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Results for “Plant Biology”

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bioRxiv2017

Time-resolved analyses of elemental distribution and concentration in living plants: An example using manganese toxicity in cowpea leaves

O_LIKnowledge of elemental distribution and concentration within plant tissues is crucial in the understanding of almost every process that occurs within plants. However, analytical limitations have hindered the microscopic determination of changes over time in the location and concentration of nutrients and contaminants in living plant tissues.\nC_LIO_LIWe developed a novel method using synchrotron-based micro X-ray fluorescence (-XRF) that allows for laterally-resolved, multi-element, kinetic analyses of plant leaf tissues in vivo. To test the utility of this approach, we examined changes in the accumulation of Mn in unifoliate leaves of 7-d-old cowpea (Vigna unguiculata) plants grown for 48 h at 0.2 and 30 M Mn in solution.\nC_LIO_LIRepeated -XRF scanning did not damage leaf tissues demonstrating the validity of the method. Exposure to 30 M Mn for 48 h increased the initial number of small spots of localized high Mn and their concentration rose from 40 to 670 mg Mn kg-1 fresh mass. Extension of the two-dimensional -XRF scans to a three-dimensional geometry provided further assessment of Mn localization and concentration.\nC_LIO_LIThis method shows the value of synchrotron-based -XRF analyses for time-resolved in vivo analysis of elemental dynamics in plant sciences.\nC_LI

plant biology↗

bioRxiv2017

The transcriptional dynamics of two filmy ferns from Hymenophyllaceae with different niche preferences unravel key aspects of their desiccation tolerance and vertical distribution along host trees

Ferns from the Hymenophyllaceae family are one of the main components of the epiphytic species diversity in the Chilean temperate rain forest. Having membranous fronds of a single layer of cells, they show a poikilohydric strategy most typical from bryophytes. Although Hymenophyllaceae species shows the ability to tolerate desiccation, there are interspecific differences in their water loss kinetic. Counter-intuitively, those species that have rapid desiccation kinetic are able to reach higher host height and tolerate higher light exposure and vapor pressure deficit. Therefore, what are the mechanisms (constitutive and/or induced) responsible of the desiccation tolerance in this fern family? As this primitive fern family is closely related with mosses, it can be hypothesized that desiccation tolerance in this particular group would be associated with constitutive features rather than induced responses during dehydration. However, the inter-specific differences in water loss and vertical distribution would be associated to different degrees of induction either within the dehydration or rehydration phases. We applied an ecophysiological transcriptomic approach to study the dynamic of gene expression in two species of filmy ferns with contrasting desiccation kinetics and vertical distribution on the host tree. Our analysis identified commonalities and differences in gene regulation, and key genes correlated with the fronds hydration state, providing the patterns of gene expression responding to microenvironmental signals and behind the physiology of their resurrection strategy

plant biology↗

bioRxiv2017

Alleviation of Al toxicity by Si is associated with the formation of Al-Si complexes in root tissues of sorghum

Silicon is reported to reduce the toxic effects of Al on root elongation but the in planta mechanism by which this occurs remains unclear. Using seedlings of soybean (Glycine max) and sorghum (Sorghum bicolor), we examined the effect of up to 2 mM Si on root elongation rate (RER) in Al-toxic nutrient solutions. Synchrotron-based low energy X-ray fluorescence (LEXRF) was then used for the in situ examination of the distribution of Al and Si within cross-sections cut from the apical tissues of sorghum roots. The addition of Si potentially increased RER in Al-toxic solutions, with RER being up to ca. 0.3 mm h-1 (14 %) higher for soybean and ca. 0.2 mm h-1 (17 %) higher for sorghum relative to solutions without added Si. This improvement in RER could not be attributed to a change in Al-chemistry of the bulk nutrient solution, nor was it due to a change in the concentration of Al within the apical (0-10 mm) root tissues. Using LEXRF to examine sorghum, it was demonstrated that in roots exposed to both Al and Si, much of the Al was co-located with Si in the mucigel and outer apoplast. These observations suggest that Si reduces the toxicity of Al in planta through formation of Al-Si complexes in mucigel and outer cellular tissues, thereby decreasing the binding of Al to the cell wall where it is known to inhibit wall loosening as required for cell elongation.

