RESUMEN
A multi-step phosphorelay system is the main conduit of cytokinin signal transduction. However, several groups of additional factors that also play a role in this signaling pathway have been found-among them the Cytokinin Response Factors (CRFs). In a genetic screen, CRF9 was identified as a regulator of the transcriptional cytokinin response. It is mainly expressed in flowers. Mutational analysis indicates that CRF9 plays a role in the transition from vegetative to reproductive growth and silique development. The CRF9 protein is localized in the nucleus and functions as a transcriptional repressor of Arabidopsis Response Regulator 6 (ARR6)-a primary response gene for cytokinin signaling. The experimental data suggest that CRF9 functions as a repressor of cytokinin during reproductive development.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Citocininas/metabolismo , Factor IX/genética , Flores/metabolismo , Regulación de la Expresión Génica de las PlantasRESUMEN
Cytokinin has strong connections to development and a growing role in the abiotic stress response. Here we show that CYTOKININ RESPONSE FACTOR 2 (CRF2) is additionally involved in the salt (NaCl) stress response. CRF2 promoter-GUS expression indicates CRF2 involvement in the response to salt stress as well as the previously known cytokinin response. Interestingly, CRF2 mutant seedlings are quite similar to the wild type (WT) under non-stressed conditions yet have many distinct changes in response to salt stress. Cytokinin levels measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS) that increased in the WT after salt stress are decreased in crf2, potentially from CRF2 regulation of cytokinin biosynthesis genes. Ion content measured by inductively coupled plasma optical emission spectrometry (ICP-OES) was increased in the WT for Na, K, Mn, Ca and Mg after salt stress, whereas the corresponding Ca and Mg increases are lacking in crf2. Many genes examined by RNA-seq analysis were altered transcriptionally by salt stress in both the WT and crf2, yet interestingly approximately one-third of salt-modified crf2 transcripts (2655) showed unique regulation. Different transcript profiles for salt stress in crf2 compared with the WT background was further supported through an examination of co-expressed genes by weighted gene correlation network analysis (WGCMA) and principal component analysis (PCA). Additionally, Gene Ontology (GO) enrichment terms found from salt-treated transcripts revealed most photosynthesis-related terms as only being affected in crf2, leading to an examination of chlorophyll levels and the efficiency of photosystem II (via the ratio of variable fluorescence to maximum fluorescence, Fv /Fm ) as well as physiology after salt treatment. Salt stress-treated crf2 plants had both reduced chlorophyll levels and lower Fv /Fm values compared with the WT, suggesting that CRF2 plays a role in the modulation of salt stress responses linked to photosynthesis.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Clorofila/metabolismo , Cromatografía Liquida , Citocininas/metabolismo , Regulación de la Expresión Génica de las Plantas , Estrés Salino , Estrés Fisiológico , Espectrometría de Masas en TándemRESUMEN
Tobacco etch virus (TEV; genus Potyvirus) is flexuous rod shaped with a single molecule of single-stranded RNA and causes serious yield losses in species in the Solanaceae. Three TEV strains (HAT, Mex21, and N) are genetically distinct and cause different disease symptoms in plants. Here, a transcriptomic RNA sequencing approach was taken for each TEV strain to evaluate gene expression of the apical stem segment of pepper plants during two stages of disease development. Distinct profiles of Differentially Expressed Genes (DEGs) were identified for each TEV strain. DEG numbers increased with degree of symptom severity: 24 from HAT, 1190 from Mex21, and 4010 from N. At 7 days post-inoculation (dpi), when systemic symptoms were similar, there were few DEGs for HAT- and Mex21-infected plants, whereas N-infected plants had 2516 DEGs. DEG patterns from 7 to 14 dpi corresponded to severity of disease symptoms: milder disease with smaller DEG changes for HAT and Mex21 and severe disease with larger DEG changes for N. Strikingly, in each of these comparisons, there are very few overlapping DEGs among the TEV strains, including no overlapping DEGs between all three strains at 7 or 14 dpi.
