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1.
Methods Mol Biol ; 2354: 143-154, 2021.
Article in English | MEDLINE | ID: mdl-34448159

ABSTRACT

The potato is among the most important food crops in the world and of incalculable value for global food security. In 2012, the crop area for potato in Northern and Western Europe reached almost 1 million ha and a production of over 37 million tons with an average yield between 18 and 45 tons/ha. However, current potato production is put in jeopardy by a number of important biotic stress factors including late blight (Phytophthora infestans), which was responsible for the disastrous Irish potato famine during 1843-1845. P. infestans shows a remarkable capacity for adaptation with respect to host genotype and applied fungicides. This has made disease management to become more and more difficult and put substantial emphasis on gaining more detailed insight into the molecular bases of plant pathogen interactions, in order to find more sophisticated ways for biological pest control. The plant hormones jasmonic acid (JA) and salicylic acid (SA) play central roles in the regulation of plant responses to biotic foes. In addition, other phytohormones including auxins and abscisic acid (ABA) have also been associated with plant defense responses. For this reason, the parallel analysis of multiple plant hormones in small tissue amounts represents an important field of research in contemporary plant sciences. Here, we describe a highly sensitive and accurate method for the quantitative analysis of ABA, JA, SA, and indole-3-acetic acid in potato plants by gas chromatography-coupled tandem mass spectrometry (GC-MS/MS).


Subject(s)
Solanum tuberosum , Abscisic Acid , Gas Chromatography-Mass Spectrometry , Plant Diseases , Plant Growth Regulators , Salicylic Acid , Stress, Physiological , Tandem Mass Spectrometry
2.
Biomolecules ; 11(8)2021 08 13.
Article in English | MEDLINE | ID: mdl-34439873

ABSTRACT

The diversification of land plants largely relies on their ability to cope with constant environmental fluctuations, which negatively impact their reproductive fitness and trigger adaptive responses to biotic and abiotic stresses. In this limiting landscape, cumulative research attention has centred on deepening the roles of major phytohormones, mostly auxins, together with brassinosteroids, jasmonates, and abscisic acid, despite the signaling networks orchestrating the crosstalk among them are so far only poorly understood. Accordingly, this review focuses on the Arabidopsis Amidase Signature (AS) superfamily members, with the aim of highlighting the hitherto relatively underappreciated functions of AMIDASE1 (AMI1) and FATTY ACID AMIDE HYDROLASE (FAAH), as comparable coordinators of the growth-defense trade-off, by balancing auxin and ABA homeostasis through the conversion of their likely bioactive substrates, indole-3-acetamide and N-acylethanolamine.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Ethanolamines/metabolism , Indoleacetic Acids/metabolism , Amidohydrolases/metabolism , Gene Expression Regulation, Plant
3.
Int J Mol Sci ; 22(4)2021 Feb 18.
Article in English | MEDLINE | ID: mdl-33670805

ABSTRACT

The major auxin, indole-3-acetic acid (IAA), is associated with a plethora of growth and developmental processes including embryo development, expansion growth, cambial activity, and the induction of lateral root growth. Accumulation of the auxin precursor indole-3-acetamide (IAM) induces stress related processes by stimulating abscisic acid (ABA) biosynthesis. How IAM signaling is controlled is, at present, unclear. Here, we characterize the ami1rooty double mutant, that we initially generated to study the metabolic and phenotypic consequences of a simultaneous genetic blockade of the indole glucosinolate and IAM pathways in Arabidopsisthaliana. Our mass spectrometric analyses of the mutant revealed that the combination of the two mutations is not sufficient to fully prevent the conversion of IAM to IAA. The detected strong accumulation of IAM was, however, recognized to substantially impair seed development. We further show by genome-wide expression studies that the double mutant is broadly affected in its translational capacity, and that a small number of plant growth regulating transcriptional circuits are repressed by the high IAM content in the seed. In accordance with the previously described growth reduction in response to elevated IAM levels, our data support the hypothesis that IAM is a growth repressing counterpart to IAA.


