GARNet Research Roundup: June 4th

This weeks GARNet Research Roundup begins with a paper from researchers at the University of Dundee, James Hutton Institute, Durham University and the University of Glasgow that characterises a functional role for alternative splicing during the cold response. Second is a paper from Newcastle University that investigates the role of the OXI1 kinase during aphid predation. Third is a paper that includes University of Bristol co-authors that looks at strigolactone signaling in moss whilst the fourth paper from researchers at Leeds and QMUL studies the role of ascorbate during photosynthesis. The final paper from Warwick and York uses gene expression data from pathogen-infected plants to generate a model for predicting a strategy for synthetic engineering of the defence response.


Calixto CPG, Guo W, James AB, Tzioutziou NA, Entizne JC, Panter PE, Knight H, Nimmo H, Zhang R, Brown JWS (2018) Rapid and dynamic alternative splicing impacts the Arabidopsis cold response transcriptome. Plant Cell doi: 10.1105/tpc.18.00177.

www.plantcell.org/content/early/2018/05/15/tpc.18.00177.long

Open Access

Cristiane Calixto and Wenbin Guo work with John Brown at University of Dundee and the James Hutton Institute and in this large-scale biology paper they characterise the role of alternative splicing (AS) during a stress response. RNAseq was performed on plants exposed to cold stress and they showed that hundreds of genes undergo AS just a few hours after temperature decrease and that this response is sensitive to small changes. The authors propose that AS is a mechanism to fine-tune changes in thermo-plasticity of gene expression and in addition they investigate the activity of the novel splicing factor U2B”-LIKE.

Christiane will discuss this research at the upcoming GARNet2018 meeting held at the University of York in September 2018.


Shoala T, Edwards MG, Knight MR, Gatehouse AMR. OXI1 kinase plays a key role in resistance of Arabidopsis towards aphids (Myzus persicae) (2018) Transgenic Res. doi: 10.1007/s11248-018-0078-x.

Open Access

This work is led by Tahsin Shoala in the lab of Angharad Gatehouse at Newcastle University and demonstrates a novel role for MAPK cascades in resistance to aphid predation. They investigate mutants in OXI1 kinase, a gene that activates MAPK signaling and demonstrate a reduction in the aphid population build-up. Furthermore they show that the effect of OXI works through a mechanism that involves callose deposition, demonstrated as oxi1 mutants lack the upregulation of a set of β-1,3-glucanase genes following predation.


Lopez-Obando M, de Villiers R, Hoffmann B, Ma L, de Saint Germain A, Kossmann J, Coudert Y, Harrison CJ, Rameau C, Hills P, Bonhomme S (2018) Physcomitrella patens MAX2 characterization suggests an ancient role for this F-box protein in photomorphogenesis rather than strigolactone signalling. New Phytol. doi: 10.1111/nph.15214

GARNet committee member Jill Harrison is a co-author on this paper that is led by Mauricio Lopez‐Obando working at Université Paris-Saclay. In Physcomitrella patens development they investigate the role of the moss ortholog of the Arabidopsis strigolactone signaling mutant MAX2. Previous work had shown that moss does response to SL signaing but they find that although Ppmax2 mutants showed defects in early development and photomorphogenesis they do not show changes in the SL response. Fascinatingly this indicates that the molecular components that control SL signaling have diverged in vascular plants and seemingly co-opted a role for MAX2 that was previously not required in mosses.


https://academic.oup.com/jxb/article/69/11/2823/4991886

Plumb W, Townsend AJ, Rasool B, Alomrani S, Razak N, Karpinska B, Ruban AV, Foyer CH. Ascorbate-mediated regulation of growth, photoprotection and photoinhibition in Arabidopsis thaliana (2018) J Exp Bot. doi: 10.1093/jxb/ery170

William Plumb (Leeds) and Alexandra Townsend (QMUL) are the lead authors on this study that investigates the importance of ascorbate during photosynthesis. In this work they analysed the growth of ascorbate synthesis mutants that are smaller and have less biomass than wildtype plants. However these plants have normal levels of non-photoinhibiton, allowing the authors to conclude that ascorbate is needed for growth but not photoprotection.