Asunto(s)
Capsicum/genética , Capsicum/virología , Perfilación de la Expresión Génica , Tallos de la Planta/virología , Potyvirus/patogenicidad , Transcriptoma , Capsicum/anatomía & histología , Enfermedades de las Plantas/virología , Hojas de la Planta/genética , Hojas de la Planta/virología , Tallos de la Planta/genética , Potyvirus/clasificación , Potyvirus/genética , Potyvirus/crecimiento & desarrolloRESUMEN
Cytokinin response factors (CRFs) are transcription factors that are involved in cytokinin (CK) response, as well as being linked to abiotic stress tolerance. In particular, oxidative stress responses are activated by Clade III CRF members, such as AtCRF6. Here we explored the relationships between Clade III CRFs and oxidative stress. Transcriptomic responses to oxidative stress were determined in two Clade III transcription factors, Arabidopsis AtCRF5 and tomato SlCRF5. AtCRF5 was required for regulated expression of >240 genes that are involved in oxidative stress response. Similarly, SlCRF5 was involved in the regulated expression of nearly 420 oxidative stress response genes. Similarities in gene regulation by these Clade III members in response to oxidative stress were observed between Arabidopsis and tomato, as indicated by Gene Ontology term enrichment. CK levels were also changed in response to oxidative stress in both species. These changes were regulated by Clade III CRFs. Taken together, these findings suggest that Clade III CRFs play a role in oxidative stress response as well as having roles in CK signaling.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Citocininas , Solanum lycopersicum , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulación de la Expresión Génica de las Plantas , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Estrés Oxidativo , Proteínas de Plantas/metabolismoRESUMEN
Cytokinin signaling by way of a multistep two-component signaling pathway has been highly examined over the last 20 years with a lot of attention to how its signaling components regulate essential growth and developmental processes from seedlings to senescence. Most studies have focused on Angiosperm plants, particularly Arabidopsis, with very little attention across the rest of the plant kingdom, making it difficult to know if cytokinin-regulated developmental processes are have remained evolutionarily conserved or distinct among different plant groups. Here the prevalence of cytokinin signaling components throughout plants, algae, and other organisms are examined. Particular examination is paid to how minimal sets of these base signaling components are related to development in Bryophytes, a primary group of study outside of Angiosperms. General comparisons of the role of signaling components between these distant groups suggest some evolutionary conservation of function has been maintained across plants as a whole. This review also highlights the need to investigate how cytokinin signaling components, recently identified in every plant group, contribute to development; this will give higher resolution to our understanding of cytokinin roles in development across plant taxa. Also revealed is a strong need to further pursue how cytokinin signaling components recently identified in every plant group contribute to development in order to further dissect these connections.
Asunto(s)
Citocininas/metabolismo , Regulación de la Expresión Génica de las Plantas/genética , Magnoliopsida/metabolismo , Transducción de SeñalRESUMEN
Cytokinins (CKs) are well-known as a class of phytohormones capable of delaying senescence in detached leaves. However, CKs are often treated as a monolithic group of compounds even though dozens of CK species are present in plants with varied degrees of reported biological activity. One specific type of CK, isopentenyladenine base (iP), has been demonstrated as having roles in delaying leaf senescence, inhibition of root growth, and promoting shoot regeneration. However, its N-glucosides isopentenyladenine-7- and -9-glucoside (iP7G, iP9G) have remained understudied and thought of as inactive cytokinins for several decades, despite their relatively high concentrations in plants such as the model species Arabidopsis thaliana. Here we show that iP and one of its glucosides, iP9G, are capable of delaying senescence in leaves, though the glucosides having little to no activity in other bioassays. Additionally, we performed the first transcriptomic study of iP-delayed cotyledon senescence which shows that iP is capable of upregulating photosynthetic genes and downregulating catabolic genes in detached cotyledons. Transcriptomic analysis also shows iP9G has limited effects on gene expression, but that the few affected genes are CK-related and are similar to those seen from iP effects during senescence as seen for the type-A response regulator ARR6. These findings suggest that iP9G functions as an active CK during senescence.
