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1.
Mol Cell ; 70(1): 136-149.e7, 2018 04 05.
Artigo em Inglês | MEDLINE | ID: mdl-29625034

RESUMO

Insect herbivory causes severe damage to plants and threatens the world's food production. During evolutionary adaptation, plants have evolved sophisticated mechanisms to rapidly accumulate a key defense hormone, jasmonate (JA), that triggers plant defense against herbivory. However, little is known about how plants initially activate JA biosynthesis at encounter with herbivory. Here, we uncover that a novel JAV1-JAZ8-WRKY51 (JJW) complex controls JA biosynthesis to defend against insect attack. In healthy plants, the JJW complex represses JA biosynthesis to restrain JA at a low basal level to ensure proper plant growth. When plants are injured by insect attack, injury rapidly triggers calcium influxes to activate calmodulin-dependent phosphorylation of JAV1, which disintegrates JJW complex and activates JA biosynthesis, giving rise to the rapid burst of JA for plant defense. Our findings offer new insights into the highly sophisticated defense systems evolved by plants to defend against herbivory.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Cálcio/metabolismo , Calmodulina/metabolismo , Proteínas Correpressoras/metabolismo , Ciclopentanos/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Oxilipinas/metabolismo , Folhas de Planta/enzimologia , Plantas Geneticamente Modificadas/enzimologia , Spodoptera/fisiologia , Fatores de Transcrição/metabolismo , Animais , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Sinalização do Cálcio , Calmodulina/genética , Proteínas Correpressoras/genética , Regulação da Expressão Gênica de Plantas , Herbivoria , Peptídeos e Proteínas de Sinalização Intracelular/genética , Complexos Multiproteicos , Fosforilação , Folhas de Planta/genética , Plantas Geneticamente Modificadas/genética , Fatores de Transcrição/genética
2.
Plant J ; 118(5): 1619-1634, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38456566

RESUMO

The plant cuticle is composed of cuticular wax and cutin polymers and plays an essential role in plant tolerance to diverse abiotic and biotic stresses. Several stresses, including water deficit and salinity, regulate the synthesis of cuticular wax and cutin monomers. However, the effect of wounding on wax and cutin monomer production and the associated molecular mechanisms remain unclear. In this study, we determined that the accumulation of wax and cutin monomers in Arabidopsis leaves is positively regulated by wounding primarily through the jasmonic acid (JA) signaling pathway. Moreover, we observed that a wound- and JA-responsive gene (CYP96A4) encoding an ER-localized cytochrome P450 enzyme was highly expressed in leaves. Further analyses indicated that wound-induced wax and cutin monomer production was severely inhibited in the cyp96a4 mutant. Furthermore, CYP96A4 interacted with CER1 and CER3, the core enzymes in the alkane-forming pathway associated with wax biosynthesis, and modulated CER3 activity to influence aldehyde production in wax synthesis. In addition, transcripts of MYC2 and JAZ1, key genes in JA signaling pathway, were significantly reduced in cyp96a4 mutant. Collectively, these findings demonstrate that CYP96A4 functions as a cofactor of the alkane synthesis complex or participates in JA signaling pathway that contributes to cuticular wax biosynthesis and cutin monomer formation in response to wounding.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Ciclopentanos , Sistema Enzimático do Citocromo P-450 , Regulação da Expressão Gênica de Plantas , Lipídeos de Membrana , Oxilipinas , Folhas de Planta , Ceras , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/enzimologia , Ceras/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Sistema Enzimático do Citocromo P-450/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Oxilipinas/metabolismo , Ciclopentanos/metabolismo , Lipídeos de Membrana/metabolismo , Folhas de Planta/metabolismo , Folhas de Planta/genética , Transdução de Sinais , Epiderme Vegetal/metabolismo , Epiderme Vegetal/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/genética , Carbono-Carbono Liases , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos
3.
Plant J ; 116(5): 1325-1341, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37596913