Foo M, Gherman I, Zhang P, Bates DG, Denby K (2018) A Framework for Engineering Stress Resilient Plants using Genetic Feedback Control and Regulatory Network Rewiring. ACS Synth Biol. doi: 10.1021/acssynbio.8b00037
Mathias Foo and Iulia Gherman (University of Warwick) are lead authors on work that analyses gene expression data taken from Botrytis cinerea-infected Arabidopsis. They have identified a network of genes involved in the defence response. They validate their model against previously obtained time series data and then perturb the model in two differences ways, focused on the transcription factor CHE. This analysis demonstrates the potential of combining feedback control theory with synthetic engineering in order to generate plants that are resistant to biotic stress.

https://pubs.acs.org/doi/10.1021/acssynbio.8b00037

GARNet Research Roundup: May 17th

This weeks GARNet research roundup includes six excellent papers investigating many aspects of Arabidopsis cell biology. Firstly Eirini Kaiserli from Glasgow introduces a novel regulator of blue-light signaling. Second is a paper that analyses the circadian clock with single cell resolution and is led by James Locke (SLCU) and Anthony Hall (Earlham Institute). The next two papers investigate different aspects of hormone signaling, with Keith Lindsey’s group at Durham looking at the relationshop between the HYDRA protein and the auxin response whilst Ottoline Leysers group in Cambridge looks at the link between auxin and cytokinin during shoot growth. The fifth paper from Phillip Mullineaux (University of Essex) provides a genome-wide analysis into the role of HEAT SHOCK TRANSCRIPTION FACTORA1b protein. The final paper from the lab of Piers Hemsley (James Hutton Institute, University of Dundee) should be of interest to many plant molecular biologists as they assess the functional significance of different epitope tags.


Perrella G, Davidson MLH, O’Donnell L, Nastase AM, Herzyk P, Breton G, Pruneda-Paz JL, Kay SA, Chory J, Kaiserli E (2018) ZINC-FINGER interactions mediate transcriptional regulation of hypocotyl growth in Arabidopsis. Proc Natl Acad Sci U S A.. pii: 201718099. doi: 10.1073/pnas.1718099115

Open Access

Eirini Kaiserli (University of Glasgow) leads this study that identifies the ZINC-FINGER HOMEODOMAIN 10 (ZFHD10) as a novel regulator of light signaling. ZFHD10 physically interacts with TANDEM ZINC-FINGER PLUS3 (TZP) and these proteins coassociate at promotors that are blue-light regulated. These results reveal of novel mechanism of action for the key multiple signal integrator TZP in the light regulated growth of Arabidopsis hypocotyls.

Eirini discusses this paper on the GARNet YouTube channel.


Gould PD, Domijan M, Greenwood M, Tokuda IT, Rees H, Kozma-Bognar L, Hall AJ, Locke JC (2018). Coordination of robust single cell rhythms in the Arabidopsis circadian clock via spatial waves of gene expression. Elife. 26;7. pii: e31700. doi: 10.7554/eLife.31700 Open Access

https://elifesciences.org/articles/31700

This paper is led by James Locke (SLCU) and Anthony Hall (Earlham) and investigates the circadian clock at single cell resolution. They use Arabidopsis plants grown in constant environmental conditions to show two desynchronised yet robust single cell oscillations that move both up and down the root. Their results indicate that the clock shows cell-to-cell coupling and they they modeled this relationship to recapitulate the observed waves of activity. Overall their results are suggestive of multiple coordination points for the Arabidopsis clock, which is different from the mammalian system of regulation.


http://dev.biologists.org/content/145/10/dev160572

Short E, Leighton M, Imriz G, Liu D, Cope-Selby N, Hetherington F, Smertenko A, Hussey PJ, Topping JF, Lindsey K (2018) Epidermal expression of a sterol biosynthesis gene regulates root growth by a non-cell autonomous mechanism in Arabidopsis. Development . pii: dev.160572. doi: 10.1242/dev.160572 Open Access

This collaboration between the research groups of Keith Lindsey and Patrick Hussey at the University of Durham investigates the role of the HYDRA1 (HYD1) sterol Δ8-Δ7 isomerase in epidermal patterning. This tissue shows highest HYD1 expression and hyd mutants have major root patterning defects. Tissue-specific expression of HYD1 indicates that it is involved with some type of non-cell autonomous signaling. Analysis of PIN1 and PIN2 protein expression suggests that auxin might be this functional signal


http://www.plantphysiol.org/content/early/2018/05/01/pp.17.01691.long

Waldie T, Leyser O (2018) Cytokinin targets auxin transport to promote shoot branching. Plant Physiol. 2018 May 1. pii: pp.01691.2017. doi: 10.1104/pp.17.01691.Open Access

This study from the lab of Ottoline Leyser (SLCU) investigates the integration between the plant hormones cytokinin and auxin. They investigate the role of cytokinin in shoot branching through analysis of Arabidopsis Response Regulators (ARRs) mutants. They show arr mutant phenotypes correlate with changes in stem auxin transport mediated by the PIN3, PIN4 and PIN7 efflux carriers, the expression of each respond to cytokinin signaling. Overall this study identifies a novel alternative pathway by which cytokinin impacts bud outgrowth through alterations in auxin transport.


