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Allele specific expression analysis identifies regulatory variation associated with stress-related genes in the Mexican highland maize landrace Palomero Toluqueno

BackgroundGene regulatory variation has been proposed to play an important role in the adaptation of plants to environmental stress. In the central highlands of Mexico, farmer selection has generated a unique group of maize landraces adapted the challenges of the highland niche. In this study, gene expression in Mexican highland maize and a reference maize breeding line were compared to identify evidence of regulatory variation in stress-related genes. It was hypothesised that local adaptation in Mexican highland maize would be associated with a transcriptional signature observable even under benign conditions.\n\nMethodsAllele specific expression analysis was performed using the seedling-leaf transcriptome of an F1 individual generated from the cross between the highland adapted Mexican landrace Palomero Toluqueno and the reference line B73, grown under benign conditions. Results were compared with a published dataset describing the transcriptional response of B73 seedlings to cold, heat, salt and UV treatments.\n\nResultsA total of 2386 genes were identified to show allele specific expression. Of these, 277 showed an expression difference between Palomero Toluqueno and B73 alleles that mirrored the response of B73 cold, heat, salt and/or UV treatments, and, as such, were considered to display a constitutive stress response. Constitutive stress response candidates included genes associated with plant hormone signaling and a number of transcription factors. Construction of a gene co-expression network revealed further signaling and stress-related genes to be among the potential targets of the transcription factors candidates.\n\nDiscussionConstitutive activation of responses may represent the best strategy when stresses are severe but predictable. Expression differences observed here between PT and B73 alleles indicate the presence of cis-acting regulatory variation linked to stress-related genes in PT. Considered alongside gene annotation and population data, allele specific expression analysis of plants grown under benign conditions provides an attractive strategy to identify functional variation potentially linked to local adaptation.

plant biology

Combining semi-automated image analysis techniques with machine learning algorithms to accelerate large scale genetic studies

BackgroundGenetic analyses of plant root system development require large datasets of extracted architectural traits. To quantify such traits from images of root systems, researchers often have to choose between automated tools (that are prone to error and extract only a limited number of architectural traits) or semi-automated ones (that are highly time consuming).\n\nFindingsWe trained a Random Forest algorithm to infer architectural traits from automatically-extracted image descriptors. The training was performed on a subset of the dataset, then applied to its entirety. This strategy allowed us to (i) decrease the image analysis time by 73% and (ii) extract meaningful architectural traits based on image descriptors. We also show that these traits are sufficient to identify Quantitative Trait Loci that had previously been discovered using a semi-automated method.\n\nConclusionsWe have shown that combining semi-automated image analysis with machine learning algorithms has the power to increase the throughput in large scale root studies. We expect that such an approach will enable the quantification of more complex root systems for genetic studies. We also believe that our approach could be extended to other area of plant phenotyping.

plant biology

Deriving C4 photosynthesis parameters by fitting intensive A/Ci curves

Measurements of photosynthetic assimilation rate as a function of intercellular CO2 (A/Ci curves) are widely used to estimate photosynthetic parameters for C3 species, yet few parameters have been reported for C4 plants, because of a lack of estimation methods. Here, we extend the framework of widely-used estimation methods for C3 plants to build estimation tools by exclusively fitting intensive A/Ci curves (6-8 more sampling points) for C4 using three versions of photosynthesis models with different assumptions about carbonic anhydrase processes and ATP distribution. We use simulation-analysis, out-of-sample tests, existing in vitro measurements and chlorophyll-fluorescence-measurements to validate the new estimation methods. Of the five/six photosynthetic parameters obtained, sensitivity analyses show that maximal-Rubisco-carboxylation-rate, electron-transport-rate, maximal-PEP-carboxylation-rate and carbonic-anhydrase were robust to variation in the input parameters, while day-respiration and mesophyll-conductance varied. Our method provides a way to estimate carbonic anhydrase activity, a new parameter, from A/Ci curves, yet also shows that models that do not explicitly consider carbonic anhydrase yield approximate results. The two photosynthesis models, differing in whether ATP could freely transport between RuBP and PEP regeneration processes yielded consistent results under high light, but they may diverge under low light intensities. Modeling results show selection for Rubisco of low specificity and high catalytic rate, low leakage of bundle sheath and high PEPC affinity, which may further increase C4 efficiency.

