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1.
New Phytol ; 243(4): 1361-1373, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38934066

RESUMEN

Posttranslational modification of multiple ABA signaling components is an essential process for the adaptation and survival of plants under stress conditions. In our previous study, we established that the pepper group A PP2C protein CaAITP1, one of the core components of ABA signaling, undergoes ubiquitination mediated by the RING-type E3 ligase CaAIRE1. In this study, we discovered an additional form of regulation mediated via the SUMOylation of CaAITP1. Pepper plants subjected to drought stress were characterized by reductions in both the stability and SUMOylation of CaAITP1 protein. Moreover, we identified a SUMO protease, Capsicum annuum DeSUMOylating Isopeptidase 2 (CaDeSI2), as a new interacting partner of CaAITP1. In vitro and in vivo analyses revealed that CaAITP1 is deSUMOylated by CaDeSI2. Silencing of CaDeSI2 in pepper plants led to drought-hypersensitive and ABA-hyposensitive phenotypes, whereas overexpression of CaDeSI2 in transgenic Arabidopsis plants resulted in the opposite phenotypes. Importantly, we found that the CaAITP1 protein was stabilized in response to the silencing of CaDeSI2, and CaDeSI2 and CaAITP1 co-silenced pepper plants were characterized by drought-tolerant phenotypes similar to those observed in CaAITP1-silenced pepper. Collectively, our findings indicate that CaDeSI2 reduces the stability of CaAITP1 via deSUMOylation, thereby positively regulating drought tolerance.


Asunto(s)
Ácido Abscísico , Capsicum , Sequías , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas , Estrés Fisiológico , Capsicum/genética , Capsicum/fisiología , Capsicum/enzimología , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Ácido Abscísico/metabolismo , Estrés Fisiológico/genética , Sumoilación , Plantas Modificadas Genéticamente , Arabidopsis/genética , Arabidopsis/fisiología , Silenciador del Gen , Fosfoproteínas Fosfatasas/metabolismo , Fosfoproteínas Fosfatasas/genética , Unión Proteica , Estabilidad Proteica , Fenotipo
2.
Plant Cell Environ ; 47(4): 1319-1333, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38221841

RESUMEN

Controlling protein stability or degradation via the ubiquitin-26S proteasome system is a crucial mechanism in plant cellular responses to stress conditions. Previous studies have revealed that the pepper FANTASTIC FOUR-like gene, CaFAF1, plays a positive role in salt tolerance and that, in this process, CaFAF1 protein degradation is delayed. Here, we sought to isolate the E3 ligases potentially responsible for modulating CaFAF1 protein stability in response to salt stress. The pepper RING-type E3 ligase CaFIRF1 (Capsicum  annuum  FAF1  Interacting  RING  Finger protein  1) was found to interact with and ubiquitinate CaFAF1, leading to the degradation of CaFAF1 proteins. In response to high-salt treatments, CaFIRF1-silenced pepper plants exhibited tolerant phenotypes. In contrast, co-silencing of CaFAF1 and CaFIRF1 led to increased sensitivity to high-salt treatments, revealing that CaFIRF1 functions upstream of CaFAF1. A cell-free degradation analysis showed that high-salt treatment suppressed CaFAF1 protein degradation via the 26S proteasome pathway, in which CaFIRF1 is functionally involved. In addition, an in vivo ubiquitination assay revealed that CaFIRF1-mediated ubiquitination of CaFAF1 proteins was reduced by high-salt treatment. Taken together, these findings suggest that the degradation of CaFAF1 mediated by CaFIRF1 has a critical role in pepper plant responses to high salinity.


Asunto(s)
Ácido Abscísico , Ubiquitina-Proteína Ligasas , Ubiquitina-Proteína Ligasas/genética , Ubiquitina-Proteína Ligasas/metabolismo , Ácido Abscísico/metabolismo , Estrés Fisiológico/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estrés Salino , Plantas Modificadas Genéticamente/metabolismo , Regulación de la Expresión Génica de las Plantas
3.
Artículo en Inglés | MEDLINE | ID: mdl-38189362

