RESUMEN
In a consequence of global warming, grapevine trunk diseases (GTDs) have become a pertinent problem to viticulture, because endophytic fungi can turn necrotrophic upon host stress killing the plant. In Neofusicoccum parvum Bt-67, plant-derived ferulic acid makes the fungus release Fusicoccin aglycone triggering plant cell death. Now, we show that the absence of ferulic acid lets the fungus secrete 4-hydroxyphenylacetic acid (4-HPA), mimicking the effect of auxins on grapevine defence and facilitating fungal spread. Using Vitis suspension cells, we dissected the mode of action of 4-HPA during defence triggered by the bacterial cell-death elicitor, harpin. Early responses (cytoskeletal remodelling and calcium influx) are inhibited, as well as the expression of Stilbene Synthase 27 and phytoalexin accumulation. In contrast to other auxins, 4-HPA quells transcripts for the auxin conjugating GRETCHEN HAGEN 3. We suggest that 4-HPA is a key component of the endophytic phase of N. parvum Bt-67 preventing host cell death. Therefore, our study paves the way to understand how GTDs regulate their latent phase for successful colonisation, before turning necrotrophic and killing the vines.
RESUMEN
Plant responses to salt exposure involve large reconfigurations of hormonal pathways that orchestrate physiological changes towards tolerance. Jasmonate (JA) hormones are essential to withstand biotic and abiotic assaults, but their roles in salt tolerance remain unclear. Here we describe the dynamics of JA metabolism and signaling in root and leaf tissue of rice, a plant species that is highly exposed and sensitive to salt. Roots activate the JA pathway in an early pulse, while the second leaf displays a biphasic JA response with peaks at 1 h and 3 d post-exposure. Based on higher salt tolerance of a rice JA-deficient mutant (aoc), we examined, through kinetic transcriptome and physiological analysis, the salt-triggered processes that are under JA control. Profound genotype-differential features emerged that could underlie the observed phenotypes. Abscisic acid (ABA) content and ABA-dependent water deprivation responses were impaired in aoc shoots. Moreover, aoc accumulated more Na+ in roots, and less in leaves, with reduced ion translocation correlating with root derepression of the HAK4 Na+ transporter gene. Distinct reactive oxygen species scavengers were also stronger in aoc leaves, along with reduced senescence and chlorophyll catabolism markers. Collectively, our results identify contrasted contributions of JA signaling to different sectors of the salt stress response in rice.
Asunto(s)
Oryza , Tolerancia a la Sal , Oryza/metabolismo , Estrés Salino , Oxilipinas/metabolismo , Ácido Abscísico/metabolismo , Ciclopentanos/metabolismo , Estrés Fisiológico , Regulación de la Expresión Génica de las Plantas , Raíces de Plantas/metabolismoRESUMEN
BACKGROUND: The annual yield losses caused by the Rice Blast Fungus, Magnaporthe oryzae, range to the equivalent for feeding 60 million people. To ward off infection by this fungus, rice has evolved a generic basal immunity (so called compatible interaction), which acts in concert with strain-specific defence (so-called incompatible interaction). The plant-defence hormone jasmonic acid (JA) promotes the resistance to M. oryzae, but the underlying mechanisms remain elusive. To get more insight into this open question, we employ the JA-deficient mutants, cpm2 and hebiba, and dissect the JA-dependent defence signalling in rice for both, compatible and incompatible interactions. RESULTS: We observe that both JA-deficient mutants are more susceptible to M. oryzae as compared to their wild-type background, which holds true for both types of interactions as verified by cytological staining. Secondly, we observe that transcripts for JA biosynthesis (OsAOS2 and OsOPR7), JA signalling (OsJAZ8, OsJAZ9, OsJAZ11 and OsJAZ13), JA-dependent phytoalexin synthesis (OsNOMT), and JA-regulated defence-related genes, such as OsBBTI2 and OsPR1a, accumulate after fungal infection in a pattern that correlates with the amplitude of resistance. Thirdly, induction of defence transcripts is weaker during compatible interaction. CONCLUSION: The study demonstrates the pivotal role of JA in basal immunity of rice in the resistance to M. oryzae in both, compatible and incompatible interactions.
