Статті в журналах з теми "Mangrove plants, Effect of salt on"

ABSTRACT The mangrove forest is important in shoreline stabilization and as a nursery for many open ocean species. Complex adaptations of the plants for existence in anaerobic conditions and salt water make the mangrove forest highly vulnerable to oil pollution. Clearly, advance protection of mangrove forests is the optimal solution to minimize oil pollution damage. The more difficult question remains: What can be done to minimize the impact of oil that has entered fringe mangrove forests? Active flushing of mangroves and passive oil collection by absorbents are moderately successful in minimizing oil contact with mangroves.

Mangrove forests are increasingly threatened by plant invasions worldwide, but some mangrove species are invasive and threaten salt marsh and native mangrove ecosystems. The southern coast of China is invaded by the cordgrass Spartina alterniflora, and the mangrove Sonneratia apetala, providing a model system for studying the processes and mechanisms through which non-native species establish and spread. We used a transplant experiment to test the overarching hypothesis that native herbivores and plants provided biotic resistance against invasion by S. apetala, and that the importance of these factors would vary geographically. Survival of transplanted mangrove seedlings was lower in Zhangjiang Estuary (23°55′ N) than in Leizhou Bay (20°56′ N), and varied with species and habitats. S. apetala had higher survival and growth rates than native mangroves at both sites, and S. apetala grew taller than the S. alterniflora canopy at Leizhou Bay in only two growing seasons. In contrast, native mangroves grew poorly in S. alterniflora. Grazing by rodents and insects suppressed the growth and survival of Kandelia obovata and Avicennia marina in Zhangjiang Estuary and Leizhou Bay, but had little effect on S. apetala. Competition with vegetation (S. alterniflora and native mangroves) exacerbated the reduced survival of native mangroves, and these effects varied across study sites. Low survival of non-native S. apetala in vegetated habitats at Zhangjiang Estuary was likely due to a synergistic effect of low winter temperatures and low light intensity. Escape from herbivory (the opposite to biotic resistance) and fast growth may drive the quick expansion of non-native S. apetala in China. Rapid encroachment of S. apetala may transform the native mangrove forests and monospecific intertidal Spartina grasslands into non-native mangrove forests in the southern coast of China.

ABSTRACT As part of an ongoing study of the long-term effects of the Zoe Colocotroni oil spill of March 17, 1973, in Bahia Sucia, Puerto Rico, the effects of the remaining oil on the red mangrove trees in the impact area were investigated. This study involved four sampling trips to the spill site and a reference area outside the spill zone between April 1979 and April 1981. The present study was based on the observation that stressed mangrove trees in the heavily oil-affected areas had a similar appearance to trees exhibiting stress due to hypersalinity in unoiled areas. The working hypothesis was that petroleum hydrocarbons induce stress in salt-excluding plants such as red mangroves by disrupting the ability of the roots to exclude ions from seawater. Measurements of sodium (the principal seawater cation) and potassium (a major physiological ion) were made on leaf samples taken from trees from oil-affected areas and reference areas. Sediment core samples were taken from the root zone of the trees sampled at each site and analyzed for hydrocarbons by gravimetric and gas chromatographic methods. The results show a relationship between sediment hydrocarbon concentration and the ratio of sodium to potassium for mangrove leaves sampled at each site. The results show that for trees exposed at the root zone to the least weathered oil, the values of this ratio for the leaves had the largest values, reflecting an oil-induced impairment of the salt (Na) exclusion mechanism. This approach to measuring the physiological health of mangrove trees at an oil spill site offers a potentially useful means of documenting oil stress and recovery from oil stress in salt-excluding halophytes. Because oil stress in mangroves appears to be a root membrane-directed effect, there may be a “window in time” between initial oil impact and plant damage, as oil penetrates the sediments, during which mitigation measures could be taken.

BACKGROUND: Mangrove forest is a typical forest found along the coast or river mouth which is affected by tides and salinity. Although polyisoprenoid was widespread in the plant kingdom, the physiological roles of these compounds are not well understood, especially from mangrove plants. It is therefore essential to characterize the polyisoprenoid content under abiotic stress. AIM: This study aimed to determine the effect of salinity and subsequent fresh water change on polyisoprenoids concentration in Bruguiera cylindrica seedlings. METHODS: Bruguiera cylindrica planted in a greenhouse for three months under various salinity concentrations. After three months grew under variable salinity, these seedlings were then divided into two treatment groups, and grown for another three months: one continuously in a salt solution and another in fresh water to relieve salt stress. The leaves and roots of B. cylindrica seedlings were harvested after six months of cultivation. The leaves and roots of B. cylindrica seedlings were extracted for polyisoprenoids content and composition analyzed using two-dimensional thin layer chromatography. RESULTS: Polyisoprenoids composition under salinity and subsequent fresh water with dominating dolichols (more than 90%) were found in leaves and roots of B. cylindrica seedlings referring type I of polyisoprenoid composition. The carbon chain length of dolichols located in the leaves and roots were ranging from C75–C100 and C75–C105, respectively. CONCLUSION: Dolichol dominated over polyprenol both in B. cylindrical leaves and roots under salinity and subsequent relief supported the previous finding on the predominance dolichols over polyprenols in mangrove plants. The present study suggested the significance of dolichols in the adaptation to cope with salt stress and or water stress.

Mangrove ecosystems are vulnerable to rising sea levels as the plants are exposed to high salinity and tidal submergence. The ways in which these plants respond to varying salinities, immersion depths, and levels of light irradiation are poorly studied. To understand photosynthesis in response to salinity and submergence in mangroves acclimated to different tidal elevations, two-year-old seedlings of two native mangrove species, Kandelia obovata and Rhizophora stylosa, were treated at different salinity concentrations (0, 10, and 30 part per thousand, ppt) with and without immersion conditions under fifteen photosynthetic photon flux densities (PPFD μmol photon·m−2·s−1). The photosynthetic capacity and the chlorophyll fluorescence (ChlF) parameters of both species were measured. We found that under different PPFDs, electron transport rate (ETR) induction was much faster than photosynthetic rate (Pn) induction, and Pn was restricted by stomatal conductance (Gs). The Pn of the immersed K. obovata plants increased, indicating that this species is immersed-tolerant, whereas the Pn level of the R. stylosa plants is salt-tolerant with no immersion. All of the plants treated with 30 ppt salinity exhibited lower Pn but higher non-photochemical quenching (NPQ) and heat quenching (D) values, followed by increases in the excess energy and photoprotective effects. Since NPQ or D can be easily measured in the field, these values provide a useful ecological monitoring index that may provide a reference for mangrove restoration, habitat creation, and ecological monitoring.

Salt stress is a major increasing threat to global agriculture. Pongamia (Millettia pinnata), a semi-mangrove, is a good model to study the molecular mechanism of plant adaptation to the saline environment. Calcium signaling pathways play critical roles in the model plants such as Arabidopsis in responding to salt stress, but little is known about their function in Pongamia. Here, we have isolated and characterized a salt-responsive MpCML40, a calmodulin-like (CML) gene from Pongamia. MpCML40 protein has 140 amino acids and is homologous with Arabidopsis AtCML40. MpCML40 contains four EF-hand motifs and a bipartite NLS (Nuclear Localization Signal) and localizes both at the plasma membrane and in the nucleus. MpCML40 was highly induced after salt treatment, especially in Pongamia roots. Heterologous expression of MpCML40 in yeast cells improved their salt tolerance. The 35S::MpCML40 transgenic Arabidopsis highly enhanced seed germination rate and root length under salt and osmotic stresses. The transgenic plants had a higher level of proline and a lower level of MDA (malondialdehyde) under normal and stress conditions, which suggested that heterologous expression of MpCML40 contributed to proline accumulation to improve salt tolerance and protect plants from the ROS (reactive oxygen species) destructive effects. Furthermore, we did not observe any measurable discrepancies in the development and growth between the transgenic plants and wild-type plants under normal growth conditions. Our results suggest that MpCML40 is an important positive regulator in response to salt stress and of potential application in producing salt-tolerant crops.

Mangroves, which dominate tropical intertidal zones and estuaries, are salt-tolerant plants that reproduce through propagules, which are little reproductive units. Because mangroves can grow in seawater, microorganisms capable of interacting with the host and exerting beneficial effects under salt stress should be present. Three mangrove areas in Saudi Arabia yielded sixteen bacterial and actinobacterial species (Thuwal, Jazan, and Farasan islands). The microbial profile of the examined regions was analysed using a 16S rRNA-based metagenomics technique. Based on 16s rRNA and phylogenetic analysis, the selected isolates were identified. The bacterial extracts were tested for antibacterial activity and pharmacological characteristics. The 16S RNA-based metagenomics revealed that eight phyla of bacteria were found in the same region: Bacteroidetes, Firmicutes, Chloroflexi, Gemmatimonadetes, Proteobacteria, Acidobacteria, Actinobacteria, and Planctomycetes. Bacteroidetes, was the most common phyla, followed by Firmicutes and Proteobacteria. The Bacillus genus was found in the majority of samples, followed by Streptomyces. The pharmacological characteristics of the bacterial extracts were assessed. Antibacterial testing revealed that 13 of 16 bacterial extracts were having antimicrobial activities against at least one of the microorganisms examined.

