An official website of the United States government
Summary Abscisic acid (ABA) receptors belong to theSTARTdomain superfamily, which encompasses ligand‐binding proteins present in all kingdoms of life.STARTdomain proteins contain a central binding pocket that, depending on the protein, can couple ligand binding to catalytic, transport or signaling functions. In Arabidopsis, the best characterizedSTARTdomain proteins are the 14PYR/PYL/RCAR ABAreceptors, while the other members of the superfamily do not have assigned ligands. To address this, we used affinity purification of biotinylated proteins expressed transiently inNicotiana benthamianacoupled to untargetedLC‐MSto identify candidate binding ligands. We optimized this method usingABA–PYLinteractions and show thatABAco‐purifies with wild‐typePYL5 but not a binding site mutant. TheKdofPYL5 forABAis 1.1 μm, which suggests that the method has sufficient sensitivity for many ligand–protein interactions. Using this method, we surveyed a set of 37STARTdomain‐related proteins, which resulted in the identification of ligands that co‐purified withMLBP1 (At4G01883) orMLP165 (At1G35260). Metabolite identification and the use of authentic standards revealed thatMLBP1 binds to monolinolenin, which we confirmed using recombinantMLBP1. Monolinolenin also co‐purified withMLBP1 purified from transgenic Arabidopsis, demonstrating that the interaction occurs in a native context. Thus, deployment of this relatively simple method allowed us to define a protein–metabolite interaction and better understand protein–ligand interactions in plants.
more »
« less
Arabidopsis bioinformatics resources: The current state, challenges, and priorities for the future
Abstract Effective research, education, and outreach efforts by theArabidopsis thalianacommunity, as well as other scientific communities that depend on Arabidopsis resources, depend vitally on easily available and publicly‐shared resources. These resources include reference genome sequence data and an ever‐increasing number of diverse data sets and data types.TAIR(The Arabidopsis Information Resource) and Araport (originally named the Arabidopsis Information Portal) are community informatics resources that provide tools, data, and applications to the more than 30,000 researchers worldwide that use in their work either Arabidopsis as a primary system of study or data derived from Arabidopsis. Four years after Araport's establishment, theIAICheld another workshop to evaluate the current status of Arabidopsis Informatics and chart a course for future research and development. The workshop focused on several challenges, including the need for reliable and current annotation, community‐defined common standards for data and metadata, and accessible and user‐friendly repositories/tools/methods for data integration and visualization. Solutions envisioned included (a) a centralized annotation authority to coalesce annotation from new groups, establish a consistent naming scheme, distribute this format regularly and frequently, and encourage and enforce its adoption. (b) Standards for data and metadata formats, which are essential, but challenging when comparing across diverse genotypes and in areas with less‐established standards (e.g., phenomics, metabolomics). Community‐established guidelines need to be developed. (c) A searchable, central repository for analysis and visualization tools. Improved versioning and user access would make tools more accessible. Workshop participants proposed a “one‐stop shop” website, an Arabidopsis “Super‐Portal” to link tools, data resources, programmatic standards, and best practice descriptions for each data type. This must have community buy‐in and participation in its establishment and development to encourage adoption.