Asunto(s)
Ascomicetos , Magnaporthe , Oryza , Ciclopentanos/farmacología , Oryza/microbiología , Enfermedades de las Plantas/microbiología , Resistencia a la Enfermedad/genética , Regulación de la Expresión Génica de las PlantasRESUMEN
C-repeat binding factors (CBFs) are crucial transcriptional activators in plant responses to low temperature. CBF4 differs in its slower, but more persistent regulation and its role in cold acclimation. Cold acclimation has accentuated relevance for tolerance to late spring frosts as they have become progressively more common, as a consequence of blurred seasonality in the context of global climate change. In the current study, we explore the functions of CBF4 from grapevine, VvCBF4. Overexpression of VvCBF4 fused to GFP in tobacco BY-2 cells confers cold tolerance. Furthermore, this protein shuttles from the cytoplasm to the nucleus in response to cold stress, associated with an accumulation of transcripts for other CBFs and the cold responsive gene, ERD10d. This response differs for chilling as compared to freezing and is regulated differently by upstream signalling involving oxidative burst, proteasome activity and jasmonate synthesis. The difference between chilling and freezing is also seen in the regulation of the CBF4 transcript in leaves from different grapevines differing in their cold tolerance. Therefore, we propose the quality of cold stress is transduced by different upstream signals regulating nuclear import and, thus, the transcriptional activation of grapevine CBF4.
Asunto(s)
Regulación de la Expresión Génica de las Plantas , Complejo de la Endopetidasa Proteasomal , Aclimatación/genética , Transporte Activo de Núcleo Celular , Frío , Congelación , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Complejo de la Endopetidasa Proteasomal/metabolismoRESUMEN
The transition to terrestrial plants was accompanied by a progressive loss of microtubule minus-end-directed dynein motors. Instead, the minus-end-directed class-XIV kinesins expanded considerably, likely related to novel functions. One of these motors, OsDLK (Dual Localisation Kinesin from rice), decorates cortical microtubules but moves into the nucleus in response to cold stress. This analysis of loss-of-function mutants in rice indicates that OsDLK participates in cell elongation during development. Since OsDLK harbours both a nuclear localisation signal and a putative leucin zipper, we asked whether the cold-induced import of OsDLK into the nucleus might correlate with specific DNA binding. Conducting a DPI-ELISA screen with recombinant OsDLKT (lacking the motor domain), we identified the Opaque2 motif as the most promising candidate. This motif is present in the promoter of NtAvr9/Cf9, the tobacco homologue of Cold-Box Factor 4, a transcription factor involved in cold adaptation. A comparative study revealed that the cold-induced accumulation of NtAvr9/Cfp9 was specifically quelled in transgenic BY-2 cells overexpressing OsDLK-GFP. These findings are discussed as a working model, where, in response to cold stress, OsDLK partitions from cortical microtubules at the plasma membrane into the nucleus and specifically modulates the expression of genes involved in cold adaptation.
Asunto(s)
Cinesinas , Oryza , Núcleo Celular/metabolismo , Dineínas/metabolismo , Cinesinas/genética , Microtúbulos/metabolismo , Oryza/genética , Oryza/metabolismoRESUMEN
Grapevine trunk diseases have devastating consequences on vineyards worldwide. European wild grapevines (Vitis vinifera subs. sylvestris) from the last viable population in Germany along the Rhine river showed variable degrees of resistance against Neofusicoccum parvum (strain Bt-67), a fungus associated with Botryosphaeriaceae-related dieback. Representative genotypes from different subclades of this population were mapped with respect to their ability to induce wood necrosis, as well as their defence responses in a controlled inoculation system. The difference in colonization patterns could be confirmed by cryo-scanning electron microscopy, while there was no relationship between vessel diameter and infection success. Resistant lines accumulated more stilbenes, that were in addition significantly partitioned to nonglycosylated viniferin trimers. By contrast, the susceptible genotypes accumulated less stilbenes with a significantly higher proportion of glycosylated piceid. We suggest a model in which in the resistant genotypes phenylpropanoid metabolism is channelled rapidly and specifically to the bioactive stilbenes. Our study specifies a resistant chemotype against grapevines trunk diseases and paves a way to breed for resistance against grapevine Botryosphaeriaceae-related dieback.