plant biology

ATP sensing in living plant cells reveals tissue gradients and stress dynamics of energy physiology

Growth and development of plants is ultimately driven by light energy captured through photosynthesis. ATP acts as universal cellular energy cofactor fuelling all life processes, including gene expression, metabolism, and transport. Despite a mechanistic understanding of ATP biochemistry, ATP dynamics in the living plant have been largely elusive. Here we establish live MgATP2- assessment in plants using the fluorescent protein biosensor ATeam1.03-nD/nA. We generate Arabidopsis sensor lines and investigate the sensor in vitro under conditions appropriate for the plant cytosol. We establish an assay for ATP fluxes in isolated mitochondria, and demonstrate that the sensor responds rapidly and reliably to MgATP2- changes in planta. A MgATP2- map of the Arabidopsis seedling highlights different MgATP2- concentrations between tissues and in individual cell types, such as root hairs. Progression of hypoxia reveals substantial plasticity of ATP homeostasis in seedlings, demonstrating that ATP dynamics can be monitored in the living plant.\n\nOne-sentence SummarySensing of MgATP2- by fluorimetry and microscopy allows dissection of ATP fluxes of isolated organelles, and dynamics of cytosolic MgATP2- in vivo.\n\nFunding AgenciesThis work was supported by the Deutsche Forschungsgemeinschaft (DFG) through the Emmy-Noether programme (SCHW1719/1-1; M.S. and GR4251/1-1; C.G.), the Research Training Group GRK 2064 (M.S.; A.J.M.), the Priority Program SPP1710 (A.J.M.) and a grant (SCHW1719/5-1; M.S.) as part of the package PAK918. The Seed Fund grant CoSens from the Bioeconomy Science Center, NRW (A.J.M.; M.S.) is gratefully acknowledged. The scientific activities of the Bioeconomy Science Center were financially supported by the Ministry of Innovation, Science and Research within the framework of the NRW Strategieprojekt BioSC (No. 313/323-400-002 13). A.Co. received funding by the Ministero dellIstruzione, dellUniversita e della Ricerca through the FIRB 2010 programme (RBFR10S1LJ_001) and Piano di Sviluppo di Ateneo 2015 (Universita degli Studi di Milano). M.Z. received funding by the Ministero dellIstruzione, dellUniversita e della Ricerca (Italy) through the PRIN 2010 programme (PRIN2010CSJX4F). S.W. and T.N. received travel support by the Deutscher Akademischer Austauschdienst (DAAD). V.D.C. was supported by the European Social Fund, Operational Programme 2007/2013, and an Erasmus+ Traineeship grant. M.D.F was supported by The Human Frontier Science Program (RPG0053/2012), and the Leverhulme Foundation (RPG-2015-437). I.M.M. was supported by a grant from the Danish Council for Independent Research - Natural Sciences. V.C.P. was supported by the Innovation and Technology Fund (Funding Support to Partner State Key Laboratories in Hong Kong) of the HKSAR.\n\nAbbreviationsAAC - ADP/ATP carrier; AK - adenylate kinase; cAT - carboxyatractyloside; CCCP - carbonyl cyanide m-chlorophenyl hydrazone; CFP - cyan fluorescent protein; CLSM - confocal laser scanning microscopy; ETC - electron transport chain; FRET - Forster Resonance Energy Transfer; LSFM - light sheet fluorescence microscopy.