RESUMO

Sensing of environmental challenges, such as mechanical injury, by a single plant tissue results in the activation of systemic signaling, which attunes the plant's physiology and morphology for better survival and reproduction. As key signals, both calcium ions (Ca2+ ) and hydrogen peroxide (H2 O2 ) interplay with each other to mediate plant systemic signaling. However, the mechanisms underlying Ca2+ -H2 O2 crosstalk are not fully revealed. Our previous study showed that the interaction between glycolate oxidase and catalase, key enzymes of photorespiration, serves as a molecular switch (GC switch) to dynamically modulate photorespiratory H2 O2 fluctuations via metabolic channeling. In this study, we further demonstrate that local wounding induces a rapid shift of the GC switch to a more interactive state in systemic leaves, resulting in a sharp decrease in peroxisomal H2 O2 levels, in contrast to a simultaneous outburst of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase-derived apoplastic H2 O2 . Moreover, the systemic response of the two processes depends on the transmission of Ca2+ signaling, mediated by glutamate-receptor-like Ca2+ channels 3.3 and 3.6. Mechanistically, by direct binding and/or indirect mediation by some potential biochemical sensors, peroxisomal Ca2+ regulates the GC switch states in situ, leading to changes in H2 O2 levels. Our findings provide new insights into the functions of photorespiratory H2 O2 in plant systemic acclimation and an optimized systemic H2 O2 signaling via spatiotemporal interplay between the GC switch and NADPH oxidases.


Assuntos
Oxirredutases do Álcool , Plantas , Catalase/metabolismo , Plantas/metabolismo , Oxirredutases do Álcool/metabolismo , Receptores de Glutamato , Peróxido de Hidrogênio/metabolismo
4.
Plant Mol Biol ; 114(1): 6, 2024 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-38265739

RESUMO

tRNA-derived small RNAs (tsRNAs), a new category of regulatory small non-coding RNA existing in almost all branches of life, have recently attracted broad attention. Increasing evidence has shown that tsRNAs are not random degradation debris of tRNAs, but products cleaved by specific endoribonucleases, with versatile functions in response to various developmental and environmental cues. However, it is still unclear about the diversity, biogenesis and function of tsRNAs in plants. In this study, we comprehensively profiled 10-60 nts small RNAs in Arabidopsis thaliana leaf with or without wounding stress and identified four 16 nts tiny tRFs (tRNA-derived fragments) sharply increased after wounding, namely tRF5'Ala. Notably, genetic, biochemical and bioinformatic data indicated that RNS2, a member of class II RNase T2 enzymes, was the main endoribonuclease responsible for the biogenesis of tRF5'Ala. Moreover, tRF5'Ala was highly abundant and conserved in Arabidopsis and rice pollen. However, tRF5'Ala did not associate with AGO 1 in vivo or display any inhibitory effect on the translation of a luciferase mRNA in vitro. Altogether, our study highlights the discovery of a novel class of tiny tsRNAs drastically increased under wounding stress as well as their generation by RNS2, which provides a new insight into tsRNAs research in plants.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Ribonucleases , Biologia Computacional , RNA , RNA de Transferência , Proteínas de Arabidopsis/genética , Ribonucleases/genética
5.
Plant Cell Physiol ; 65(4): 660-670, 2024 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-38195149

RESUMO

In response to both biotic and abiotic stresses, vascular plants transmit long-distance Ca2+ and electrical signals from localized stress sites to distant tissues through their vasculature. Various models have been proposed for the mechanisms underlying the long-distance signaling, primarily centered around the presence of vascular bundles. We here demonstrate that the non-vascular liverwort Marchantia polymorpha possesses a mechanism for propagating Ca2+ waves and electrical signals in response to wounding. The propagation velocity of these signals was approximately 1-2 mm s-1, equivalent to that observed in vascular plants. Both Ca2+ waves and electrical signals were inhibited by La3+ as well as tetraethylammonium chloride, suggesting the crucial importance of both Ca2+ channel(s) and K+ channel(s) in wound-induced membrane depolarization as well as the subsequent long-distance signal propagation. Simultaneous recordings of Ca2+ and electrical signals indicated a tight coupling between the dynamics of these two signaling modalities. Furthermore, molecular genetic studies revealed that a GLUTAMATE RECEPTOR-LIKE (GLR) channel plays a central role in the propagation of both Ca2+ waves and electrical signals. Conversely, none of the three two-pore channels were implicated in either signal propagation. These findings shed light on the evolutionary conservation of rapid long-distance Ca2+ wave and electrical signal propagation involving GLRs in land plants, even in the absence of vascular tissue.