Facultative halophyte Kandelia obovata plants were exposed to mild (1.5% NaCl) and severe (3% NaCl) salt stress with or without sodium nitroprusside (SNP; 100 µM; a NO donor), hemoglobin (Hb, 100 µM; a NO scavenger), or Nω-nitro-L-arginine methyl ester (L-NAME, 100 µM; a NO synthase inhibitor). The plants were significantly affected by severe salt stress. They showed decreases in seedling growth, stomatal conductance, intercellular CO2 concentration, SPAD value, photosynthetic rate, transpiration rate, water use efficiency, and disrupted antioxidant defense systems, overproduction of reactive oxygen species, and visible oxidative damage. Salt stress also induced ion toxicity and disrupted nutrient homeostasis, as indicated by elevated leaf and root Na+ contents, decreased K+ contents, lower K+/Na+ ratios, and decreased Ca contents while increasing osmolyte (proline) levels. Treatment of salt-stressed plants with SNP increased endogenous NO levels, reduced ion toxicity, and improved nutrient homeostasis while further increasing Pro levels to maintain osmotic balance. SNP treatment also improved gas exchange parameters and enhanced antioxidant enzymes’ activities (catalase, ascorbate peroxidase, monodehydroascorbate reductase, and dehydroascorbate reductase). Treatment with Hb and l-NAME reversed these beneficial SNP effects and exacerbated salt damage, confirming that SNP promoted stress recovery and improved plant growth under salt stress.

As a dominant mangrove species, Kandelia obovata is distributed in an intertidal marsh with an active H2S release. Whether H2S participates in the salt tolerance of mangrove plants is still ambiguous, although increasing evidence has demonstrated that H2S functions in plant responses to multiple abiotic stresses. In this study, NaHS was used as an H2S donor to investigate the regulatory mechanism of H2S on the salt tolerance of K. obovata seedlings by using a combined physiological and proteomic analysis. The results showed that the reduction in photosynthesis (Pn) caused by 400 mM of NaCl was recovered by the addition of NaHS (200 μM). Furthermore, the application of H2S enhanced the quantum efficiency of photosystem II (PSII) and the membrane lipid stability, implying that H2S is beneficial to the survival of K. obovata seedlings under high salinity. We further identified 37 differentially expressed proteins by proteomic approaches under salinity and NaHS treatments. Among them, the proteins that are related to photosynthesis, primary metabolism, stress response and hormone biosynthesis were primarily enriched. The physiological and proteomic results highlighted that exogenous H2S up-regulated photosynthesis and energy metabolism to help K. obovata to cope with high salinity. Specifically, H2S increased photosynthetic electron transfer, chlorophyll biosynthesis and carbon fixation in K. obovata leaves under salt stress. Furthermore, the abundances of other proteins related to the metabolic pathway, such as antioxidation (ascorbic acid peroxidase (APX), copper/zinc superoxide dismutase (CSD2), and pancreatic and duodenal homeobox 1 (PDX1)), protein synthesis (heat-shock protein (HSP), chaperonin family protein (Cpn) 20), nitrogen metabolism (glutamine synthetase 1 and 2 (GS2), GS1:1), glycolysis (phosphoglycerate kinase (PGK) and triosephosphate isomerase (TPI)), and the ascorbate–glutathione (AsA–GSH) cycle were increased by H2S under high salinity. These findings provide new insights into the roles of H2S in the adaptations of the K. obovata mangrove plant to high salinity environments.

Abstract L-asparaginase is an enzyme that converts L-asparagine to L-aspartate and ammonia. L-asparaginase is recommended as medical treatment of Acute Lymphoblastic Leukemia (ALL) and other malignant cancers. Bacteria are the most effective source of L-asparaginase as they can easily be cultured so that the enzyme can be extracted and purified. The commercial L-asparaginase is now available from bacterial but has many side effects for the patients. Therefore, the alternative source of this enzyme is highly necessary to be explored for a more effective and safer future production of L-asparaginase. For this reason, this study was carried out to investigate the endophytic bacteria producing L-asparaginase from mangrove Rhizophora mucronata. The samples of the mangrove plants, i.e., roots, stems, and leaves, were surface sterilized with alcohol and sodium hypochlorite. Endophytic bacteria were screened for L-asparaginase production using the rapid plate assay on Minimal Salt Medium with L-Asparagine as a substrate. Asparaginase-producing endophytes were detected by a pink zone formation on the agar, indicating the hydrolysis of asparagine into aspartic acid and ammonia, which changed the phenol red dye indicator from yellow (acidic conditions) to pink (alkaline conditions). The results revealed that out of the mangroves taken from the seaside, i.e., between sea and land, there were six bacterial colonies from the roots, 12 bacterial colonies from the stems, and 20 bacterial colonies from the leaves, each showing positive L-asparaginase. The isolated bacteria had varied colony morphology and different ability to produce the L-asparaginase enzyme.

Sea level rise and the increasing landward intrusion of storm surges pose the threat of replacement of salinity-intolerant vegetation of important coastal habitats by salinity-tolerant vegetation. Therefore, a means is needed to better understand the processes that influence this vegetation shift and to aid in the management of coastal resources. For this purpose, a hydrology–salinity–vegetation model known as MANTRA was developed by coupling a spatially explicit model (MANHAM) for simulation of vegetation community dynamics along coastal salinity gradients with SUTRA, a USGS groundwater flow and transport model. MANTRA has been used to project possible future changes in Coot Bay Hammock in southern Florida under conditions of gradually rising sea level and storm surges. The simulation study concluded that feasibility exists of a regime shift from hardwood hammocks to mangroves subject to a few conditions, namely severe damage to the existing hammock after a storm surge and a sufficiently persistent high salinity condition and high input of mangrove seedlings. Early detection of salinity stress in vegetation may facilitate sustainable conservation measures being applied. It has been shown that the δ18O value of water in the xylem of trees can be used as a surrogate for salinity in the rooting zone of plants, which is difficult to measure directly. Hence, the model MANTRA is revised into MANTRA-O18 by including the δ18O of the tree xylem dynamics. A simulation study by MANTRA-O18 shows that effects of increasing salinization can be detected many years before the salinity-intolerant trees are threatened with replacement.

Naturally grown two-month-old seedlings of Avicennia marina contain high concentrations of Na+ and Cl-.+ Our NMR studies revealed an accumulation of glycinebetaine, asparagine and stachyose in A. marina. The highest concentration of glycinebetaine was observed in young leaves, while the distribution of stachyose was restricted in stems and roots. A sparagine comprised more than 96% of total free amino acids in roots and 84% in leaves from two-year-old plants. Little or no accumulation of proline or polyols, which are proposed as compatible solutes in other plants, could be detected in A. marina. The activities of phosphofructokinase, pyrophosphate:fructose-6-phosphate 1-phosphotransferase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase (decarboxylating), phosphoenolpyruvate carboxylase and NAD:malate dehydrogenase from young leaves of A. marina were inhibited by NaCl, while the activity of fructose-1,6-bisphosphate aldolase was activated by 50-200 m M NaCl. There was little or no effect of high concentrations (up to 500 mᴍ ) of glycinebetaine on the activities of any of these enzymes. No significant protection by glycinebetaine was detected against NaCl inhibition of these enzymatic activities. Based on these results, possible mechanisms for the salt-resistance of A. marina cells are discussed.

The effects of organic osmolytes synthesised and accumulated by red algae from mangrove habitats were investigated on the in vitro activities of two major enzymes, one of the citric acid cycle (malate dehydrogenase, MDH) and one of the oxidative pentose phosphate pathway (glucose-6- phosphate dehydrogenase, G6PDH). These enzymes were extracted from the mangrove algae Bostrychia tenella, Caloglossa leprieurii, Catenella nipae and Stictosiphonia hookeri. In each case, activity of the enzymes was inhibited with increasing NaCl concentrations up to 600 mM . In contrast, equimolar concentrations of mannitol (the major osmolyte in C. leprieurii), sorbitol (the major osmolyte in B. Tenella and S. hookeri) and a heteroside mixture (of which floridoside is the major osmolyte in C. nipae) did not inhibit enzyme function. Dulcitol, the second most important organic osmolyte in B. tenella, exerted no negative effect at its maximum solubility of 180 rnM on the salt-sensitive MDH. These data are all consistent with the proposed function of these organic compounds as compatible solutes.

Phytophthora species are potent pathogens that can devastate terrestrial plants, causing billions of dollars of damage yearly to agricultural crops and harming fragile ecosystems worldwide. Yet, virtually nothing is known about the distribution and pathogenicity of their marine relatives. This is surprising, as marine plants form vital habitats in coastal zones worldwide (i.e. mangrove forests, salt marshes, seagrass beds), and disease may be an important bottleneck for the conservation and restoration of these rapidly declining ecosystems. We are the first to report on widespread infection of Phytophthora and Halophytophthora species on a common seagrass species, Zostera marina (eelgrass), across the northern Atlantic and Mediterranean. In addition, we tested the effects of Halophytophthora sp. Zostera and Phytophthora gemini on Z. marina seed germination in a full-factorial laboratory experiment under various environmental conditions. Results suggest that Phytophthora species are widespread as we found these oomycetes in eelgrass beds in six countries across the North Atlantic and Mediterranean. Infection by Halophytophthora sp . Zostera, P. gemini , or both, strongly affected sexual reproduction by reducing seed germination sixfold. Our findings have important implications for seagrass ecology, because these putative pathogens probably negatively affect ecosystem functioning, as well as current restoration and conservation efforts.