plant biology↗

bioRxiv2017

Extremely flexible infection programs in a fungal plant pathogen

Filamentous plant pathogens exhibit extraordinary levels of genomic variability that is proposed to facilitate rapid adaptation to changing host environments. However, the impact of genomic variation on phenotypic differentiation in pathogen populations is largely unknown. Here, we address the extent of variability in infection phenotypes of the hemibiotrophic wheat pathogen Zymoseptoria tritici by studying three field isolates collected in Denmark, Iran, and the Netherlands. These three isolates differ extensively in genome structure and gene content, but produce similar disease symptoms in the same susceptible wheat cultivar. Using advanced confocal microscopy, staining of reactive oxygen species, and comparative analyses of infection stage-specific RNA-seq data, we demonstrate considerable variation in the temporal and spatial course of infection of the three isolates. Based on microscopic observation, we determined four core infection stages: establishment, biotrophic growth, lifestyle transition, and necrotrophic growth and asexual reproduction. Comparative analyses of the fungal transcriptomes, sequenced for every infection stage, revealed that the gene expression profiles of the isolates differed significantly, and 20% of the genes are differentially expressed between the three isolates during infection. The genes exhibiting isolate-specific expression patterns are enriched in genes encoding effector candidates that are small, secreted, cysteine-rich proteins and putative virulence determinants. Moreover, the differentially expressed genes were located significantly closer to transposable elements, which are enriched for the heterochromatin-associated histone marks H3K9me3 and H3K27me3 on the accessory chromosomes. This observation indicates that transposable elements and epigenetic regulation contribute to the infection-associated transcriptional variation between the isolates. Our findings illustrate how high genetic diversity in a pathogen population can result in highly differentiated infection and expression phenotypes that can support rapid adaptation in changing environments. Furthermore, our study reveals an exceptionally high extent of plasticity in the infection program of an important wheat pathogen and shows a substantial redundancy in infection-related gene expression.\n\nAuthor summaryZymoseptoria tritici is a pathogen that infects wheat and induces necrosis in leaf tissue. Z. tritici field populations exhibit high levels of genetic diversity, and here we addressed the consequences of this diversity on infection phenotypes. We conducted a detailed comparison of the infection processes of three Z. tritici isolates collected in Denmark, the Netherlands, and Iran. We inoculated leaves of a susceptible wheat cultivar and monitored development of disease symptoms and infection structures in leaf tissue by confocal microscopy. The three isolates exhibited highly differentiated spatial and temporal patterns of infection, although quantitative disease was similar. Furthermore, more than 20% of the genes were differentially expressed in the three isolates during wheat infection. Variation in gene expression is particularly associated with transposable elements, suggesting a role of epigenetic regulation in transcriptional variation among the three isolates. Finally, we find that genes encoding putative virulence determinants were enriched among the differentially expressed genes, suggesting that each of the three Z. tritici isolates utilizes different strategies to manipulate host defenses. Our results emphasize that phenotypic diversity plays an important role in pathogen populations and should be considered when developing crop protection strategies.

plant biology↗

bioRxiv2018

Synergistic binding of bHLH transcription factors to the promoter of the maize NADP-ME gene used in C4 photosynthesis is based on an ancient code found in the ancestral C3 state

C4 photosynthesis has evolved repeatedly from the ancestral C3 state to generate a carbon concentrating mechanism that increases photosynthetic efficiency. This specialised form of photosynthesis is particularly common in the PACMAD clade of grasses, and is used by many of the worlds most productive crops. The C4 cycle is accomplished through cell-type specific accumulation of enzymes but cis-elements and transcription factors controlling C4 photosynthesis remain largely unknown. Using the NADP-Malic Enzyme (NADP-ME) gene as a model we aimed to better understand molecular mechanisms associated with the evolution of C4 photosynthesis. Two basic Helix-Loop-Helix (bHLH) transcription factors, ZmbHLH128 and ZmbHLH129, were shown to bind the C4 NADP-ME promoter from maize. These proteins form heterodimers and ZmbHLH129 impairs trans-activation by ZmbHLH128. Electrophoretic mobility shift assays indicate that a pair of cis-elements separated by a seven base pair spacer synergistically bind either ZmbHLH128 or ZmbHLH129. This pair of cis-elements is found in both C3 and C4 species of the PACMAD clade. Our analysis is consistent with this cis-element pair originating from a single motif present in the ancestral C3 state. We conclude that C4 photosynthesis has co-opted an ancient C3 regulatory code built on G-box recognition by bHLH to regulate the NADP-ME gene. More broadly, our findings also contribute to the understanding of gene regulatory networks controlling C4 photosynthesis.

plant biology↗

bioRxiv2017

LRX- and FER-dependent extracellular sensing coordinates vacuolar size for cytosol homeostasis

