and hormone signalling. Previous work showed that GIGANTEA (GI) is necessary to maintain sucrose-dependent circadian oscillations in the dark. This extension of that work demonstrates that sucrose stabilises GI by altering the activity of the ZEITLUPE (ZTL) F-box protein, which is surprisingly dependent on the key ethylene signalling intermediate CONSTITUTIVE TRIPLE RESPONSE 1 (CTR1). Over recent times the role of ethylene on control of the circadian clock has been overlooked but this study shows that the hormone can shorten the circadian period through the activity of sucrose and GI. This study integrates another signal into our understanding of the exquisite regulatory relationships that control clock function.


Please subscribe to the GARNet iTunes channel!

Arabidopsis Research Roundup: March 6th.

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Published on: March 6, 2017

This weeks Arabidopsis Research Roundup includes four papers that focus on different aspects of plant cell biology. Firstly Ian Henderson’s research group in Cambridge defines the role of a critical component that determines crossover frequency in plants and other eukaryotes. Secondly Karl Oparka (Edinburgh) leads a broad collaboration that defines the mechanism of unloading of solutes and macromolecules from the root phloem. Thirdly Keith Lindsey (Durham) has developed a model that describes how auxin patterns the Arabidopsis root. Finally Mike Blatt (Glasgow) is part of a group that uses Arabidopsis as a framework for the study of ABA-signaling during stomatal movement in ferns.


Ziolkowski PA, Underwood CJ, Lambing C, Martinez-Garcia M, Lawrence EJ, Ziolkowska L, Griffin C, Choi K, Franklin FC, Martienssen RA, Henderson IR (2017) Natural variation and dosage of the HEI10 meiotic E3 ligase control Arabidopsis crossover recombination. Genes Dev

http:/​/​dx.​doi.​org/10.1101/gad.295501.116

Open Access

GARNet committee member Ian Henderson (University of Cambridge) leads this work that features collaborators from the UK, US and Poland. They use an experimental technique that allows facile analysis of recombination rates alongside a study of Arabidopsis natural variation to isolate a QTL that is critical for maintaining the correct number of crossovers during meiosis. This HEI10 gene codes for an E3 ligase (the targets of which are currently unknown) whose copy number is a key component in the control of recombination rate. Hei10 mutants have less crossovers whilst plants with extra copies of HEI10 have an increased number, especially in sub-telomeric regions of the genome. HEI10 is a highly conserved protein, demonstrating its important role to ensure appropriate levels of recombination throughout the evolution of eukaryotes.

Ian kindly takes ten minutes to discuss this paper with GARNet on our YouTube Channel.


Ross-Elliott TJ, Jensen KH, Haaning KS, Wager BM, Knoblauch J, Howell AH, Mullendore DL, Monteith AG, Paultre D, Yan D, Otero-Perez S, Bourdon M, Sager R, Lee JY, Helariutta Y, Knoblauch M, Oparka KJ (2017) Phloem unloading in Arabidopsis roots is convective and regulated by the phloem-pole pericycle. Elife.

http:/​/​dx.​doi.​org/10.7554/eLife.24125

Open Access

Karl Oparka (University of Edinburgh) is the corresponding author of this study that includes researchers from the UK, US and Denmark. Movement of solutes and macromolecules through the plant phloem is key for the correct distribution of nutrients allowing for optimal growth. In this paper they discover that unloading of molecules from the phloem occurs via a set of specialized funnel plasmodesmata that link the phloem to adjacent pericycle cells. Remarkably they find that whereas solutes are constantly unloaded, larger proteins are released through these plasmodesmata in discrete pulses, which they describe as ‘batch unloading’. Overall this study provides evidence of a major role for the phloem-pericycle cells in the process of moving essential nutrients from the phloem into surrounding tissues.


Moore S, Liu J, Zhang X, Lindsey K (2017) A recovery principle provides insight into auxin pattern control in the Arabidopsis root. Sci Rep. http:/​/​dx.​doi.​org/10.1038/srep43004

Open Access

The work comes from the lab of Keith Lindsey (University of Durham) and developes a data-driven model that predicts the role of auxin patterning in the recovery of an Arabidopsis root following a perturbation of polar auxin transport. They demonstrate three main principles that define the role of auxin influx and efflux carriers in this process and also provide experimental validation for their predictions.


Cai S, Chen G, Wang Y, Huang Y, Marchant B, Wang Y, Yang Q, Dai F, Hills A, Franks PJ, Nevo E, Soltis D, Soltis P, Sessa E, Wolf PG, Xue D, Zhang G, Pogson BJ, Blatt MR, Chen ZH (2017)

Evolutionary Conservation of ABA Signaling for Stomatal Closure in Ferns Plant Physiol

http:/​/​dx.​doi.​org/10.1104/pp.16.01848

Open Access

Mike Blatt (University of Glasgow) is a co-author on this global study that looks into the evolution of ABA-signaling in the control of stomatal closure. Although this study is focused on this process in ferns they build their findings on the analysis of transcriptional networks from Arabidopsis. Ultimately they find that the evolution of ABA-controlled guard cells movements are important in the adaptation of ferns to a terrestrial environment.

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