RESUMEN
Cytokinins (CKs) are a class of phytohormones affecting many aspects of plant growth and development. In the complex process of CK homeostasis in plants, N-glucosylation represents one of the essential metabolic pathways. Its products, CK N7- and N9-glucosides, have been largely overlooked in the past as irreversible and inactive CK products lacking any relevant physiological impact. In this work, we report a widespread distribution of CK N-glucosides across the plant kingdom proceeding from evolutionary older to younger plants with different proportions between N7- and N9-glucosides in the total CK pool. We show dramatic changes in their profiles as well as in expression levels of the UGT76C1 and UGT76C2 genes during Arabidopsis ontogenesis. We also demonstrate specific physiological effects of CK N-glucosides in CK bioassays including their antisenescent activities, inhibitory effects on root development, and activation of the CK signaling pathway visualized by the CK-responsive YFP reporter line, TCSv2::3XVENUS. Last but not least, we present the considerable impact of CK N7- and N9-glucosides on the expression of CK-related genes in maize and their stimulatory effects on CK oxidase/dehydrogenase activity in oats. Our findings revise the apparent irreversibility and inactivity of CK N7- and N9-glucosides and indicate their involvement in CK evolution while suggesting their unique function(s) in plants.
Asunto(s)
Citocininas/genética , Evolución Molecular , Glucósidos/genética , Glucosiltransferasas/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Regulación de la Expresión Génica de las Plantas/genética , Oxidorreductasas/genética , Reguladores del Crecimiento de las Plantas/genética , Reguladores del Crecimiento de las Plantas/metabolismo , Zea mays/genética , Zea mays/metabolismoRESUMEN
Cytokinins are multifaceted plant hormones that play crucial roles in plant interactions with the environment. Modulations in cytokinin metabolism and signaling have been successfully used for elevating plant tolerance to biotic and abiotic stressors. Here, we analyzed Arabidopsis thaliana response to INhibitor of CYtokinin DEgradation (INCYDE), a potent inhibitor of cytokinin dehydrogenase. We found that at low nanomolar concentration, the effect of INCYCE on seedling growth and development was not significantly different from that of trans-Zeatin treatment. However, an alteration in the spatial distribution of cytokinin signaling was found at low micromolar concentrations, and proteomics analysis revealed a significant impact on the molecular level. An in-depth proteome analysis of an early (24 h) response and a dose-dependent response after 168 h highlighted the effects on primary and secondary metabolism, including alterations in ribosomal subunits, RNA metabolism, modulations of proteins associated with chromatin, and the flavonoid and phenylpropanoid biosynthetic pathway. The observed attenuation in stress-response mechanisms, including abscisic acid signaling and the metabolism of jasmonates, could explain previously reported positive effects of INCYDE under mild stress conditions.
RESUMEN
Cytokinin is a phytohormone involved in the regulation of diverse developmental and physiological processes in plants. Its potential in biotechnology and for development of higher-yield and more resilient plants has been recognized, yet the molecular mechanisms behind its action are far from understood. In this report, the roots of barley seedlings were explored as a new source to reveal as yet unknown cytokinin-responsive proteins for crop improvement. Here we found significant differences reproducibly observed for 178 proteins, for which some of the revealed cytokinin-responsive pathways were confirmed in metabolome analysis, including alterations phenylpropanoid pathway, amino acid biosynthesis and ROS metabolism. Bioinformatics analysis indicated a significant overlap between cytokinin response and response to abiotic stress. This was confirmed by comparing proteome and metabolome profiles in response to drought, salinity or a period of temperature stress. The results illustrate complex abiotic stress response in the early development of model crop plant and confirm an extensive crosstalk between plant hormone cytokinin and response to temperature stimuli, water availability or salinity stress.