Assuntos
Sinalização do Cálcio , Cálcio , Marchantia , Marchantia/fisiologia , Marchantia/genética , Marchantia/metabolismo , Cálcio/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Lantânio/farmacologia , Receptores de Glutamato/metabolismo , Receptores de Glutamato/genética , Canais de Cálcio/metabolismo , Canais de Cálcio/genética , Tetraetilamônio/farmacologia , Canais de Potássio/metabolismo , Canais de Potássio/genética
6.
Planta ; 259(6): 152, 2024 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-38735012

RESUMO

MAIN CONCLUSION: Overexpression of Artemisia annua jasmonic acid carboxyl methyltransferase (AaJMT) leads to enhanced artemisinin content in Artemisia annua. Artemisinin-based combination therapies remain the sole deterrent against deadly disease malaria and Artemisia annua remains the only natural producer of artemisinin. In this study, the 1101 bp gene S-adenosyl-L-methionine (SAM): Artemisia annua jasmonic acid carboxyl methyltransferase (AaJMT), was characterised from A. annua, which converts jasmonic acid (JA) to methyl jasmonate (MeJA). From phylogenetic analysis, we confirmed that AaJMT shares a common ancestor with Arabidopsis thaliana, Eutrema japonica and has a close homology with JMT of Camellia sinensis. Further, the Clustal Omega depicted that the conserved motif I, motif III and motif SSSS (serine) required to bind SAM and JA, respectively, are present in AaJMT. The relative expression of AaJMT was induced by wounding, MeJA and salicylic acid (SA) treatments. Additionally, we found that the recombinant AaJMT protein catalyses the synthesis of MeJA from JA with a Km value of 37.16 µM. Moreover, site-directed mutagenesis of serine-151 in motif SSSS to tyrosine, asparagine-10 to threonine and glutamine-25 to histidine abolished the enzyme activity of AaJMT, thus indicating their determining role in JA substrate binding. The GC-MS analysis validated that mutant proteins of AaJMT were unable to convert JA into MeJA. Finally, the artemisinin biosynthetic and trichome developmental genes were upregulated in AaJMT overexpression transgenic lines, which in turn increased the artemisinin content.


Assuntos
Acetatos , Artemisia annua , Artemisininas , Ciclopentanos , Metiltransferases , Oxilipinas , Filogenia , Artemisia annua/genética , Artemisia annua/enzimologia , Artemisia annua/metabolismo , Ciclopentanos/metabolismo , Ciclopentanos/farmacologia , Artemisininas/metabolismo , Oxilipinas/metabolismo , Oxilipinas/farmacologia , Metiltransferases/metabolismo , Metiltransferases/genética , Acetatos/farmacologia , Acetatos/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Regulação da Expressão Gênica de Plantas , Ácido Salicílico/metabolismo
7.
Plant Cell Environ ; 47(6): 2206-2227, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38481105

RESUMO

Terpenoids are defense metabolites that are induced upon infection or wounding. However, their role in systemic-induced resistance (SIR) is not known. Here, we explored the role of terpenoids in this phenomenon at a very early stage in the interaction between Austrian pine and the tip blight and canker pathogen Diplodia pinea. We induced Austrian pine saplings by either wounding or inoculating the lower stems with D. pinea. The seedlings were then challenged after 12 h, 72 h, or 10 days with D. pinea on the stem 15 cm above the induction. Lesion lengths and terpenoids were quantified at both induction and challenge locations. Key terpenoids were assayed for antifungal activity in in vitro bioassays. SIR increased with time and was correlated with the inducibility of several compounds. α-Pinene and a cluster of ß-pinene, limonene, benzaldehyde, dodecanol, and n-dodecyl acrylate were positively correlated with SIR and were fungistatic in vitro, while other compounds were negatively correlated with SIR and appeared to serve as a carbon source for D. pinea. This study shows that, overall, terpenoids are involved in SIR in this system, but their role is nuanced, depending on the type of induction and time of incubation. We hypothesize that some, such as α-pinene, could serve in SIR signaling.