Cellular elongation requires the defined coordination of intra- and extracellular processes. The vacuole is the biggest plant organelle and its dimension has a role in limiting cell expansion (Lofke et al., 2015; Scheuring et al., 2016). We reveal that the increase in vacuolar occupancy enables cellular elongation with relatively little enlargement of the cytosole. It remains, however, completely unknown how the vacuolar size is coordinated with other growth-relevant processes. Intriguingly, we show that extracellular constraints impact on the intracellular expansion of the vacuole. The underlying cell wall sensing mechanism requires the interaction of the extracellular leucine-rich repeat extensin (LRX) with the receptor-like kinase Feronia (FER). Our data suggests that LRX links the plasma membrane localised FER with the cell wall, allowing this module to jointly sense and convey extracellular signals to the underlying cell. This mechanism coordinates cell wall acidification/loosening with the increase in vacuolar size, contributing cytosol homeostasis during plant cell expansion.

plant biology↗

bioRxiv2017

BRUTUS-LIKE proteins moderate the transcriptional response to iron deficiency in roots

Iron is an essential micronutrient but in excess is toxic inside cells. Under iron deficiency, the expression of iron uptake genes is increased, but it is not known how the transcriptional response is controlled to avoid uptake of too much iron. The hemerythrin E3 ligases BRUTUS (BTS) and BTS-LIKE (BTSL) have previously been identified as negative regulators of the iron deficiency response. Our phylogenetic analysis indicated that BTSL proteins are present in dicotyledonous plants only and form a separate clade from BTS homologs. BTSL1 and BTSL2 in Arabidopsis thaliana are in a network with nearly all iron uptake genes, whereas BTS is in a shoot-specific network. BTSL1 and BTSL2 are expressed predominantly in the root epidermis and cortex, separate from BTS in the root stele, shoot and embryos. Mutant analysis identified BTSL2 as the dominant paralog of the otherwise redundant BTSL genes. The btsl double mutant had increased protein levels of FIT, the FER-like Iron deficiency-induced Transcription factor, and failed to switch off the transcriptional response upon iron resupply, leading to dramatic iron accumulation in roots and shoots. Protein interaction between the C-terminus of BTSL proteins and FIT indicate that FIT is a direct target for degradation. Taken together, our studies show that BTSL1 and BTSL2 control iron uptake in the epidermis and cortex, upstream of BTS in the vasculature and leaves.

plant biology↗

bioRxiv2017

VlbZIP30 of grapevine functions in drought tolerance via the abscisic acid core signaling pathway

Drought stress limits the growth and development of grapevines, thereby reducing productivity, but the mechanisms by which grapevines respond to drought stress remain largely uncharacterized. Here, we characterized a group A bZIP gene from Kyoho grapevine, VlbZIP30, which was shown to be induced by abscisic acid (ABA) and dehydration stress. Overexpression of VlbZIP30 in transgenic Arabidopsis enhanced dehydration tolerance during seed germination, and in the seedling and adult stages. Transcriptome analysis revealed that a major proportion of ABA- and/or drought-responsive genes are transcriptionally regulated by VlbZIP30 during ABA or mannitol treatment at the cotyledon greening stage. We identified an A. thaliana G-box motif (CACGTG) and a potential grapevine G-box motif (MCACGTGK) in the promoters of the 39 selected A. thaliana genes up-regulated in the transgenic plants and in the 35 grapevine homologs, respectively. Subsequently, using two grapevine-related databases, we found that 74% and 84% (a total of 27 genes) of the detected grapevine genes were significantly up-regulated by ABA and drought stress, respectively, suggesting that these 27 genes involve in ABA or dehydration stress and may be regulated by VlbZIP30 in grapevine. We propose that VlbZIP30 functions as a positive regulator of drought-responsive signaling in the ABA core signaling pathway.\n\nHighlightVlbZIP30 positively regulate plant drought tolerance through regulated the expression of 27 grapevine candidate genes via G-box cis-element (MCACGTGK) in ABA signaling pathway.

plant biology↗

bioRxiv2017

A Genetic Pathway Composed of EDT1/HDG11, ERECTA, and E2Fa Loci Regulates Water Use Efficiency by Modulating Stomatal Density