RESUMEN
Cytokinin is an indispensable phytohormone responsible for physiological processes ranging from root development to leaf senescence. The term "cytokinin" refers to several dozen adenine-derived compounds occurring naturally in plants. Cytokinins (CKs) can be divided into various classes and forms; base forms are generally considered to be active while highly abundant cytokinin-N-glucosides (CKNGs), composed of a CK base irreversibly conjugated to a glucose molecule, are considered inactive. However, results from early CK studies suggest CKNGs do not always lack activity despite the perpetuation over several decades in the literature that they are inactive. Here we show that exogenous application of trans-Zeatin-N-glucosides (tZNGs, a specific class of CKNGs) to Arabidopsis results in CK response comparable to the application of an active CK base. These results are most apparent in senescence assays where both a CK base (tZ) and tZNGs (tZ7G, tZ9G) delay senescence in cotyledons. Further experiments involving root growth and shoot regeneration revealed tZNGs do not always have the same effects as tZ, and have largely distinct effects on the transcriptome and proteome. These data are in contrast to previous reports of CKNGs being inactive and raise questions about the function of these compounds as well as their mechanism of action.
Asunto(s)
Arabidopsis/crecimiento & desarrollo , Citocininas/metabolismo , Glucósidos/metabolismo , Raíces de Plantas/crecimiento & desarrollo , Zeatina/metabolismo , Arabidopsis/metabolismo , Reguladores del Crecimiento de las Plantas , Raíces de Plantas/metabolismoRESUMEN
Cytokinin (CK) is a plant hormone crucial to plant development and growth. Cytokinin Response Factor 6 (CRF6) is a CK-induced transcription factor that is part of the CK signaling cascade. While the role of CRF6 has been examined in oxidative stress response, there has been surprisingly little investigation of CRF6 in the context of CK signaling, including identifying CK-regulated targets of CRF6. Here, we conduct a transcriptomic study of Arabidopsis examining the CRF6 mutant (crf6) in the presence and absence of CK, revealing 163 downstream CRF6-dependent CK-regulated differentially expressed genes (DEGs). 15.3% of these DEGS were found as overlapping with larger number of standardly identified CK-regulated DEGs, suggesting that CRF6 is involved in regulating a subset of downstream CK responses through these gene targets. The general transcriptional regulation of CRF6-dependent CK-regulated DEGs indicates that CRF6 may function as a negative regulator of CK response. We investigated one subset of CRF6 CK-dependent targets (SKOR, HAK5, and NRT1. 5) involved in an underexamined functional role of CK response: the uptake and transportation of potassium. To determine how CK and CRF6 are involved in potassium acquisition and distribution, ionomic and physiological experiments were conducted on plants grown in media with sufficient and deficient potassium concentrations and in the presence and absence of CK. In order to investigate how CK alone affects potassium transport, similar experiments were performed on skor, hak5, and nrt1.5 mutant lines of these CRF6-dependent CK-regulated targets. These findings indicate novel connections between CK and potassium transport, which appear to be regulated in a CRF6-dependent manner.
RESUMEN
Hydrogen peroxide promotes seed germination, but the molecular mechanisms underlying this process are unclear. This study presents the results of eggplant (Solanum melongena) germination analyses conducted at two different temperatures and follows the effect of hydrogen peroxide treatment on seed germination and the seed proteome. Hydrogen peroxide was found to promote eggplant germination in a way not dissimilar to that of increased temperature stimuli. LC-MS profiling detected 729 protein families, 77 of which responded to a temperature increase or hydrogen peroxide treatment. These differentially abundant proteins were found to be involved in a number of processes, including protein and amino acid metabolism, carbohydrate metabolism, and the glyoxylate cycle. There was a very low overlap between hydrogen peroxide and temperature-responsive proteins, highlighting the differences behind the seemingly similar outcomes. Furthermore, the observed changes from the seed proteome indicate that hydrogen peroxide treatment diminished the seed endogenous hydrogen peroxide pool and that a part of manifested positive hydrogen peroxide effect might be related to altered sensitivity to abscisic acid.