Albihlal WS, Irabonosi O, Blein T, Persad R, Chernukhin I, Crespi M, Bechtold U, Mullineaux PM (2018) Arabidopsis Heat Shock Transcription FactorA1b regulates multiple developmental genes under benign and stress conditions. J Exp Bot. doi: 10.1093/jxb/ery142 Open Access

Phillip Mullineaux (University of Essex) leads this collaboration with French colleagues in a study that investigates the genome-wide targets of the HEAT SHOCK TRANSCRIPTION FACTORA1b (HSFA1b) protein. Under non-stress ad heat-stress conditions they showed that 1000s of genes are differentially expressed with a smaller proportion of genes showing different levels of direct interaction. The indirect targets of HSFA1b are regulated through a network of 27 transcription factors and they also provide evidence for the role of hundreds of natural antisense non-coding RNA in the regulation of HSFA1b targets. Overall they show that HSFA1b is a key regulator of environmental cues to regulate both developmental genes and those involved in stress tolerance.


Hurst CH, Turnbull D, Myles SM, Leslie K, Keinath NF, Hemsley PA (2018) Variable effects of C-terminal tags on FLS2 function – not all epitope tags are created equal. Plant Physiol. doi: 10.1104/pp.17.01700 Open Access

This study from the Hemsley lab (James Hutton Institute, University of Dundee) is a cautionary tale on the use and interpretation of results obtained from experiments with commonly-used epitope tags. They assessed the activity of plants containing transgenic FLS2 proteins, which is a receptor-like kinase (RLKs) involved in the defence response. They show that various FLS2 C-terminal epitope fusions reveal highly variable and unpredictable outputs, indicating that the presence of different tags significantly alters protein function. This finding might require a reassessment of many experiments that rely on interpreting the function of epitope-tagged proteins and has significant for many if not all plant molecular biologists.

GARNet Research Roundup: April 27th

This weeks GARNet research roundup features four papers that include Malcolm Bennett (University of Nottingham) as an author. The first three are linked manuscripts that investigate the role of auxin on root hair development that is controlled by varying phosphate levels. Ranjan Swarup provides an audio summary of two of these papers on the GARNet YouTube and podcast feeds.

The fourth paper from Nottingham is a collaboration with GARNet PI Jim Murray (Cardiff University) that characterises the STM gene network and its influence on meristem development.

The fifth paper from the lab of Paul Dupree in Cambridge characterises the stem transcriptome whilst the next paper from Iain Johnston and George Bassel (University of Birmingham) identifies a bet-hedging network that influences seed germination. The final paper features Seth Davies (University of York) as a co-author and investigates the impact of changes in circadian rhythms on short architecture.


Researchers at the University of Nottingham are involved in three back-to-back papers that add a mechanistic framework to the relationship between phosphate and auxin signaling in root hairs.

Bhosale R, Giri J, Pandey BK, Giehl RFH, Hartmann A, Traini R, Truskina J, Leftley N, Hanlon M, Swarup K, Rashed A, Voß U, Alonso J, Stepanova A, Yun J, Ljung K, Brown KM, Lynch JP, Dolan L, Vernoux T, Bishopp A, Wells D, von Wirén N, Bennett MJ, Swarup R (2018) A mechanistic framework for auxin dependent Arabidopsis root hair elongation to low external phosphate. Nat Commun. 9(1):1409. doi: 10.1038/s41467-018-03851-3

https://www.nature.com/articles/s41467-018-03851-3

Open Access

The first paper is led by Ranjan Swarup and defines components of auxin biosynthetic, transport and signaling pathways that are involved in the change root hair development in response to different phosphate concentrations in Arabidopsis. Ranjan discusses this paper on YouTube.

Giri J, Bhosale R, Huang G, Pandey BK, Parker H, Zappala S, Yang J, Dievart A, Bureau C, Ljung K, Price A, Rose T, Larrieu A, Mairhofer S, Sturrock CJ, White P, Dupuy L, Hawkesford M, Perin C, Liang W, Peret B, Hodgman CT, Lynch J, Wissuwa M, Zhang D, Pridmore T, Mooney SJ, Guiderdoni E, Swarup R, Bennett MJ (2018). Rice auxin influx carrier OsAUX1 facilitates root hair elongation in response to low external phosphate. Nat Commun. 9(1):1408. doi: 10.1038/s41467-018-03850-4

https://www.nature.com/articles/s41467-018-03850-4

Open Access

This paper led by Malcolm Bennett uses a rice model to investigate the relationship between auxin and root hair elongation in response to low concentrations of phosphate. They show rice aux1 mutants have significant changes in root architecture.