Subject(s)
Gene Regulatory Networks , Indoleacetic Acids/metabolism , Organelle Biogenesis , Ribosomes/metabolism , Arabidopsis/embryology , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis Proteins/chemistry , Arabidopsis Proteins/metabolism , Germination , Indoleacetic Acids/chemistry , Metabolic Networks and Pathways , Models, Molecular , Mutation/genetics , Phenotype , Protein Biosynthesis/genetics , Reproducibility of Results , Seeds/metabolism , Transcription, Genetic
4.
J Exp Bot ; 72(2): 459-475, 2021 02 02.
Article in English | MEDLINE | ID: mdl-33068437

ABSTRACT

The evolutionary success of plants relies to a large extent on their extraordinary ability to adapt to changes in their environment. These adaptations require that plants balance their growth with their stress responses. Plant hormones are crucial mediators orchestrating the underlying adaptive processes. However, whether and how the growth-related hormone auxin and the stress-related hormones jasmonic acid, salicylic acid, and abscisic acid (ABA) are coordinated remains largely elusive. Here, we analyse the physiological role of AMIDASE 1 (AMI1) in Arabidopsis plant growth and its possible connection to plant adaptations to abiotic stresses. AMI1 contributes to cellular auxin homeostasis by catalysing the conversion of indole-acetamide into the major plant auxin indole-3-acetic acid. Functional impairment of AMI1 increases the plant's stress status rendering mutant plants more susceptible to abiotic stresses. Transcriptomic analysis of ami1 mutants disclosed the reprogramming of a considerable number of stress-related genes, including jasmonic acid and ABA biosynthesis genes. The ami1 mutants exhibit only moderately repressed growth but an enhanced ABA accumulation, which suggests a role for AMI1 in the crosstalk between auxin and ABA. Altogether, our results suggest that AMI1 is involved in coordinating the trade-off between plant growth and stress responses, balancing auxin and ABA homeostasis.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Abscisic Acid , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Gene Expression Regulation, Plant , Indoleacetic Acids , Plant Growth Regulators
5.
Plant Cell Environ ; 43(2): 315-326, 2020 02.
Article in English | MEDLINE | ID: mdl-31600827

ABSTRACT

Permeability is a crucial trait that affects seed longevity and is regulated by different polymers including proanthocyanidins, suberin, cutin and lignin located in the seed coat. By testing mutants in suberin transport and biosynthesis, we demonstrate the importance of this biopolymer to cope with seed deterioration. Transcriptomic analysis of cog1-2D, a gain-of-function mutant with increased seed longevity, revealed the upregulation of several peroxidase genes. Reverse genetics analysing seed longevity uncovered redundancy within the seed coat peroxidase gene family; however, after controlled deterioration treatment, seeds from the prx2 prx25 double and prx2 prx25 prx71 triple mutant plants presented lower germination than wild-type plants. Transmission electron microscopy analysis of the seed coat of these mutants showed a thinner palisade layer, but no changes were observed in proanthocyanidin accumulation or in the cuticle layer. Spectrophotometric quantification of acetyl bromide-soluble lignin components indicated changes in the amount of total polyphenolics derived from suberin and/or lignin in the mutant seeds. Finally, the increased seed coat permeability to tetrazolium salts observed in the prx2 prx25 and prx2 prx25 prx71 mutant lines suggested that the lower permeability of the seed coats caused by altered polyphenolics is likely to be the main reason explaining their reduced seed longevity.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Peroxidases/metabolism , Seeds/metabolism , Transcription Factors/metabolism , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Cell Wall/metabolism , Gene Expression Profiling , Gene Expression Regulation, Plant , Germination/genetics , Germination/physiology , Lignin , Lipid Metabolism , Lipids , Membrane Lipids , Mutation , Peroxidases/genetics , Proanthocyanidins , Seeds/genetics
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