more »
« less
- Award ID(s):
- 1750698
- PAR ID:
- 10082774
- Publisher / Repository:
- Wiley Blackwell (John Wiley & Sons)
- Date Published:
- Journal Name:
- Plant Direct
- Volume:
- 3
- Issue:
- 1
- ISSN:
- 2475-4455
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
More Like this
-
-
Abstract ARGONAUTES are the central effector proteins ofRNAsilencing which bind target transcripts in a smallRNA‐guided manner.Arabidopsis thalianahas 10ARGONAUTE(AGO) genes, with specialized roles inRNA‐directedDNAmethylation, post‐transcriptional gene silencing, and antiviral defense. To better understand specialization amongAGOgenes at the level of transcriptional regulation we tested a library of 1497 transcription factors for binding to the promoters ofAGO1,AGO10, andAGO7using yeast 1‐hybrid assays. A ranked list of candidateDNA‐bindingTFs revealed binding of theAGO7promoter by a number of proteins in two families: the miR156‐regulatedSPLfamily and the miR319‐regulatedTCPfamily, both of which have roles in developmental timing and leaf morphology. Possible functions forSPLandTCPbinding are unclear: we showed that these binding sites are not required for the polar expression pattern ofAGO7, nor for the function ofAGO7in leaf shape. NormalAGO7transcription levels and function appear to depend instead on an adjacent 124‐bp region. Progress in understanding the structure of this promoter may aid efforts to understand how the conservedAGO7‐triggeredTAS3pathway functions in timing and polarity.more » « less
-
Summary In flowering plants, cell–cell communication plays a key role in reproductive success, as both pollination and fertilization require pathways that regulate interactions between many different cell types. Some of the most critical of these interactions are those between the pollen tube (PT) and the embryo sac, which ensure the delivery of sperm cells required for double fertilization. Synergid cells function to attract thePTthrough secretion of small peptides and inPTreception via membrane‐bound proteins associated with the endomembrane system and the cell surface. While many synergid‐expressed components regulatingPTattraction and reception have been identified, few tools exist to study the localization of membrane‐bound proteins and the components of the endomembrane system in this cell type. In this study, we describe the localization and distribution of seven fluorescent markers that labelled components of the secretory pathway in synergid cells ofArabidopsis thaliana. These markers were used in co‐localization experiments to investigate the subcellular distribution of the twoPTreception componentsLORELEI, aGPI‐anchored surface protein, andNORTIA, aMILDEW RESISTANCE LOCUSO protein, both found within the endomembrane system of the synergid cell. These secretory markers are useful tools for both reproductive and cell biologists, enabling the analysis of membrane‐associated trafficking within a haploid cell actively involved in polar transport.more » « less
-
ABSTRACT Necrotrophic pathogens cause serious threats to agricultural crops, and understanding the resistance genes and their genetic networks is key to breeding new plant cultivars with better resistance traits. AlthoughAlternaria alternatacauses black spot in important leafy brassica vegetables, and leads to significant loss of yield and food quality, little is known about plant–A. alternatainteractions. In this study, we used a unique and large collection of single, double and triple mutant lines of defence metabolite regulators inArabidopsisto explore how these transcription factors and their epistatic networks may influenceA. alternatainfections. This identified nine novel regulators and 20 pairs of epistatic interactions that modulateArabidopsisplants' defence responses toA. alternatainfection. We further showed that the glucosinolate 4‐methoxy‐indol‐3‐ylmethyl is the only glucosinolate consistently responsive toA. alternatainfection in Col‐0 ecotype. With the further exploration of the regulators and the genetic networks on modulating the accumulation of glucosinolates underA. alternatainfection, an inverted triangle regulatory model was proposed forArabidopsisplants' defence responses at a metabolic level and a phenotypic level.more » « less
-
Abstract AimThe International Tree‐Ring Data Bank (ITRDB) is the most comprehensive database of tree growth. To evaluate its usefulness and improve its accessibility to the broad scientific community, we aimed to: (a) quantify its biases, (b) assess how well it represents global forests, (c) develop tools to identify priority areas to improve its representativity, and d) make available the corrected database. LocationWorldwide. Time periodContributed datasets between 1974 and 2017. Major taxa studiedTrees. MethodsWe identified and corrected formatting issues in all individual datasets of theITRDB. We then calculated the representativity of theITRDBwith respect to species, spatial coverage, climatic regions, elevations, need for data update, climatic limitations on growth, vascular plant diversity, and associated animal diversity. We combined these metrics into a global Priority Sampling Index (PSI) to highlight ways to improveITRDBrepresentativity. ResultsOur refined dataset provides access to a network of >52 million growth data points worldwide. We found, however, that the database is dominated by trees from forests with low diversity, in semi‐arid climates, coniferous species, and in western North America. Conifers represented 81% of theITRDBand even in well‐sampled areas, broadleaves were poorly represented. OurPSIstressed the need to increase the database diversity in terms of broadleaf species and identified poorly represented regions that require scientific attention. Great gains will be made by increasing research and data sharing in African, Asian, and South American forests. Main conclusionsThe extensive data and coverage of theITRDBshow great promise to address macroecological questions. To achieve this, however, we have to overcome the significant gaps in the representativity of theITRDB. A strategic and organized group effort is required, and we hope the tools and data provided here can guide the efforts to improve this invaluable database.more » « less
