Astaxanthin attenuates cigarette smoke-induced small airway remodeling via the AKT1 signaling pathway.

BACKGROUND: Astaxanthin (AXT) is a keto-carotenoid with a variety of biological functions, including antioxidant and antifibrotic effects. Small airway remodeling is the main pathology of chronic obstructive pulmonary disease (COPD) and is caused by epithelial-to-mesenchymal transition (EMT) and fibroblast differentiation and proliferation. Effective therapies are still lacking. This study aimed to investigate the role of AXT in small airway remodeling in COPD and its underlying mechanisms. METHODS: First, the model of COPD mice was established by cigarette smoke (CS) exposure combined with intraperitoneal injection of cigarette smoke extract (CSE). The effects of AXT on the morphology of CS combined with CSE -induced emphysema, EMT, and small airway remodeling by using Hematoxylin-eosin (H&E) staining, immunohistochemical staining, and western blot. In addition, in vitro experiments, the effects of AXT on CSE induced-EMT and fibroblast function were further explored. Next, to explore the specific mechanisms underlying the protective effects of AXT in COPD, potential targets of AXT in COPD were analyzed using network pharmacology. Finally, the possible mechanism was verified through molecular docking and in vitro experiments. RESULTS: AXT alleviated pulmonary emphysema, EMT, and small airway remodeling in a CS combined with CSE -induced mouse model. In addition, AXT inhibited the EMT process in airway cells and the differentiation and proliferation of fibroblasts. Mechanistically, AXT inhibited myofibroblast activation by directly binding to and suppressing the phosphorylation of AKT1. Therefore, our results show that AXT protects against small airway remodeling by inhibiting AKT1. CONCLUSIONS: The present study identified and illustrated a new food function of AXT, indicating that AXT could be used in the therapy of COPD-induced small airway remodeling.

The Role of SHP2 in Advancing COPD: Insights into Oxidative Stress, Endoplasmic Reticulum Stress, and Pyroptosis.

Background: Chronic Obstructive Pulmonary Disease (COPD) is characterized by airflow limitation and inflammation resulting from genetic and environmental factors, notably cigarette smoke. Pyroptosis, a cell death process, is implicated in COPD, but its mechanisms are unclear. SHP2, a phosphatase, modulates inflammatory pathways, suggesting a role in COPD pathogenesis and potential therapeutic avenues. Objective: This study investigates the mechanism by which SHP2 regulates cell pyroptosis in bronchial epithelial cells in COPD patients. Methods: In this prospective study, we employed in vivo and in vitro models to investigate the mechanisms underlying COPD progression. Hematoxylin and eosin (H&E) staining were utilized to assess the morphological changes characteristic of COPD. Electron microscopy enabled precise quantification of pyroptotic bodies to highlight cellular changes associated with COPD pathogenesis. Immunofluorescence analysis facilitated the measurement of protein fluorescence intensity, allowing for the assessment of inflammatory responses within bronchial epithelial cells. Additionally, Western blot analysis was conducted to evaluate the expression levels of key pathway proteins involved in COPD progression. Results: In the COPD model, lesions worsened in SHP2-KD mice compared to SHP2-NC. Western blot results showed increased p22, p47, p-IRE1α, XBP1, STING, p-TBK1, NLRP3, Caspase1, and IL-1ß expression levels in both in vivo and in vitro models. Transmission electron microscopy revealed more pyroptotic bodies in SHP2-KD+CSE than in SHP2-NC+CSE. Immunofluorescence demonstrated significantly higher NLRP3 and GSDMD fluorescence intensities in SHP2-KD+CSE versus SHP2-NC+CSE. Additionally, Western blot analysis indicated increased expression of Bax, Caspase3, Caspase8, and Caspase9 proteins in the in vitro model. No differences were observed between SHP2 NC and SHP2-KD groups without CSE stimulation in immunofluorescence, electron microscopy, and Western blot findings in the cellular model. Conclusions: SHP2 promotes COPD progression by inducing oxidative stress, endoplasmic reticulum stress, and pyroptosis.

Characteristics of SlCML39, a Tomato Calmodulin-like Gene, and Its Negative Role in High Temperature Tolerance of Arabidopsis thaliana during Germination and Seedling Growth.

