NADPH oxidase-dependent H2 O2 production mediates salicylic acid-induced salt tolerance in mangrove plant Kandelia obovata by regulating Na+ /K+ and redox homeostasis.

Salicylic acid (SA) is known to enhance salt tolerance in plants. However, the mechanism of SA-mediated response to high salinity in halophyte remains unclear. Using electrophysiological and molecular biological methods, we investigated the role of SA in response to high salinity in mangrove species, Kandelia obovata, a typical halophyte. Exposure of K. obovata roots to high salinity resulted in a rapid increase in endogenous SA produced by phenylalanine ammonia lyase pathway. The application of exogenous SA improved the salt tolerance of K. obovata, which depended on the NADPH oxidase-mediated H2 O2 . Exogenous SA and H2 O2 increased Na+ efflux and reduced K+ loss by regulating the transcription levels of Na+ and K+ transport-related genes, thus reducing the Na+ /K+ ratio in the salt-treated K. obovata roots. In addition, exogenous SA-enhanced antioxidant enzyme activity and its transcripts, and the expressions of four genes related to AsA-GSH cycle as well, then alleviated oxidative damages in the salt-treated K. obovata roots. However, the above effects of SA could be reversed by diphenyleneiodonium chloride (the NADPH oxidase inhibitor) and paclobutrazol (a SA biosynthesis inhibitor). Collectively, our results demonstrated that SA-induced salt tolerance of K. obovata depends on NADPH oxidase-generated H2 O2 that affects Na+ /K+ and redox homeostasis in response to high salinity.

Structural, developmental and functional analyses of leaf salt glands of mangrove recretohalophyte Aegiceras corniculatum.

Salt secretion is an important strategy used by the mangrove plant Aegiceras corniculatum to adapt to the coastal intertidal environment. However, the structural, developmental and functional analyses on the leaf salt glands, particularly the salt secretion mechanism, are not well documented. In this study, we investigated the structural, developmental and degenerative characteristics and the salt secretion mechanisms of salt glands to further elucidate the mechanisms of salt tolerance of A. corniculatum. The results showed that the salt gland cells have a large number of mitochondria and vesicles, and plenty of plasmodesmata as well, while chloroplasts were found in the collecting cells. The salt glands developed early and began to differentiate at the leaf primordium stage. We observed and defined three stages of salt gland degradation for the first time in A. corniculatum, where the secretory cells gradually twisted and wrinkled inward and collapsed downward as the salt gland degeneration increased and the intensity of salt gland autofluorescence gradually diminished. In addition, we found that the salt secretion rate of the salt glands increased when the treated concentration of NaCl increased, reaching the maximum at 400 mM NaCl. The salt-secreting capacity of the salt glands of the adaxial epidermis is significantly greater than that of the abaxial epidermis. The real-time quantitative PCR results indicate that SAD2, TTG1, GL2 and RBR1 may be involved in regulating the development of the salt glands of A. corniculatum. Moreover, Na+/H+ antiporter, H+-ATPase, K+ channel and Cl- channel may play important roles in the salt secretion of salt glands. In sum mary, this study strengthens the understanding of the structural, developmental and degenerative patterns of salt glands and salt secretion mechanisms in mangrove recretohalophyte A. corniculatum, providing an important reference for further studies at the molecular level.

RNA-sequencing transcriptome analysis of Avicennia marina (Forsk.) Vierh. leaf epidermis defines tissue-specific transcriptional response to salinity treatment.

Avicennia marina (Forsk.) Vierh. is a typical mangrove plant. Its epidermis contains salt glands, which can secrete excess salts onto the leaf surfaces, improving the salt tolerance of the plants. However, knowledge on the epidermis-specific transcriptional responses of A. marina to salinity treatment is lacking. Thus, physiological and transcriptomic techniques were applied to unravel the salt tolerance mechanism of A. marina. Our results showed that 400 mM NaCl significantly reduced the plant height, leaf area, leaf biomass and photosynthesis of A. marina. In addition, 1565 differentially expressed genes were identified, of which 634 and 931 were up- and down-regulated. Based on Kyoto Encyclopedia of Genes and Genomes metabolic pathway enrichment analysis, we demonstrated that decreased gene expression, especially that of OEE1, PQL2, FDX3, ATPC, GAPDH, PRK, FBP and RPE, could explain the inhibited photosynthesis caused by salt treatment. Furthermore, the ability of A. marina to cope with 400 mM NaCl treatment was dependent on appropriate hormone signalling and potential sulfur-containing metabolites, such as hydrogen sulfide and cysteine biosynthesis. Overall, the present study provides a theoretical basis for the adaption of A. marina to saline habitats and a reference for studying the salt tolerance mechanism of other mangrove plants.