Assuntos
Ascomicetos , Pinus , Doenças das Plantas , Terpenos , Terpenos/metabolismo , Terpenos/farmacologia , Pinus/metabolismo , Pinus/microbiologia , Pinus/efeitos dos fármacos , Doenças das Plantas/microbiologia , Ascomicetos/fisiologia , Resistência à Doença , Plântula/metabolismo , Plântula/efeitos dos fármacos
8.
J Exp Bot ; 75(1): 334-349, 2024 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-37708289

RESUMO

The carnivorous plants in the order Caryophyllales co-opted jasmonate signalling from plant defence to botanical carnivory. However, carnivorous plants have at least 11 independent origins, and here we ask whether jasmonate signalling has been co-opted repeatedly in different evolutionary lineages. We experimentally wounded and fed the carnivorous plants Sarracenia purpurea (order Ericales), Cephalotus follicularis (order Oxalidales), Drosophyllum lusitanicum (order Caryophyllales), and measured electrical signals, phytohormone tissue level, and digestive enzymes activity. Coronatine was added exogenously to confirm the role of jasmonates in the induction of digestive process. Immunodetection of aspartic protease and proteomic analysis of digestive fluid was also performed. We found that prey capture induced accumulation of endogenous jasmonates only in D. lusitanicum, in accordance with increased enzyme activity after insect prey or coronatine application. In C. follicularis, the enzyme activity was constitutive while in S. purpurea was regulated by multiple factors. Several classes of digestive enzymes were identified in the digestive fluid of D. lusitanicum. Although carnivorous plants from different evolutionary lineages use the same digestive enzymes, the mechanism of their regulation differs. All investigated genera use jasmonates for their ancient role, defence, but jasmonate signalling has been co-opted for botanical carnivory only in some of them.


Assuntos
Planta Carnívora , Carnivoridade , Proteômica
9.
J Exp Bot ; 2024 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-38329465

RESUMO

During land plant evolution, the number of genes encoding for components of the thiol redox regulatory network and the generator systems of reactive oxygen species (ROS) expanded, tentatively indicating a role in tailored environmental acclimatization. This hypothesis has been validated experimentally and theoretically during the last decades. Recent developments of dynamic roGFP-based in vivo sensors for H2O2 and the redox potential of the glutathione pool paved the way for dissecting the kinetics changes in these decisive parameters in response to environmental stressors. The versatile cellular redox sensory and response regulatory system monitors alterations in redox metabolism and controls the activity of redox target proteins, and thereby affects most, if not all, cellular processes ranging from transcription to translation and metabolism. This review exemplarily describes the role of the redox- and ROS-dependent regulatory network in realising the proper response to diverse environmental stresses. The selected case studies concern different environmental challenges, namely excess excitation energy, the heavy metal cadmium and the metalloid arsenic, nitrogen, or phosphate shortage as examples for nutrient deficiency, wounding, and nematode infestation. Each challenge affects the redox regulatory and ROS network, but the present state of knowledge also pinpoints to pressing open questions concerning the translation of redox regulation to environmental acclimatization.

10.
J Chem Ecol ; 2024 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-38900391

RESUMO

Leaf-cutting ants (Formicidae; Atta spp., Acromyrmex spp.) cut off pieces of leaves and other plant tissue and feed it to their symbiotic fungi. As this foraging behavior poses an imminent threat to agriculture, leaf-cutting ants are considered as pests of huge ecologically and economically importance. Consequently, research on leaf-cutting ants focused on their foraging decisions and interactions with their cultivated symbiotic fungi, whereas their effect on the attacked plants, apart from the loss of plant tissue, remains largely unknown. In this study, we investigated the consequences of an attack by leaf-cutting ants and analyzed the plants' defense responses in comparison to chewing caterpillars and mechanical damage. We found that an attack by leaf-cutting ants induces the production of jasmonates in several host and non-host plant species (Arabidopsis thaliana, Vicia faba, Phaseolus lunatus, Tococa quadrialata). Additionally, we showed in the natural host plant lima bean (P. lunatus) that leaf-cutting ant damage immediately leads to the emission of typical herbivory-induced plant volatiles, including green leaf volatiles and terpenoids. Further data exploration revealed clear differences in the defense-related phytohormone profile in plant species of Neotropical and Eurasian origin. Taken together, we show that leaf-cutting ant infestation and their way of clipping the plants' tissues induce jasmonate and jasmonates-mediated responses and do not differ from those to mechanical injury or larval feeding.