Asunto(s)
Estilbenos , Vitis , Ascomicetos , Alemania , Fitomejoramiento , Enfermedades de las Plantas , Estilbenos/farmacología , Vitis/genéticaRESUMEN
The detyrosination/retyrosination cycle is the most common post-translational modification of α-tubulin. Removal of the conserved C-terminal tyrosine of α-tubulin by a still elusive tubulin tyrosine carboxypeptidase, and religation of this tyrosine by a tubulin tyrosine ligase (TTL), are probably common to all eukaryotes. Interestingly, for plants, the only candidates qualifying as potential TTL homologs are the tubulin tyrosine ligase-like 12 proteins. To get insight into the biological functions of these potential TTL homologs, we cloned the rice TTL-like 12 protein (OsTTLL12) and generated overexpression OsTTLL12-RFP lines in both rice and tobacco BY-2 cells. We found, unexpectedly, that overexpression of this OsTTLL12-RFP increased the relative abundance of detyrosinated α-tubulin in both coleoptile and seminal root, correlated with more stable microtubules. This was independent of the respective orientation of cortical microtubule, and followed by correspondingly changing growth of coleoptiles and seminal roots. A perturbed organization of phragmoplast microtubules and disoriented cell walls were further characteristics of this phenotype. Thus, the elevated tubulin detyrosination in consequence of OsTTLL12 overexpression affects structural and dynamic features of microtubules, followed by changes in the axiality of cell plate deposition and, consequently, plant growth.
Asunto(s)
Microtúbulos/metabolismo , Nicotiana/metabolismo , Oryza/metabolismo , Tubulina (Proteína)/metabolismo , Carboxipeptidasas/genética , Carboxipeptidasas/metabolismo , Oryza/genética , Péptido Sintasas/genética , Péptido Sintasas/metabolismo , Tubulina (Proteína)/genéticaRESUMEN
Grapevine trunk diseases (GTDs) are progressively affecting vineyard longevity and productivity worldwide. To be able to understand and combat these diseases, we need a different concept of the signals exchanged between the grapevine and fungi than the well-studied pathogen-associated molecular pattern and effector concepts. We screened extracts from fungi associated with GTDs for their association with basal defence responses in suspension cells of grapevine. By activity-guided fractionation of the two selected extracts, O-methylmellein was identified as a candidate modulator of grapevine immunity. O-Methylmellein could not induce immune responses by itself (i.e. does not act as an elicitor), but could amplify some of the defence responses triggered by the bacterial elicitor flg22, such as the induction level of defence genes and actin remodelling. These findings show that Eutypa lata, exemplarily selected as an endophytic fungus linked with GTDs, can secrete compounds that act as amplifiers of basal immunity. Thus, in addition to elicitors that can trigger basal immunity, and effectors that down-modulate antibacterial basal immunity, once it had been activated, E. lata seems to secrete a third type of chemical signal that amplifies basal immunity and may play a role in the context of consortia of mutually competing microorganisms.
Asunto(s)
Vitis , Ascomicetos , Hongos , Enfermedades de las Plantas , Inmunidad de la PlantaRESUMEN
BACKGROUND: Productivity of important crop rice is greatly affected by salinity. The plant hormone jasmonate plays a vital role in salt stress adaptation, but also evokes detrimental side effects if not timely shut down again. As novel strategy to avoid such side effects, OsJAZ8, a negative regulator of jasmonate signalling, is expressed under control of the salt-inducible promoter of the transcription factor ZOS3-11, to obtain a transient jasmonate signature in response to salt stress. To modulate the time course of jasmonate signalling, either a full-length or a dominant negative C-terminally truncated version of OsJAZ8 driven by the ZOS3-11 promoter were expressed in a stable manner either in tobacco BY-2 cells, or in japonica rice. RESULTS: The transgenic tobacco cells showed reduced mortality and efficient cycling under salt stress adaptation. This was accompanied by reduced sensitivity to Methyl jasmonate and increased responsiveness to auxin. In the case of transgenic rice, the steady-state levels of OsJAZ8 transcripts were more efficiently induced under salt stress compared to the wild type, this induction was more pronounced in the dominant-negative OsJAZ8 variant. CONCLUSIONS: The result concluded that, more efficient activation of OsJAZ8 was accompanied by improved salt tolerance of the transgenic seedlings and demonstrates the impact of temporal signatures of jasmonate signalling for stress tolerance.
