Abstract Alternative splicing (AS) of pre-mRNAs in plants is an important mechanism of gene regulation in environmental stress tolerance but plant signals involved are essentially unknown. Pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) is mediated by mitogen-activated protein kinases and the majority of PTI defense genes are regulated by MPK3, MPK4 and MPK6. These responses have been mainly analyzed at the transcriptional level, however many splicing factors are direct targets of MAPKs. Here, we studied alternative splicing induced by the PAMP flagellin in Arabidopsis. We identified 506 PAMP-induced differentially alternatively spliced (DAS) genes. Although many DAS genes are targets of nonsense-mediated degradation (NMD), only 19% are potential NMD targets. Importantly, of the 506 PAMP-induced DAS genes, only 89 overlap with the set of 1849 PAMP-induced differentially expressed genes (DEG), indicating that transcriptome analysis does not identify most DASevents. Global DAS analysis of mpk3 , mpk4 , and mpk6 mutants revealed that MPK4 is a key regulator of PAMP-induced differential splicing, regulating AS of a number of splicing factors and immunity-related protein kinases, such as the calcium-dependent protein kinase CPK28, the cysteine-rich receptor like kinases CRK13 and CRK29 or the FLS2 co-receptor SERK4/BKK1. These data suggest that MAP kinase regulation of splicing factors is a key mechanism in PAMP-induced AS regulation of PTI. Significance statement Alternative pre-mRNA splicing (AS) affects plant responses to environmental stresses. So far, however, the regulation of AS is little understood. Here, we studied AS induced by the pathogen-associated molecular pattern (PAMP) flagellin in Arabidopsis. We identified 506 differentially alternatively spliced (DAS) genes, 89 of which overlap with the 1849 DEG, indicating that the majority of DAS events go undetected by common transcriptome analysis. PAMP-triggered immunity is mediated by mitogen-activated protein kinases. Global DAS analysis of MAPK mutants revealed that MPK4 is a key regulator of AS by affecting splicing factors and a number of important protein kinases involved in immunity. Since PAMP-triggered phosphorylation of several splicing factors is directly mediated by MAPKs, we discovered a key mechanism of AS regulation.
In Arabidopsis thaliana, HEAT SHOCK TRANSCRIPTION FACTORA1b (HSFA1b) controls resistance to environmental stress and is a determinant of reproductive fitness by influencing seed yield. To understand how HSFA1b achieves this, we surveyed its genome-wide targets (ChIP-seq) and its impact on the transcriptome (RNA-seq) under non-stress (NS), heat stress (HS) in the wild type, and in HSFA1b-overexpressing plants under NS. A total of 952 differentially expressed HSFA1b-targeted genes were identified, of which at least 85 are development associated and were bound predominantly under NS. A further 1780 genes were differentially expressed but not bound by HSFA1b, of which 281 were classified as having development-associated functions. These genes are indirectly regulated through a hierarchical network of 27 transcription factors (TFs). Furthermore, we identified 480 natural antisense non-coding RNA (cisNAT) genes bound by HSFA1b, defining a further mode of indirect regulation. Finally, HSFA1b-targeted genomic features not only harboured heat shock elements, but also MADS box, LEAFY, and G-Box promoter motifs. This revealed that HSFA1b is one of eight TFs that target a common group of stress defence and developmental genes. We propose that HSFA1b transduces environmental cues to many stress tolerance and developmental genes to allow plants to adjust their growth and development continually in a varying environment.
In this paper, we propose a novel and efficient two-stage variable selection approach for sparse GLARMA models, which are pervasive for modelling discrete-valued time series. Our approach consists in iteratively combining the estimation of the autoregressive moving average (ARMA) coefficients of GLARMA models with regularized methods designed for performing variable selection in regression coefficients of Generalized Linear Models (GLM). We first establish the consistency of the ARMA part coefficient estimators in a specific case. Then, we explain how to efficiently implement our approach. Finally, we assess the performance of our methodology using synthetic data, compare it with alternative methods and illustrate it on an example of real-world application. Our approach, which is implemented in the GlarmaVarSel R package, is very attractive since it benefits from a low computational load and is able to outperform the other methods in terms of recovering the non-null regression coefficients.
Rotation-invariant descriptions are required in many 3D volumetric image analysis tasks. The histogram-of-oriented-gradient (HOG) is widely used in 2D images and proves to be a very robust local description. This paper concentrates on how to use the HOG feature in 3D volumetric images when rotation-invariance is concerned. This is challenging because of the complexity of 3D rotations. We present a decent solution based on the spherical harmonics theory which is an effective tool for analysing 3D rotations, together with the spherical tensor operations which explore high order tensor information in spherical coordinates. The design is quite general and could be used for different applications. It achieves high scores on Princeton Shape Benchmark and SHREC 2009 Generic Shape Benchmark, and also produces promising results when applying on biological microscopy images.