RESUMEN

Two Gram-stain-negative, obligately aerobic, motile rod bacteria, designated as G2-5T and G20-9T, exhibiting catalase- and oxidase-positive activities, were isolated from the phycosphere of a Chondrus species, a marine red alga. Strain G2-5T exhibited optimal growth at 30 °C and pH 5.0-6.0 and in the presence of 0.5-1.0% NaCl. In contrast, strain G20-9T demonstrated optimal growth at 25 °C and pH 6.0 and in the presence of 0.5-1.5% NaCl. Both strains contained ubiquinone-10, summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c), C18 : 0 and 11-methyl-C18 : 1 ω7c, and diphosphatidylglycerol and phosphatidylglycerol as the major respiratory isoprenoid quinone, cellular fatty acids and polar lipids, respectively. The genomic DNA G+C contents were 57.2 mol% for strain G2-5T and 57.5 mol% for strain G20-9T. Strains G2-5T and G20-9T exhibited 98.2 % 16S rRNA gene sequence similarity, along with 82.3 % average nucleotide identity (ANI) and 25.0 % digital DNA-DNA hybridization (dDDH) values, indicating that they represent different species. Phylogenetic analyses based on both 16S rRNA gene and genome sequences revealed that strains G2-5T and G20-9T formed distinct phylogenic lineages within the genus Devosia. Strains G2-5T and G20-9T were most closely related to Devosia limi DSM 17137T and Devosia beringensis S02T with 97.7 and 96.9 % 16S rRNA gene sequence similarities, respectively. The ANI and dDDH values between strains G2-5T and G20-9T and other Devosia species were lower than 73.9 and 19.2 %, respectively, suggesting that they constitute novel species within the genus Devosia. Based on their distinct phenotypic, chemotaxonomic, and molecular characteristics, strains G2-5T and G20-9T represent two novel species of the genus Devosia, for which the names Devosia rhodophyticola sp. nov. (G2-5T=KACC 22601T=JCM 35404T) and Devosia algicola sp. nov. (G20-9T=KACC 22650T=JCM 35405T) are proposed, respectively.


Asunto(s)
Gammaproteobacteria , Rhodophyta , Composición de Base , Ácidos Grasos/química , Filogenia , ARN Ribosómico 16S/genética , Cloruro de Sodio , Análisis de Secuencia de ADN , ADN Bacteriano/genética , Técnicas de Tipificación Bacteriana , Nucleótidos
4.
Plant Cell Environ ; 46(7): 2061-2077, 2023 07.
Artículo en Inglés | MEDLINE | ID: mdl-37128851

RESUMEN

Abscisic acid (ABA) signalling triggers drought resistance mediated by SNF1-related kinase 2s (SnRK2s), which transmits stress signals through the phosphorylation of several downstream factors. However, these kinases and their downstream targets remain elusive in pepper plants. This study aimed to isolate interacting partners of CaSnRK2.6, a homologue of Arabidopsis SnRK2.6/OST1. Among the candidate proteins, we identified a homeodomain-leucine zipper (HD-Zip) class II protein and named it CaHAT1 (Capsicum annuum homeobox ABA signalling related- transcription factor 1). CaHAT1-silenced pepper and -overexpression (OE) transgenic Arabidopsis plants were generated to investigate the in vivo function of CaHAT1 in drought response. Following the application of drought stress, CaHAT1-silenced pepper plants exhibited drought-sensitive phenotypes with reduced ABA-mediated stomatal closure and lower expression of stress-responsive genes compared with control plants. In contrast, CaHAT1-OE transgenic Arabidopsis plants showed the opposite phenotypes, including increased drought resistance and ABA sensitivity. CaHAT1, particularly its N-terminal consensus sequences, was directly phosphorylated by CaSnRK2.6. Furthermore, CaSnRK2.6 kinase activity and CaSnRK2.6-mediated CaHAT1 phosphorylation levels were enhanced by treatment with ABA and drought stress. Taken together, our results indicated that CaHAT1, which is the target protein of CaSnRK2.6, is a positive regulator of drought stress response. This study advances our understanding of CaHAT1-CaSnRK2.6 mediated defence mechanisms in pepper plants against drought stress.