plant biology

Cyclic electron flow in Chlamydomonas reinhardtii

Cyclic electron flow (CEF), one of the major alternative electron transport pathways to the primary linear electron flow (LEF) in chloroplasts has been discovered in the middle of the last century. It is defined as a return of the reductants from the acceptor side of the Photosystem I (PSI) to the pool of its donors via the cytochrome b6f, and has proven essential for photosynthesis. However, despite many efforts aimed at its characterisation, the pathway and regulation of CEF remain equivocal, and its physiological significance remains to be properly defined. Here we use novel spectroscopic approaches to measure CEF in transitory conditions in the green alga Chlamydomonas reinhardtii. We show that CEF operates at the same maximal rate regardless of the oxygen concentration, and that the latter influences LEF, rather than CEF in vivo, which questions the recent hypotheses about the CEF supercomplex formation. We further reveal that the pathways proposed for CEF in the literature are inconsistent with the kinetic information provided by our measurements. We finally provide cues on the regulation of CEF by light.

plant biology

Varying water deficit stress (WDS) tolerance in grain amaranths involves multifactorial shifts in WDS-related responses

In this study, water deficit stress (WDS)-tolerance in several cultivars of grain amaranth species (Amaranthus hypochondriacus [Ahypo], A. cruentus [Acru] and A. caudatus [Acau]), in addition to A. hybridus (Ahyb), an ancestral amaranth, was examined. Ahypo was the most WDS-tolerant species, whereas Acau and Ahyb were WDS-sensitive. Data revealed that the differential WDS tolerance observed was multifactorial. It involved increased proline and raffinose (Raf) in leaves and/ or roots. Higher foliar Raf coincided with induced Galactinol synthase 1 (AhGolS1) and Raffinose synthase (AhRafS) expression. Unknown compounds, possibly larger RFOs, also accumulated in leaves of WDS-tolerant amaranths, which had high Raf/ Verbascose ratios. Distinct nonstructural carbohydrate (NSC) accumulation patterns were observed in tolerant species under WDS and recovery, such as: i) high Hex/ Suc ratios in roots coupled to increased cell wall and vacuolar invertase and sucrose synthase activities; ii) a severer depletion of starch reserves; iii) lower NSC content in leaves, and iv) higher basal hexose levels in roots which further increased under WDS. WDS-marker gene expression patterns proposed a link between amaranths WDS tolerance and abscisic acid-dependent signaling. Results obtained also suggest that AhTRE, AhTPS9, AhTPS11, AhGolS1 and AhRafS are reliable gene markers of WDS tolerance in amaranth.\n\nHighlightDifferential water deficit stress tolerance in grain amaranths and their ancestor, Amaranthus hybridus, is a multifactorial process involving various biochemical changes and modified expression patterns of key stress-related genes.

plant biology

Elevated ozone reduces photosynthetic carbon gain by accelerating leaf senescence of inbred and hybrid maize in a genotype-specific manner

Exposure to elevated tropospheric ozone concentration ([O3]) accelerates leaf senescence in many C3 crops. However, the effects of elevated [O3] on C4 crops including maize (Zea mays L.) are poorly understood in terms of physiological mechanism and genetic variation in sensitivity. Using Free Air gas Concentration Enrichment (FACE), we investigated the photosynthetic response of 18 diverse maize inbred and hybrid lines to season-long exposure to elevated [O3] (~100 nL L-1) in the field. Gas exchange was measured on the leaf subtending the ear throughout the grain filling period. On average over the lifetime of the leaf, elevated [O3] led to reductions in photosynthetic CO2 assimilation of both inbred (-22%) and hybrid (-33%) genotypes. There was significant variation among both inbred and hybrid lines in the sensitivity of photosynthesis to elevated [O3], with some lines showing no change in photosynthesis at elevated [O3]. Based on analysis of inbred line B73, the reduced CO2 assimilation at elevated [O3] was associated with accelerated senescence decreasing photosynthetic capacity, and not altered stomatal limitation. These findings across diverse maize genotypes could advance the development of more ozone tolerant maize, and provide experimental data for parameterization and validation of studies modeling how O3 impacts crop performance.