Calmodulin-like (CML) proteins are primary calcium sensors and function in plant growth and response to stress stimuli. However, so far, the function of plant CML proteins, including tomato, is still unclear. Previously, it was found that a tomato (Solanum lycopersicum) CML, here named SlCML39, was significantly induced by high temperature (HT) at transcription level, but its biological function is scarce. In this study, the characteristics of SlCML39 and its role in HT tolerance were studied. SlCML39 encodes a protein of 201 amino acids containing four EF hand motifs. Many cis-acting elements related to plant stress and hormone response appear in the promoter regions of SlCML39. SlCML39 is mainly expressed in the root, stem, and leaf and can be regulated by HT, cold, drought, and salt stresses as well as ABA and H2O2. Furthermore, heterologous overexpression of SlCML39 reduces HT tolerance in Arabidopsis thaliana at the germination and seedling growth stages. To better understand the molecular mechanism of SlCML39, the downstream gene network regulated by SlCML39 under HT was analyzed by RNA-Seq. Interestingly, we found that many genes involved in stress responses as well as ABA signal pathway are down-regulated in the transgenic seedlings under HT stress, such as KIN1, RD29B, RD26, and MAP3K18. Collectively, these data indicate that SlCML39 acts as an important negative regulator in response to HT stress, which might be mediated by the ABA signal pathway.

Genome-Wide Identification and Transcriptional Expression Analysis of Annexin Genes in Capsicum annuum and Characterization of CaAnn9 in Salt Tolerance.

Annexin (Ann) is a polygenic, evolutionarily conserved, calcium-dependent and phospholipid-binding protein family, which plays key roles in plant growth, development, and stress response. However, a comprehensive understanding of CaAnn genes of pepper (Capsicum annuum) at the genome-wide level is limited. Based on the available pepper genomic information, we identified 15 members of the CaAnn gene family. Phylogenetic analysis showed that CaAnn proteins could be categorized into four different orthologous groups. Real time quantitative RT-PCR analysis showed that the CaAnn genes were tissue-specific and were widely expressed in pepper leaves after treatments with cold, salt, and drought, as well as exogenously applied MeJA and ABA. In addition, the function of CaAnn9 was further explored using the virus-induced gene silencing (VIGS) technique. CaAnn9-silenced pepper seedlings were more sensitive to salt stress, reflected by the degradation of chlorophyll, the accumulation of reactive oxygen species (ROS), and the decrease of antioxidant defense capacity. This study provides important information for further study of the role of pepper CaAnn genes and their coding proteins in growth, development, and environmental responses.

Proteomic analysis by iTRAQ-PRM provides integrated insight into mechanisms of resistance in pepper to Bemisia tabaci (Gennadius).

BACKGROUND: The Bemisia tabaci is a major leaf feeding insect pest to pepper (Capsicum annuum), causing serious damage to pepper growth and yield. It is particularly important to study the mechanism of pepper resistance to B. tabaci, and to breed and promote the varieties of pepper resistant to B. tabaci. However, very limited molecular mechanism is available about how plants perceive and defend themselves from the destructive pest. Proteome technologies have provided an idea method for studying plant physiological processes in response to B. tabaci. RESULTS: Here, a highly resistant genotype and a highly susceptible genotype were exposed to B. tabaci feeding for 48 h to explore the defense mechanisms of pepper resistance to B. tabaci. The proteomic differences between both genotypes were compared using isobaric tag for relative and absolute quantification (iTRAQ). The quantitative data were validated by parallel reaction monitoring (PRM). The results showed that 37 differential abundance proteins (DAPs) were identified in the RG (resistant genotype), while 17 DAPs were identified in the SG (susceptible genotype) at 48 h after B. tabaci feeding. 77 DAPs were identified when comparing RG with SG without feeding. The DAP functions were determined for the classification of the pathways, mainly involved in redox regulation, stress response, protein metabolism, lipid metabolism and carbon metabolism. Some candidate DAPs are closely related to B. tabaci resistance such as annexin D4-like (ANN4), calreticulin-3 (CRT3), heme-binding protein 2-like (HBP1), acidic endochitinase pcht28-like (PR3) and lipoxygenase 2 (LOX2). CONCLUSIONS: Taken together, this study indicates complex resistance-related events in B. tabaci interaction, provides novel insights into the molecular mechanism underlying the response of plant to B. tabaci, and identifies some candidate proteins against B. tabaci attack.