Asunto(s)
Ácido Abscísico , Arabidopsis , Ácido Abscísico/metabolismo , Arabidopsis/metabolismo , Sequías , Genes Homeobox , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Transducción de Señal , Plantas Modificadas Genéticamente/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Estrés Fisiológico , Regulación de la Expresión Génica de las Plantas
6.
Artículo en Inglés | MEDLINE | ID: mdl-35704462

RESUMEN

A Gram-stain-positive, facultative aerobic, oxidase-negative, catalase-positive, non-sporulating, and non-motile bacterium, which degraded benzophenone-3, was isolated from stream sediment collected in the Republic of Korea and designated as strain S2-17T. Cells of this strain were rod-shaped during the early growth phase but became coccoid after the late exponential growth phase. Bacterial growth was observed at 15-37 °C (optimum, 25-30 °C) and pH 6.0-9.5 (optimum, pH 7.5-8.5) and in the presence of 0-9.0 % (w/v) NaCl (optimum, 0-1.0 %). Menaquinone-8 (H2) was the sole isoprenoid quinone, and C16 : 0, C17 : 1 ω8c, summed feature 3 (comprising C16 : 1 ω7c/C16 : 1 ω6c) and C18 : 1 ω9c were the major fatty acids. The cell wall of strain S2-17T contained meso-diaminopimelic acid, and arabinose, galactose and mycolic acid were found in whole-cell hydrolysates, suggesting a chemotype IV cell wall. The G+C content of the genome was 65.6 mol%. Phylogenetic analyses revealed that strain S2-17T formed a phyletic lineage within the genus Rhodococcus and was most closely related to Rhodococcus jostii DSM 44719T (99.2 % 16S rRNA gene sequence similarity). Average nucleotide identity and digital DNA-DNA hybridization values between strain S2-17T and R. jostii DSM 44719T were 82.6 and 26.5 %, respectively, indicating differences between the species. Based on its phenotypic, chemotaxonomic and molecular features, strain S2-17T represents a novel species of the genus Rhodococcus, for which the name Rhodococcus oxybenzonivorans sp. nov. is proposed. The type strain is S2-17T (=KACC 19281T=JCM 32046T).


Asunto(s)
Rhodococcus , Ríos , Técnicas de Tipificación Bacteriana , Composición de Base , Benzofenonas , ADN Bacteriano/genética , Ácidos Grasos/química , Fosfolípidos/química , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN
7.
Artículo en Inglés | MEDLINE | ID: mdl-35119982

RESUMEN

A Gram-stain-negative, catalase- and oxidase-positive and aerobic bacterium, designated strain R798T, was isolated from soil in South Korea. Cells were motile rods by means of a single polar flagellum. Growth of strain R798T was observed at 15-35 °C (optimum, 25-30 °C), pH 5.0-8.0 (optimum, 6.0) and 0-1.5 % NaCl (optimum, 0.3 %). Strain R798T contained ubiquinone-8 as the sole isoprenoid quinone, summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c) and C16 : 0 as the major fatty acids and phosphatidylglycerol and phosphatidylethanolamine as the major polar lipids. The DNA G+C content of strain R798T calculated from the whole genome sequence was 63.3 mol%. Phylogenetic analyses based on the 16S rRNA gene and whole genome sequences revealed that strain R798T formed a distinct phyletic lineage within the genus Massilia. Strain R798T was most closely related to Massilia eurypsychrophila B528-3T with a 98.0 % 16S rRNA gene sequence similarity. Average nucleotide identity and digital DNA-DNA hybridization values between strain R798T and the type strain of M. eurypsychrophila were 79.2 and 22.7 %, respectively. Based on the phenotypic, chemotaxonomic and molecular analyses, strain R798T represents a novel species of the genus Massilia, for which the name Massilia soli sp. nov. is proposed. The type strain is R798T (=KACC 22114T=JCM 34601T).


Asunto(s)
Oxalobacteraceae/clasificación , Filogenia , Microbiología del Suelo , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Ácidos Grasos/química , Oxalobacteraceae/aislamiento & purificación , Fosfolípidos/química , ARN Ribosómico 16S/genética , República de Corea , Análisis de Secuencia de ADN , Ubiquinona/química
8.
Plant Signal Behav ; 16(12): 1974725, 2021 12 02.
Artículo en Inglés | MEDLINE | ID: mdl-34658295