The distribution of vegetation communities on floodplains within Kakadu National Park, in tropical northern Australia, is related to micro-topography and, therefore, water depth and duration of flooding. Floodplains of the Kakadu Region, because of their proximity to the coast, are most vulnerable to the impacts of climate change, with saltwater intrusion, as a result of sea-level rise, being a serious risk. Our main objectives were to determine the variability of the distribution of plant communities on the floodplains and understand the potential risk of increased saltwater intrusion to these communities. We present data on the natural salinity-tolerance range of selected floodplain plants and discuss the likely effects of saltwater intrusion on floodplain plant distributions and productivity. The results of change analysis using high spatial-resolution satellite data showed the importance of the variation of water availability in determining patterns of plant communities. Hydrodynamic modelling suggests that sea level rises will result in 40% of the floodplain transformed into saline habitats by 2070. The most obvious effect of this would be the conversion of the freshwater vegetation to salt-tolerant mangroves and other salt-marsh plants, with a concomitant change in animals and their use of these areas.

Root-associated microbial communities are very important for biogeochemical cycles in wetland ecosystems and help to elaborate the mechanisms of plant invasions. In the estuary of Jiulong River (China), Spartina alterniflora has widely invaded Kandelia obovata-dominated habitats, offering an opportunity to study the influence of root-associated bacteria. The community structures of endophytic and rhizosphere bacteria associated with selected plant species were investigated using the barcoded Illumina paired-end sequencing technique. The diversity indices of bacteria associated with the roots of S. alterniflora were higher than those of the transition stands and K. obovata monoculture. Using principal coordinate analysis with UniFrac metrics, the comparison of β-diversity showed that all samples could be significantly clustered into 3 major groups, according to the bacteria communities of origin. Four phyla, namely Proteobacteria, Bacteroidetes, Chloroflexi, and Firmicutes, were enriched in the rhizoplane of both salt marsh plants, while they shared higher abundances of Cyanobacteria and Proteobacteria among endophytic bacteria. Members of the phyla Spirochaetes and Chloroflexi were found among the endophytic bacteria of S. alterniflora and K. obovata, respectively. One of the interesting findings was that endophytes were more sensitive in response to plant invasion than were rhizosphere bacteria. With linear discriminate analysis, we found some predominant rhizoplane and endophytic bacteria, including Methylococcales, Pseudoalteromonadacea, Clostridium, Vibrio, and Desulfovibrio, which have the potential to affect the carbon, nitrogen, and sulfur cycles. Thus, the results provide clues to the isolation of functional bacteria and the effects of root-associated microbial groups on S. alterniflora invasions.

The functional diversity of the New Caledonian mangrove sediments was examined, observing the distribution of fungal dye-decolorizing peroxidases (DyPs), together with the complete biochemical characterization of the main DyP. Using a functional metabarcoding approach, the diversity of expressed genes encoding fungal DyPs was investigated in surface and deeper sediments, collected beneath either Avicennia marina or Rhizophora stylosa trees, during either the wet or the dry seasons. The highest DyP diversity was observed in surface sediments beneath the R. stylosa area during the wet season, and one particular operational functional unit (OFU1) was detected as the most abundant DyP isoform. This OFU was found in all sediment samples, representing 51–100% of the total DyP-encoding sequences in 70% of the samples. The complete cDNA sequence corresponding to this abundant DyP (OFU 1) was retrieved by gene capture, cloned, and heterologously expressed in Pichia pastoris. The recombinant enzyme, called DyP1, was purified and characterized, leading to the description of its physical–chemical properties, its ability to oxidize diverse phenolic substrates, and its potential to decolorize textile dyes; DyP1 was more active at low pH, though moderately stable over a wide pH range. The enzyme was very stable at temperatures up to 50 °C, retaining 60% activity after 180 min incubation. Its ability to decolorize industrial dyes was also tested on Reactive Blue 19, Acid Black, Disperse Blue 79, and Reactive Black 5. The effect of hydrogen peroxide and sea salt on DyP1 activity was studied and compared to what is reported for previously characterized enzymes from terrestrial and marine-derived fungi.

Mangroves form extensive, unique communities in tropical coastal areas and tidal lowlands, dominating 60–75% of tropical shorelines. The aim of the present review is to summarize the current knowledge concerning the mechanisms underlying the most striking feature of these plants—their unique ability to obtain water from the surrounding sea. Mangroves are thought to accomplish this by rejecting potentially harmful salts. Some species actively excrete those salts leaking into the plant by means of specialized salt glands in their leaves. Mangroves are rooted in anaerobic soils, a condition giving rise to the spectacular aerial roots, such as pneumatophores and stilt roots, characteristic of mangroves, that provide oxygen to submerged tissues.We shall also discuss recent studies that have focused on physiological issues in mangroves, such as oscillatory behavior of their stomata, the structure and function of salt glands, and the compatible solutes in their leaves, which balance the osmotic pressure of the seawater. Salinity effects on their germination, distribution, CO2assimilation, respiration, and the functioning of some of their enzymes have also been examined.Finally, we shall draw attention to open questions related to the salt and water regime of mangroves and the underlying mechanisms responsible for their remarkable success in a hostile environment.

Mangrove forests are a group of plants that grow along sub-tropical coastlines that have a special function in an environment containing salt and landscapes such as beaches with anaerobic soil reactions. In general, mangrove forests can be defined as forest types that grow in tidal areas (especially on sheltered beaches, lagoons, and river estuaries) that are flooded by tidal and free from submerged entanglement where the mangrove ecosystem is tolerant of salt.

The leaf ultrastructure of mangrove Kandelia candel (L.) Druce planted in pots under different salinity conditions was compared under a transmission electron microscope (TEM). The results showed that the plasmalemma in plants grown in salinity conditions of 0&permil; treatment (control) and 25&permil; treatment was tightly combined, while in plants with salinity of 50&permil; treatment, the plasmalemma crimpled remarkably and plasmolysis occurred. The nucleus and its two-layer membranes were obvious in control plants. In the case of 25&permil; treatment, the membrane breakdown was observed, nucleoplasm dispersed in cytoplasm, and the electron density of cells was lower than that in control plants. In plants treated with 50&permil; salinity the nucleus collapsed and no structure of the nucleus could be observed. As far as chloroplasts in control plants were concerned, they were oblong with a typical arrangement of grana and stroma thylakoids and one or two grains of starch. However, the chloroplasts in plants treated with 25&permil; salinity were swelling and usually contained more grains of starch and few plastoglobuli. Most chloroplasts had a reduced number of grana, particularly of thylakoids in grana as compared with control plants. The chloroplasts of plants treated with <br />50&permil; salinity had a considerably reduced system of grana and stroma thylakoids, and sometimes they were even defor-<br />med morphologically. They were mixed-up and contained more grains of starch and plastoglobuli. The indistinct structure of mitochondrial cristae was observed only in plants treated with 50&permil; salinity. These showed that mitochondria are cell organs less sensitive to hypersaline conditions than chloroplasts and nucleus, and it was deduced that respiration was more conservative to an environment change than photosynthesis.

Coastal ecosystems have high plant levels, for example mangroves and seagrasses. Mangrove forest is a type of forest located in tidal areas, especially on protected beaches, lagoons, river estuaries that are inundated and free from inundation at low tide, whose plant communities tolerate salt. Gedangan Village is one of the villages in Purwodadi District, which has mangrove areas in Purworejo Regency. This study aims to determine the types of mangrove plants that are useful the Gedangan Village, Purwodadi District, Purworejo Regency, Central Java as medicinal products. The study was conducted using roaming method in the form of observations or field observations in the mangrove area of Gedangan Village, Purwodadi District, Purworejo Regency, Central Java. Based on the research, there were eight (8) types of mangrove plants that were found as medicinal plants in Gedangan Village, namely Rhizophora mucronata, Sonneratia alba, Calotropis gigantea, Nypa fruticans, Acanthus ilicifolius, Hibiscus tiliaceus, Ipomoea pescaprae, and Wedelia biflora. Traditionally, these mangrove species can be used as a medicine items for beri-beri, hepatitis, ulcers, wounds, diarrhea, fever, antibacterial, anti-inflammatory, dizziness, asthma, bronchitis, dyspepsia, leprosy, tumors, diabetes, stomach ache, toothache, thrush, tuberculosis, muscle aches, and eczema.