Chloroplast-localized BICAT proteins shape stromal calcium signals and are required for efficient photosynthesis.

The photosynthetic machinery of plants must be regulated to maximize the efficiency of light reactions and CO2 fixation. Changes in free Ca2+ in the stroma of chloroplasts have been observed at the transition between light and darkness, and also in response to stress stimuli. Such Ca2+ dynamics have been proposed to regulate photosynthetic capacity. However, the molecular mechanisms of Ca2+ fluxes in the chloroplasts have been unknown. By employing a Ca2+ reporter-based approach, we identified two chloroplast-localized Ca2+ transporters in Arabidopsis thaliana, BICAT1 and BICAT2, that determine the amplitude of the darkness-induced Ca2+ signal in the chloroplast stroma. BICAT2 mediated Ca2+ uptake across the chloroplast envelope, and its knockout mutation strongly dampened the dark-induced [Ca2+ ]stroma signal. Conversely, this Ca2+ transient was increased in knockout mutants of BICAT1, which transports Ca2+ into the thylakoid lumen. Knockout mutation of BICAT2 caused severe defects in chloroplast morphology, pigmentation and photosynthetic light reactions, rendering bicat2 mutants barely viable under autotrophic growth conditions, while bicat1 mutants were less affected. These results show that BICAT transporters play a role in chloroplast Ca2+ homeostasis. They are also involved in the regulation of photosynthesis and plant productivity. Further work will be required to reveal whether the effect on photosynthesis is a direct result of their role as Ca2+ transporters.

The Tomato Mitogen-Activated Protein Kinase SlMPK1 Is as a Negative Regulator of the High-Temperature Stress Response.

High-temperature (HT) stress is a major environmental stress that limits plant growth and development. MAPK cascades play key roles in plant growth and stress signaling, but their involvement in the HT stress response is poorly understood. Here, we describe a 47-kD MBP-phosphorylated protein (p47-MBPK) activated in tomato (Solanum lycopersicum) leaves under HT and identify it as SlMPK1 by tandem mass spectrometry analysis. Silencing of SlMPK1 in transgenic tomato plants resulted in enhanced tolerance to HT, while overexpression resulted in reduced tolerance. Proteomic analysis identified a set of proteins involved in antioxidant defense that are significantly more abundant in RNA interference-SlMPK1 plants than nontransgenic plants under HT stress. RNA interference-SlMPK1 plants also showed changes in membrane lipid peroxidation and antioxidant enzyme activities. Furthermore, using yeast two-hybrid screening, we identified a serine-proline-rich protein homolog, SlSPRH1, which interacts with SlMPK1 in yeast, in plant cells, and in vitro. We demonstrate that SlMPK1 can directly phosphorylate SlSPRH1. Furthermore, the serine residue serine-44 of SlSPRH1 is a crucial phosphorylation site in the SlMPK1-mediated antioxidant defense mechanism activated during HT stress. We also demonstrate that heterologous expression of SlSPRH1 in Arabidopsis (Arabidopsis thaliana) led to a decrease in thermotolerance and lower antioxidant capacity. Taken together, our results suggest that SlMPK1 is a negative regulator of thermotolerance in tomato plants. SlMPK1 acts by regulating antioxidant defense, and its substrate SlSPRH1 is involved in this pathway.

A comprehensive meta-analysis of plant morphology, yield, stay-green, and virus disease resistance QTL in maize (Zea mays L.).

MAIN CONCLUSION: The meta-QTL and candidate genes will facilitate the elucidation of molecular bases underlying agriculturally important traits and open new avenues for functional markers development and elite alleles introgression in maize breeding program. A large number of QTLs attributed to grain productivity and other agriculturally important traits have been identified and deposited in public repositories. The integration of fruitful QTL becomes a major issue in current plant genomics. To this end, we first collected QTL for six agriculturally important traits in maize, including yield, plant height, ear height, leaf angle, stay-green, and maize rough dwarf disease resistance. The meta-analysis method was then employed to retrieve 113 meta-QTL. Additionally, we also isolated candidate genes for target traits by the bioinformatic technique. Several candidates, including some well-characterized genes, GA3ox2 for plant height, lg1 and lg4 for leaf angle, zfl1 and zfl2 for flowering time, were co-localized with established meta-QTL intervals. Intriguingly, in a relatively narrow meta-QTL region, the maize ortholog of rice yield-related gene GW8/OsSPL16 was believed to be a candidate for yield. Leveraging results presented in this study will provide further insights into the genetic architecture of maize agronomic traits. Moreover, the meta-QTL and candidate genes reported here could be harnessed for the enhancement of stress tolerance and yield performance in maize and translation to other crops.