Dindas J, Scherzer S, Roelfsema MRG, von Meyer K, Müller HM, Al-Rasheid KAS, Palme K, Dietrich P, Becker D, Bennett MJ, Hedrich R (2018) AUX1-mediated root hair auxin influx governs SCF(TIR1/AFB)-type Ca(2+) signaling. Nat Commun. 9(1):1174. doi: 10.1038/s41467-018-03582-5

Open Access

The third paper includes Malcolm Bennett as a co-author and investigates how the auxin transport and signaling pathways stimulate calcium signaling during root hair elongation.


Scofield S, Murison A, Jones A, Fozard J, Aida M, Band LR, Bennett M, Murray JAH (2018) Coordination of meristem and boundary functions by transcription factors in the SHOOT MERISTEMLESS regulatory network. Development pii: dev.157081. doi: 10.1242/dev.157081

http://dev.biologists.org/content/early/2018/04/12/dev.157081.long

Open Access
GARNet PI Jim Murray is the corresponding author on this manuscript that is led by Dr Simon Scofield and includes collaborators from the University of Nottingham. They have explored the gene regulatory network that is regulated by the key meristem identity gene SHOOT MERISTEMLESS (STM). This network includes an over-representation of transcription factor families, each of which have distinct roles in meristem development. They use in planta experimentation and in silico modeling to investigate the relationship between STM and CUC1 in more detail. Overall this study confirms that STM is a central regulator of shoot meristem function.


Faria-Blanc N, Mortimer JC, Dupree P (2018) A Transcriptomic Analysis of Xylan Mutants Does Not Support the Existence of a Secondary Cell Wall Integrity System in Arabidopsis. Front Plant Sci. 9:384. doi: 10.3389/fpls.2018.00384

Open Access

Paul Dupree (University of Cambridge) is the corresponding author of this study that uses a range of Arabidopsis cell wall mutants to investigate the stem transcriptome. In plants with defects in xylan synthesis the authors found surprisingly few transcriptional changes. This indicates that once plants have committed to a terminal secondary cell wall program there is little need for transcriptional changes even after cell wall damage.


Johnston IG, Bassel GW (2018) Identification of a bet-hedging network motif generating noise in hormone concentrations and germination propensity in Arabidopsis. J R Soc Interface. 15(141). pii: 20180042. doi: 10.1098/rsif.2018.0042.

Open Access
Iain Johnston (University of Birmingham) leads this study that has identified a bet-hedging network that links hormone signaling during Arabidopsis germination. This type of network allows plants to more easily adapt to varying environmental conditions but can preclude maximum growth. In this system the network is based upon the regulation of ABA synthesis, activity and degradation, which is maintained at a constant mean level even though it exhibits significant noise. They investigate the parameters that might be tweaked to reduce variation in germination rate and therefore might be targets for modification in order to maximise responses under particular environmental conditions.

George Bassel who is a co-author on this paper will be speaking at the GARNet2018 Meeting in York in September.

http://rsif.royalsocietypublishing.org/content/15/141/20180042.long

Rubin MJ, Brock MT, Baker RL, Wilcox S, Anderson K, Davis SJ, Weinig C (2018) Circadian rhythms are associated with shoot architecture in natural settings. New Phytol. doi: 10.1111/nph.15162.

Seth Davies (University of York) is a co-author on this study that assesses the effect of circadian rhythms on aerial phenotypes that lead to fruit production in field grown Arabidopsis. This was assessed over two growing seasons and they show that variation in clock function significantly impacts shoot architecture.

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: March 2nd.

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Published on: March 2, 2018

The first two papers in this weeks Arabidopsis Research Roundup investigate different aspects of the plants response to temperature fluctuations. Firstly Lars Ostergaard (JIC) looks at the influence of temperature in the control of fruit dehiscence whilst Phil Wigge (SLCU) investigates crosstalk between chloroplast and nuclear signaling.

The third paper from Ian Henderson (University of Cambridge) studies the genetic elements that control rates of meiotic recombination. The next paper from the University of Leeds looks at the potential of using MET1 in the induction of novel epi-alleles whilst the penultimate paper includes the GARNet PI Jim Murray (Cardiff University) as a co-author and defines the role of CYCD7;1 in guard cell formation.