Improvement of crop drought resistance and water use efficiency (WUE) has been a major endeavor in agriculture. ERECTA is the first identified major effector of water use efficiency. However, the underlying molecular mechanism is not well understood. Here, we report a genetic pathway, composed of EDT1/HDG11, ERECTA, and E2Fa loci, which regulates water use efficiency by modulating stomatal density. The HD-START transcription factor EDT1/HDG11 transcriptionally activates ERECTA expression by binding to an HD cis-element in the ERECTA promoter. ERECTA in turn relies on E2Fa to control the expression of cell-cycle related genes and the transition from mitosis to endocycle, which leads to increased nuclear DNA content in leaf cells, and therefore increased cell size and decreased stomatal density. The decreased stomatal density improves plant WUE. Our study demonstrates the EDT1/HDG11-ERECTA-E2Fa genetic pathway that reduces stomatal density by increasing cell size, providing a new avenue to improve WUE of crops.

plant biology↗

bioRxiv2017

An epigenetic breeding system in soybean for increased yield and stability

Epigenetic variation has been associated with a wide range of adaptive phenotypes in plants, but there exist few direct means for exploiting this variation. RNAi suppression of the plant-specific gene, MutS HOMOLOG1 (MSH1), in multiple plant species produces a range of developmental changes accompanied by modulation of defense, phytohormone, and abiotic stress response pathways. This msh1-conditioned developmental reprogramming is retained independent of transgene segregation, giving rise to transgene-null memory effects. An isogenic memory line crossed to wild type produces progeny families displaying increased variation in adaptive traits that respond to selection. This study investigates amenability of the MSH1 system for inducing epigenetic variation in soybean that may be of value agronomically. We developed epi-line populations by crossing with msh1-acquired soybean memory lines. Derived soybean epi-lines showed increase in variance for multiple yield-related traits including pods per plant, seed weight, and maturity time in both greenhouse and field trials. Selected epi-F2:4 and epi-F2:5 lines showed an increase in seed yield over wild type. By epi-F2:6, we observed a return of MSH1-derived enhanced growth back to wild type levels. Epi-populations also showed evidence of reduced epitype-by-environment (e x E) interaction, indicating higher yield stability. Transcript profiling of the soybean epi-lines identified putative signatures of enhanced growth behavior across generations. Genes related to cell cycle, abscisic acid biosynthesis, and auxin-response, particularly SMALL AUXIN UP RNAs (SAURs), were differentially expressed in epi-F2:4 lines that showed increased yield when compared to epi-F2:6. These data support the potential of msh1-derived epigenetic variation in plant breeding for enhanced yield and yield stability.

plant biology↗

bioRxiv2017

Exploring the Genetics of Lesion and Nodal Resistance in Pea (Pisum sativum L.) to Sclerotinia sclerotiorum Using Genome-wide Association Studies and RNA-Seq

The disease white mold caused by the fungus Sclerotinia sclerotiorum is a significant threat to pea production and improved resistance to this disease is needed. Nodal resistance in plants is a phenomenon where a fungal infection is prevented from passing through a node and the infection is limited to an internode region. Nodal resistance has been observed in some pathosystems such as the pea (Pisum sativum L.)-S. sclerotiorum pathosystem. Other than nodal resistance, different pea lines display different levels of stem lesion size restriction, referred to as lesion resistance. It is unclear whether the genetics of lesion resistance and nodal resistance are identical or different. This study applied genome-wide association studies (GWAS) and RNA-Seq to understand the genetic makeup of these two types of resistance. The time series RNA-Seq experiment consisted of two pea lines (the susceptible Lifter and the partially resistant PI 240515), two treatments (mock samples and S. sclerotiorum inoculated samples), and three time points (12, 24, and 48 hours post-inoculation). Integrated results from GWAS and RNA-Seq analyses identified different redox-related transcripts for lesion and nodal resistances. A transcript encoding a glutathione S-transferase was the only shared resistance source for both phenotypes. There were more leucine rich-repeat containing transcripts found for lesion resistance, while different candidate resistance transcripts such as a VQ motif-containing protein and a myo-inositol oxygenase were found for nodal resistance. This study demonstrated the robustness of combining GWAS and RNA-Seq for identifying white mold resistance in pea, and results suggest different genetics underlying lesion and nodal resistance.

plant biology↗

bioRxiv2017

Bundle-sheath cells are internal leaf "water valves" controlled via xylem acidification by H+-ATPase