11.
J Plant Res ; 2024 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-38536536

RESUMO

Pollination in Marantaceae is mediated by an explosive style movement. Before release, style tension is held by the hooded staminode. When a pollinator touches the trigger appendage of the hooded staminode the latter deforms and the style rapidly curls upwards. This movement has been interpreted as a turgor movement by some authors, but recent studies clearly indicate that setup, hold and release of tension are purely mechanical processes. However, in view of the high diversity of hooded staminodes, the question arises what keeps the tension in species with very thin staminodes. To test the holding mechanisms, we conducted mechanical and physico-chemical release experiments in four species with robust and four species with thin hooded staminodes in their natural tropical environment. We found almost the same response of all species to mechanical treatments, but species-specific reactions to different physico-chemical conditions. This indicates that style release follows the same mechanical principles in all species, but that the sensitivity of the explosive movement depends on material properties like tissue thickness and turgescence. As to the holding mechanisms, we found different degrees of floral synorganization. The hood of the hooded staminode formerly interpreted as an important holding structure does not play a noteworthy role. Instead, the basal plate of the hooded staminode antagonises the pressure of the style head against the holding point of the hooded staminode in species with robust hooded staminodes and well-developed basal plates. In some species with a thin hooded staminode, the latter is closely attached to the style and most likely stabilises tension by adhesive forces. In another species, a morphologically analogous structure adopts the function of the basal plate. We conclude that the holding mechanism of the style tension diversified during the evolution of Marantaceae whereas the release mechanism itself has been conserved throughout the family.

12.
Int J Mol Sci ; 25(11)2024 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-38892085

RESUMO

In wounded Arabidopsis thaliana leaves, four 13S-lipoxygenases (AtLOX2, AtLOX3, AtLOX4, AtLOX6) act in a hierarchical manner to contribute to the jasmonate burst. This leads to defense responses with LOX2 playing an important role in plant resistance against caterpillar herb-ivory. In this study, we sought to characterize the impact of AtLOX2 on wound-induced phytohormonal and transcriptional responses to foliar mechanical damage using wildtype (WT) and lox2 mutant plants. Compared with WT, the lox2 mutant had higher constitutive levels of the phytohormone salicylic acid (SA) and enhanced expression of SA-responsive genes. This suggests that AtLOX2 may be involved in the biosynthesis of jasmonates that are involved in the antagonism of SA biosynthesis. As expected, the jasmonate burst in response to wounding was dampened in lox2 plants. Generally, 1 h after wounding, genes linked to jasmonate biosynthesis, jasmonate signaling attenuation and abscisic acid-responsive genes, which are primarily involved in wound sealing and healing, were differentially regulated between WT and lox2 mutants. Twelve h after wounding, WT plants showed stronger expression of genes associated with plant protection against insect herbivory. This study highlights the dynamic nature of jasmonate-responsive gene expression and the contribution of AtLOX2 to this pathway and plant resistance against insects.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Ciclopentanos , Regulação da Expressão Gênica de Plantas , Lipoxigenase , Oxilipinas , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Lipoxigenase/metabolismo , Lipoxigenase/genética , Oxilipinas/metabolismo , Ciclopentanos/metabolismo , Transcriptoma , Ácido Salicílico/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Folhas de Planta/genética , Folhas de Planta/metabolismo , Mutação , Perfilação da Expressão Gênica , Lipoxigenases
13.
Int J Mol Sci ; 25(13)2024 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-39000244