Leaf sodium homeostasis controlled by salt gland is associated with salt tolerance in mangrove plant Avicennia marina.

Avicennia marina, a mangrove plant growing in coastal wetland habitats, is frequently affected by tidal salinity. To understand its salinity tolerance, the seedlings of A. marina were treated with 0, 200, 400 and 600 mM NaCl. We found the whole-plant dry weight and photosynthetic parameters increased at 200 mM NaCl but decreased over 400 mM NaCl. The maximum quantum yield of primary photochemistry (Fv/Fm) significantly decreased at 600 mM NaCl. Transmission electron microscopy observations showed high salinity caused the reduction in starch grain size, swelling of the thylakoids and separation of the granal stacks, and even destruction of the envelope. In addition, the dense protoplasm and abundant mitochondria in the secretory and stalk cells, and abundant plasmodesmata between salt gland cells were observed in the salt glands of the adaxial epidermis. At all salinities, Na+ content was higher in leaves than in stems and roots; however, Na+ content increased in the roots while it remained at a constant level in the leaves over 400 mM NaCl treatment, due to salt secretion from the salt glands. As a result, salt crystals on the leaf adaxial surface increased with salinity. On the other hand, salt treatment increased Na+ and K+ efflux and decreased H+ efflux from the salt glands by the non-invasive micro-test technology, although Na+ efflux reached the maximum at 400 mM NaCl. Further real-time quantitative PCR analysis indicated that the expression of Na+/H+ antiporter (SOS1 and NHX1), H+-ATPase (AHA1 and VHA-c1) and K+ channel (AKT1, HAK5 and GORK) were up-regulated, and only the only Na+ inward transporter (HKT1) was down-regulated in the salt glands enriched adaxial epidermis of the leaves under 400 mM NaCl treatment. In conclusion, salinity below 200 mM NaCl was beneficial to the growth of A. marina, and below 400 mM, the salt glands could excrete Na+ effectively, thus improving its salt tolerance.

Hydrogen sulfide upregulates the alternative respiratory pathway in mangrove plant Avicennia marina to attenuate waterlogging-induced oxidative stress and mitochondrial damage in a calcium-dependent manner.

Hydrogen sulfide (H2 S) is considered to mediate plant growth and development. However, whether H2 S regulates the adaptation of mangrove plant to intertidal flooding habitats is not well understood. In this study, sodium hydrosulfide (NaHS) was used as an H2 S donor to investigate the effect of H2 S on the responses of mangrove plant Avicennia marina to waterlogging. The results showed that 24-h waterlogging increased reactive oxygen species (ROS) and cell death in roots. Excessive mitochondrial ROS accumulation is highly oxidative and leads to mitochondrial structural and functional damage. However, the application of NaHS counteracted the oxidative damage caused by waterlogging. The mitochondrial ROS production was reduced by H2 S through increasing the expressions of the alternative oxidase genes and increasing the proportion of alternative respiratory pathway in the total mitochondrial respiration. Secondly, H2 S enhanced the capacity of the antioxidant system. Meanwhile, H2 S induced Ca2+ influx and activated the expression of intracellular Ca2+ -sensing-related genes. In addition, the alleviating effect of H2 S on waterlogging can be reversed by Ca2+ chelator and Ca2+ channel blockers. In conclusion, this study provides the first evidence to explain the role of H2 S in waterlogging adaptation in mangrove plants from the mitochondrial aspect.

Physiological and transcriptomic analysis dissects the molecular mechanism governing meat quality during postmortem aging in Hu sheep (Ovis aries).

Introduction: Hu sheep, known for its high quality and productivity, lack fundamental scientific research in China. Methods: This study focused on the effects of 24 h postmortem aging on the meat physiological and transcriptomic alteration in Hu sheep. Results: The results showed that the 24 h aging process exerts a substantial influence on the mutton color, texture, and water content as compared to untreated group. Transcriptomic analysis identified 1,668 differentially expressed genes. Functional enrichment analysis highlighted the importance of glycolysis metabolism, protein processing in endoplasmic reticulum, and the FcγR-mediated phagocytosis pathway in mediating meat quality modification following postmortem aging. Furthermore, protein-protein interaction analysis uncovered complex regulatory networks involving glycolysis, the MAPK signaling pathway, protein metabolism, and the immune response. Discussion: Collectively, these findings offer valuable insights into the molecular mechanisms underlying meat quality changes during postmortem aging in Hu sheep, emphasizing the potential for improving quality control strategies in mutton production.