O_LIThe leaf vascular bundle sheath cells (BSCs) that tightly envelop the leaf veins, are a selective and dynamic barrier to xylem-sap water and solutes radially entering the mesophyll cells. Under normal conditions, xylem-sap pH of <6 is presumably important for driving and regulating the transmembranal solute transport. Having discovered recently a differentially high expression of a BSCs proton pump, AHA2, we now test the hypothesis that it regulates this pH and leaf radial water fluxes. C_LIO_LIWe monitored the xylem-sap pH in the veins of detached leaves of WT Arabidopsis, AHA mutants, and aha2 mutants complemented with AHA2 gene solely in BSCs. We tested an AHA inhibitor and stimulator, and different pH buffers. We monitored their impact on the xylem-sap pH and the whole leaf hydraulic conductance (Kleaf), and the effect of pH on the water osmotic permeability (Pf) of isolated BSCs protoplasts. C_LIO_LIOur results demonstrated that AHA2 is necessary for xylem-sap acidification, and in turn, for elevating Kleaf. Conversely, knocking out AHA2 alkalinized the xylem-sap. Also, elevating xylem sap pH to 7.5 reduced Kleaf and elevating external pH to 7.5 decreased the BSCs Pf. C_LIO_LIAll these demonstrate a causative link between AHA2 activity in BSCs and leaf radial water conductance. C_LI One-sentence summaryBundle-sheath cells can control the leaf hydraulic conductance by proton-pump-regulated xylem sap pH

plant biology↗

bioRxiv2017

Trait components of whole plant water use efficiency are defined by unique, environmentally responsive genetic signatures in the model C4 grass Setaria

Plant growth and water use are interrelated processes influenced by the genetic control of both plant morphological and biochemical characteristics. Improving plant water use efficiency (WUE) to sustain growth in different environments is an important breeding objective that can improve crop yields and enhance agricultural sustainability. However, genetic improvements of WUE using traditional methods have proven difficult due to low throughput and environmental heterogeneity encountered in field settings. To overcome these limitations the study presented here utilizes a high-throughput phenotyping platform to quantify plant size and water use of an interspecific Setaria italica x Setaria viridis recombinant inbred line population at daily intervals in both well-watered and water-limited conditions. Our findings indicate that measurements of plant size and water use in this system are strongly correlated; therefore, a linear modeling approach was used to partition this relationship into predicted values of plant size given water use and deviations from this relationship at the genotype level. The resulting traits describing plant size, water use and WUE were all heritable and responsive to soil water availability, allowing for a genetic dissection of the components of plant WUE under different watering treatments. Linkage mapping identified major loci underlying two different pleiotropic components of WUE. This study indicates that alleles controlling WUE derived from both wild and domesticated accessions of the model C4 species Setaria can be utilized to predictably modulate trait values given a specified precipitation regime.

plant biology↗

bioRxiv2018

Mapping and dynamics of regulatory DNA during seed development

The genome is reprogrammed during development to produce diverse cell types, largely through altered expression and activity of key transcription factors. The accessibility and critical functions of epidermal cells have made them a model for connecting transcriptional events to development in a range of model systems. In Arabidopsis thaliana and many other plants, fertilization triggers differentiation of specialized epidermal seed coat cells that have a unique morphology caused by large extracellular deposits of pectin. Here, we used DNase I-seq to generate regulatory landscapes of A. thaliana seeds at two critical time points in seed coat maturation, enriching for seed coat cells with the INTACT method. We found over 3000 developmentally dynamic regulatory DNA elements and explored their relationship with nearby gene expression. The dynamic regulatory elements were enriched for motifs for several transcription factors families; most notably the TCP family at the earlier time point and the MYB family at the later one. To assess the extent to which the observed regulatory sites in seeds added to previously known regulatory sites in A. thaliana, we compared our data to 11 other data sets generated with seven-day-old seedlings for diverse tissues and conditions. Surprisingly, over a quarter of the regulatory, i.e. accessible, bases observed in seeds were novel. Notably, in this comparison, development exerted a stronger effect on the plant regulatory landscape than extreme environmental perturbations, highlighting the importance of extending studies of regulatory landscapes to other tissues and cell types during development.

plant biology↗

bioRxiv2017

Multiple strategies for heat adaptation in rice endosperms revealed by on-site cell-specific analysis

Plant cells have multiple strategies to adapt to environmental stresses. Rice endosperms form chalkiness in a part of the tissue under heat conditions during the grain-filling stage, although nitrogen supply reduces chalky rice. Air spaces formed in the cells cause an irregular light reflection and create chalkiness, yet what exactly occurs remains unclear at cell level. Through on-site cell-specific analysis, we show that heat-treated cells adjust osmotically and retard protein synthesis to preserve protein storage vacuoles in the cytosol, resulting in air space formation. Application of nitrogen enhances heat tolerance to sustain protein body and amyloplast development during strong osmotic adjustment, which diminishes air spaces to avoid chalkiness. Hence, we conclude that rice endosperm cells could alter organelle compartments spatially during the heat adaptation, depending on the available nitrogen level. Our findings provide new insight into the cellular mechanism of rice chalky formation as a strategy for heat acclimation.