RESUMEN

Deubiquitination, a type of post-translational modification, cleaves ubiquitin from target proteins, thereby regulating their stability or activity. Deubiquitination enzymes, ubiquitin-specific proteases (UBP/USP), have been reported to be involved in numerous cellular processes in plants, including meristem development, circadian clock regulation, and immunity. In contrast to model plants, however, the functions of UBP in other higher plants remain poorly understood. Here, we isolated a deubiquitination enzyme, ubiquitin-specific protease 12 (NbUBP12), from Nicotiana benthamiana, which shows high sequence homology with the core enzyme regions of UBP12 from other plants. Quantitative reverse-transcription PCR analysis revealed that NbUBP12 gene expression was significantly induced after drought treatment, and its level was higher in seed than in other tissues. Using a virus-induced gene silencing technique, we generated NbUBP12-silenced tobacco plants to analyze NbUBP12 gene function in response to drought stress and found that compared with control plants, NbUBP12-silenced plants exhibited a lower survival rate after exposure to drought stress. In addition, they were characterized by lower leaf surface temperatures and larger stomatal pore size following abscisic acid (ABA) treatment. On the basis of these observations, we suggest that NbUBP12 is involved in modulating drought resistance in N. benthamiana, which is associated with ABA-mediated stomatal closure.


Asunto(s)
Sequías , Endopeptidasas , Nicotiana , Ácido Abscísico/metabolismo , Ácido Abscísico/farmacología , Regulación de la Expresión Génica de las Plantas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Estrés Fisiológico/genética , Proteasas Ubiquitina-Específicas/genética , Proteasas Ubiquitina-Específicas/metabolismo
9.
Plant J ; 107(4): 1148-1165, 2021 08.
Artículo en Inglés | MEDLINE | ID: mdl-34145668

RESUMEN

Abscisic acid (ABA) is a plant hormone that activates adaptive mechanisms to environmental stress conditions. Plant adaptive mechanisms are complex and highly modulated processes induced by stress-responsive proteins; however, the precise mechanisms by which these processes function under adverse conditions remain unclear. Here, we isolated CaUBP12 (Capsicum annuum ubiquitin-specific protease 12) from pepper (C. annuum) leaves. We show that CaUBP12 expression is significantly induced after exposure to abiotic stress treatments. We conducted loss-of-function and gain-of-function genetic studies to elucidate the biological functions of CaUBP12 in response to ABA and dehydration stress. CaUBP12-silenced pepper plants and CaUBP12-overexpressing Arabidopsis plants displayed dehydration-sensitive and dehydration-tolerant phenotypes, respectively; these phenotypes were characterized by regulation of transpirational water loss and stomatal aperture. Under dehydration stress conditions, CaUBP12-silenced pepper plants and CaUBP12-overexpressing Arabidopsis plants exhibited lower and higher expression levels of stress-related genes, respectively, than the control plants. We isolated a CaUBP12 interaction protein, CaSnRK2.6, which is a homolog of Arabidopsis OST1; degradation of this protein was partially inhibited by CaUBP12. Similar to CaUBP12-silenced pepper plants and CaUBP12-overexpressing Arabidopsis plants, CaSnRK2.6-silenced pepper plants and CaSnRK2.6-overexpressing Arabidopsis displayed dehydration-sensitive and dehydration-tolerant phenotypes, respectively. Our findings suggest that CaUBP12 positively modulates the dehydration stress response by suppressing CaSnRK2.6 protein degradation.


Asunto(s)
Capsicum/fisiología , Deshidratación/genética , Proteínas de Plantas/metabolismo , Proteasas Ubiquitina-Específicas/metabolismo , Ácido Abscísico/farmacología , Arabidopsis/genética , Arabidopsis/fisiología , Proteínas de Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Germinación/efectos de los fármacos , Mutación , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente , Proteínas Quinasas/genética , Estabilidad Proteica , Semillas/efectos de los fármacos , Semillas/fisiología , Proteasas Ubiquitina-Específicas/genética
10.
Curr Genomics ; 22(1): 4-15, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-34045920

RESUMEN

Under drought stress, plants have developed various mechanisms to survive in the reduced water supply, of which the regulation of stress-related gene expression is responsible for several transcription factors. The basic leucine zippers (bZIPs) are one of the largest and most diverse transcription factor families in plants. Among the 10 Arabidopsis bZIP groups, group A bZIP transcription factors function as a positive or negative regulator in ABA signal transduction and drought stress response. These bZIP transcription factors, which are involved in the drought response, have also been isolated in various plant species such as rice, pepper, potato, and maize. Recent studies have provided substantial evidence that many bZIP transcription factors undergo the post-translational modifications, through which the regulation of their activity or stability affects plant responses to various intracellular or extracellular stimuli. This review aims to address the modulation of the bZIP proteins in ABA signaling and drought responses through phosphorylation, ubiquitination and sumoylation.