Skin lesion classification based on two-modal images using a multi-scale fully-shared fusion network.

BACKGROUND AND OBJECTIVE: Due to the complexity of skin lesion features, computer-aided diagnosis of skin diseases based on multi-modal images is considered a challenging task. Dermoscopic images and clinical images are commonly used to diagnose skin diseases in clinical scenarios, and the complementarity of their features promotes the research of multi-modality classification in the computer-aided diagnosis field. Most current methods focus on the fusion between modalities and ignore the complementary information within each of them, which leads to the loss of the intra-modality relation. Multi-modality models for integrating features both within single modalities and across multiple modalities are limited in the literature. Therefore, a multi-modality model based on dermoscopic and clinical images is proposed to address this issue. METHODS: We propose a Multi-scale Fully-shared Fusion Network (MFF-Net) that gathers features of dermoscopic images and clinical images for skin lesion classification. In MFF-Net, the multi-scale fusion structure combines deep and shallow features within individual modalities to reduce the loss of spatial information in high-level feature maps. Then Dermo-Clinical Block (DCB) integrates the feature maps from dermoscopic images and clinical images through channel-wise concatenation and using a fully-shared fusion strategy that explores complementary information at different stages. RESULTS: We validated our model on a four-class two-modal skin diseases dataset, and proved that the proposed multi-scale structure, the fusion module DCBs, and the fully-shared fusion strategy improve the performance of MFF-Net independently. Our method achieved the highest average accuracy of 72.9% on the 7-point checklist dataset, outperforming the state-of-the-art single-modality and multi-modality methods with an accuracy boost of 7.1% and 3.4%, respectively. CONCLUSIONS: The multi-scale fusion structure demonstrates the significance of intra-modality relations between clinical images and dermoscopic images. The proposed network combined with the multi-scale structure, DCBs, and the fully-shared fusion strategy, can effectively integrate the features of the skin lesions across the two modalities and achieved a promising accuracy among different skin diseases.

Functional insights of plant bcl-2-associated ahanogene (BAG) proteins: Multi-taskers in diverse cellular signal transduction pathways.

Bcl-2-associated athanogene (BAG) gene family is a highly conserved molecular chaperone cofactor in evolution from yeast to humans and plants playing important roles in a variety of signal pathways. Plant BAG proteins have special structures, especially those containing CaM-binding IQ motifs which are unique to plants. While early studies focused more on the structure and physiological function of plant BAGs, recent studies have revealed many novel functional mechanisms involved in multiple cellular processes. How to achieve signal specificity has become an interesting topic of plant BAG research. In this review, we have provided a historic view of plant BAG research and summarized recent advances in the establishment of BAG as essential components in normal plant growth, environmental stress response, and plant immunity. Based on the relationship between BAG proteins and their newly interacting proteins, this review highlights the functional mechanisms of various cellular signals mediated by plant BAGs. Future work needs to focus on the post-translational modification of BAG proteins, and on understanding how specificity is achieved among BAG signaling pathways.

A critical review on plant annexin: Structure, function, and mechanism.

Plant annexins are evolutionary conserved protein family widely exist in almost all plant species, characterized by a shorter N-terminal region and four conservative annexin repeats. Plant annexins have Ca2+ channel-regulating activity and peroxidase as well as ATPase/GTPase activities, which give annexins functional specificity. They are widely involved in regulating diverse aspects of biochemical and cellular processes, plant growth and development, and responses to biotic and abiotic environmental stresses. Though many studies have reviewed the function of annexins, great progress have been made in the study of plant annexins recently. In this review, we outline the current understanding of basic properties of plant annexins and summarize the emerging advances in understanding the functional roles of annexins in plants and highlight the regulation mechanisms of annexin protein in response to stress especially to salt and cold stress. The interesting questions related to plant annexin that remain to be further elucidated are also discussed.