Asunto(s)
Proteínas Co-Represoras/metabolismo , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Plantas Tolerantes a la Sal/metabolismo , Proteínas Co-Represoras/genética , Proteínas Co-Represoras/fisiología , Ciclopentanos/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Oryza/genética , Oxilipinas/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/fisiología , Plantas Modificadas Genéticamente/genética , Estrés Salino , Plantas Tolerantes a la Sal/genética , Transducción de Señal , Nicotiana/genéticaRESUMEN
Fatty acid hydroperoxides can generate short-chained volatile aldehydes that may participate in plant defence. A grapevine hydroperoxide lyase (VvHPL1) clustering to the CYP74B class was functionally characterized with respect to a role in defence. In grapevine leaves, transcripts of this gene accumulated rapidly to high abundance in response to wounding. Cellular functions of VvHPL1 were investigated upon heterologous expression in tobacco BY-2 cells. A C-terminal green fluorescent protein (GFP) fusion of VvHPL1 was located in plastids. The overexpression lines were found to respond to salinity stress or the bacterial elicitor harpin by increasing cell death. This signal-dependent mortality response was mitigated either by addition of exogenous jasmonic acid or by treatment with diphenyleneiodonium (DPI), an inhibitor of NADPH oxidases. By feeding different substrates to recombinantly expressed enzyme, VvHPL1 could also be functionally classified as true 13-HPL. The cognate products generated by this 13-HPL were cis-3-hexenal and trans-2-hexenal. Using a GFP-tagged actin marker line, one of these isomeric products, cis-3-hexenal, was found specifically to elicit a rapid disintegration of actin filaments. This response was not only observed in the heterologous system (tobacco BY-2), but also in a grapevine cell strain expressing this marker, as well as in leaf discs from an actin marker grape used as a homologous system. These results are discussed in the context of a role for VvHPL1 in a lipoxygenase-dependent signalling pathway triggering cell death-related defence that bifurcates from jasmonate-dependent basal immunity.
Asunto(s)
Actinas/metabolismo , Aldehído-Liasas/genética , Muerte Celular/fisiología , Sistema Enzimático del Citocromo P-450/genética , Proteínas de Plantas/genética , Vitis/fisiología , Compuestos Orgánicos Volátiles/metabolismo , Aldehído-Liasas/metabolismo , Sistema Enzimático del Citocromo P-450/metabolismo , Hojas de la Planta/metabolismo , Proteínas de Plantas/metabolismo , Vitis/genéticaRESUMEN
Cytoplasmic male sterile (CMS) rice has been widely used for hybrid rice seed production in China. However, CMS rice suffers from undesirable flowering habits including scattered floret opening time (FOT), which causes different FOTs among parental rice plants and greatly reduces hybrid rice seed production. Little is known about the mechanism of scattered FOT in CMS rice. Our results demonstrate that scattered FOT in CMS rice Zhenshan 97A (ZS97A) resulted from the lack of a driving force to open florets, which was directly caused by retarded lodicule expansion. Our results indicate that retarded lodicule expansion in ZS97A was caused by reduced water accumulation due to retarded accumulation of osmotic regulation substances (ORSs). Further, the retardation in accumulation of ORSs and water were caused by jasmonic acid (JA) deficiency, resulting from down-regulation of OsAOC expression. Applying JA restored scattered FOT in ZS97A by promoting ORS and water accumulation, and inducing the expansion of the lodicules. Taken together, JA deficiency inhibited lodicule expansion by retarding the accumulation of ORSs and water, leading to scattered FOT in CMS rice ZS97A.