Asunto(s)
Germinación/efectos de los fármacos , Peróxido de Hidrógeno/farmacología , Proteínas de Plantas/metabolismo , Solanum melongena/fisiología , Metabolismo de los Hidratos de Carbono/efectos de los fármacos , Cromatografía Liquida , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Espectrometría de Masas , Solanum melongena/efectos de los fármacos , Estrés Fisiológico/efectos de los fármacos , TemperaturaRESUMEN
Cytokinin Response Factors (CRFs) are a family of transcription factors which make up a side branch of the classical cytokinin two-component signaling pathway. CRFs were originally identified and have been primarily studied in Arabidopsis thaliana, although orthologs have be found throughout all land plants. Research into the evolution of CRFs as sub-group members of the larger APETALA2/Ethylene Response Factor (AP2/ERF) family has yielded interesting and useful insights related to the functional roles of CRFs in plants. Recent studies of CRFs suggest that these transcription factors are a lot more than just a group of cytokinin related genes and play important roles in both plant development and environmental stress response. This review focuses on recent advances in understanding the roles of CRFs beyond cytokinin, in reproductive development and abiotic stress response, as well as to other environmental cues.
Asunto(s)
Fenómenos Fisiológicos de las Plantas/genética , Proteínas de Plantas/genética , Factores de Transcripción/genética , Citocininas/metabolismo , Proteínas de Plantas/metabolismo , Reproducción/genética , Estrés Fisiológico/genética , Factores de Transcripción/metabolismoRESUMEN
Expression patterns of genes are controlled by short regions of DNA in promoter regions known as cis-regulatory elements. How expression patterns change due to alterations in cis-regulatory elements in the context of gene duplication are not well studied in plants. Over 300 promoter sequences from a small, well-conserved family of plant transcription factors known as Cytokinin Response Factors (CRFs) were examined for conserved motifs across several known clades present in Angiosperms. General CRF and lineage specific motifs were identified. Once identified, significantly enriched motifs were then compared to known transcription factor binding sites to elucidate potential functional roles. Additionally, presence of similar motifs shows that levels of conservation exist between different CRFs across land plants, likely occurring through processes of neo- or sub-functionalization. Furthermore, significant patterns of motif conservation are seen within and between CRF clades suggesting cis-regulatory regions have been conserved throughout CRF evolution.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Linaje de la Célula , Citocininas/metabolismo , Regulación de la Expresión Génica de las Plantas , Regiones Promotoras Genéticas , Secuencias Reguladoras de Ácidos Nucleicos , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Proteínas de Arabidopsis/genética , Citocininas/genética , FilogeniaRESUMEN
Cytokinin Response Factor (CRF) genes are a subgroup of AP2/ERF domain-containing transcription factors that are defined by the CRF domain, from which five clades of CRF genes have been identified. Clade III CRFs are strongly induced by cytokinin, as well as other abiotic stress factors, such as oxidative stress. While this appears well studied for the Clade III CRFs in Arabidopsis and tomato, there have been almost no studies done outside of these model systems. This study expands upon that and represents the first CRF research in the Sunflower family, Asteraceae. Fifty Asterid Clade III CRF protein sequences were examined, and novel Clade III CRF C-terminus motifs were identified. Clade III CRF genes of Marshallia mohrii and M. caespitosa were assembled from genome-skimming and transcriptomic data. Expression experiments were conducted on M. caespitosa to test responsiveness to both cytokinin and oxidative stress. Low levels of basal expression for the McCRF1 were found to be strongly induced in both treatment groups. These are the first experiments to show regulation of a nuclear gene in a Marshallia species, and these results suggest there is broad conservation in the sequence, form, and regulation of Clade III CRF genes and proteins.