RESUMO

Cutaneous wounds, both acute and chronic, begin with loss of the integrity, and thus barrier function, of the skin. Surgery and trauma produce acute wounds. There are 22 million surgical procedures per year in the United States alone, based on data from the American College of Surgeons, resulting in a prevalence of 6.67%. Acute traumatic wounds requiring repair total 8 million per year, 2.42% or 24.2 per 1000. The cost of wound care is increasing; it approached USD 100 billion for just Medicare in 2018. This burden for wound care will continue to rise with population aging, the increase in metabolic syndrome, and more elective surgeries. To heal a wound, an orchestrated, evolutionarily conserved, and complex series of events involving cellular and molecular agents at the local and systemic levels are necessary. The principal factors of this important function include elements from the neurological, cardiovascular, immune, nutritional, and endocrine systems. The objectives of this review are to provide clinicians engaged in wound care and basic science researchers interested in wound healing with an updated synopsis from recent publications. We also present data from our primary investigations, testing the hypothesis that cannabidiol can alter cutaneous wound healing and documenting their effects in wild type (C57/BL6) and db/db mice (Type 2 Diabetes Mellitus, T2DM). The focus is on the potential roles of the endocannabinoid system, cannabidiol, and the important immune-regulatory wound cytokine IL-33, a member of the IL-1 family, and connective tissue growth factor, CTGF, due to their roles in both normal and abnormal wound healing. We found an initial delay in the rate of wound closure in B6 mice with CBD, but this difference disappeared with time. CBD decreased IL-33 + cells in B6 by 70% while nearly increasing CTGF + cells in db/db mice by two folds from 18.6% to 38.8% (p < 0.05) using a dorsal wound model. We review the current literature on normal and abnormal wound healing, and document effects of CBD in B6 and db/db dorsal cutaneous wounds. CBD may have some beneficial effects in diabetic wounds. We applied 6-mm circular punch to create standard size full-thickness dorsal wounds in B6 and db/db mice. The experimental group received CBD while the control group got only vehicle. The outcome measures were rate of wound closure, wound cells expressing IL-33 and CTGF, and ILC profiles. In B6, the initial rate of wound closure was slower but there was no delay in the time to final closure, and cells expressing IL-33 was significantly reduced. CTGF + cells were higher in db/bd wounds treated with CBD. These data support the potential use of CBD to improve diabetic cutaneous wound healing.


Assuntos
Canabidiol , Pele , Cicatrização , Cicatrização/efeitos dos fármacos , Animais , Canabidiol/farmacologia , Canabidiol/uso terapêutico , Humanos , Pele/metabolismo , Pele/efeitos dos fármacos , Camundongos , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/tratamento farmacológico
14.
Plant Cell Physiol ; 2023 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-37927069

RESUMO

Wounding caused by insects or abiotic factors such as wind and hail can cause severe stress for plants. Intrigued by the observation that wounding induces expression of genes involved in surface wax synthesis in a jasmonoyl-isoleucine (JA-Ile)-independent manner, the role of wax biosynthesis and respective genes upon wounding was investigated. Wax, a lipid-based barrier, protects plants both from environmental threats as well as from an uncontrolled loss of water. Its biosynthesis is described to be regulated by abscisic acid (ABA), whereas the main wound-signal is the hormone JA-Ile. We show in this study, that genes coding for enzymes of surface wax synthesis are induced upon wounding in Arabidopsis thaliana leaves in a JA-Ile-independent but ABA-dependent manner. Furthermore, the ABA-dependent transcription factor MYB96 is a key regulator of wax biosynthesis upon wounding. On the metabolite level, wound-induced wax accumulation is strongly reduced in JA-Ile-deficient plants, but this induction is only slightly decreased in ABA-reduced plants. To further analyze the ABA-dependent wound response, we conducted wounding experiments in high humidity. They show that high humidity prevents the wound-induced wax accumulation in A. thaliana leaves. Together the data presented in this study show that wound-induced wax accumulation is JA-Ile-dependent on the metabolite level, but the expression of genes coding for enzymes of wax synthesis is regulated by ABA.