Cadmium promotes the absorption of ammonium in hyperaccumulator Solanum nigrum L. mediated by ammonium transporters and aquaporins.

Cadmium (Cd) is a toxic heavy metal affecting the normal growth of plants. Nitrate (NO3-) and ammonium (NH4+) are the primary forms of inorganic nitrogen (N) absorbed by plants. However, the mechanism of N absorption and regulation under Cd stress remains unclear. This study found that: (1) Cd treatment affected the biomass, root length, and Cd2+ flux in Solanum nigrum seedling roots. Specifically, 50 µM Cd significantly inhibited NO3- influx while increased NH4+ influx compared with 0 and 5 µM Cd treatments measured by non-invasive micro-test technology. (2) qRT-PCR analysis showed that 50 µM Cd inhibited the expressions of nitrate transporter genes, SnNRT2;4 and SnNRT2;4-like, increased the expressions of ammonium transporter genes, SnAMT1;2 and SnAMT1;3, in the roots. (3) Under NH4+ supply, 50 µM Cd significantly induced the expressions of the aquaporin genes, SnPIP1;5, SnPIP2;7, and SnTIP2;1. Our results showed that 50 µM Cd stress promoted NH4+ absorption by up-regulating the gene expressions of NH4+ transporter and aquaporins, suggesting that high Cd stress can affect the preference of N nutrition in S. nigrum.

Exogenous hydrogen sulfide mediates Na+ and K+ fluxes of salt gland in salt-secreting mangrove plant Avicennia marina.

Hydrogen sulfide (H2S), is a crucial biological player in plants. Here, we primarily explored the interaction between sodium hydrosulfide (NaHS, a H2S donor) and the fluxes of Na+ and K+ from the salt glands of mangrove species Avicennia marina (Forsk.) Vierh. with non-invasive micro-test technology (NMT) and quantitative real-time PCR (qRT-PCR) approaches under salinity treatments. The results showed that under 400-mM NaCl treatment, the addition of 200-µM NaHS markedly increased the quantity of salt crystals in the adaxial epidermis of A. marina leaves, accompanied by an increase in the K+/Na+ ratio. Meanwhile, the endogenous content of H2S was dramatically elevated in this process. The NMT result revealed that the Na+ efflux was increased from salt glands, whereas K+ efflux was decreased with NaHS application. On the contrary, the effects of NaHS were reversed by H2S scavenger hypotaurine (HT), and DL-propargylglycine (PAG), an inhibitor of cystathionine-γ-lyase (CES, a H2S synthase). Moreover, enzymic assay revealed that NaHS increased the activities of plasma membrane and tonoplast H+-ATPase. qRT-PCR analysis revealed that NaHS significantly increased the genes transcript levels of tonoplast Na+/H+ antiporter (NHX1), plasma membrane Na+/H+ antiporter (SOS1), plasma membrane H+-ATPase (AHA1) and tonoplast H+-ATPase subunit c (VHA-c1), while suppressed above-mentioned gene expressions by the application of HT and PAG. Overall, H2S promotes Na+ secretion from the salt glands of A. marina by up-regulating the plasma membrane and tonoplast Na+/H+ antiporter and H+-ATPase.

Effects of shrimp pond effluent on functional traits and functional diversity of mangroves in Zhangjiang Estuary.

In recent years, the scale of shrimp ponds has rapidly increased adjacent to mangrove forests. Discharge of shrimp pond effluent has led to degradation of the surrounding environment and reduction of biodiversity in the estuary. But it remains poorly understood how shrimp pond effluent affects functional traits and functional diversity of mangroves. We sampled roots, stems and leaves of Kandelia obovata and other mangrove plants, as well as sediments and pore water from shrimp pond effluent polluted area (P) and clean area (control area, C) in Zhangjiang Estuary in southeast coast of China. Twenty plant functional traits and six functional diversity indices were analyzed to explore the effects of shrimp pond effluent on individual plants and mangrove communities. The results showed that the discharge of shrimp pond effluent significantly affected the nutrient content in soils and pore water, for example, sediment NH4+ and NO3- concentration increased from 0.26 ± 0.06 to 0.77 ± 0.29 mg/g and from 0.05 ± 0.03 to 0.16 ± 0.05 mg/g, respectively, when comparing the C and P site. Furthermore, some mangrove plant functional traits such as plant height, diameter at breast height, canopy thickness and specific leaf area were significantly increased by the effluent discharge. Functional diversity in the polluted area reduced as a whole compared to the control area. In particular, ammonium and nitrate nitrogen input is the main reason to induce the changes of plant functional traits and functional diversity. Besides, the community structure changed from functional differentiation to functional convergence after shrimp pond effluent discharge. In addition, the long-term shrimp pond effluent discharge may lead to the ecological strategy shift of K. obovata, while different organs may adopt different ways of nutrient uptake and growth strategies in the face of effluent disturbance. In conclusion, pollution from shrimp pond does affect the functional traits of mangrove plants and functional diversity of mangrove community. These results provide strong evidence to assess the impact of effluent discharges on mangrove plants and provide theoretical basis for conservation and sustainable development of mangroves.