Asunto(s)
Ciclopentanos/metabolismo , Flores/fisiología , Oryza/metabolismo , Oxilipinas/metabolismo , Quimera , Ciclopentanos/farmacología , Flores/genética , Regulación de la Expresión Génica de las Plantas , Oryza/efectos de los fármacos , Oryza/fisiología , Oxilipinas/farmacología , Infertilidad Vegetal , Agua/metabolismoRESUMEN
Induced defenses play a key role in plant resistance against leaf feeders. However, very little is known about the signals that are involved in defending plants against root feeders and how they are influenced by abiotic factors. We investigated these aspects for the interaction between rice (Oryza sativa) and two root-feeding insects: the generalist cucumber beetle (Diabrotica balteata) and the more specialized rice water weevil (Lissorhoptrus oryzophilus). Rice plants responded to root attack by increasing the production of jasmonic acid (JA) and abscisic acid, whereas in contrast to in herbivore-attacked leaves, salicylic acid and ethylene levels remained unchanged. The JA response was decoupled from flooding and remained constant over different soil moisture levels. Exogenous application of methyl JA to the roots markedly decreased the performance of both root herbivores, whereas abscisic acid and the ethylene precursor 1-aminocyclopropane-1-carboxylic acid did not have any effect. JA-deficient antisense 13-lipoxygenase (asLOX) and mutant allene oxide cyclase hebiba plants lost more root biomass under attack from both root herbivores. Surprisingly, herbivore weight gain was decreased markedly in asLOX but not hebiba mutant plants, despite the higher root biomass removal. This effect was correlated with a herbivore-induced reduction of sucrose pools in asLOX roots. Taken together, our experiments show that jasmonates are induced signals that protect rice roots from herbivores under varying abiotic conditions and that boosting jasmonate responses can strongly enhance rice resistance against root pests. Furthermore, we show that a rice 13-lipoxygenase regulates root primary metabolites and specifically improves root herbivore growth.
Asunto(s)
Ciclopentanos/metabolismo , Herbivoria , Oryza/metabolismo , Oryza/parasitología , Oxilipinas/metabolismo , Raíces de Plantas/parasitología , Transducción de Señal , Animales , Biomasa , Escarabajos/fisiología , Resistencia a la Enfermedad , Regulación de la Expresión Génica de las Plantas , Silenciador del Gen , Proteínas de Plantas/metabolismo , Raíces de Plantas/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Sacarosa/metabolismo , AguaRESUMEN
Stilbene synthase is a key enzyme for the production of the phytoalexin resveratrol. Some clones of Vitis sylvestris, a wild European grapevine species which is almost extinct, have been shown to accumulate more resveratrol in response to different forms of stress. In the current study, we asked whether the induction of stilbene synthase transcripts in Hoe29, one of the V. sylvestris clones with elevated stilbene inducibility, might result from the elevated induction of the transcription factor MYB14. The MYB14 promoter of Hoe29 and of Ke83 (a second stilbene-inducible genotype) harboured distinct regions and were applied to a promoter-reporter system. We show that stilbene synthase inducibility correlates with differences in the induction of MYB14 transcripts for these two genotypes. Both alleles were induced by UV in a promoter-reporter assay, but only the MYB14 promoter from Hoe29 was induced by flg22, consistent with the stilbene synthase expression of the donor genotypes, where both respond to UV but only Hoe29 is responsive to Plasmopara viticola during defence. We mapped upstream signals and found that a RboH-dependent oxidative burst, calcium influx, a MAPK cascade, and jasmonate activated the MYB14 promoter, whereas salicylic acid was ineffective. Our data suggest that the Hoe29 allele of the MYB14 promoter has potential as a candidate target for resistance breeding.