plant biology↗

bioRxiv2017

Blue Light Negatively Regulates Tolerance to Phosphate Deficiency in Arabidopsis

Plants have evolved mechanisms to improve utilization efficiency or acquisition of inorganic phosphate (Pi) in response to Pi deficiency, such as altering root architecture, secreting acid phosphatases, and activating the expression of genes related to Pi uptake and recycling. Although many genes responsive to Pi starvation have been identified, transcription factors that affect tolerance to Pi deficiency have not been well characterized. We show here that defect in the ELONGATED HYPOCOTYL 5 (HY5) transcription factor gene results in tolerance to Pi deficiency in Arabidopsis. The primary root length of hy5 was only slightly inhibited under Pi deficient condition and its fresh weight was significantly higher than that of wild type. The Pi deficiency-tolerant phenotype of hy5 was similarly observed when grown on the medium without Pi. In addition, a double mutant, hy5slr1, without lateral roots also showed tolerance to phosphate deficiency, indicating that the tolerance of hy5 does not result from increase of external Pi uptake and may be related to internal Pi utilization or recycling. Moreover, we found that blue light negatively regulates tolerance to Pi-deficiency and that hy5 exhibits tolerance to Pi deficiency due to blockage of blue-light responses. Collectively, this study points out light quality may play an important role in the regulation of internal Pi recycling and utilization efficiency. Also, it may contribute to reducing Pi fertilizer requirements in plants through a proper illumination.

plant biology↗

bioRxiv2017

Fluorescent reporter lines for auxin and cytokinin signalling in barley (Hordeum vulgare)

The phytohormones auxin and cytokinin influence the development and maintenance of plant stem cell niches. Although barley (Hordeum vulgare) is the fourth most abundant cereal crop plant, the knowledge about these important phytohormones in regard to the root and shoot stem cell niche in barley is still negligible. In this study, we analyse the influence of auxin and cytokinin on the barley root meristem and present reporter lines to describe the auxin and cytokinin signalling output. Application of high concentrations of auxin and cytokinin to barley seedlings had a negative influence on barley root and meristem growth. The expression of the cytokinin reporter TCSn revealed that cytokinin signalling mostly takes place in the stele cells proximal to the QC and in the differentiated root cap cells, but can additionally be activated in the root stem cell niche by cytokinin application. Analysing signalling targets of auxin showed that a homologue of AtPLT1, HvPLT1, is expressed in a similar way as AtPLT1 in Arabidopsis, in particular in the QC and the surrounding cells. Furthermore, a homologue of the auxin PIN transporters PIN1, HvPIN1, was expressed in the root and the shoot meristem and polarly localizes to the plasma membrane. Its expression is regulated by cytokinin and the intracellular localisation is affected by BFA. With this study, we provide a valuable tool set of fluorescent barley reporter lines for auxin and cytokinin.

plant biology↗

bioRxiv2017

Synthetic hormone-responsive transcription factors can monitor and re-program plant development

Developmental programs continuously sculpt plant morphology to meet environmental challenges, and these same programs have been manipulated to increase agricultural productivity1,2. Small molecule phytohormones act as signals within these programs creating chemical circuitry3 that, in many cases, has been represented in mathematical models4,5. To date, model validation and forward engineering of plant morphology has been largely restricted to adding or subtracting genes, as more nuanced tools to modulate key control parameters identified by such models in vivo are severely limited6,7. Here, we use Arabidopsis thaliana to validate a novel set of synthetic and modular hormone activated Cas9-based repressors (HACRs) that respond to three phytohormones: auxin, gibberellins and jasmonates. We demonstrate that HACRs can regulate genes in response to both exogenous hormone treatments, as well as in response to local differences in endogenous hormone levels associated with developmental events. We further show that HACRs can be used to reprogram the agriculturally relevant traits of shoot branching and phyllotaxy by tuning canalization strength, a critical control parameter predicted by mathematical models. By deploying a HACR to re-parameterize the threshold for induction of the auxin transporter PIN-FORMED1 (PIN1), we observed a decrease in shoot branching and phyllotactic noise as predicted by existing models4,5. The approach described here provides a framework for improved mapping of developmental circuitry, as well as a means to better leverage model predictions to engineer development.

plant biology↗