Ibañez, C.; Poeschl, Y.; Peterson, T.; Bellstädt, J.; Denk, K.; Gogol-Döring, A.; Quint, M.; Delker, C. Ambient temperature and genotype differentially affect developmental and phenotypic plasticity in Arabidopsis thaliana BMC Plant Biol 17, 114, (2017) DOI: 10.1186/s12870-017-1068-5
BackgroundGlobal increase in ambient temperatures
constitute a significant challenge to wild and cultivated plant species.
Forward genetic analyses of individual temperature-responsive traits
have resulted in the identification of several signaling and response
components. However, a comprehensive knowledge about temperature
sensitivity of different developmental stages and the contribution of
natural variation is still scarce and fragmented at best.ResultsHere, we
systematically analyze thermomorphogenesis throughout a complete life
cycle in ten natural Arabidopsis thaliana accessions grown under long
day conditions in four different temperatures ranging from 16 to 28 °C.
We used Q10, GxE, phenotypic divergence and correlation analyses to
assess temperature sensitivity and genotype effects of more than 30
morphometric and developmental traits representing five phenotype
classes. We found that genotype and temperature differentially affected
plant growth and development with variing strengths. Furthermore,
overall correlations among phenotypic temperature responses was
relatively low which seems to be caused by differential capacities for
temperature adaptations of individual
accessions.ConclusionGenotype-specific temperature responses may be
attractive targets for future forward genetic approaches and
accession-specific thermomorphogenesis maps may aid the assessment of
functional relevance of known and novel regulatory components.
Hoehenwarter, W.; Mönchgesang, S.; Neumann, S.; Majovsky, P.; Abel, S.; Müller, J. Comparative expression profiling reveals a role of the root apoplast in local phosphate response BMC Plant Biol 16 , 106, (2016) DOI: 10.1186/s12870-016-0790-8
BackgroundPlant adaptation to limited phosphate availability
comprises a wide range of responses to conserve and remobilize internal
phosphate sources and to enhance phosphate acquisition. Vigorous
restructuring of root system architecture provides a developmental
strategy for topsoil exploration and phosphate scavenging. Changes in
external phosphate availability are locally sensed at root tips and
adjust root growth by modulating cell expansion and cell division. The
functionally interacting Arabidopsis genes, LOW PHOSPHATE RESPONSE 1 and
2 (LPR1/LPR2) and PHOSPHATE DEFICIENCY RESPONSE 2 (PDR2), are key
components of root phosphate sensing. We recently demonstrated that the
LOW PHOSPHATE RESPONSE 1 - PHOSPHATE DEFICIENCY RESPONSE 2 (LPR1-PDR2)
module mediates apoplastic deposition of ferric iron (Fe3+) in the
growing root tip during phosphate limitation. Iron deposition coincides
with sites of reactive oxygen species generation and triggers cell wall
thickening and callose accumulation, which interfere with cell-to-cell
communication and inhibit root growth.ResultsWe took advantage of
the opposite phosphate-conditional root phenotype of the phosphate
deficiency response 2 mutant (hypersensitive) and low phosphate response
1 and 2 double mutant (insensitive) to investigate the phosphate
dependent regulation of gene and protein expression in roots using
genome-wide transcriptome and proteome analysis. We observed an
overrepresentation of genes and proteins that are involved in the
regulation of iron homeostasis, cell wall remodeling and reactive oxygen
species formation, and we highlight a number of candidate genes with a
potential function in root adaptation to limited phosphate availability.
Our experiments reveal that FERRIC REDUCTASE DEFECTIVE 3 mediated,
apoplastic iron redistribution, but not intracellular iron uptake and
iron storage, triggers phosphate-dependent root growth modulation. We
further highlight expressional changes of several cell wall-modifying
enzymes and provide evidence for adjustment of the pectin network at
sites of iron accumulation in the root.ConclusionOur study
reveals new aspects of the elaborate interplay between phosphate
starvation responses and changes in iron homeostasis. The results
emphasize the importance of apoplastic iron redistribution to mediate
phosphate-dependent root growth adjustment and suggest an important role
for citrate in phosphate-dependent apoplastic iron transport. We
further demonstrate that root growth modulation correlates with an
altered expression of cell wall modifying enzymes and changes in the
pectin network of the phosphate-deprived root tip, supporting the
hypothesis that pectins are involved in iron binding and/or phosphate
Halim, V.A.; Altmann, S.; Ellinger, D.; Eschen-Lippold, L.; Miersch, O.; Scheel, D.; Rosahl, S. PAMP-induced defense responses in potato require both salicylic acid and jasmonic acid Plant Journal 57, 230 - 242, (2009) DOI: 10.1111/j.1365-313X.2008.03688.x
To elucidate the molecular mechanisms underlying pathogen-associated molecular pattern (PAMP)-induced defense responses in potato (Solanum tuberosum), the role of the signaling compounds salicylic acid (SA) and jasmonic acid (JA) was analyzed. Pep-13, a PAMP from Phytophthora, induces the accumulation of SA, JA and hydrogen peroxide, as well as the activation of defense genes and hypersensitive-like cell death. We have previously shown that SA is required for Pep-13-induced defense responses. To assess the importance of JA, RNA interference constructs targeted at the JA biosynthetic genes, allene oxide cyclase and 12- oxophytodienoic acid reductase, were expressed in transgenic potato plants. In addition, expression of the F-box protein COI1 was reduced by RNA interference. Plants expressing the RNA interference constructs failed to accumulate the respective transcripts in response to wounding or Pep-13 treatment, neither did they contain significant amounts of JA after elicitation. In response to infiltration of Pep-13, the transgenic plants exhibited a highly reduced accumulation of reactive oxygen species as well as reduced hypersensitive cell death. The ability of the JA-deficient plants to accumulate SA suggests that SA accumulation is independent or upstream of JA accumulation. These data show that PAMP responses in potato require both SA and JA and that, in contrast to Arabidopsis, these compounds act in the same signal transduction pathway. Despite their inability to fully respond to PAMP treatment, the transgenic RNA interference plants are not altered in their basal defense against Phytophthora infestans.