11.
J Exp Bot ; 72(12): 4520-4534, 2021 05 28.
Artículo en Inglés | MEDLINE | ID: mdl-33837765

RESUMEN

Plants have developed defense mechanisms to survive in extreme environmental conditions. Abscisic acid (ABA) is a key phytohormone associated with plant adaptation to environmental stress. In this study, we isolated and functionally characterized the pepper RING-type E3 ligase CaAIRE1 (Capsicum annuum ABA Induced RING-type E3 ligase 1) containing the C3HC4-type RING domain. CaAIRE1 was induced by ABA and drought, and CaAIRE1 had E3 ligase activity. CaAIRE1-silenced pepper and CaAIRE1-overexpressing Arabidopsis presented drought-sensitive and drought-tolerant phenotypes, respectively, which were accompanied by altered transpiration water loss and ABA sensitivity. Moreover, we found that CaAIRE1 interacts with and ubiquitinates the pepper type 2C protein phosphatase, CaAITP1 (Capsicum annuum CaAIRE1 Interacting Target Phosphatase 1). A cell-free degradation assay with CaAIRE1-silenced peppers and CaAIRE1-overexpressing Arabidopsis plants revealed that the CaAITP1 protein level was negatively modulated by the expression level of CaAIRE1. In contrast to CaAIRE1, CaAITP1-silenced pepper showed ABA-sensitivity phenotypes. CaAITP1-overexpressing Arabidopsis plants were the most insensitive phenotypes to ABA compared with the wild type and other pepper PP2C-overexpressing plants. Taken together, our data indicate that CaAITP1 plays a major role as a negative modulator in ABA signaling, and CaAIRE1 regulates the ABA signaling and drought response through modulation of CaAITP1 stability.


Asunto(s)
Ácido Abscísico , Proteínas de Arabidopsis , Proteínas de Arabidopsis/genética , Capsicum , Sequías , Regulación de la Expresión Génica de las Plantas , Fosfoproteínas Fosfatasas/genética , Fosfoproteínas Fosfatasas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Transducción de Señal , Estrés Fisiológico , Ubiquitina-Proteína Ligasas/genética , Ubiquitina-Proteína Ligasas/metabolismo
12.
J Nanosci Nanotechnol ; 20(1): 42-47, 2020 01 01.
Artículo en Inglés | MEDLINE | ID: mdl-31383138

RESUMEN

There is an urgent need for in situ methods for detecting environmental pollution quickly and accurately. With the development of nanotechnology, a huge potential has been created for the design of highly sensitive sensors with low energy consumption and low costs. If a composite material constructed with carbon nanotubes is used as an electrode in contact with a contaminant, this material undergoes an oxidation-reduction reaction with the contaminant that allows the electrode to function as an electrochemical sensor. This study involved the application of multi-walled carbon nanotubes and modified working electrodes constructed with multi-walled carbon nanotube composites (Ag- and ZnO-multi-walled carbon nanotubes) as electrochemical sensors. These electrodes have good response speed and sensitivity at low concentrations, and they are reusable. To lower the price of these sensors, our goal was to maximize their sensitivity by using the low-cost multiwalled carbon nanotubes in conjunction with silver electroless plating of the multi-walled carbon nanotubes and multi-walled carbon nanotube composites.

13.
Plant J ; 100(3): 473-486, 2019 11.
Artículo en Inglés | MEDLINE | ID: mdl-31436880

RESUMEN

Recent work has established a core ABA signaling pathway in which A-type PP2C protein phosphatases act as central negative modulators. Although ABA signaling inhibits PP2C activity through ABA-receptor complex, it remains unknown if other mechanisms exist to modulate the level of PP2Cs. Here, we identified a RING domain ubiquitin E3 ligase, PIR1 (PP2CA interacting RING finger protein 1), that interacted with PP2CA. Of the two splicing isoforms, PIR1.2 was isolated from leaf tissue. The PIR1.2 exhibited E3 ligase activity and determined PP2CA stability in the presence of ABA. Consistent with the conclusion that PIR1 promotes ABA signaling by removing PP2CA, a negative modulator, the pir1 knockout mutant displayed an ABA-hyposensitive phenotype. We further showed that PIR2, the closest homologue of PIR1.2, also interacted with PP2CA. Although the pir2 knockout mutant did not display altered ABA response, the pir1-1/pir2 double mutant became more insensitive to ABA than the wild-type or pir1-1 and pir2 single mutants. Using a cell-free degradation assay, ABA promoted degradation of PP2CA, however, such degradation was delayed when incubated with protein extract prepared from the pir1-1/pir2 double mutant. Our data suggest that PIR1 and PIR2 positively modulate ABA signaling by targeting PP2CA for degradation.