Ammonium has stronger Cd detoxification ability than nitrate by reducing Cd influx and increasing Cd fixation in Solanum nigrum L.

Cadmium (Cd) is a harmful heavy metal that affects the growth and development of plants. Nitrogen (N) is an essential nutrient for plants, and appropriate N management can improve Cd tolerance. The aim of our study was to explore the effects of different forms of N on the molecular and physiological responses of the hyperaccumulator Solanum nigrum to Cd toxicity. Measurement of biomass, photosynthetic parameters, and Cd2+ fluxes using non-invasive micro-test technique, Cd fluorescent dying, biochemical methods and quantitative real-time PCR analysis were performed in our study. Our results showed that ammonium (NH4+) has stronger Cd detoxification ability than nitrate (NO3-), which are likely attributed to the following three reasons: (1) NH4+ decreased the influx and accumulation of Cd2+ by regulating the transcription of Cd transport-related genes; (2) the ameliorative effects of NH4+ were accompanied by the increased retention of Cd in the cell walls of roots; and (3) NH4+ up-regulated SnExp expression.

Source or sink? A study on the methane flux from mangroves stems in Zhangjiang estuary, southeast coast of China.

Mangrove ecosystems are an important component of "blue carbon". However, it is not clear whether the stems play roles in the CH4 budget of mangrove ecosystems. This study investigated the CH4 emission from mangrove stems and its potential driving factors. We set up six sample plots in the Zhangjiang Estuary National Mangrove Nature Reserve, where Kandelia obovata, Avicennia marina and Aegiceras corniculata are the main mangrove tree species. Soil properties such as total carbon content, redox potential and salinity were determined in each plot. The dynamic chamber method was used to measure mangrove stems and soil CH4 fluxes. Combined field survey results with Principal Component Analysis (PCA) of soil properties, we divided the six plots into two sites (S1 and S2) to perform statistical analyses of stem CH4 fluxes. Then the CH4 fluxes from mangrove tree stems and soil were further scaled up to the ecosystem level through the mapping model. Under different backgrounds of soil properties, salinity and microbial biomass carbon were the main factors modified soil CH4 fluxes in the two sites, and further affected the stem CH4 fluxes of mangroves. The soil of both sites are sources of CH4, and the soil CH4 emission of S2 was about twice higher than that of S1. Results of upscaling model showed that mangrove stems in S1 were CH4 sinks with -105.65 g d-1. But stems in S2 were CH4 sources around 1448.24 g d-1. Taken together, our results suggested that CH4 emission from mangrove soils closely depends on soils properties. And mangrove stems were found to act as both CH4 sources and CH4 sinks depend on soil CH4 production. Therefore, when calculating the CH4 budget of the mangrove ecosystem, the contribution of mangrove plant stems cannot be ignored.

Proteomic analysis reveals the protective role of exogenous hydrogen sulfide against salt stress in rice seedlings.

Hydrogen sulfide (H2S) is an important gaseous signal molecule which participates in various abiotic stress responses. However, the underlying mechanism of H2S associated salt tolerance remains elusive. In this study, sodium hydrosulfide (NaHS, donor of H2S) was used to investigate the protective role of H2S against salt stress at the biochemical and proteomic levels. Antioxidant activity and differentially expressed proteins (DEPs) of rice seedlings treated by NaCl or/and exogenous H2S were investigated by the methods of biochemical approaches and comparative proteomic analysis. The protein-protein interaction (PPI) analysis was used for understanding the interaction networks of stress responsive proteins. In addition, relative mRNA levels of eight selected identified DEPs were analyzed by quantitative real-time PCR. The result showed that H2S alleviated oxidative damage caused by salt stress in rice seedling. The activities of some antioxidant enzymes and glutathione metabolism were mediated by H2S under salt stress. Proteomics analyses demonstrated that NaHS regulated antioxidant related proteins abundances and affected related enzyme activities under salt stress. Proteins related to light reaction system (PsbQ domain protein, plastocyanin oxidoreductase iron-sulfur protein), Calvin cycle (phosphoglycerate kinase, sedoheptulose-1,7-bisphosphatase precursor, ribulose-1,5-bisphosphate carboxylase/oxygenase) and chlorophyll biosynthesis (glutamate-1-semialdehyde 2,1-aminomutase, coproporphyrinogen III oxidase) are important for NaHS against salt stress. ATP synthesis related proteins, malate dehydrogenase and 2, 3-bisphosphoglycerate-independent phosphoglycerate mutase were up-regulated by NaHS under salt stress. Protein metabolism related proteins and cell structure related proteins were recovered or up-regulated by NaHS under salt stress. The PPI analysis further unraveled a complicated regulation network among above biological processes to enhance the tolerance of rice seedling to salt stress under H2S treatment. Overall, our results demonstrated that H2S takes protective roles in salt tolerance by mitigating oxidative stress, recovering photosynthetic capacity, improving primary and energy metabolism, strengthening protein metabolism and consolidating cell structure in rice seedlings.