Levy, M.; Wang, Q.; Kaspi, R.; Parrella, M.P.; Abel, S. Arabidopsis IQD1, a novel calmodulin-binding nuclear protein, stimulates glucosinolate accumulation and plant defense Plant Journal 43, 79 - 96, (2005) DOI: 10.1111/j.1365-313X.2005.02435.x
Glucosinolates are a class of secondary metabolites with important roles in plant defense and human nutrition. To uncover regulatory mechanisms of glucosinolate production, we screened Arabidopsis thaliana T-DNA activation-tagged lines and identified a high-glucosinolate mutant caused by overexpression of IQD1 (At3g09710). A series of gain- and loss-of-function IQD1 alleles in different accessions correlates with increased and decreased glucosinolate levels, respectively. IQD1 encodes a novel protein that contains putative nuclear localization signals and several motifs known to mediate calmodulin binding, which are arranged in a plant-specific segment of 67 amino acids, called the IQ67 domain. We demonstrate that an IQD1-GFP fusion protein is targeted to the cell nucleus and that recombinant IQD1 binds to calmodulin in a Ca2+-dependent fashion. Analysis of steady-state messenger RNA levels of glucosinolate pathway genes indicates that IQD1 affects expression of multiple genes with roles in glucosinolate metabolism. Histochemical analysis of tissue-specific IQD1::GUS expression reveals IQD1 promoter activity mainly in vascular tissues of all organs, consistent with the expression patterns of several glucosinolate-related genes. Interestingly, overexpression of IQD1 reduces insect herbivory, which we demonstrated in dual-choice assays with the generalist phloem-feeding green peach aphid (Myzus persicae), and in weight-gain assays with the cabbage looper (Trichoplusia ni), a generalist-chewing lepidopteran. As IQD1 is induced by mechanical stimuli, we propose IQD1 to be novel nuclear factor that integrates intracellular Ca2+ signals to fine-tune glucosinolate accumulation in response to biotic challenge.
Abel, S.; Savchenko, T.; Levy, M. Genome-wide comparative analysis of the <em>IQD</em> gene families in <em>Arabidopsis thaliana</em> and Oryza sativa BMC Evolutionary Biology 5, 72 (1-25), (2005)
We identified and analyzed 33 and 29 IQD1-like genes in Arabidopsis thaliana and Oryza sativa, respectively. The encoded IQD proteins contain a plant-specific domain of 67 conserved amino acid residues, referred to as the IQ67 domain, which is characterized by a unique and repetitive arrangement of three different calmodulin recruitment motifs, known as the IQ, 1-5-10, and 1-8-14 motifs. We demonstrated calmodulin binding for IQD20, the smallest IQD protein in Arabidopsis, which consists of a C-terminal IQ67 domain and a short N-terminal extension. A striking feature of IQD proteins is the high isoelectric point (~10.3) and frequency of serine residues (~11%). We compared the Arabidopsis and rice IQD gene families in terms of gene structure, chromosome location, predicted protein properties and motifs, phylogenetic relationships, and evolutionary history. The existence of an IQD-like gene in bryophytes suggests that IQD proteins are an ancient family of calmodulin-binding proteins and arose during the early evolution of land plants. Comparative phylogenetic analyses indicate that the major IQD gene lineages originated before the monocot-eudicot divergence. The extant IQD loci in Arabidopsis primarily resulted from segmental duplication and reflect preferential retention of paralogous genes, which is characteristic for proteins with regulatory functions. Interaction of IQD1 and IQD20 with calmodulin and the presence of predicted calmodulin binding sites in all IQD family members suggest that IQD proteins are a new class of calmodulin targets. The basic isoelectric point of IQD proteins and their frequently predicted nuclear localization suggest that IQD proteins link calcium signaling pathways to the regulation of gene expression. Our comparative genomics analysis of IQD genes and encoded proteins in two model plant species provides the first step towards the functional dissection of this emerging family of putative calmodulin targets.