Asunto(s)
Ácido Abscísico/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimología , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Complejo de la Endopetidasa Proteasomal/metabolismo , Transducción de Señal , Empalme Alternativo , Arabidopsis/genética , Arabidopsis/fisiología , Proteínas de Arabidopsis/genética , Técnicas de Inactivación de Genes , Péptidos y Proteínas de Señalización Intercelular/genética , Isoenzimas , Fenotipo , Fosfoproteínas Fosfatasas/genética , Fosfoproteínas Fosfatasas/metabolismo , Hojas de la Planta/enzimología , Hojas de la Planta/genética , Hojas de la Planta/fisiología , Proteolisis , Dominios RING Finger , Semillas/enzimología , Semillas/genética , Semillas/fisiología , Ubiquitina-Proteína Ligasas/genética , Ubiquitina-Proteína Ligasas/metabolismo
14.
Plant J ; 96(2): 452-467, 2018 10.
Artículo en Inglés | MEDLINE | ID: mdl-30051516

RESUMEN

Abscisic acid (ABA) is a plant hormone that plays a key role in the environmental stress response, especially the induction of ABA-responsive and stress-responsive genes and modulation of the stomatal aperture in response to drought stress. Here, we identified CaDILZ1 (Capsicum annuum Drought-Induced Leucine Zipper 1) belonging to subgroup D of the bZIP protein family; gene functions of this family in response to ABA and drought signaling still remain unknown. CaDILZ1 expression was significantly induced in pepper leaves after exposure to ABA, drought, and NaCl. The CaDILZ1 protein localized in the nucleus of plant cells. In response to drought stress, CaDILZ1-silenced pepper and CaDILZ1-overexpressing Arabidopsis plants exhibited drought-sensitive and drought-tolerant phenotypes, respectively, via altered ABA content, stomatal closure, and expression of ABA-responsive and drought-responsive marker genes. We isolated the RING finger protein CaDSR1 (Capsicum annuum Drought Sensitive RING finger protein 1), which interacted with CaDILZ1 in the nucleus. The CaDSR1 protein exhibited E3 ligase activity and promoted CaDILZ1 degradation via the 26S proteasome pathway. Under drought stress conditions, CaDSR1-silenced pepper and CaDSR1-overexpressing Arabidopsis plants exhibited contrasting phenotypes to those of CaDILZ1-silenced pepper and CaDILZ1-overexpressing Arabidopsis plants. Taken together, our data suggest that CaDSR1 and CaDILZ1 function in ABA-mediated drought stress signaling in pepper plants.


Asunto(s)
Ácido Abscísico/metabolismo , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/metabolismo , Capsicum/genética , Capsicum/fisiología , Reguladores del Crecimiento de las Plantas/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/genética , Capsicum/enzimología , Sequías , Silenciador del Gen , Hojas de la Planta/enzimología , Hojas de la Planta/genética , Hojas de la Planta/fisiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Complejo de la Endopetidasa Proteasomal/metabolismo , Transducción de Señal , Estrés Fisiológico , Ubiquitina-Proteína Ligasas/genética
15.
Plant Cell Environ ; 41(7): 1593-1604, 2018 07.
Artículo en Inglés | MEDLINE | ID: mdl-29574779

RESUMEN

Abscisic acid (ABA) is major plant hormone involved in regulating abiotic stress responses. Several studies have established that an ABA-signalling transduction pathway-from ABA perception to response-functions in plant cells. The group A PP2Cs constitute core components of ABA signalling, and they negatively regulate ABA signalling and stress responses. Recent studies have identified and functionally analysed regulators of PP2C activity; however, the precise regulatory mechanisms remain unclear. In the present study, we used a yeast 2-hybrid (Y2H) screening analysis to identify the DEAD-box RNA helicase RH8, which interacted with PP2CA in the nucleus. rh8 knockout mutants exhibited ABA hyposensitivity and drought-susceptible phenotypes characterized by high levels of transpirational water loss via reduced stomatal closure and decreased leaf temperatures. However, rh8/pp2ca double mutants showed ABA hypersensitivity and drought-tolerant phenotypes, indicating that RH8 and PP2CA function in the same ABA-signalling pathway in the drought stress response; moreover, RH8 functions upstream of PP2CA. In vitro phosphatase and kinase assays revealed that RH8 inhibits PP2CA phosphatase activity. Our data indicate that RH8 and its interacting partner PP2CA modulate the drought stress response via ABA-dependent signalling.