plant biology

Development of Gene-Specific Real-Time PCR Screening Method for Detection of cp4-epsps Gene in GM Crops

Among the genetically modified (GM) crops that are being approved for commercialization, herbicide resistant crops, especially those harboring cp4-epsps, have a considerable contribution. Gene-specific methods can be used to screen the presence of GMOs. To establish an effective qualitative and quantitative screening method, a set of primers were designed considering the cp4-epsps sequence. The specificity, the limit of detection, the efficiency, and the ability to quantify the GMO content were tested in GM cotton, soybean, and canola events. The results demonstrated that the primers can specifically detect cp4-epsps GM crops. The limit of detection was found to be 0.4 ng /l DNA per PCR reaction with the ability to detect 1-16 copies of the haploid genome of each GM event. The efficiency of this screening method (which was 94-110 % with an R2 higher than 0.96) indicated that these new primers can be applied to the screening of GM samples that contain the cp4-epsps gene. Also, the gene-specific real-time PCR screening method could be successfully developed for qualification of different types of GM cotton, soybean and canola events with the construction of a serial dilution ranging from 10 % to 1 %.

plant biology

Natural Genetic Variation Shapes Root System Responses to Phytohormones in Arabidopsis

Plants adjust their architecture by modulating organ growth. This ability is largely dependent on phytohormones. While many genes involved in phytohormone pathways have been identified, it remains unclear to which extent and how these pathways are modulated in non-reference strains and whether this is relevant for local adaptation. Here we assess variation of root traits in response to perturbations of the auxin, cytokinin, and abscisic acid pathways in 192 Arabidopsis accessions. We identify common response patterns, uncover the extent of their modulation by specific genotypes, and find that the Col-0 reference accession is not a good representative of the species in this regard. We conduct GWAS and identify 114 significant associations, most of them relating to ABA treatment. The numerous ABA candidate genes are not enriched for known ABA associated genes indicating that we largely uncovered unknown players. We then study two associated regions in detail and identify the CRF3 gene as a modulator of multiple hormone pathways. Finally, we show that natural variation in root traits is significantly associated with climate parameters relevant for local adaption in Arabidopsis thaliana and that, in particular, ABA regulated lateral root traits are likely to be relevant for adaptation to soil moisture.\n\nAuthor SummaryThe root system is a key component for plant survival and productivity. Apart from anchoring the plant, its architecture determines which parts of the soil are foraged for water and nutrients, and it serves as an interface for interaction with microbes and other soil organisms. Plant hormones play crucial roles in the development of root system architecture and its plasticity. However, while there is substantial natural variation of root architectures, it is not clear to which extent genetic variation in hormone related pathways contribute. Using the model species Arabidopsis thaliana we quantitatively explore the breadth of natural variation in plant hormone responses to three major plant hormones: auxin, cytokinin, and abscisic acid. The Col-0 reference strain can be quite different from a large proportion of the natural accessions of the species, illustrating a severe caveat of relying on a single reference strain in model species and drawing generalizations from it. Using GWAS, we further identify a large number of loci underlying the variation of responses to plant hormones, in particular to abscisic acid, find links between local adaptation and root responses to hormones, and finally using mutants for GWAS candidate genes, identify novel players involved in regulating hormone responses.

plant biology

Correlated evolution of self and interspecific incompatibly across the range of a Texas wildflower