Asunto(s)
Alelos , Inmunidad de la Planta/genética , Proteínas de Plantas/genética , Factores de Transcripción/genética , Vitis/genética , Vitis/inmunología , Genotipo , Modelos Biológicos , Compuestos Onio/farmacología , Oomicetos/fisiología , Oomicetos/efectos de la radiación , Inmunidad de la Planta/efectos de los fármacos , Proteínas de Plantas/metabolismo , Regiones Promotoras Genéticas/genética , Factores de Transcripción/metabolismo , Rayos Ultravioleta , Vitis/microbiología , Vitis/efectos de la radiaciónRESUMEN
Salinity stress represents a global constraint for rice, the most important staple food worldwide. Therefore the role of the central stress signal jasmonate for the salt response was analysed in rice comparing the responses to salt stress for two jasmonic acid (JA) biosynthesis rice mutants (cpm2 and hebiba) impaired in the function of ALLENE OXIDE CYCLASE (AOC) and their wild type. The aoc mutants were less sensitive to salt stress. Interestingly, both mutants accumulated smaller amounts of Na(+) ions in their leaves, and showed better scavenging of reactive oxygen species (ROS) under salt stress. Leaves of the wild type and JA mutants accumulated similar levels of abscisic acid (ABA) under stress conditions, and the levels of JA and its amino acid conjugate, JA-isoleucine (JA-Ile), showed only subtle alterations in the wild type. In contrast, the wild type responded to salt stress by strong induction of the JA precursor 12-oxophytodienoic acid (OPDA), which was not observed in the mutants. Transcript levels of representative salinity-induced genes were induced less in the JA mutants. The absence of 12-OPDA in the mutants correlated not only with a generally increased ROS-scavenging activity, but also with the higher activity of specific enzymes in the antioxidative pathway, such as glutathione S-transferase, and fewer symptoms of damage as, for example, indicated by lower levels of malondialdehyde. The data are interpreted in a model where the absence of OPDA enhanced the antioxidative power in mutant leaves.
Asunto(s)
Oxidorreductasas Intramoleculares/genética , Oryza/enzimología , Reguladores del Crecimiento de las Plantas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Cloruro de Sodio/farmacología , Ácido Abscísico/metabolismo , Ciclopentanos/metabolismo , Ácidos Grasos Insaturados/metabolismo , Regulación de la Expresión Génica de las Plantas , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Oxidorreductasas Intramoleculares/metabolismo , Isoleucina/análogos & derivados , Isoleucina/metabolismo , Peroxidación de Lípido , Oryza/efectos de los fármacos , Oryza/genética , Oryza/fisiología , Oxilipinas/metabolismo , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/enzimología , Hojas de la Planta/genética , Hojas de la Planta/fisiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/enzimología , Raíces de Plantas/genética , Raíces de Plantas/fisiología , Plantones/efectos de los fármacos , Plantones/enzimología , Plantones/genética , Plantones/fisiología , Estrés FisiológicoRESUMEN
Two photomorphogenic mutants of rice, coleoptile photomorphogenesis 2 (cpm2) and hebiba, were found to be defective in the gene encoding allene oxide cyclase (OsAOC) by map-based cloning and complementation assays. Examination of the enzymatic activity of recombinant GST-OsAOC indicated that OsAOC is a functional enzyme that is involved in the biosynthesis of jasmonic acid and related compounds. The level of jasmonate was extremely low in both mutants, in agreement with the fact that rice has only one gene encoding allene oxide cyclase. Several flower-related mutant phenotypes were observed, including morphological abnormalities of the flower and early flowering. We used these mutants to investigate the function of jasmonate in the defence response to the blast fungus Magnaporthe oryzae. Inoculation assays with fungal spores revealed that both mutants are more susceptible than wild-type to an incompatible strain of M. oryzae, in such a way that hyphal growth was enhanced in mutant tissues. The level of jasmonate isoleucine, a bioactive form of jasmonate, increased in response to blast infection. Furthermore, blast-induced accumulation of phytoalexins, especially that of the flavonoid sakuranetin, was found to be severely impaired in cpm2 and hebiba. Together, the present study demonstrates that, in rice, jasmonate mediates the defence response against blast fungus.