Ticconi, C.A.; Delatorre, C.A.; Lahner, B.; Salt, D.E.; Abel, S. Arabidopsis <span style="font-style: italic;">pdr2</span> reveals a phosphate-sensitive checkpoint in root development Plant Journal 37, 801 - 814, (2004)
Wong, L.M.; Abel, S.; Shen, N.; de la Foata, M.; Mal, Y.; Theologis, A. Differential activation of the primary auxin response genes, PS-IAA4/5 and PS-IAA6, during early plant development. Plant Journal 9, 587-599, (1996)
The plant growth hormone auxin typified by indoleacetic acid (IAA) transcriptionally activates early genes in pea, PS-IAA4/5 and PS-IAA6, that are members of a multigene family encoding short-lived nuclear proteins. To gain first insight into the biological role of PS-IAA4/5 and PSIAA6, promoter-β-glucuronidase (GUS) gene fusions were constructed and their expression during early development of transgenic tobacco seedlings was examined. The comparative analysis reveals spatial and temporal expression patterns of both genes that correlate with cells, tissues, and developmental processes known to be affected by auxin. GUS activity in seedlings of both transgenic lines is located in the root meristem, sites of lateral root initiation and in hypocotyls undergoing rapid elongation. In addition, mutually exclusive cell-specific expression is evident. For instance, PS-IAA4/5GUS but not PS-IAA6GUS is expressed in root vascular tissue and in guard cells, whereas only PS-IAA6GUS activity is detectable in glandular trichomes and redistributes to the elongating side of the hypocotyl upon gravitropic stimulation. Expression of PS-IAA4/5 and PS-IAA6 in elongating, dividing, and differentiating cell types indicates multiple functions during development. The common and yet distinct activity patterns of both genes suggest a combinatorial code of spatio-temporal co-expression of the various PS-IAA4/ 5-like gene family members in plant development that may mediate cell-specific responses to auxin.
Abel, S.; Theologis, A. A polymorphic bipartite motif signals nuclear targeting of early auxin- inducible proteins related to PS-IAA4 from pea (Pisum sativum) Plant Journal 8, 87-96, (1995)
The plant hormone, indoleacetic acid (IAA), transcriptionally activates two early genes in pea, PS-IAA4/5 and PS-IAA6, that encode short-lived nuclear proteins. The identification of the nuclear localization signals (NLS) in PS-IAA4 and PS-IAA6 using progressive deletion analysis and site-directed mutagenesis is reported. A C-terminal SV40-type NLS is sufficient to direct the β-glucuronidase reporter to the nucleus of transiently transformed tobacco protoplasts, but is dispensible for nuclear localization of both proteins. The dominant and essential NLS in PS-IAA4 and PS-IAA6 overlap with a bipartite basic motif which is polymorphic and conserved in related proteins from other plant species, having the consensus sequence (KKNEK)KR-X(2471)-(RSXRK)/(RK/RK). Both basic elements of this motif in PS-IAA4, (KR-X41-RSYRK), function interdependently as a bipartite NLS. However, in PS-IAA6 (KKNEKKR-X36-RKK) the upstream element of the corresponding motif contains additional basic residues which allow its autonomous function as an SV40-type monopartite NLS. The spacer-length polymorphism, X(2470), in respective bipartite NLS peptides of several PS-IAA4-like proteins from Arabidopsis thaliana does not affect nuclear targeting function. The structural and functional variation of the bipartite basic motif in PS-IAA4-like proteins supports the proposed integrated consensus of NLS.
Abel, S.; Theologis, A. Transient transformation of Arabidopsis leaf protoplasts: a versatile experimental system to study gene expression Plant Journal 5, 421-427, (1994)
An improved protocol is reported to isolate and transiently transform mesophyll protoplasts of Arabidopsis thaliana. Transfected leaf protoplasts support high levels of expression of the bacterial reporter gene coding for β-glucuronidase (GUS), under the control of the cauliflower mosaic virus (CaMV) 35S promoter. Transient expression of GUS activity was monitored spectrophotometrically and reached a maximum between 18 and 48 h after polyethylene glycol (PEG)-mediated DNA uptake. Histochemical staining for GUS activity revealed reproducible transformation frequencies between 40 and 60%, based on the number of protoplasts survived. To demonstrate the applicability of the transient expression system, the subcellular localization of GUS proteins tagged with different nuclear polypeptides was studied in transfected mesophyll protoplasts, revealing nuclear compartmentalization of the chimeric GUS enzymes. Furthermore, Arabidopsis mesophyll protoplasts support auxin-mediated induction of chloramphenicol acetyl-transferase (CAT) activity when transfected with a transcriptional fusion between the CAT reporter gene and the early auxin-inducible PS-IAA4/5 promoter. Hence, the method allows in vivo analysis of promoter activity and subcellular localization of fusion proteins in a homologous transformation system.