<p><strong>Herbs are plants with moist stems that grow wild along roadsides or on the unmaintained ground. Herbs are more commonly associated with plants that are harmful to civilization, even though they have numerous uses in life, including traditional medicine, ornamental plants, and contextual instructional resources. People are unaware of herbs and plants categorized as herbs, which is one of the reasons why they are considered hazardous plants. A mangrove forest is a type of forest with a tidal environment; plants that may grow in mangrove forests are characteristic vegetation, notably salt-tolerant plants. The goal of this research was to describe the plant variety in the Muara Aluh-Aluh Mangrove Area. The data was gathered in the 500-meter-long Muara Aluh-Aluh Mangrove Area, which was divided into ten locations separated by 50 meters. A plot of 1 m x 1 m with as many as 4 pieces was made at each place using the line transect method and oriented diagonally. The quantity and varieties of herbaceous plants found in the plot were counted as samples. According to the findings of the study on herbaceous plant diversity in the Muara Aluh-Aluh Mangrove area, there are 21 different varieties of herbaceous plants with a diversity index of 2.66, putting them in the medium diversity category.</strong></p><p><em><strong>Keywords</strong> -<strong> </strong>Diversity, Herbs, Mangroves, Muara Aluh-Aluh</em></p>

Salt marshes and mangroves are currently being affected by rising temperatures. Mangroves thrive below −29° N latitude in Florida, USA, and have a low tolerance for extreme cold events, whereas salt marshes dominate further north. One potential effect of climate change is a reduction in the frequency of extreme cold events, which may lead to mangrove expansion into salt marsh systems. Our research identified sediment proxy indicators of salt marsh and mangrove environments. These indicators were applied to soil cores from intertidal wetlands near the current northern limit of mangrove presence on the east coast of Florida, to determine if mangrove expansion into salt marsh environments has precedence in the deeper past. Our findings suggest that mangrove and salt marsh sediments can be distinguished using a combination of stable carbon isotope ratios of sedimentary organic matter and macroscopic plant fragments, and our results showed that a mangrove stand that we cored established only recently. This result is consistent with other work in the southeastern United States that suggests that mangroves established at the current boreal limit only recently after the end of the Little Ice Age, and that the current mangrove expansion may be fueled by anthropogenic climate change.

The mangrove woodland is a distinct bush or brine habitat characterized by a coastal sedimentation environment in which fine sediment (often rich in organic matter) accumulates in areas protected from high energy waves. Mangrove forests thrive on the coasts of tropical and subtropical regions, including Indonesia. Mangrove forest is a complex ecosystem that has a high diversity of plants, microorganisms, and animals. One of them is the Oil Mangrove (R. apiculata) which grows well on the coast of Lampung. This plant is hard, rich in tannins, and dense, mainly used to make charcoal and firewood. This plant traditionally used to treat diarrhea and nausea. Mangrove plants are tolerant to high salt levels, this special trait is due to the presence of secondary metabolites produced in response to various environmental stresses. Flavonoid compounds, alkaloids, terpenoids and steroids are secondary metabolites produced by mangrove plants. Secondary metabolite compounds from mangroves have bioactivity such as antidiabetic, antimicrobial, antioxidant and anticancer. Exploration of secondary metabolites from mangroves, especially oil mangroves locally named as Bakau Minyak, which are widely grown in the Lempasing area, coastal Lampung for the development of medicinal compounds, has not been intensely reported. This investigation aims to study the potential phytochemicals profile of R. apiculata’s leaf as an antioxidant

Prawn fisheries are among the main sources of income in several tropical countries, where juveniles of many species inhabit estuarine wetlands. Although plants in these wetlands are considered to be essential food sources for juvenile prawns, some studies suggest that wetland producers are of limited importance. In the present study, δ13C and δ15N were used to identify differences in diet between penaeid species and size classes, and to determine if terrestrial wetland producers are important for nutrition. Two estuarine floodplain pools were sampled: one surrounded by mangroves and one surrounded by salt marsh. There were differences in diet between species and size classes. As mangrove δ13C (–29.7 to –26.3‰) was very different from salt marsh δ13C (–16.3 to –15.4‰), the importance of these producers was examined by comparing the isotopic composition of the prawns between sites and by using the IsoSource model. Although aquatic sources were the most important, salt marsh grass was also a significant contributor, supporting the hypothesis that these wetland producers are important for juvenile prawn nutrition. There was no evidence that mangrove material was of major importance for any species, suggesting that mangrove productivity is not the primary reason for the occurrence of penaeid prawns in mangrove habitats.

Elongated and spiral &beta;-1,3-glucan (callose) fibers were obtained by new factors from protoplasts cultured in liquid medium from suspension cultured cells of two salt-tolerant mangrove species; Avicennia alba and Bruguiera sexangula. Differences in salt factor for protoplast-fiber formation were compared with those of the callose fibers developed from protoplasts of non-mangrove tree plants, Larix leptolepis and Betula platyphylla, which high concentrations of divalent cations, Mg2+ (50 mM) or Ca2+ (100 mM), were stimulatory. In the halophilic A. alba protoplasts, whose cell division was stimulated by up to 400 mM NaCl, addition of Mg2+, Ca2+, K+ ions inhibited protoplast-fiber formation. In B. sexangula, protoplast-fibers were rapidly and efficiently formed only by another new factor, electric cell fusion treatment of protoplasts. Spiral fibers developed from mangrove protoplasts were detected under an inverted microscope, and their specific blue-green color for callose after staining with Aniline Blue dye was detected under a fluorescence microscope. Enzymatic certification of callose was further performed with laminarinase, specific for callose, in comparison with cellulase CBH1, specific for cellulose. Differences in sub-structures, fibrils and sub-fibrils of two mangrove protoplast-fibers were analyzed using laser confocal scanning microscopy, atomic force microscopy and image J analysis. Tube-like fine structure was observed using transmission electron microscopy in single protoplast-fiber of B. sexangula selected with a micromanipulator.

This review explores effects of elevated CO2 concentrations on growth in relation to water use and salt balance of halophytic and non-halophytic species. Under saline conditions, the uptake and distribution of sodium and chloride must be regulated to protect sensitive metabolic sites from salt toxicity. Salt-tolerant species exclude most of the salt from the transpiration stream, but the salt flux from a highly saline soil is still considerable. To maintain internal ion concentrations within physiologically acceptable levels, the salt influx to leaves must match the capacities of leaves for salt storage and/or salt export by either retranslocation or secretion from glands. Hence the balance between carbon gain and the expenditure of water in association with salt uptake is critical to leaf longevity under saline conditions. Indeed, one of the striking features of halophytic vegetation, such as mangroves, is the maintenance of high water use efficiencies coupled with relatively low rates of water loss and growth. These low evaporation rates are further reduced under elevated CO2 conditions. This, with increased growth, leads to even higher water use efficiency. Leaves of plants grown under elevated CO2 conditions might be expected to contain lower salt concentrations than those grown under ambient CO2 if salt uptake is coupled with water uptake. However, salt concentrations in shoot tissues are similar in plants grown under ambient and elevated CO2 conditions despite major differences in water use efficiency. This phenomenon occurs in C3 halophytes and in both C3 and C2 non-halophytes. These results imply shoot/root communication in regulation of the salt balance to adjust to environmental factors affecting the availability of water and ions at the roots (salinity) and those affecting carbon gain in relation to water loss at the leaves (atmospheric concentrations of water vapour and carbon dioxide).

In the Cananeia region of southeastern Brazil, Spartina alterniflora marshes colonize tidal flats fringing mangrove woodlands and displaying a zonation typical of monocultures. The pattern observed can be explained by the combined effects of organism resistance to emersion and physical dependence on the plants as habitat. In this context, it is interesting to quantify the aggregation index for the dominant species associated with the salt marsh. A tool which enables us to do it is Taylor´s power law, which combines the mean and the variance distributions of species in a known area. From August 1988 to January 1989, ten random samples were taken monthly from the lower and upper marshes using a 20 cm diameter corer (0.03 m2) at a depth of 10 cm. The five most representative species of the system were selected for further analysis, and for each of these, Taylor´s power law parameters were calculated. Epifaunal species present aggregation indexes approaching randomness. The aggregation indexes for the infaunal species were observed to have consistently high values even in clearly different conditions of population density and availability of organic matter. The smaller number of infauna forms in the lower marsh as compared to the upper marsh does not point to a competitive disadvantage since there is no alteration in b values. For the infauna species only, the value of a shows a sharp decrease from the lower to the upper marsh. Cálculo de los parámetros a y b de la Ley de Poder de Taylor para especies macrobentónicas de marisma En la región de Cananeia al SE de Brasil, el pasto Spartina alterniflora coloniza los bajos de marea de las marismas que rodean los manglares, mostrando una zonación típica de monocultivos. El patrón observado se puede explicar por el efecto combinado de la resistencia de los organismos a la exposición al aire y a la dependencia física de las plantas como hábitat. En este contexto, es interesante cuantificar el índice de agregación para las especies dominantes asociadas a la marisma. Una herramienta que nos permite hacerlo es la Ley de Poder de Taylor, la cual combina las distribuciones promedio y de la varianza de las especies en un área conocida. De agosto de 1988 a enero de 1989, diez muestras mensuales fueron tomadas aleatoriamente de la marisma superior e inferior mediante un nucleador de 20 cm de diámetro (0.03 m2) a una profundidad de 10 cm. Las cinco especies más representativas del sistema fueron elegidas para un análisis posterior y se calcularon los parámetros de la Ley de Poder de Taylor a cada una de ellas. Las especies de epifauna presentaron un índice de agregación cercano a la aleatoriedad. Los índices de agregación de las especies de la endofauna mostraron consistentemente altos valores, incluso en diferentes condiciones de densidad de población y de disponibilidad de materia orgánica. El reducido número de formas de endofauna en la marisma inferior, comparada con la superior, no apunta a una desventaja competitiva, ya que no hay alteraciones en los valores de b. Solo para las especies de endofauna, el valor de a mostró un agudo decremento desde la marisma inferior hacia la superior.