Asunto(s)
Asteraceae/metabolismo , Citocininas/metabolismo , Filogenia , Proteínas de Plantas/metabolismo , Regiones no Traducidas 5'/genética , Secuencia de Aminoácidos , Asteraceae/genética , Secuencia Conservada/genética , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Sistemas de Lectura Abierta/genética , Proteínas de Plantas/química , Polimorfismo de Nucleótido Simple/genéticaRESUMEN
Cytokinins are well-known to be involved in processes responsible for plant growth and development. More recently, these hormones have begun to be associated with stress responses as well. However, it is unclear how changes in cytokinin biosynthesis, signaling, or transport relate to stress effects. This study examines in parallel how two different stresses, salt, and oxidative stress, affect changes in both cytokinin levels and whole plant transcriptome response. Solanum lycopersicum seedlings were given a short-term (6 hr) exposure to either salt (150 mM NaCl) or oxidative (20 mM H2O2) stress and then examined to determine both changes in cytokinin levels and transcriptome. LC-MS/MS was used to determine the levels of 22 different types of cytokinins in tomato plants including precursors, active, transported, and conjugated forms. When examining cytokinin levels we found that salt treatment caused an increase in both active and inactive cytokinin levels and oxidative stress caused a decrease in these levels. RNA-sequencing analyses of these same stress-treated tissues revealed 6,643 significantly differentially expressed genes (DEGs). Although many DEGs are similar between the two stresses, approximately one-third of the DEGs in each treatment were unique to that stress. Several cytokinin-related genes were among the DEGs. Examination of photosystem II efficiency revealed that cytokinins affect physiological response to stress in tomato, further validating the changes in cytokinin levels seen in planta.
RESUMEN
Cytokinin Response Factors (CRFs) are AP2/ERF transcription factors involved in cytokinin signal transduction. CRF proteins consist of a N-terminal dimerization domain (CRF domain), an AP2 DNA-binding domain, and a clade-specific C-terminal region of unknown function. Using a series of sequential deletions in yeast-2-hybrid assays, we provide evidence that the C-terminal region of Arabidopsis CRF5 can confer transactivation activity. Although comparative analyses identified evolutionarily conserved protein sequence within the C-terminal region, deletion experiments suggest that this transactivation domain has a partially redundant modular structure required for activation of target gene transcription.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Factores de Transcripción/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/fisiología , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Dominios Proteicos/genética , Dominios Proteicos/fisiología , Factores de Transcripción/genéticaRESUMEN
Cytokinin is a phytohormone that is well known for its roles in numerous plant growth and developmental processes, yet it has also been linked to abiotic stress response in a less defined manner. Arabidopsis (Arabidopsis thaliana) Cytokinin Response Factor 6 (CRF6) is a cytokinin-responsive AP2/ERF-family transcription factor that, through the cytokinin signaling pathway, plays a key role in the inhibition of dark-induced senescence. CRF6 expression is also induced by oxidative stress, and here we show a novel function for CRF6 in relation to oxidative stress and identify downstream transcriptional targets of CRF6 that are repressed in response to oxidative stress. Analysis of transcriptomic changes in wild-type and crf6 mutant plants treated with H2O2 identified CRF6-dependent differentially expressed transcripts, many of which were repressed rather than induced. Moreover, many repressed genes also show decreased expression in 35S:CRF6 overexpressing plants. Together, these findings suggest that CRF6 functions largely as a transcriptional repressor. Interestingly, among the H2O2 repressed CRF6-dependent transcripts was a set of five genes associated with cytokinin processes: (signaling) ARR6, ARR9, ARR11, (biosynthesis) LOG7, and (transport) ABCG14. We have examined mutants of these cytokinin-associated target genes to reveal novel connections to oxidative stress. Further examination of CRF6-DNA interactions indicated that CRF6 may regulate its targets both directly and indirectly. Together, this shows that CRF6 functions during oxidative stress as a negative regulator to control this cytokinin-associated module of CRF6-dependent genes and establishes a novel connection between cytokinin and oxidative stress response.
Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Citocininas/metabolismo , Perfilación de la Expresión Génica/métodos , Regulación de la Expresión Génica de las Plantas/genética , Estrés Oxidativo , Factores de Transcripción/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Clorofila/química , Clorofila/metabolismo , Fluorescencia , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Ontología de Genes , Peróxido de Hidrógeno/farmacología , Mutación , Oxidantes/farmacología , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Plantas Modificadas Genéticamente , Unión Proteica , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Plantones/genética , Plantones/metabolismo , Factores de Transcripción/metabolismo , Técnicas del Sistema de Dos HíbridosRESUMEN
KEY MESSAGE: Cytokinin response factor 4 (CRF4) shows a short-term induction by cold (4 °C) that appears to play a role in non-acclimated freezing tolerance as seen in mutant and overexpression lines. Responses to abiotic stresses, such as cold stress, are critical to plant growth and optimal production. Examination of Arabidopsis cytokinin response factors (CRFs) showed transcriptional induction after exposure to cold (4 °C). In particular, CRF4 was strongly induced in both root and shoot tissues. As CRF4 is one of several CRFs not transcriptionally regulated by cytokinin, we further investigated its response to cold. Peak CRF4 induction occurred 6 h post cold exposure, after which expression was maintained at moderately elevated levels during extended cold and subsequent treatment recovery. Examination of CRF4 mutant and overexpression lines under standard (non-cold) conditions revealed little difference from WT. One exception was a small, but significant increase in primary root growth of overexpression plants (CRF4OX). Under cold conditions, the only phenotype observed was a reduction in the rate of germination of CRF4OX seeds. The pattern of CRF4 expression along with the lack of strong phenotype at 4 °C led us to hypothesize that cold induction of CRF4 could play a role in short-term cold acclimation leading to increased freeze tolerance. Examination of CRF4OX and crf4 plants exposed to freezing temperatures revealed mutants lacking expression of CRF4 were more sensitive to freezing, while CRF4OXs with increased levels CRF4 levels were more tolerant. Altered transcript expression of CBF and COR15a cold signaling pathway genes in crf4 mutant and overexpression lines suggest that CRF4 may be potentially connected to this pathway. Overall this indicates that CRF4 plays an important role in both cold response and freezing stress.
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Proteínas de Arabidopsis/genética , Frío , Congelación , Regulación de la Expresión Génica de las Plantas , Factores de Transcripción/genética , Aclimatación/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Mutación , Raíces de Plantas/genética , Brotes de la Planta/genética , Plantas Modificadas Genéticamente , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transducción de Señal/genéticaRESUMEN
Reniform nematode is a semi-endoparasitic nematode species causing significant yield loss in numerous crops, including cotton (Gossypium hirsutum L.). An RNA-sequencing analysis was conducted to measure transcript abundance in reniform nematode susceptible (DP90 & SG747), resistant (BARBREN-713), and hypersensitive (LONREN-1) genotypes of cotton (Gossypium hirsutum L.) with and without reniform nematode infestation. Over 90 million trimmed high quality reads were assembled into 84,711 and 80, 353 transcripts using the G. arboreum and the G. raimondii genomes as references. Many transcripts were significantly differentially expressed between the three different genotypes both prior to and during nematode pathogenesis, including transcripts corresponding to the gene ontology categories of cell wall, hormone metabolism and signaling, redox reactions, secondary metabolism, transcriptional regulation, stress responses, and signaling. Further analysis revealed that a number of these differentially expressed transcripts mapped to the G. raimondii and/or the G. arboreum genomes within 1 megabase of quantitative trait loci that had previously been linked to reniform nematode resistance. Several resistance genes encoding proteins known to be strongly linked to pathogen perception and resistance, including LRR-like and NBS-LRR domain-containing proteins, were among the differentially expressed transcripts mapping near these quantitative trait loci. Further investigation is required to confirm a role for these transcripts in reniform nematode susceptibility, hypersensitivity, and/or resistance. This study presents the first systemic investigation of reniform nematode resistance-associated genes using different genotypes of cotton. The candidate reniform nematode resistance-associated genes identified in this study can serve as the basis for further functional analysis and aid in further development of reniform a nematode resistant cotton germplasm.