Overexpression of a Bcl-2-associated athanogene SlBAG9 negatively regulates high-temperature response in tomato.

The Bcl-2-associated athanogene (BAG) gene is a multi-functional family of co-chaperones regulator, modulating plant stress response. Our previous study revealed that the SlBAG9 of tomato (Solanum lycopersicum) had the higher expression level induced by high-temperature (HT) at the transcriptional and protein levels, but its biological function was still unclear. Here, we conducted an in-depth analysis of SlBAG9. SlBAG9 protein was not located in the mitochondria but in the cytoplasm and nucleus. Many cis-acting elements involved in plant stress and hormone responses were located in the promoter regions of SlBAG9 including heat-shock element (HSE1). The ß-glucuronidase (GUS) histochemical analysis showed that SlBAG9 promoter could drive GUS gene expression in transiently transformed Nicotiana tabacum leaves under non-inducing condition and HSE1 is critical for HT-induced GUS activity under HT. The transcription of SlBAG9 was expressed in different organs and was regulated by HT, cold, drought, and salt stresses as well as exogenous abscisic acid (ABA) and H2O2. To further elucidate SlBAG9 function in response to HT, the transgenic tomato plants overexpressing SlBAG9 were developed. Compared to the wild-type plants, SlBAG9-overexpressing plants exhibited more sensitivity to HT stress, reflected by the burning symptoms, the degradation of chlorophyll, and the reduction of photosynthetic rates. Additionally, SlBAG9-overexpressing lines showed higher accumulation of lipid peroxidation production (MDA) and H2O2, but lower activities of superoxide dismutase, catalase, and peroxidase. Therefore, it is speculated that SlBAG9 plays a negative role in thermotolerance probably by inhibition of antioxidant enzyme system leading to the oxidative damage, consequently aggravating the HT-caused injury phenotype.

Genome-Wide Identification of the Bcl-2 Associated Athanogene (BAG) Gene Family in Solanum lycopersicum and the Functional Role of SlBAG9 in Response to Osmotic Stress.

The Bcl-2-associated athanogene (BAG) proteins are a family of multi-functional group of co-chaperones regulators, modulating diverse processes from plant growth and development to stress response. Here, 10 members of SlBAG gene family were identified based on the available tomato (Solanum lycopersicum) genomic information and named as SlBAG1-10 according to their chromosomal location. All SlBAG proteins harbor a characteristic BAG domain, categorized into two groups, and SlBAG4, SlBAG7, and SlBAG9 of group I contain a plant-specific isoleucine glutamine (IQ) calmodulin-binding motif located in the N terminus. The quantitative real-time PCR expression analysis revealed that these SlBAG genes had organ-specific expression patterns and most SlBAG genes were differentially expressed in multiple abiotic stresses including drought, salt, high temperature, cold, and cadmium stress as well as abscisic acid and H2O2. In addition, heterologous overexpression of SlBAG9 increased the sensitivity of Arabidopsis to drought, salt, and ABA during seed germination and seedling growth. The decreased tolerance may be due to the downregulation of stress-related genes expression and severe oxidative stress. The expression levels of some stress and ABA-related genes, such as ABI3, RD29A, DREB2A, and P5CS1, were significantly inhibited by SlBAG9 overexpression under osmotic stress. Meanwhile, the overexpression of SlBAG9 inhibited the expression of FSD1 and CAT1 under stress conditions and the decreased levels of superoxide dismutase and catalase enzyme activities were detected accompanying the trends in the expression of both genes, which resulted in H2O2 accumulation and lipid peroxidation. Taken together, these findings lay a foundation for the future study of the biological function of SlBAG genes in tomato.

The role of plant-specific VQ motif-containing proteins: An ever-thickening plot.

VQ proteins are a class of plant-specific proteins containing the conserved motif FxxhVQxhTG（h denotes hydrophobic residues and x represents any amino acid）and are named VQ for the V and Q residues. By analyzing the structure of VQ members it was found that most VQ genes do not contain introns and the number of encoded amino acids is less than 300 aa. A majority of VQ proteins are located in the nucleus. Accumulated evidence has highlighted the importance of VQ proteins mainly participating in signal pathways through interacting with partners (eg. WRKYs and MAPKs) to regulate plant growth and development and respond to biotic and abiotic stresses. This review primarily focuses on the structure of VQ members in plant kingdom and the biological function and the mechanism of VQ protein action, and discusses recent advances in understanding the pivotal role of VQ-motif, which provides a solid foundation for further exploration on VQ proteins.