Asunto(s)
Ciclopentanos/metabolismo , Oxidorreductasas Intramoleculares/metabolismo , Magnaporthe/patogenicidad , Oryza/enzimología , Oryza/metabolismo , Oxilipinas/metabolismo , Oxidorreductasas Intramoleculares/genética , Oryza/genética , Oryza/microbiologíaRESUMEN
The bioactive form of jasmonate is the conjugate of the amino acid isoleucine (Ile) with jasmonic acid (JA), which is biosynthesized in a reaction catalysed by the GH3 enzyme JASMONATE RESISTANT 1 (JAR1). We examined the biochemical properties of OsJAR1 and its involvement in photomorphogenesis of rice (Oryza sativa). OsJAR1 has a similar substrate specificities as its orthologue in Arabidopsis. However, osjar1 loss-of-function mutants did not show as severe coleoptile phenotypes as the JA-deficient mutants coleoptile photomorphogenesis 2 (cpm2) and hebiba, which develop long coleoptiles in all light qualities we examined. Analysis of hormonal contents in the young seedling stage revealed that osjar1 mutants are still able to synthesize JA-Ile conjugate in response to blue light, suggesting that a redundantly active enzyme can conjugate JA and Ile in rice seedlings. A good candidate for this enzyme is OsJAR2, which was found to be able to catalyse the conjugation of JA with Ile as well as with some additional amino acids. In contrast, if plants in the vegetative stage were mechanically wounded, the content of JA-Ile was severely reduced in osjar1, demonstrating that OsJAR1 is the most important JA-Ile conjugating enzyme in the wounding response during the vegetative stage.
Asunto(s)
Ciclopentanos/metabolismo , Isoleucina/análogos & derivados , Luz , Oryza/metabolismo , Oryza/efectos de la radiación , Proteínas de Plantas/metabolismo , Transducción de Señal/efectos de la radiación , Vías Biosintéticas/genética , Vías Biosintéticas/efectos de la radiación , Etiolado/efectos de la radiación , Regulación de la Expresión Génica de las Plantas/efectos de la radiación , Isoleucina/metabolismo , Morfogénesis/efectos de la radiación , Mutación/genética , Oryza/genética , Oxilipinas/metabolismo , Fenotipo , Proteínas de Plantas/genética , Plantones/metabolismo , Plantones/efectos de la radiación , Transducción de Señal/genética , Especificidad por Sustrato/efectos de la radiación , Transcripción Genética/efectos de la radiaciónRESUMEN
Defence to pathogens must be specific. In the past, we have dissected early signalling deployed by bacterial elicitors in a grapevine cell system. In the current work, we asked, how defence of fungi differs. Fungal diseases of grapevine pose great challenges for global viticulture and require massive plant protection measures. Plant cells are able to sense chitin, a central component of fungal cell walls and respond by activation of basal defence. We, therefore mapped early defence responses evoked by chitosan, a chitin fragment able to bind to chitin receptors. We found an activation of calcium influx, monitored by extracellular alkalinisation due to a co-transport of protons, remodelling of actin (but not of microtubules), and the activation of transcripts for phytoalexin synthesis, jasmonate-signalling, salicylate signalling, and chitinase. Interestingly, Gadolinium, an inhibitor of calcium influx, can inhibit extracellular alkalinisation in response to chitosan, while the induction of the phytoalexin synthesis transcripts was specifically promoted. In contrast, both DMSO and benzyl alcohol, compounds known to modulate membrane fluidity, partially inhibited the transcript responses to chitosan. We discuss these data with a model, where chitosan deploys signalling culminating in activation of defence related transcripts, but at the same time activates calcium influx that negatively feeds back on the same signal chain, which might be a mechanism to achieve a temporal signature that is rapid, but transient.