In the Cananeia region of southeastern Brazil, Spartina alterniflora marshes colonize tidal flats fringing mangrove woodlands and displaying a zonation typical of monocultures. The pattern observed can be explained by the combined effects of organism resistance to emersion and physical dependence on the plants as habitat. In this context, it is interesting to quantify the aggregation index for the dominant species associated with the salt marsh. A tool which enables us to do it is Taylor´s power law, which combines the mean and the variance distributions of species in a known area. From August 1988 to January 1989, ten random samples were taken monthly from the lower and upper marshes using a 20 cm diameter corer (0.03 m2) at a depth of 10 cm. The five most representative species of the system were selected for further analysis, and for each of these, Taylor´s power law parameters were calculated. Epifaunal species present aggregation indexes approaching randomness. The aggregation indexes for the infaunal species were observed to have consistently high values even in clearly different conditions of population density and availability of organic matter. The smaller number of infauna forms in the lower marsh as compared to the upper marsh does not point to a competitive disadvantage since there is no alteration in b values. For the infauna species only, the value of a shows a sharp decrease from the lower to the upper marsh. Cálculo de los parámetros a y b de la Ley de Poder de Taylor para especies macrobentónicas de marisma En la región de Cananeia al SE de Brasil, el pasto Spartina alterniflora coloniza los bajos de marea de las marismas que rodean los manglares, mostrando una zonación típica de monocultivos. El patrón observado se puede explicar por el efecto combinado de la resistencia de los organismos a la exposición al aire y a la dependencia física de las plantas como hábitat. En este contexto, es interesante cuantificar el índice de agregación para las especies dominantes asociadas a la marisma. Una herramienta que nos permite hacerlo es la Ley de Poder de Taylor, la cual combina las distribuciones promedio y de la varianza de las especies en un área conocida. De agosto de 1988 a enero de 1989, diez muestras mensuales fueron tomadas aleatoriamente de la marisma superior e inferior mediante un nucleador de 20 cm de diámetro (0.03 m2) a una profundidad de 10 cm. Las cinco especies más representativas del sistema fueron elegidas para un análisis posterior y se calcularon los parámetros de la Ley de Poder de Taylor a cada una de ellas. Las especies de epifauna presentaron un índice de agregación cercano a la aleatoriedad. Los índices de agregación de las especies de la endofauna mostraron consistentemente altos valores, incluso en diferentes condiciones de densidad de población y de disponibilidad de materia orgánica. El reducido número de formas de endofauna en la marisma inferior, comparada con la superior, no apunta a una desventaja competitiva, ya que no hay alteraciones en los valores de b. Solo para las especies de endofauna, el valor de a mostró un agudo decremento desde la marisma inferior hacia la superior.

C2H2 zinc finger proteins (ZFPs) play important roles in plant development and response to abiotic stresses, and have been studied extensively. However, there are few studies on ZFPs in mangroves and mangrove associates, which represent a unique plant community with robust stress tolerance. MpZFP1, which is highly induced by salt stress in the mangrove associate Millettia pinnata, was cloned and functionally characterized in this study. MpZFP1 protein contains two zinc finger domains with conserved QALGGH motifs and targets to the nucleus. The heterologous expression of MpZFP1 in Arabidopsis increased the seeds’ germination rate, seedling survival rate, and biomass accumulation under salt stress. The transgenic plants also increased the expression of stress-responsive genes, including RD22 and RD29A, and reduced the accumulation of reactive oxygen species (ROS). These results indicate that MpZFP1 is a positive regulator of plant responses to salt stress due to its activation of gene expression and efficient scavenging of ROS.

Sunderban, the mangroves in the Ganga-Brahmaputra-Meghna delta shared between India and Bangladesh are the largest coastal wetland system, well known for their floral diversity. Vegetation survey and analysis was carried out at 42 forest sites randomly selected in Sundarban to record 30 species of mangrove plants belonging to 18 families. Change dynamics of mangrove forests was also studied taking Champion and Seth (1968) forest type classification as a baseline to record striking changes in forest composition. Significant changes were observed in vegetation composition at sites previously demarcated under a particular sub-type. Palm swamp with predominantly Phoenix paludosa have converted into mangrove scrub with mixed species composition. Some new species have emerged in pure mangrove forest (4B/TS2) which has acquired character of salt water mixed forest (Heretiera). These composition and structure changes of Sundarban forests may be attributed climatic change and anthropogenic pressure.

Mangroves are salt tolerant plants inhabiting saline environment. Multiple factors contribute to their salt tolerance and we need multifaceted approach to reveal the mechanisms of salt tolerance in the plant. In the present study, leaves of the mangrove, Rhizophora mucronata grown in the presence and absence of salt were used, free amino acids and the expression of selected genes were analyzed. Chromatographic technique showed the accumulation of free amino acids like proline, glycine, aspartic acid, valine, leucine and glutamic acid in the presence of salt. RNA was isolated from the leaf sample and cDNA was synthesized. Gene specific primers were designed and standardized. Among the genes studied (P5CS, BADH, NHX1), Betaine Aldehyde dehydrogenase (BADH) gene was found to be expressed.

Mangroves are salt-tolerant plant species that grow in coastal saline water and are adapted to harsh environmental conditions. In this study, we de novo assembled and functionally annotated the transcriptome of Rhizophora stylosa, the widely distributed mangrove from the largest mangrove family (Rhizophoraceae). The final transcriptome consists of 200,491 unigenes with an average length, and N50 of 912.7 and 1334 base pair, respectively. We then compared the genome-wide expression profiles between the two morphologically distinct natural populations of this species growing under different levels of salinity depending on their distance from the ocean. Among the 200,491 unigenes, 40,253 were identified as differentially expressed between the two populations, while 15,741 and 24,512 were up- and down-regulated, respectively. Functional annotation assigned thousands of upregulated genes in saline environment to the categories related to abiotic stresses such as response to salt-, osmotic-, and oxidative-stress. Validation of those genes may contribute to a better understanding of adaptation in mangroves species. This study reported, for the first time, the transcriptome of R. stylosa, and the dynamic of it in response to salt stress and provided a valuable resource for elucidation of the molecular mechanism underlying the salt stress response in mangroves and other plants that live under stress.

Mangrove is a green plants tolerant of salt water, which grows mainly along the sheltered coastal areas, especially along the bay or in estuaries. Final Disposal (TPA) Rubbish Suwung which located in the village Pedungan South Denpasar District is the rubbish dumps originating from the city of Denpasar and Badung. There is a natural mangrove vegetation in the river TPA. Mertsari area which located in the village of Sanur, West Denpasar District is a mangrove planting area and tourism destination on the Mertasari Beach. The purpose of this research are as follows: (1) To identify the content of nitrate and phosphate in mangrove sediments, (2) To determine the density of mangrove in Region Mertasari and TPA Suwung River Flow and (3) To describe the mangrove density is linkage with nitrate and phosphate mangrove sediments. The method used on this research is linear regression. Nitrate and phosphate sediments of mangroves in TPA Suwung River Flow and Region Mertasari range of 0.04 ppm - 79.034 ppm. The average density results of the mangrove tree level, saplings and seedlings in different locations ranged 0.01 ind / m2 - 0.32 ind / m2. The river's flow TPA Suwung produce a simple linear regression calculation of nitrate mangrove sediments density y = -0,002x + 0,288 (R2 ) 10,1 %, and the calculation of the density sedimentary phosphate mangrove produce y = 0,007x + 0,125 (R2) 6,1 %. The results of simple linear regression calculation of nitrate sediments density Mertasari mangrove area is y = -0,002x+ 0,537 R2 = 4,7%, and the calculation of the density sedimentary phosphate mangrove produce y = -0,038x + 0,777 (R2) 63,7 %.

Their high adaptability to difficult coastal conditions makes mangrove trees a valuable resource and an interesting model system for understanding the molecular mechanisms underlying stress tolerance and adaptation of plants to the stressful environmental conditions. In this study, we used RNA sequencing (RNA-Seq) for de novo assembling and characterizing the Bruguiera gymnorhiza (L.) Lamk leaf transcriptome. B. gymnorhiza is one of the most widely distributed mangrove species from the biggest family of mangroves; Rhizophoraceae. The de novo assembly was followed by functional annotations and identification of individual transcripts and gene families that are involved in abiotic stress response. We then compared the genome-wide expression profiles between two populations of B. gymnorhiza, growing under different levels of stress, in their natural habitats. One population living in high salinity environment, in the shore of the Pacific Ocean- Japan, and the other population living about one kilometre farther from the ocean, and next to the estuary of a river; in less saline and more brackish condition. Many genes involved in response to salt and osmotic stress, showed elevated expression levels in trees growing next to the ocean in high salinity condition. Validation of these genes may contribute to future salt-resistance research in mangroves and other woody plants. Furthermore, the sequences and transcriptome data provided in this study are valuable scientific resources for future comparative transcriptome research in plants growing under stressful conditions.