Mitogen-activated protein kinase action in plant response to high-temperature stress: a mini review.

In recent years, extreme weather events such as high temperature (HT) are becoming more frequent. HT has become one of the main environmental factors affecting crop growth and development. In nature, plant cells initiate corresponding tolerant mechanisms by sensing and transducing HT signals. The mitogen-activated protein kinase (MAPK) cascade is widely involved in the signal transduction of plants to various environmental stresses. MAPK-mediated HT responses have attracted more and more attention. We herein focus on the current state of knowledge of MAPK in the plant under HT stress and summarize the mechanisms of MAPK in HT response from Ca2+ signal, reactive oxygen species (ROS) signal, heat shock transcription factor and heat shock protein, antioxidant system, and the direct downstream targets of MAPK. This review encapsulates the known plant MAPK cascade and provides prospects for ongoing research on HT response.

A novel mitogen-activated protein kinase gene in maize (Zea mays), ZmMPK3, is involved in response to diverse environmental cues.

In search for components of mitogen-activated protein kinase (MAPK) cascades in maize (Zea mays) involved in response to abscisic acid (ABA) stimulus, a novel MAPK gene, ZmMPK3, from ABA-treated maize leaves cDNA was isolated and characterized. The full length of the ZmMPK3 gene is 1 520 bp and encodes a 376 amino acid protein with a predicted molecular mass of 43.5 kD and a pI of 5.83. ZmMPK3 contains all 11 MAPK conserved subdomains and the phosphorylation motif TEY. Amino acid sequence alignment revealed that ZmMPK3 shared high identity with group-A MAPK in plants. A time course (30-360 min) experiment using a variety of signal molecules and stresses revealed that the transcripts level of ZmMPK3 accumulated markedly and rapidly when maize seedlings were subjected to exogenous signaling molecules: ABA, H2O2, jasmonic acid and salicylic acid, various abiotic stimuli such as cold, drought, ultraviolet light, salinity, heavy metal and mechanical wounding. Its transcription was also found to be tissue-specific regulated. Here, we show that ABA and H2O2 induced a significant increase in the ZmMPK3 activity using immunoprecipitation and in-gel kinase assay. Furthermore, the results showed that the ZmMPK3 protein is localized mainly to the nucleus. These results suggest that the ZmMPK3 may play an important role in response to environmental stresses.

Transcriptome -wide modulation combined with morpho-physiological analyses of Typha orientalis roots in response to lead challenge.

Lead (Pb) is a common pollutant in many environments, including in the soil, water, and/or air. Typha orientalis Presl, a large emergent aquatic plant, has been reported to function as a Pb-tolerant and Pb-accumulating plant; however, very little molecular information regarding the tolerance of T. orientalis towards Pb is known. In this study, Pb accumulation and key factors involved in the Pb stress response at different Pb concentrations were investigated. Pb was primarily accumulated in the roots and was mainly located in the cell wall and membrane systems. Differentially expressed genes (DEGs) were identified in T. orientalis roots after Pb exposure via RNA-seq analyses. In the 0.10 mM and 0.25 mM Pb2+-treated groups, a total of 3275 DEGs were detected relative to the control. Many of these genes were associated with oxidation-reduction processes, metal transport, protein kinase/phosphorylation, and DNA binding transcription factors, which were shown to be Pb-responsive DEGs. Mapping Kyoto Encyclopedia of Genes and Genomes (KEGG) database, "phenylpropanoid biosynthesis" was analyzed as the major pathway of the important modules of overlapping DEGs of 0.10 mM and 0.25 mM Pb2+ treatments. Furthermore, a lead response gene named ToLR1 with unknown function was of particular interest. The full-length of ToLR1 sequence was cloned using rapid amplification of cDNA ends (RACE) and overexpressed in Arabidopsis thaliana, which resulted in enhanced resistance to Pb stress. This is the first report providing genomic information detailing Pb responsive genes in T. orientalis. Moreover, this study provides novel insights into the molecular mechanisms underlying the response of T. orientalis and other accumulators towards Pb stress. The key genes identified in this study may serve as potential targets for genetic engineering targeting phytoremediation.