Asunto(s)
Quitosano , Vitis , Calcio/metabolismo , Actinas/metabolismo , Vitis/metabolismo , Quitosano/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMEN
COI1-mediated perception of jasmonate is critical for plant development and responses to environmental stresses. Monocots such as rice have two groups of COI genes due to gene duplication: OsCOI1a and OsCOI1b that are functionally equivalent to the dicotyledons COI1 and OsCOI2 whose function remains unclear. In order to assess the function of OsCOI2 and its functional redundancy with COI1 genes, we developed a series of rice mutants in the 3 genes OsCOI1a, OsCOI1b and OsCOI2 by CRISPR Cas9-mediated editing and characterized their phenotype and responses to jasmonate. Characterization of OsCOI2 uncovered its important roles in root, leaf and flower development. In particular, we show that crown root growth inhibition by jasmonate relies on OsCOI2 and not on OsCOI1a nor on OsCOI1b, revealing a major function for the non-canonical OsCOI2 in jasmonate-dependent control of rice root growth. Collectively, these results point to a specialized function of OsCOI2 in the regulation of plant development in rice and indicate that sub-functionalisation of jasmonate receptors has occurred in the monocot phylum.
Asunto(s)
Oryza , Oryza/genética , Ciclopentanos , Duplicación de Gen , Inhibición PsicológicaRESUMEN
Salt stress is a major constraint for many crop plants, such as the moderately salt-sensitive economically important fruit crop grapevine. Plants have evolved different strategies for protection against salinity and drought. Jasmonate signalling is a central element of both biotic and abiotic stress responses. To discriminate stress quality, there must be cross-talk with parallel signal chains. Using two grapevine cell lines differing in salt tolerance, the response of jasmonate ZIM/tify-domain (JAZ/TIFY) proteins (negative regulators of jasmonate signalling), a marker for salt adaptation Na(+)/H(+) EXCHANGER (NHX1), and markers for biotic defence STILBENE SYNTHASE (StSy) and RESVERATROL SYNTHASE (RS) were analysed. It is shown that salt stress signalling shares several events with biotic defence including activity of a gadolinium-sensitive calcium influx channel (monitored by apoplastic alkalinization) and transient induction of JAZ/TIFY transcripts. Exogenous jasmonate can rescue growth in the salt-sensitive cell line. Suppression of jasmonate signalling by phenidone or aspirin blocks the induction of JAZ/TIFY transcripts. The rapid induction of RS and StSy characteristic for biotic defence in grapevine is strongly delayed in response to salt stress. In the salt-tolerant line, NHX1 is induced and the formation of reactive oxygen species, monitored as stress markers in the sensitive cell line, is suppressed. The data are discussed in terms of a model where salt stress signalling acts as a default pathway whose readout is modulated by a parallel signal chain triggered by biotic factors downstream of jasmonate signalling.
Asunto(s)
Ciclopentanos/farmacología , Oxilipinas/farmacología , Reguladores del Crecimiento de las Plantas/farmacología , Proteínas de Plantas/metabolismo , Transducción de Señal/efectos de los fármacos , Cloruro de Sodio/farmacología , Vitis/fisiología , Secuencia de Aminoácidos , Aspirina/farmacología , Secuencia de Bases , Calcio/metabolismo , Sequías , Gadolinio/farmacología , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Concentración de Iones de Hidrógeno , Modelos Biológicos , Datos de Secuencia Molecular , Filogenia , Hojas de la Planta , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente , Pirazoles/farmacología , Especies Reactivas de Oxígeno/metabolismo , Tolerancia a la Sal , Análisis de Secuencia de ADN , Estrés Fisiológico , Vitis/efectos de los fármacos , Vitis/crecimiento & desarrolloRESUMEN
Global warming is predicted to change the growth conditions for plants and crops in regions at high latitudes (>60° N), including the Arctic. This will be accompanied by alterations in the composition of natural plant and pest communities, as herbivorous arthropods will invade these regions as well. Interactions between previously non-overlapping species may occur and cause new challenges to herbivore attack. However, plants growing at high latitudes experience less herbivory compared to plants grown at lower latitudes. We hypothesize that this finding is due to a gradient of constitutive chemical defense towards the Northern regions. We further hypothesize that higher level of defensive compounds is mediated by higher level of the defense-related phytohormone jasmonate. Because its biosynthesis is light dependent, Arctic summer day light conditions can promote jasmonate accumulation and, hence, downstream physiological responses. A pilot study with bilberry (Vaccinium myrtillus) plants grown under different light regimes supports the hypothesis.