ABSTRACT Mangroves are dynamic and unique inter-tidal ecosystems, common in tropical and subtropical coastal environments. They are among the world’s most productive ecosystems and are important in protecting coasts from erosion by fierce tides, in promoting the diversity of marine organisms and fisheries by contributing a quantity of food and providing favourable habitats for animals. These economic uses of mangroves indicate that they play an important role in the lives and economies in the coastal regions of different countries. Mangrove forests are under immense threat worldwide due to their multiple economic uses and alterations of freshwater inflows by various upstream activities in catchment areas. Mangrove plants with unique adaptations play a crucial role in sustaining life in mangrove forests. Their reproductive biology is central to understanding the structural and functional components of mangrove forests. The success of sexual reproduction and subsequent population expansion in mangrove plants is linked to flowering timings, pollinators and tidal currents. Viviparous and cryptoviviparous plants are true mangroves while non-viviparous ones are mangrove associates. The dispersal propagule is seedling in viviparous and non-viviparous plants while it is seed in nonviviparous plants. In this study, viviparous and crypto-viviparous species were included for study. These species are self-compatible, self-pollinating and also cross-pollinating; such a breeding system is a requirement for the success of sexual reproduction and subsequent build up and expansion of population. They are entomophilous in the study region. The viviparous plants include Ceriops tagal, C. decandra, Rhizophora apiculata, R. mucronata, Bruguiera gymnorrhiza and B. cylindrica. The non-viviparous plants include Avicennia alba, A. marina, A. officinalis, Aegiceras corniculatum and Aegialitis rotundifolia. Sexual reproduction and regeneration events are annual in these plants and are dependent on local insects, tidal currents and nutrient content in estuarine environment. In recent times, erratic and insufficient rainfall together with industrial pollutants released into rivers is causing negative effects on the growth, development and regeneration of mangrove flora. In effect, there is a gradual decrease in mangrove cover. Added to this is continuous exploitation of mangrove plants for fuel wood, creation of shelters for cattle and changes for industrial establishments and aquaculture development in estuarine regions. As a consequence, the existing mangrove cover is struggling to survive and also not in a position to support local needs and provide livelihood opportunities through fishery resources. Further, reduced mangrove cover is showing catastrophic effects on fishing communities who live along the shore line during the period of cyclonic surges and tsunami events.

Salt treatment (50 mM NaCl) reduced plant growth of loquat (Eribotria japonica Lindl.) (by up to 40%) but not that of anger (Cydonia oblonga Mill.). Salt stress induced a strong leaf Na+ accumulation in both species. However, the observed increase in leaf Cl– level was higher in loquat (13-fold) than in anger plants (3.8-fold). Addition of Ca2+ (25 mM) significantly reduced Na+ and Cl– concentrations in both salt-treated species. In anger leaves, calcium addition to the nutrient media did not change the leaf calcium contents in salt-treated or untreated plants, this value being lower in salt-treated plants. However, in loquat plants, an increase in leaf Ca2+ was observed after the calcium addition. Surprisingly, an increase in Ca2+ concentration was also observed in salt-treated loquat plants. In general, anger plants had higher constitutive antioxidant enzyme levels in both control and salt-treated plants. Salt stress did not change antioxidant enzyme levels in loquat plants. A similar effect was observed in anger plants, but in this case a 2-fold induction of monodehydroascorbate reductase (MDHAR) activity was observed.In both species, salinity produced an oxidative stress, indicated by an increase in lipid peroxidation, this value being much higher in loquat (83%) than in anger (40%) plants. In salt-treated plants, Ca2+ addition provided some protection to the membranes, because the increases observed in thiobarbituric-acid-reactive substances (TBARS) were not significant.In contrast, in control plants Ca2+ treatments increased glutathione reductase (GR) and decreased catalase activity in anger, but increased MDHAR, dehydroascorbate reductase (DHAR), GR and superoxide dismutase (SOD) in loquat plants. In salt-treated plants, Ca2+ additions decreased catalase (CAT) and ascorbate peroxidase (APX) for anger and raised DHAR, GR and SOD for loquat. However, the mechanism by which Ca2+ regulates antioxidant enzymes remains to be determined.These results suggest that anger plants have a higher capacity to scavenge AOS, both under saline and non-saline conditions. Accordingly, and related to the smaller Cl– increase observed, anger plants are more salt-tolerant, at least partly owing to the higher antioxidant enzyme levels observed.