White lupin ( Lupinus albus L. ) is a legume that produces seeds recognized for their high protein content and good nutritional value (lowest glycemic index of all grains, high dietary fiber content, and zero gluten or starch) 1–5 . White lupin can form nitrogen-fixing nodules but has lost the ability to form mycorrhizal symbiosis with fungi 6 . Nevertheless, its root system is well adapted to poor soils: it produces cluster roots, constituted of dozens of determinate lateral roots that improve soil exploration and phosphate remobilization 7 . As phosphate is a limited resource that comes from rock reserves 8 , the production of cluster roots is a trait of interest to improve fertilizers efficiency. Using long reads sequencing technologies, we provide a high-quality genome sequence of a modern variety of white lupin (2n=50, 451 Mb), as well as de novo assemblies of a landrace and a wild relative. We describe how domestication impacted soil exploration capacity through the early establishment of lateral and cluster roots. We identify the APETALA2 transcription factor LaPUCHI-1 , homolog of the Arabidopsis morphogenesis coordinator 9 , as a potential regulator of this trait. Our high-quality genome and companion genomic and transcriptomic resources enable the development of modern breeding strategies to increase and stabilize yield and to develop new varieties with reduced allergenic properties (caused by conglutins 10 ), which would favor the deployment of this promising culture.
Abstract In plants, aberrant RNAs produced by endogenous genes or transgenes are normally degraded by the nuclear and cytosolic RNA quality control (RQC) pathways. Under certain biotic or abiotic stresses, RQC is impaired, and aberrant RNAs are converted into siRNAs that initiate post-transcriptional gene silencing (PTGS) in the cytosol. How aberrant RNAs are selected and brought to the cytoplasm is not known. Here we show that the RNA-binding protein SUPPRESSOR OF GENE SILENCING (SGS)3 shuttles between the cytosol and the nucleus where it associates with the ISWI-like CHROMATIN REMODELER (CHR)11. CHR11 binds to transgenes prone to trigger PTGS. Knocking down CHR11 and its paralog CHR17 strongly reduces transgene PTGS, suggesting that SGS3 recruitment by CHR11/17 facilitates PTGS initiation. CHR11 is also enriched at endogenous protein-coding genes (PCGs) producing nat-siRNAs and va-siRNAs under biotic or abiotic stresses, and this production is reduced in chr11 chr17 double mutants at genome-wide level. Moreover, impairing CHR11 and CHR17 rescues the lethal phenotype caused by the massive production of siRNAs from PCGs in RQC-deficient mutants. We propose that SGS3 recruitment by CHR11/17 allows exporting RNAs to the cytosol to initiate the production of siRNAs. One Sentence Summary The plant PTGS component SGS3 is recruited by the chromatin remodeler CHR11 to select nuclear RNAs for export to the cytosol and siRNA production.
Leaves show a wide range of shapes that results from the combinatory variations of two main parameters: the relative duration of the morphogenetic phase and the pattern of dissection of the leaf margin. To further understand the mechanisms controlling leaf shape, we have studied the interactions between several loci leading to increased dissection of the Arabidopsis leaf margins. Thus, we have used (i) mutants in which miR164 regulation of the CUC2 gene is impaired, (ii) plants overexpressing miR319/miRJAW that down-regulates multiple TCP genes and (iii) plants overexpressing the STIMPY/WOX9 gene. Through the analysis of their effects on leaf shape and KNOX I gene expression, we show that these loci act in different pathways. We show, in particular, that they have synergetic effects and that plants combining two or three of these loci show dramatic modifications of their leaf shapes. Finally, we present a working model for the role of these loci during leaf development.
Legume plants can acquire mineral nitrogen (N) either through their roots or via a symbiotic interaction with N-fixing rhizobia bacteria housed in root nodules. To identify shoot-to-root systemic signals acting in Medicago truncatula plants at N deficit or N satiety, plants were grown in a split-root experimental design in which either high or low N was provided to half of the root system, allowing the analysis of systemic pathways independently of any local N response. Among the plant hormone families analyzed, the cytokinin trans-zeatin accumulated in plants at N satiety. Cytokinin application by petiole feeding led to inhibition of both root growth and nodulation. In addition, an exhaustive analysis of miRNAs revealed that miR2111 accumulates systemically under N deficit in both shoots and non-treated distant roots, whereas a miRNA related to inorganic phosphate (Pi) acquisition, miR399, accumulates in plants grown under N satiety. These two accumulation patterns are dependent on Compact Root Architecture 2 (CRA2), a receptor required for C-terminally Encoded Peptide (CEP) signaling. Constitutive ectopic expression of miR399 reduced nodule numbers and root biomass depending on Pi availability, suggesting that the miR399-dependent Pi-acquisition regulatory module controlled by N availability affects the development of the whole legume plant root system.