Asunto(s)
Ácido Abscísico/metabolismo , Proteínas de Arabidopsis/fisiología , ARN Helicasas DEAD-box/fisiología , Fosfoproteínas Fosfatasas/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Transducción de Señal , Arabidopsis/enzimología , Arabidopsis/metabolismo , Arabidopsis/fisiología , Proteínas de Arabidopsis/metabolismo , ARN Helicasas DEAD-box/metabolismo , Deshidratación , Técnicas de Inactivación de Genes , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
16.
Plant Cell Environ ; 40(10): 2359-2368, 2017 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-28742940

RESUMEN

Plant adaptive responses to abiotic stress are coordinated by restriction of plant growth and development. The plant hormone abscisic acid (ABA) is the key regulator of the response to abiotic stress, and its sensitivity determines abiotic stress tolerance levels. We previously showed that the E3 ubiquitin ligase CaAIRF1 functions as a positive regulator of ABA and drought stress via modulation of transcription and stability of the type 2C protein phosphatase CaADIP1. Here, we report the identification and functional analysis of a novel-type 2C phosphatase, CaAIPP1 (Capsicum annuum CaAIRF1 Interacting Protein Phosphatase 1). CaAIPP1 interacted with and was ubiquitinated by CaAIRF1. CaAIPP1 gene expression in pepper leaves was induced by ABA and drought. CaAIPP1 degradation was faster in crude protein extracts from ABA-treated pepper plants than in those from control plants. CaAIPP1-overexpressing plants displayed an ABA-hyposensitive phenotype during seed germination and seedling growth. Moreover, these plants exhibited a drought-sensitive phenotype characterized by high levels of transpirational water loss via decreased stomatal closure and reduced leaf temperatures. Our data indicate that CaAIPP1 is a negative regulator of the drought stress response via ABA-mediated signalling. Our findings provide a valuable insight into the plant defence mechanism that operates during drought stress.


Asunto(s)
Ácido Abscísico/metabolismo , Capsicum/enzimología , Capsicum/metabolismo , Sequías , Fosfoproteínas Fosfatasas/metabolismo , Proteínas de Plantas/metabolismo , Transducción de Señal , Estrés Fisiológico , Adaptación Fisiológica/genética , Secuencia de Aminoácidos , Arabidopsis/genética , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente , Unión Proteica , Proteolisis , Análisis de Secuencia de Proteína , Ubiquitinación
18.
Plant Cell Physiol ; 58(4): 779-788, 2017 04 01.
Artículo en Inglés | MEDLINE | ID: mdl-28339719

RESUMEN

Plants are constantly challenged by various environmental stresses, including high salinity and drought, and they have evolved defense mechanisms to counteract the deleterious effects of these stresses. The plant hormone ABA regulates plant growth and developmental processes and mediates abiotic stress responses. Here, we report the identification and characterization of a novel CaWDP1 (Capsicum annuum) protein. The expression of CaWDP1 in pepper leaves was induced by ABA, drought and NaCl treatments, suggesting its role in the abiotic stress response. CaWDP1 proteins show conserved sequence homology with other known WDP1 proteins, and they are localized in the nucleus and cytoplasm. We generated CaWDP1-silenced peppers via virus-induced gene silencing (VIGS). We evaluated the responses of these CaWDP1-silenced pepper plants and CaWDP1-overexpressing (OX) transgenic Arabidopsis plants to ABA and drought. CaWDP1-silenced pepper plants displayed enhanced tolerance to drought stress, and this was characterized by low levels of leaf water loss in the drought-treated leaves. In contrast to CaWDP1-silenced plants, CaWDP1-OX plants exhibited an ABA-hyposensitive and drought-susceptible phenotype, which was accompanied by high levels of leaf water loss, low leaf temperatures, increased stomatal pore size and low expression levels of stress-responsive genes. Our results indicate that CaWDP1, a novel pepper negative regulator of ABA, regulates the ABA-mediated defense response to drought stress.