Plant species have repeatedly evolved recognition systems between pollen and pistils that identify and reject inappropriate matings. Two of the most important systems recognize self-pollen and interspecific pollen. Outstanding questions are whether and how these two recognition systems are linked and if this association could constrain the evolution of mate choice. Our study characterizes variation in self and interspecific incompatibility in the native range of the Texas wildflower Phlox drummondii. We found quantitative variation in self-incompatibility and demonstrate that this variation is significantly correlated with variation in incompatibility with its close congener P. cuspidata. Furthermore, we find strong evidence that self and interspecific incompatibility involve common mechanisms of pollen adhesion or early pollen-tube germination. Finally, we show that P. drummondii populations that co-occur and hybridize with P. cuspidata have significantly higher interspecific incompatibility and self-incompatibility than isolated P. drummondii populations. This geographic variation suggests that the evolution of self-compatibility is constrained by selection favoring interspecific incompatibility to reduce maladaptive hybridization. To our knowledge this is the strongest evidence that a correlation between variation in self and interspecific incompatibilities could influence the evolution of pollen recognition across the range of a species.

plant biology

Variation in the oxidative burst in response to wounding and bacterial infection among native and invading genotypes of yellow starthistle (Centaurea solstitialis)

O_LIPremise of the study: Invasive plants may leave enemies behind when they colonize a new habitat, allowing selection to favor increased investment in growth and/or reproduction over defensive traits. Previous studies have identified reduced diversity of potential bacterial pathogens and evolutionary increases in growth and reproduction in invading populations of yellow starthistle (Centaurea solstitialis). This study leverages a recently developed high-throughput assay of immune function to test for evidence of a trade-off between increased growth and defense against bacterial pathogens in yellow starthistles invasion of California (USA).\nC_LIO_LIMethods: Seven bacterial strains were cultured from infected leaf tissue in the native range. Healthy leaf tissue from five native European collections and six invading collections were exposed to these native bacterial strains. A standardized assay of peroxidase activity was used measure the oxidative burst immune response to pathogen recognition by the leaf. Immune responses were compared to plant growth within and between ranges to assess evidence for a trade-off.\nC_LIO_LIKey Results: Plant genotypes from the native range demonstrated a higher immune response to bacterial strains than did invading genotypes, consistent with a trade-off with plant growth across regions. The same trade-off was also apparent across genotypes from the native range, but not across genotypes from the invaded range.\nC_LIO_LIConclusions: Our results provide evidence that increased growth in a highly invasive plant species may come at a cost to immune function, consistent with the hypothesis that escape from enemies can provide opportunities for shifts in resource allocation that favor the proliferation of non-native species.\nC_LI

plant biology

Genetic variants affecting plant size and chemical defenses jointly shape herbivory in Arabidopsis

Herbivorous insects exhibit strong feeding preferences when choosing among plant genotypes, yet experiments to map loci mediating plant susceptibility to herbivory rarely incorporate host choice. To address this gap, we applied genome-wide association (GWA) mapping to uncover genetic polymorphisms mediating damage from foraging insects (two populations of Scaptomyza flava) across a mixture of Arabidopsis thaliana genotypes in experimental enclosures. The effect of chemical defenses (glucosinolates) on herbivory depended on herbivore genotype. Unlike many studies that minimize the effects of host choice behavior, we also found a large effect of plant size on herbivory--likely through its effect on plant apparency--that was independent of herbivore genotype. These herbivory-associated loci are polymorphic at fine spatial scales, and thus have potential to shape variation in herbivory within natural populations. We also show that the polymorphism with the largest effect on herbivory underlies adaptive latitudinal variation in Arabidopsis plant size across Europe. Overall, our results provide genetic support for ecological observations that variation in both chemical defenses and non-canonical defense traits (e.g., plant size and phenology) jointly shapes plant-herbivore interactions.

plant biology

The plant circadian clock influences rhizosphere community structure and function