Unraveling hydrogen sulfide-promoted lateral root development and growth in mangrove plant Kandelia obovata: insight into regulatory mechanism by TMT-based quantitative proteomic approaches.

Mangroves are the main intertidal ecosystems with varieties of root types along the tropical and subtropical coastlines around the world. The typical characteristics of mangrove habitats, including the abundant organic matter and nutrients, as well as the strong reductive environment, are favor for the production of hydrogen sulfide (H2S). H2S, as a pivotal signaling molecule, has been evidenced in a wide variety of plant physiological and developmental processes. However, whether H2S functions in the mangrove root system establishment is not clear yet. Here, we reported the possible role of H2S in regulation of Kandelia obovata root development and growth by tandem mass tag (TMT)-based quantitative proteomic approaches coupled with bioinformatic methods. The results showed that H2S could induce the root morphogenesis of K. obovata in a dose-dependent manner. The proteomic results successfully identified 8075 proteins, and 697 were determined as differentially expressed proteins. Based on the functional enrichment analysis, we demonstrated that H2S could promote the lateral root development and growth by predominantly regulating the proteins associated with carbohydrate metabolism, sulfur metabolism, glutathione metabolism and other antioxidant associated proteins. In addition, transcriptional regulation and brassinosteroid signal transduction associated proteins also act as important roles in lateral root development. The protein-protein interaction analysis further unravels a complicated regulation network of carbohydrate metabolism, cellular redox homeostasis, protein metabolism, secondary metabolism, and amino acid metabolism in H2S-promoted root development and growth of K. obovata. Overall, our results revealed that H2S could contribute to the morphogenesis of the unique root system of mangrove plant K. obovata, and play a positive role in the adaption of mangrove plants to intertidal habitats.

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In order to reveal the photosynthetic characteristics and the mechanism about accumulation and distribution of photosynthetic assimilation products under different light environments, potted tobacco was cultivated in a climate control chamber, there were three light intensity treatments (shading: (400±15)-(500±15) µmol·m-2·s-1; natural light: (800±15)-(1000±15) µmol·m-2·s-1; high light: (1500±15)-(1800±15) µmol·m-2·s-1). The results showed that with the decrease of light intensity, the biomass and root allocation, the net photosynthetic rate (Pn), stomata conductance (gs) and transpiration rate (Tr) were decreased, but the intercellular CO2 concentration (Ci) increased. The maximum net photosynthetic rate (Amax), light saturation point (LSP), light saturation point (LCP) and dark respiration rate (Rd) reached the maximum level under high light condition while the apparent quantum efficiency was higher in low light environment. The effects of light intensity on the absorption, accumulation and distribution of 13C in tobacco were significant, with a reduced proportion of enriched 13C distributed to the root under shading. The changes in the external light environment not only significantly affected the photosynthetic cha-racteristics and biomass accumulation of tobacco leaves, but also the distribution pattern of photosynthetic carbon in tobacco plant-soil system.

The liver tissue bank and clinical database in China.

To develop a standardized and well-rounded material available for hepatology research, the National Liver Tissue Bank (NLTB) Project began in 2008 in China to make well-characterized and optimally preserved liver tumor tissue and clinical database. From Dec 2008 to Jun 2010, over 3000 individuals have been enrolled as liver tumor donors to the NLTB, including 2317 cases of newly diagnosed hepatocellular carcinoma (HCC) and about 1000 cases of diagnosed benign or malignant liver tumors. The clinical database and sample store can be managed easily and correctly with the data management platform used. We believe that the high-quality samples with detailed information database will become the cornerstone of hepatology research especially in studies exploring the diagnosis and new treatments for HCC and other liver diseases.