Green gram (Vigna radiata) is considered the chief legume in Pakistan. Thus, current study was conducted to examine the ameliorating effect of phytohormones pre-treatments under salt stress on proteome profile of green gram by sodium-dodecyl-sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The soluble green gram seedlings proteins were resolved on 4% stacking and 12% resolving gels. The SDS-PAGE resolved 24 polypeptide bands ranging from 200 to 17kDa. Among these, 12 out of 24 bands of proteins were essentials house-keeping or growth proteins of green grams. While, 120, 114.6, 51.8, 29.1, and 22.8 kDa bands were over-expressed under 50 to 350mM salt with phytohormones treatments. The others 104.5 kDa, 99.8 kDa, 95.3 kDa, 91.0 kDa, 55 kDa, 46 kDa, and 17kDa bands were related to the GAᴣ, IAA, and SA induced tolerance. Overall 120 kDa, 114.6 kDa, 104.5 kDa, 99.8, 95.3 kDa, 51.8 kDa, 29.1 kDa and 22.8kDa bands were first time identified in the current study. The information retrieved from NCBI protein database, the resolved peptides were principally belonging to 7S and 8S vicilin, 2S, 8S, 11S, and 16.5S globulins. It is determined that seed priming with SA enhanced tolerance in green gram by rapidly synthesizing stress alleviating peptides.Key word: Cluster analysis, dendrogram, mungbean, salt stress, SDS-PAGEINTRODUCTIONVarious world-wide health concerning organization recommended the use of high graded plant protein such as legumes to prevent the risk of metabolic disorder (Hou et al., 2019). Legumes are most important protein crop on the earth. Among the legumes, the green gram is the major pulses. Its seeds are rich in superior quality storage protein, which account 85% of the total protein while, another 15% have not been broadly studied (Yi-Shen et al., 2018). The soluble storage protein comprises of 60% globulins, 25% albumin and 15% prolamins. Globulins are further divided into 3.4% basic-type (7S), 7.6% legumin-type (11S), and 89% vicilin-type (8S) (Mendoza et al., 2001; Itoh et al., 2006). Other than proteins, the green gram seeds also contain starch, fiber, phenolic compound, saponins, vitamins, calcium zinc, potassium, folate, magnesium, manganese and very low in fat that made it meager man’s meat (Hou et al., 2019). It is also a good source of green manure and fodder (El-Kafafi et al., 2015). Its root has ability to fix 30 to 50 Kg/ha atmospheric nitrogen in the soil which is essential for maintaining soil fertility (Chadha, 2010). The green gram is the valuable and the major Rabi pulse crop of Pakistan. Its cultivation area in 2016-2017 was about 179,000 hectares with seed yield of 130,000 tones. In comparison during 2017-2018, it was cultivated on 161,800 hectares land with 118,800 tones seed yield (GOP, 2018). One of the reasons of this 9% decrease in both land and productivity is the shortage of irrigated land due to soil salinity. The salinity induce oxidative bust in the mungbean cells, caused by responsive oxygen species (ROS) such as hydrogen peroxide, singlet oxygen, hydroxyl radical and superoxide radical. The ROS create hindrance in various metabolic processes of plant via interacting with macromolecules like proteins (Alharby et al., 2016). However, phytohormones like gibberellic acid (GAᴣ), indole acetic acid (IAA), and salicylic acid (SA) take part in the biosynthesis of salt tolerance proteins under salinity. These salt tolerance proteins acclimate plants under salinity stress. Application of biotechnology plays a significant role in agriculture (Khan et al., 2017). Therefore, production of particular proteins under salt stress is a specific response of cell which can be analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). SDS-PAGE is the simple, valid, and cost-effective biochemical marker (Mushtaq et al., 2018). This marker has been widely used to determine the extent of evolutionary variations in crops (El-Kafafi et al., 2015).OBJECTIVES The present study was directed first time with the aim to investigate the toxic effect of sodium chloride (0-350 mM) and stress acclimation by pre-treatment of GAᴣ, IAA, and SA on the proteome profile of NM-92 cultivar of a Pakistani green gram.MATERIALS AND METHODSThe present study was replicated thrice in the plant laboratory of Department of Genetics, Faculty of Science, and University of Karachi. The seeds of mung bean cultivar NM-92 were acquired from National Agricultural Research Centre (NARC), Islamabad. These freshly collected 15 seedsˉ1 treatment / replication were divided into two sets. The first was named as sodium chloride (SC) stress treatments were imbibed in sterile distilled water (DW) whereas, second set soaked in gibberellic acid (GAᴣ) (BDH Chemicals, England), indole acetic acid (IAA) (Fluka, Switzerland), and salicylic acid (SA) (J.T. Baker, Holland) in the separate beaker for 24 hours under dark condition. After 24 hours, given ample time to both the sets at room temperature. After recovery, all 20 treatments were sown in the 150 X 30 mm sized petri-dishes containing 0, 50, 150, 250, and 350 millimolar (mM) sodium chloride solution (Fisher Scientific, UK) for 72 hours.Protein extraction: Protein extraction was done by taking 0.3g of seedlings in an ice chilled mortar and crushed by adding 600µL 0.2 M Tris-HCl buffer having pH 7.5 contained 5% SDS (w/v) and 5% 2-mercaptoethanol (v/v). The homogenate was incubated at 0oC for 30 min., boiled in the water bath for 3 min. at 100oC. Samples were centrifuged in Heraeus Biofuge D-37520, Germany for 30 min. at 8000 rpm. The protein supernatant was saved at below 0°C for quantitative and qualitative determination with minor modifications. The total soluble protein content of the samples was estimated via “Bovine Serum Albumin (BSA) standard curve” and explicit in µg protein milligramˉ1 fresh weight of mung seedlings.Bovine serum albumin standard curve (2000 μg/mL): Total protein standard curve was made by dissolving 0.05g of Bovine Serum Albumin (BSA) in 25mL of distilled water. Ten serial dilutions were made from 0.1 mL to 1mL by BSA solution then performed Lowry. A standard curve of total proteins was plotted by taking BSA absorbance at Y-axis and 2000 μg BSA / mL at X-axisSample preparation for SDS-PAGE: For qualitative assessment of total proteins; the 35μL of saved protein supernatant was combined with 15μL of sample diluting buffer (SDB). The SDB was made up of 0.0625 M Tris-HCl pH 6.8 with 2% of SDS, 10% of glycerol, 0.003% of bromophenol blue dye and 5% of 2-mercaptoethanol. Boil the 50μL protein SDB supernatant at 100oC in water bath for 3 min., centrifuged at 6000 rpm for 4 min. The supernatant was loaded on SDS-PAGE gel with the given formulae. The SDS- PAGE: Total proteins were fractionated via SDS-PAGE with 4% stacking and 12% resolving gel. The resolving gel of 12% was made by taking 6mL solution A, 1.8 mL 3 M Tris 1 M HCl buffer pH 8.8, 144μL 10% SDS, 5.74 mL sterile distilled water, 720μL 1.5% ammonium persulphate (APS) in deionized water and 10μL TEMED. While, stacking was composed of 1.25mL of solution A, 2.5mL of 0.5M Tris 1M HCl buffer pH 6.8, 100μL 10% SDS, 1.8 mL of distilled water, 500μL 1.5% APS and 12μL TEMED. Solution A was prepared by conjoining 30% acrylamide and 0.8% N, N’-methylene-bisacrylamide in deionized water. To avoid polymerization in the beaker; the prepared solution was quickly poured into the 3 mm thick gel plates after adding TEMED. The stacking was lined over resolving gel, then combs were inserted between the gel plates of SCIE-PLAS TV-100 separation system, UK, and allowed to polymerize for ½ an hour. After polymerization gel was placed in the tank which were filled with Tris-Glycine buffer (electrode buffer) pH 8.4 then combs were removed. The electrode buffer contained 0.3% Tris, 1.41% Glycine and 0.1% SDS in 2000mL d/w. The gel was pre-run for 15 min. at 60 volts and 120 mA currents. The prepared SDS-PAGE samples were loaded in wells with BlueStepTM Broad Range Protein Marker, AMRESCO, USA as standard and run at 60 volts & 120 mA for about 45 min. When samples entered in resolving gel, and then gave 100 volts and 200 mA currents for around 2.5 hours. Furthermore, electrophoresis was carried out at a constant watt.The Gel was washed with 30% ethanol on Uni Thermo Shaker NTS-1300 EYELA, Japan at the constant shaking for 30 min. Then gels were placed in 10% glacial acetic acid in 50% methanol solution (Fixative) for 24 hours. SDS Gel was stained until protein bands were visible thereat placed as 5% of Methanol in 7.5% acetic acid glacial solution to destain the bands background. SDS-PAGE stain composed of 0.125% coomassie brilliant blue R-250 dissolved in 40% of Methanol and 7% acetic acid glacial solution. The stain was stirred on Magnetic stirrer & hot plate M6/1, Germany for 6-10 hours before used. Photographs were taken by Sanyo digital camera VPC-T1284BL and bands were scored through numbering pattern. Gels preserved in 10% acetic acid solution at 4°C.Interpretation of bands and data analysis: The total soluble protein bands relative mobility calculated by below formulae and Dendrogram was constructed via SPSS v. 20Where,F=(Migrated distance of protein band)/(Migrated distance of dye front)Slop=(Log MW of protein marker lower limit band–log〖MW of protein marker upper limit band )/(RF protein marker lower limit band –RF of protein marker upper limit band)RESULTS:The total soluble proteins extracted from green gram were perceived by SDS-PAGE Blue StepTm broad range biochemical markers. The protein-based marker was used to evaluate the toxic effect of sodium chloride along with pre-treatments of GAᴣ, IAA, and SA on proteome assay. In the current work, seedlings total soluble proteome resolved 24 polypeptide bands ranging from 200 to 17.1 kDa were recognized by using SDS-PAGE. The figure 1 showed Dendrogram assay, which classified the 20 treatments of SC, GAᴣ, IAA and SA into two major clusters where, the cluster I was the largest one (figure 1). Cluster I consisted of 15 treatments that further divided into I-A, and I-B. The pre-treatments of SC50+SA, SC150+SA, SC250+SA, and SC350+IAA were grouped together into C-1 of sub-cluster I-A. The C-2 of sub-cluster I-A, pre-treatment SC350+SA was most diverse among 20 treatments. The C-1 treatments showed 99% homology when compared with each other while, it was 97% similar with C-2. The sub-cluster I-B comprised another 10 treatments, SC0+GAᴣ, SC50+GAᴣ, SC150+GAᴣ, SC250+GAᴣ, SC350+GAᴣ, SC0+IAA, SC50+IAA, SC150+IAA, SC250+IAA, and SC0+SA that were also 99% similar for total proteins. Sub-cluster I-B pre-treatments was exhibiting 94% homology with the sub-cluster I-A. The second cluster was the smallest one that was divided into two sub-clusters, II-A and II-B. The II-A was comprised of SC50, SC150, and SC250 while, sub-cluster II-B consisted of SC0 and SC350. Within each sub-cluster, pre-treatments expressed 99% homology whereas, II-A was 97 different from II-B. Furthermore, cluster I showed 75% similarities with cluster II (figure 1). The seedlings storage proteome profile of green gram was shown in table 1.The results showed that 120kDa, 114.6 kDa, 51.8 kDa, 29.1 kDa and 22.8 kDa proteins bands were not induced at 0 mM SC, GAᴣ, IAA, and SA. The table 1 depicted the presence of 120 kDa and 114.6 kDa bands only at 350 mM SC level with all phytohormones treatments. Similarly, 51.8 kDa protein bands were appearing at 150SC, 250SC and 350SC stress with phytohormones. Based on the information collected from the NCBI protein database, this peptide was related to the 8S globulin alpha subunits. The two other, 7S globulins sub-units having 29.1kDa and 22.8 kDa molecular weights bands were synthesized under 50mM, 150mM, 250mM, 350mM SC stress with phytohormones. Concerning protein polypeptide of molecular weight 104.5 kDa, 99.8 kDa, 91.0 kDa, 55.0 kDa, and 46.0 kDa, those were induced by GAᴣ, IAA and SA at 0 to 350 mM SC. While, 17kDa protein band was appearing in SA, and IAA treated samples and 95.3kDa band was only present in SA treatment. Other 12 protein bands were present in all treatments proved as house-keeping proteins of green gram (table 1).DISCUSSIONThe SDS-PAGE profiling for proteome is the reliable and applied biochemical approach that has been used as biochemical marker in various crop differentiation, and characterization. In the current study, first time SDS-PAGE was utilized to investigate the impact of GAᴣ, IAA, and SA pre-soaking on green gram under salt toxicity. The salt toxicity adversely affects all seed, seedling, and plant metabolic process (Parveen et al., 2016). At salt toxicity, the endogenous GAᴣ, IAA, and SA levels markedly decrease (El-Khallal et al., 2009). In such condition, exogenous application of GAᴣ, IAA, and SA enhance seedlings survival rate by increasing synthesis of seed storage proteins. Likewise, our Dendrogram characterization based on 20 treatments showed significant diversity under 0 to 350 mM SC stress. The salicylic acid treatments were grouped together except SC0+SA treatment, exhibiting a close relationship, which proved its acclimating role under salt stress. These findings will help plant breeder toward enhancing food quality and quantity of green gram in future breeding programme on saline sodic land.The SDS-PAGE assay revealed 200. kDa, 109.4 kDa, 77 kDa, 68 kDa, 49 kDa, 38 kDa, 33 kDa, 26 kDa, 24 kDa, 22 kDa, 21 kDa and 19 kDa fractions as essential green gram proteins. Among these, 68 kDa, 49 kDa, 33 kDa, 26 kDa, 24 kDa and 21 kDa peptides were seed biotinylated isoform protein (Riascos et al., 2009), putative NADH-ubiquinone oxidoreductase subunit H (Gostinčar et al., 2019), heat shock protein 33 (Hamidian et al., 2015), globulin protein, seed coat / maturation protein (Dhaubhadel et al., 2005), and protein for dimerization. While, 22 kDa proteins belonged to the class of prolamin alpha zein Z1C1_2, Z1C1_4, and Z1C1_8 precursors, and 19kDa peptide was related with Z1A1_2, Z1A2_2, and Z1B_6 precursors (Miclaus et al., 2011). Further, the 91 kDa peptide is sucrose synthase SS1 protein, and 77kDa protein is the NADPH-cytochrome P450 reductase (Wang et al., 2004). Also, the phosphatase-associated two other proteins having 46 and 55 kDa molecular weight were reported earlier in Mucuna pruriens. Hameed et al. (2012) and Malviya et al. (2008) found 55 and 46kDa peptides as 7S vicilin small sub-units and 17kDa as 11S globulins sub-unit in the studied Vigna radiata. Some other molecular weight proteome such as 68 kDa and 49kDa are 7S vicilin, 33kDa is 8S vicilin, 38 and 26kDa 8S globulins, 24kDa 11S globulins, and 22kDa 16.5S globulins. These proteins required for germination and seed establishment of green gram plant (Hameed et al., 2012).The vast accumulation of 23kDa and 22kDa peptides under salt stress by salicylic acid, were reported previously in the mangrove Bruguiera parviffora and Zea mays (El-Khallal et al., 2009). Correspondingly, El-Kafafi et al. (2015) reported the presence of 115kDa, 23kDa, and 22kDa bands in the salt tolerant lines of green gram. These proteomes induced under salt stress may play a pivotal part in the stress acclimation and osmotic adjustment. Similarly, the induction of 104 kDa and 100kDa MW polypeptide by SC stress in the salt tolerant genotypes of green gram indicated the functional role of phytohormones in various metabolic and defense response El-Kafafi et al. (2015); Alharby et al. (2016), El-Khallal et al. (2009), Qados (2010). Ali et al. (2007), Alharby et al. (2016), and El-Kafafi et al. (2015) observed 17kDa, 26kDa, 33kDa and 77kDa bands involving in salt tolerance and can be considered as a positive biochemical marker for salt stress. Further, 26 kDa MW peptide also functions as osmotin under the salt stress that involved in enhancing the accumulation of glycine betaine and proline in the cells. Hence, proteome assay of green gram showed that GAᴣ, IAA, and SA could regulate the expression of salt stress proteins that are anticipated to play a crucial part in the salt tolerance mechanism. Likewise, the involvement of phytohormones in the induction of changes in the proteome profile pattern was attributed to their part in managing cell division by regulating some genes of apical meristems.CONCLUSIONFinally, the results revealed the presence of the ten new bands with MW of 200kDa, 120 kDa, 114.6 kDa, 109.4kDa, 104.5kDa, 99.8kDa, 95.3kDa, 51.8kDa, 29.1kDa and 22.8kDa have not reported previously under salt stress with phytohormones treatments in green gram. Furthermore, it was observed that phytohormones alleviate the negative impact of salt stress on green gram by enhancing synthesis of salt defense polypeptides. Hence, higher accumulation of proteins was observed in salicylic acid treated seedlings. Thus, present work recommended the pre-soaking of phytohormones to overcome the toxic impact of sodium chloride on green gram. 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The above-ground biomass of three dominant salt-marsh vascular plants (Juncus kraussii, Sarcocornia quinquejlora and Sporobolus virginicus) was measured to assess both spatial and temporal variation and to provide baseline data. Additionally, the culm dynamics of the rush J. kraussii were measured so that aboveground productivity could be estimated. No distinct seasonal patterns were detected in above-ground biomass in J. kraussii. Averaged over all sites and times, the above-ground biomass of J. kraussii was 1116 g dry weight m-2. Culms are replaced annually, hence standing crop approximated annual above-ground productivity. Much of the dead aboveground biomass appears to accumulate in the upper marsh, as evidenced by the elevated nutrient and organic carbon content of the soil there relative to the sediment in the mangrove zone. Above-ground biomass of the decumbent perennial grass Sporobolus virginicus and the procumbent perennial chenopod Sarcocornia quinqueflora showed no consistent spatial or temporal trends. The above-ground standing crops of these species were about one-third that of J. kraussii.