The final paper focusses on an enzyme involved in chlorophyll biosynthesis and includes Guy Hanke (QMUL) as a co-author.


https://linkinghub.elsevier.com/retrieve/pii/S1674-2052(18)30023-6

Li XR, Deb J, Kumar SV, Østergaard L (2018) Temperature Modulates Tissue-Specification Program to Control Fruit Dehiscence in Brassicaceae. Molecular Plant doi: 10.1016/j.molp.2018.01.003 Open Access

Lars Ostergaard (John Innes Centre) is the corresponding author that continues his groups work on the function of the INDEHISCENT protein, on this occasion looking at its involvement in the link between temperature and fruit dehiscence. They show that fruit valve margin development is accelerated at higher temperatures, facilitated by the activity of IND. This activity is associated with the changes in the induction dynamics of the known thermosensory histone H2A.Z and demonstrate a molecular framework for the response to changing temperature during fruit ripening.


http://www.cell.com/cell-reports/references/S2211-1247(18)30103-7

Dickinson PJ, Kumar M, Martinho C, Yoo SJ, Lan H, Artavanis G, Charoensawan V, Schöttler MA, Bock R, Jaeger KE, Wigge PA (2018) Chloroplast Signaling Gates Thermotolerance in Arabidopsis. Cell Rep. doi: 10.1016/j.celrep.2018.01.054 Open Access

Phil Wigge (SLCU) is the corresponding author on this study of the link between light-induced chloroplast signaling and thermotolerance. A forward genetic screen allowed the authors to identify two genes that demonstrated a key role for chloroplast signaling in controlling the activity of heat shock factors (HSFs), which enable the plant to cope with temperature variations. Subsequently they show that altering the binding activities of the HSFA1a protein can mimic heat shock response independent of any changes in temperature.


Serra H, Lambing C, Griffin CH, Topp SD, Nageswaran DC, Underwood CJ, Ziolkowski PA, Séguéla-Arnaud M, Fernandes JB,, Mercier R, Henderson IR (2018) Massive crossover elevation via combination of HEI10 and recq4a recq4b during Arabidopsis meiosis. PNAS doi: 10.1073/pnas.1713071115

Ian Henderson (University of Cambridge) is the corresponding author on this collaboration with French colleagues in a study that investigates the factors that control recombination frequency in meiosis. During normal meiotic recombination the majority of double stranded breaks will not form crossovers (over 90%) so to increase this frequency they altered the active dosage of genetic elements that are either pro-crossover or anti-crossover control. This strategy results in a massive increase in crossovers and provides a genetic framework for increasing recombination, a strategy that can be critically important for increasing variation during crop breeding.


Brocklehurst S, Watson M, Carr IM, Out S, Heidmann I, Meyer P (2018) Induction of epigenetic variation in Arabidopsis by over-expression of DNA METHYLTRANSFERASE1 (MET1). PLoS One. doi: 10.1371/journal.pone.0192170 Open Access

This study from the University of Leeds is led by Peter Meyer and investigates how overexpression of the METHYLTRANSFERASE1 (MET1) gene might generate novel epi-alleles that result in altered gene expression. This strategy indeed generated novel epi-alleles that increased expression at loci encoding TEs, non-coding RNAs and protein coding genes. Importantly any altered expression can be transmitted to the next generation, independent of the presence of a MET1 expressing transgene. However the long term stability of these epi-alleles differs in an loci-specific manner.


Weimer AK, Matos JL, Sharma N, Patell F, Murray JAH, Dewitte W, Bergmann DC (2018) Lineage and stage-specific expressed CYCD7;1 coordinates the single symmetric division that creates stomatal guard cells. Development. doi: 10.1242/dev.160671

GARNet PI Jim Murray and Walter DeWitte (Cardiff University) are co-authors on this US-led study that adds complexity to our understanding of the molecular players that control guard cell specification. The authors show that the D-type cyclin CYCD7;1 is expressed during a short time window prior to the symmetry division that forms two guard cells. This activity is controlled by cell-type specific transcription factors acting in the appropriate time period.

http://dev.biologists.org/content/early/2018/02/14/dev.160671.long

Herbst J, Girke A, Hajirezaei MR, Hanke G, Grimm B (2018) Potential Roles of YCF54 and Ferredoxin-NADPH Reductase for Magnesium Protoporphyrin Monomethylester Cyclase. Plant J. doi: 10.1111/tpj.13869

Guy Hanke (QMUL) is a co-author on this German-led study that investigates an enzyme reactions that occur during chlorophyll biosynthesis. Specifically they showed that plants lacking the LCAA/YCF54 subunit of the enzyme MgProto monomethylester (MgProtoME) cyclase causes accumulation of MgProtoME and destabilization of the entire cyclase enzyme. This disrupts chlorophyll synthesis and negatively effects photosynthetic activity.

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.

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