Asunto(s)
Ácido Abscísico/metabolismo , Capsicum/fisiología , Sequías , Proteínas de Plantas/metabolismo , Ácido Abscísico/farmacología , Arabidopsis/genética , Capsicum/efectos de los fármacos , Capsicum/genética , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Germinación/efectos de los fármacos , Germinación/fisiología , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente , Salinidad , Plantones/efectos de los fármacos , Plantones/fisiología , Transducción de Señal , Estrés Fisiológico
19.
Plant Physiol ; 173(4): 2323-2339, 2017 04.
Artículo en Inglés | MEDLINE | ID: mdl-28184010

RESUMEN

Ubiquitin-mediated protein modification occurs at multiple steps of abscisic acid (ABA) signaling. Here, we sought proteins responsible for degradation of the pepper (Capsicum annuum) type 2C protein phosphatase CaADIP1 via the 26S proteasome system. We showed that the RING-type E3 ligase CaAIRF1 (Capsicum annuum ADIP1 Interacting RING Finger Protein 1) interacts with and ubiquitinates CaADIP1. CaADIP1 degradation was slower in crude proteins from CaAIRF1-silenced peppers than in those from control plants. CaAIRF1-silenced pepper plants displayed reduced ABA sensitivity and decreased drought tolerance characterized by delayed stomatal closure and suppressed induction of ABA- and drought-responsive marker genes. In contrast, CaAIRF1-overexpressing Arabidopsis (Arabidopsis thaliana) plants exhibited ABA-hypersensitive and drought-tolerant phenotypes. Moreover, in these plants, CaADIP1-induced ABA hyposensitivity was strongly suppressed by CaAIRF1 overexpression. Our findings highlight a potential new route for fine-tune regulation of ABA signaling in pepper via CaAIRF1 and CaADIP1.


Asunto(s)
Ácido Abscísico/farmacología , Capsicum/metabolismo , Sequías , Fosfoproteínas Fosfatasas/metabolismo , Proteínas de Plantas/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Adaptación Fisiológica/genética , Secuencia de Aminoácidos , Arabidopsis/enzimología , Arabidopsis/genética , Capsicum/enzimología , Capsicum/genética , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Microscopía Confocal , Fosfoproteínas Fosfatasas/genética , Reguladores del Crecimiento de las Plantas/farmacología , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente , Unión Proteica , Proteolisis/efectos de los fármacos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Homología de Secuencia de Aminoácido , Transducción de Señal , Estrés Fisiológico , Ubiquitina-Proteína Ligasas/genética
20.
Plant Mol Biol ; 91(1-2): 149-60, 2016 May.
Artículo en Inglés | MEDLINE | ID: mdl-26869261

RESUMEN

Plants are constantly challenged by various environmental stresses, including high salinity and drought, and they have evolved defense mechanisms to counteract the deleterious effects of these stresses. The plant hormone abscisic acid (ABA) regulates plant growth and developmental processes and mediates abiotic stress responses. Here, we identified the Capsicum annuum DRought Tolerance 1 (CaDRT1) gene from pepper leaves treated with ABA. CaDRT1 was strongly expressed in pepper leaves in response to environmental stresses and after ABA treatment, suggesting that the CaDRT1 protein functions in the abiotic stress response. Knockdown expression of CaDRT1 via virus-induced gene silencing resulted in a high level of drought susceptibility, and this was characterized by increased transpirational water loss via decreased stomatal closure. CaDRT1-overexpressing (OX) Arabidopsis plants exhibited an ABA-hypersensitive phenotype during the germinative, seedling, and adult stages. Additionally, these CaDRT1-OX plants exhibited a drought-tolerant phenotype characterized by low levels of transpirational water loss, high leaf temperatures, increased stomatal closure, and enhanced expression levels of drought-responsive genes. Taken together, our results suggest that CaDRT1 is a positive regulator of the ABA-mediated drought stress response.


Asunto(s)
Ácido Abscísico/metabolismo , Capsicum/metabolismo , Regulación de la Expresión Génica de las Plantas/fisiología , Proteínas de Plantas/metabolismo , Estrés Fisiológico/fisiología , Agua/metabolismo , Secuencia de Aminoácidos , Capsicum/genética , Clorofila/metabolismo , Silenciador del Gen , Germinación , Datos de Secuencia Molecular , Mutación , Filogenia , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente , ARN de Planta/genética , ARN de Planta/metabolismo , Plantones/fisiología , Cloruro de Sodio/farmacología
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