Plants alter chemical and physical properties of soil, and thereby influence rhizosphere microbial community structure. The structure of microbial communities may in turn affect plant performance. Yet, outside of simple systems with pairwise interacting partners, the plant genetic pathways that influence microbial community structure remain largely unknown, as are the performance feedbacks of microbial communities selected by the host plant genotype. We investigated the role of the plant circadian clock in shaping rhizosphere community structure and function. We performed 16S rRNA gene sequencing to characterize rhizosphere bacterial communities of Arabidopsis thaliana between day and night time points, and tested for differences in community structure between wild-type (Ws) vs. clock mutant (toc1-21, ztl-30) genotypes. We then characterized microbial community function, by growing wild-type plants in soils with an overstory history of Ws, toc1-21 or ztl-30 and measuring plant performance. We observed that rhizosphere community structure varied between day and night time points, and clock misfunction significantly altered rhizosphere communities. Finally, wild-type plants germinated earlier and were larger when inoculated with soils having an overstory history of wild-type in comparison to clock mutant genotypes. Our findings suggest the circadian clock of the plant host influences rhizosphere community structure and function.

plant biology

Metabolic process of raffinose family oligosacharrides during cold stress and recovery in cucumber leaves

Raffinose family oligosacharrides (RFOs) accumulate under stress conditions in many plants and have been suggested to act as stress protectants. To elucidate the metabolic process of RFOs under cold stress, levels of RFOs and relative carbohydrates, the expression and activities of main metabolic enzymes and their subcellular compartments were investigated during low temperature treatment and recovery period in cucumber leaves. Cold stress induced the accumulation of stachyose in vacuoles, galactinol in vacuoles and cytosols, and sucrose and raffinose in vacuoles, cytosols and chloroplasts. After cold stress removal, levels of these sugars decreased gradually in respective compartments. Among 4 galactinol synthase genes (CsGS), CsGS1 was not affected by the cold stress, while other three CsGSs were up-regulated by the low temperature. RNA levels of acid--galactosidase (GAL) 3, alkaline--galactosidase (AGA) 2 and 3, and the activities of GAL and AGA were up-regulated after cold stress removal. The GAL3 protein and GAL activity were exclusively located in the vacuole, whereas the protein of AGA2 and AGA 3 were found in the cytosol and chloroplast respectively. The results indicate that RFOs accumulated during the cold stress in different subcellular compartments in cucumber leaves could be catabolized in situ by different galactosidases after stress removal.

plant biology

Genotype, nitrogen and herbivory shape plant defense: the case of a vitamin-enriched maize

The cultivation of crops with novel traits could interfere with ecosystem services delivered by arthropods through bottom-up effects. Here we tested the hypothesis that a vitamin enriched maize (CarolightR) is similar in terms of plant-arthropod interactions to its wild type when compared in controlled environment and under field conditions. In order to assess the robustness of their equivalence we tested two nitrogen availability regimes. We used arthropod field abundance, the behavior and fitness of a keystone maize herbivore - the leafhopper Zyginidia scutellaris - and above ground chemistry of maize plants (volatile, hormone and metabolite profiling) as indicators of potential changes in plant-insect interactions. Nitrogen availability was the key driver of herbivore abundance and behavior, and determined direct and indirect chemical defense in maize plants. Both genotypes presented similar constitutive and inducible phytohormone profiles independently of the nitrogen regime. However, feeding by the herbivore suppressed the levels of JA-Ile and JA, without impairing the release of induced plant volatiles. CarolightR and M37W differed to some degree in the concentrations of phenolics (hydroxycinnamic acids and lignans) and in the abundance of a volatile compound. Overall the effect of maize genotype on the herbivores was smaller than the effect of nitrogen fertilization.\n\nHIGHLIGHTWe show the separate and interactive effects of nitrogen availability and genotype on the performance and behavior of a herbivore, and related these changes to constitutive and inducible maize defenses.