In-depth proteome analysis reveals multiple pathways involved in tomato SlMPK1-mediated high-temperature responses.

High temperature (HT) is one of the major environmental factors which limits plant growth and yield. The mitogen-activated protein kinase (MAPK) plays vital roles in environmental stress responses. However, the mechanisms triggered by MAPKs in plants in response to HT are still extremely limited. In this study, the proteomic data of differences between SlMPK1 RNA-interference mutant (SlMPK1i) and wild type and of tomato (Solanum lycopersicum) plants under HT stress using isobaric tags for relative and absolute quantitation (iTRAQ) was re-analyzed in depth. In total, 168 differently expressed proteins (DEPs) were identified in response to HT stress, including 38 DEPs only found in wild type, and 84 DEPs specifically observed in SlMPK1i after HT treatment. The majority of higher expression of 84 DEPs were annotated into photosynthesis, oxidation-reduction process, protein folding, translation, proteolysis, stress response, and amino acid biosynthetic process. More importantly, SlMPK1-mediated photosynthesis was confirmed by the physiological characterization of SlMPK1i with a higher level of photosynthetic capacity under HT stress. Overall, the results reveal a set of potential candidate proteins helping to further understand the intricate regulatory network regulated by SlMPK1 in response to HT.

Identity of an ABA-activated 46 kDa mitogen-activated protein kinase from Zea mays leaves: partial purification, identification and characterization.

Mitogen-activated protein kinase (MAPK) cascades have been shown to be important components in abscisic acid (ABA) signal transduction pathway. In this study, a 46 kDa MAPK (p46MAPK) induced by ABA was partially purified from maize (Zea mays) by Q-Sepharose FF, Phenyl-Sepharose FF, Resource Q, Mono QTM 5/50 GL, poly-L-lysine-agarose, and Superdex 75 prep-grade columns, and was identified as ZmMAPK5 (gi|4239889) by the matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry. Furthermore, the kinase showed optimal activity at pH 8.0, 30 degrees C, and 10 mM MgCl(2); the K(m) for myelin basic protein (MBP) substrate and ATP were 0.13 microg microl(-1) and 62 microM, respectively. MBP was the preferred substrate, of which the threonine residue was phosphorylated. Finally, the kinase was found to respond to diverse extracellular stimuli. These results enable us to further reveal the function of the ZmMAPK5 in ABA signaling.

Positive feedback regulation of maize NADPH oxidase by mitogen-activated protein kinase cascade in abscisic acid signalling.

In maize (Zea mays), abscisic acid (ABA)-induced H(2)O(2) production activates a 46 kDa mitogen-activated protein kinase (p46MAPK), and the activation of p46MAPK also regulates the production of H(2)O(2). However, the mechanism for the regulation of H(2)O(2) production by MAPK in ABA signalling remains to be elucidated. In this study, four reactive oxygen species (ROS)-producing NADPH oxidase (rboh) genes (ZmrbohA-D) were isolated and characterized in maize leaves. ABA treatment induced a biphasic response (phase I and phase II) in the expression of ZmrbohA-D and the activity of NADPH oxidase. Phase II induced by ABA was blocked by pretreatments with two MAPK kinase (MPKKK) inhibitors and two H(2)O(2) scavengers, but phase I was not affected by these inhibitors or scavengers. Treatment with H(2)O(2) alone also only induced phase II, and the induction was arrested by the MAPKK inhibitors. Furthermore, the ABA-activated p46MAPK was partially purified. Using primers corresponding to the sequences of internal tryptic peptides, the p46MAPK gene was cloned. Analysis of the tryptic peptides and the p46MAPK sequence indicate it is the known ZmMPK5. Treatments with ABA and H(2)O(2) led to a significant increase in the activity of ZmMPK5, although ABA treatment only induced a slight increase in the expression of ZmMPK5. The data indicate that H(2)O(2)-activated ZmMPK5 is involved in the activation of phase II in ABA signalling, but not in phase I. The results suggest that there is a positive feedback loop involving NADPH oxidase, H(2)O(2), and ZmMPK5 in ABA signalling.