15.
BMC Biotechnol ; 23(1): 32, 2023 08 28.
Artigo em Inglês | MEDLINE | ID: mdl-37641063

RESUMO

BACKGROUND: The in-vitro scratch assay is a useful method in wound healing research to assess cell migration. In this assay, a scratch is created in a confluent cell layer by mechanically removing cells through manual scraping with a sharp-edged tool. This step is traditionally done with pipette tips and is unsuitable for high-throughput assays, as the created scratches are highly variable in width and position. Commercially available solutions are often expensive, and require specific cultureware which might not be suitable for all studies. RESULTS: In this study, we have developed a flexible cell scratch device comprising a single wounding tool, a guide and an imaging template for consistent and reproducible scratch assays in 96-well plates. Our results showed that the device produced a more consistent scratch profile compared to the conventional method of using pipette tips. The imaging template also allowed operators to easily locate and image the same region of interest at different time points, which potentially could be used for other assays. CONCLUSIONS: Our flexible yet effective scratch device thus enables robust scratch assays that can be applied to different experimental needs, providing researchers with an easy and reliable tool for their studies.


Assuntos
Bioensaio , Projetos de Pesquisa , Ensaios de Triagem em Larga Escala , Cicatrização
16.
BMC Plant Biol ; 23(1): 457, 2023 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-37775771

RESUMO

BACKGROUND: Russeting is a major problem in many fruit crops. Russeting is caused by environmental factors such as wounding or moisture exposure of the fruit surface. Despite extensive research, the molecular sequence that triggers russet initiation remains unclear. Here, we present high-resolution transcriptomic data by controlled russet induction at very early stages of fruit development. During Phase I, a patch of the fruit surface is exposed to surface moisture. For Phase II, moisture exposure is terminated, and the formerly exposed surface remains dry. We targeted differentially expressed transcripts as soon as 24 h after russet induction. RESULTS: During moisture exposure (Phase I) of 'Pinova' apple, transcripts associated with the cell cycle, cell wall, and cuticle synthesis (SHN3) decrease, while those related to abiotic stress increase. NAC35 and MYB17 were the earliest induced genes during Phase I. They are therefore linked to the initial processes of cuticle microcracking. After moisture removal (Phase II), the expression of genes related to meristematic activity increased (WOX4 within 24 h, MYB84 within 48 h). Genes related to lignin synthesis (MYB52) and suberin synthesis (MYB93, WRKY56) were upregulated within 3 d after moisture removal. WOX4 and AP2B3 are the earliest differentially expressed genes induced in Phase II. They are therefore linked to early events in periderm formation. The expression profiles were consistent between two different seasons and mirrored differences in russet susceptibility in a comparison of cultivars. Furthermore, expression profiles during Phase II of moisture induction were largely identical to those following wounding. CONCLUSIONS: The combination of a unique controlled russet induction technique with high-resolution transcriptomic data allowed for the very first time to analyse the formation of cuticular microcracks and periderm in apple fruit immediately after the onset of triggering factors. This data provides valuable insights into the spatial-temporal dynamics of russeting, including the synthesis of cuticles, dedifferentiation of cells, and impregnation of cell walls with suberin and lignin.


Assuntos
Malus , Malus/metabolismo , Frutas , Transcriptoma , Lignina/metabolismo , Perfilação da Expressão Gênica
17.
Plant Cell Environ ; 46(12): 3871-3886, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37646324

RESUMO

Grafting is widely used in horticulture. Shortly after grafting, callus tissues appear at the graft interface and the vascular tissues of the scion and rootstock connect. The graft interface contains a complex mix of tissues, we hypothesised that each tissue has its own metabolic response to wounding/grafting and accumulates different metabolites at different rates. We made intact and wounded cuttings and grafts of grapevine, and then measured changes in bulk flavonoid, phenolic acid and stilbenoid concentration and used metabolite imaging to study tissue-specific responses. We show that some metabolites rapidly accumulate in specific tissues after grafting, for example, stilbene monomers accumulate in necrotic tissues surrounding mature xylem vessels. Whereas other metabolites, such as complex stilbenes, accumulate in the same tissues at later stages. We also observe that other metabolites accumulate in the newly formed callus tissue and identify genotype-specific responses. In addition, exogenous resveratrol application did not modify grafting success rate, potentially suggesting that the accumulation of resveratrol at the graft interface is not linked to graft union formation. The increasing concentration of complex stilbenes often occurs in response to plant stresses (via unknown mechanisms), and potentially increases antioxidant activity and antifungal capacities.