plant biology

Speed breeding: a powerful tool to accelerate crop research and breeding

The growing human population and a changing environment have raised significant concern for global food security, with the current improvement rate of several important crops inadequate to meet future demand [1]. This slow improvement rate is attributed partly to the long generation times of crop plants. Here we present a method called speed breeding, which greatly shortens generation time and accelerates breeding and research programs. Speed breeding can be used to achieve up to 6 generations per year for spring wheat (Triticum aestivum), durum wheat (T. durum), barley (Hordeum vulgare), chickpea (Cicer arietinum), and pea (Pisum sativum) and 4 generations for canola (Brassica napus), instead of 2-3 under normal glasshouse conditions. We demonstrate that speed breeding in fully-enclosed controlled-environment growth chambers can accelerate plant development for research purposes, including phenotyping of adult plant traits, mutant studies, and transformation. The use of supplemental lighting in a glasshouse environment allows rapid generation cycling through single seed descent and potential for adaptation to larger-scale crop improvement programs. Cost-saving through LED supplemental lighting is also outlined. We envisage great potential for integrating speed breeding with other modern crop breeding technologies, including high-throughput genotyping, genome editing, and genomic selection, accelerating the rate of crop improvement.

plant biology

Time-resolved Interaction Proteomics of the Putative Scaffold Protein GIGANTEA in Arabidopsis thaliana

The large, plant-specific protein GIGANTEA (GI) is involved in many physiological processes, mediating rhythmic, post-translational regulation in part through circadian and light regulation of GI RNA expression. GI binds several proteins implicated in the circadian clock, the control of photoperiodic flowering, and abiotic stress responses, and has co-chaperone activity. By extension, further interaction partners might mediate the less well-understood roles of GI but the number and rhythmicity of these interactors is unknown. Here, we seek potential interactors in a time-specific manner, using quantitative proteomics from a time series study of transgenic Arabidopsis thaliana plants constitutively expressing an epitope-tagged GI protein. Previously-identified, direct and indirect interactors of GI were detected but no further F-box proteins related to known GI partners ZTL/FKF1/LKP2. The predominantly non-rhythmic, interacting proteins were implicated in protein folding or degradation, metabolism and chromatin modification, including a small set of partners shared with other clock-related proteins. A transcription factor homologue that we name CYCLING DOF FACTOR 6 (CDF6) was shown to interact both with GI and the ZTL/FKF1/LKP2 proteins and to control photoperiodic flowering. Our results indicate the biochemical pathways, beyond circadian and flowering regulation, that might be affected by GIGANTEAs rhythmic, post-translational control.\n\nSignificance StatementSignificance statement of up to two sentences of no more than 75 words total;\n\nThe GIGANTEA protein of Arabidopsis was known for circadian and flowering functions, mediated by the FKF1/LKP2/ZTL family of GI-interacting, F-box proteins, then for a co-chaperone activity of unknown scope. We performed time-resolved, interaction proteomics, identifying CDF6 (At1g26790) as a morning-specific GI interactor that controls flowering time. Unlike FKF1 and CDF proteins, most of the 240 candidate partners were not rhythmically enriched. They link GI to proteostasis and metabolic functions that might mediate GIs physiological functions.

plant biology

Ancestral Reconstruction and C3 Bundle Sheath Transcript Abundance in the Paniceae Grasses Indicate the Foundations for all Three Biochemical C4 Sub-Types Were Likely Present in the Most Recent Ancestor

In C4 plants, the enzymatic machinery underpinning photosynthesis can vary, with, for example, three distinct C4 acid decarboxylases being used to release CO2 in the vicinity of RuBisCO. For decades, these decarboxylases have been used to classify C4 species into three biochemical sub-types. However, more recently the notion that C4 species mix and match C4 acid decarboxylases has increased in popularity and, as a consequence, the validity of specific biochemical sub-types has been questioned. Using five species from the grass tribe Paniceae, we show that, while in some species transcripts encoding multiple C4 acid decarboxylases accumulate, in others, transcript abundance and enzyme activity is almost entirely from one decarboxylase. In addition, the development of a bundle sheath isolation procedure for a close C3 species in the Paniceae enables the preliminary exploration of C4 sub-type evolution.

plant biology