Assuntos
Estilbenos , Vitis , Resveratrol/metabolismo , Estilbenos/metabolismo , Plantas/metabolismo , Antioxidantes/metabolismo , Vitis/fisiologia
18.
J Exp Bot ; 74(4): 1159-1161, 2023 02 13.
Artigo em Inglês | MEDLINE | ID: mdl-36779288

RESUMO

Jasmonate is a well-known defence hormone for plants, but it is also necessary for growth and development. Indeed, the identification of the COI1 receptor was based on the jasmonate-triggered response of root growth inhibition. In this special issue, a collection of review papers and two research papers discuss the current state of progress in this field, covering areas from seed germination and flowering to the Jasminum sambac genome.


Assuntos
Ciclopentanos , Oxilipinas , Ciclopentanos/farmacologia , Oxilipinas/farmacologia
19.
J Exp Bot ; 74(4): 1198-1206, 2023 02 13.
Artigo em Inglês | MEDLINE | ID: mdl-34966932

RESUMO

Plants have remarkable abilities to regenerate in response to wounding. How wounding triggers rapid signal transduction to induce a cellular response is a key topic for understanding the molecular mechanism of plant regeneration. An increasing body of evidence indicates that jasmonate, a hormone that is produced rapidly in response to wounding, plays multiple roles in different plant regeneration processes. In this review, we summarize recent advances on the roles of jasmonate in tissue repair, the formation of wound-induced callus, de novo organ regeneration, and somatic embryogenesis. Physiological and molecular analyses indicate that jasmonate can regulate stem cell activities, cell proliferation, cell fate transition, and auxin production, thereby contributing to plant regeneration. In addition, jasmonate is strictly controlled in plant cells via restriction of the jasmonate concentration and its signalling pathway in a spatial and temporal manner during regeneration. Overall, jasmonate acts as the hormone linking wounding to distinct types of regeneration in plants.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Ácidos Indolacéticos/metabolismo , Plantas/metabolismo , Hormônios/metabolismo , Regulação da Expressão Gênica de Plantas
20.
Exp Eye Res ; 234: 109599, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37488009

RESUMO

Limbal epithelial stem cells are not only critical for corneal epithelial homeostasis but also have the capacity to change from a relatively quiescent mitotic phenotype to a rapidly proliferating cell in response to population depletion following corneal epithelial wounding. Pax6+/- mice display many abnormalities including corneal vascularization and these aberrations are consistent with a limbal stem cell deficiency (LSCD) phenotype. FoxC1 has an inhibitory effect on corneal avascularity and a positive role in stem cell maintenance in many tissues. However, the role of FoxC1 in limbal epithelial stem cells remains unknown. To unravel FoxC1's role(s) in limbal epithelial stem cell homeostasis, we utilized an adeno-associated virus (AAV) vector to topically deliver human FOXC1 proteins into Pax6 +/- mouse limbal epithelium. Under unperturbed conditions, overexpression of FOXC1 in the limbal epithelium had little significant change in differentiation (PAI-2, Krt12) and proliferation (BrdU, Ki67). Conversely, such overexpression resulted in a marked increase in the expression of putative limbal epithelial stem cell markers, N-cadherin and Lrig1. After corneal injuries in Pax6 +/- mice, FOXC1 overexpression enhanced the behavior of limbal epithelial stem cells from quiescence to a highly proliferative status. Overall, the treatment of AAV8-FOXC1 may be beneficial to the function of limbal epithelial stem cells in the context of a deficiency of Pax6 function.


Assuntos
Doenças da Córnea , Epitélio Corneano , Limbo da Córnea , Animais , Humanos , Camundongos , Córnea , Doenças da Córnea/metabolismo , Desbridamento , Células Epiteliais , Epitélio Corneano/metabolismo , Limbo da Córnea/metabolismo , Células-Tronco
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