Mangroves are a diverse group of highly salt-tolerant woody plants, which grow in the inter-tidal zones in tropical and subtropical latitudes. Despite its unique services to the people, coastal and marine systems, mangroves have become one of the most rapidly disappearing ecosystems in the world. This paper reviews the available information on distribution, current status and challenges of mangroves in Sri Lanka. Today, around 160 km2 of mangrove vegetation is available in Sri Lanka and distributed mainly in Jaffna, Batticaloa, Kalpitiya, Rekawa and Trincomalee and is composed of 21 species of true mangroves and 24 species of mangrove associates. Mangroves in the island have been adversely affected due to the numerous anthropogenic activities, including land reclamation, tourism, coastal aquaculture and agriculture and other industrial activities etc. Proper conservations of mangroves are urgently required to the island to avoid further decline of mangrove ecosystem. It is imperative to evaluate policies, legal instruments and development strategies to effectively protect this valuable ecosystem.

Mangroves are well-adapted halophytes that thrive in coastal saline environments. They live under difficult environmental conditions, such as high light intensity and external salt concentrations, as well as low-oxygen environments, such as water-logged muck, that are typically inappropriate for the survival of other plants. Salinity is a major abiotic factor that affects plant growth, productivity, and dispersal in tropical and semitropical intertidal areas. Furthermore, it affects approximately 20% of all cultivable land and 50% of all irrigated land on the planet. Mangroves have developed a sophisticated salt filtration mechanism and a complicated root structure to withstand salty water exposure and tidal movement. The expression patterns of five salt tolerance genes (amFer1, amDhna, amSod1, amCat1, and amUbc2) in the Egyptian gray mangrove (Avicennia marina Forssk.) grown under different environmental conditions in South Sinai protectorates (Nabq, Ras Mohamed, Safaga, and Wadi El-Gemal), Egypt, were investigated in this study. This study aimed to assess and examine the genetic behavior of mangroves in response to salinity by using quantitative real-time PCR. Findings revealed differences in the expression patterns of the investigated genes under various conditions, showing that salinity influences plant genetic response. Ferritin gene expression was high in all locations, indicating that ferritin represents an essential component of the mangrove response mechanisms.

Aquaporins are channel proteins that facilitate the transmembrane transport of water and other small neutral molecules, thereby playing vital roles in maintaining water and nutrition homeostasis in the life activities of all organisms. Canavalia rosea, a seashore and mangrove-accompanied halophyte with strong adaptability to adversity in tropical and subtropical regions, is a good model for studying the molecular mechanisms underlying extreme saline-alkaline and drought stress tolerance in leguminous plants. In this study, a PIP2 gene (CrPIP2;3) was cloned from C. rosea, and its expression patterns and physiological roles in yeast and Arabidopsis thaliana heterologous expression systems under high salt-alkali and high osmotic stress conditions were examined. The expression of CrPIP2;3 at the transcriptional level in C. rosea was affected by high salinity and alkali, high osmotic stress, and abscisic acid treatment. In yeast, the expression of CrPIP2;3 enhanced salt/osmotic and oxidative sensitivity under high salt/osmotic and H2O2 stress. The overexpression of CrPIP2;3 in A. thaliana could enhance the survival and recovery of transgenic plants under drought stress, and the seed germination and seedling growth of the CrPIP2;3 OX (over-expression) lines showed slightly stronger tolerance to high salt/alkali than the wild-type. The transgenic plants also showed a higher response level to high-salinity and dehydration than the wild-type, mostly based on the up-regulated expression of salt/dehydration marker genes in A. thaliana plants. The reactive oxygen species (ROS) staining results indicated that the transgenic lines did not possess stronger ROS scavenging ability and stress tolerance than the wild-type under multiple stresses. The results confirmed that CrPIP2;3 is involved in the response of C. rosea to salt and drought, and primarily acts by mediating water homeostasis rather than by acting as an ROS transporter, thereby influencing physiological processes under various abiotic stresses in plants.

Aquaporins are channel proteins that facilitate the transmembrane transport of water and other small neutral molecules, thereby playing vital roles in maintaining water and nutrition homeostasis in the life activities of all organisms. Canavalia rosea, a seashore and mangrove-accompanied halophyte with strong adaptability to adversity in tropical and subtropical regions, is a good model for studying the molecular mechanisms underlying extreme saline-alkaline and drought stress tolerance in leguminous plants. In this study, a PIP2 gene (CrPIP2;3) was cloned from C. rosea, and its expression patterns and physiological roles in yeast and Arabidopsis thaliana heterologous expression systems under high salt-alkali and high osmotic stress conditions were examined. The expression of CrPIP2;3 at the transcriptional level in C. rosea was affected by high salinity and alkali, high osmotic stress, and abscisic acid treatment. In yeast, the expression of CrPIP2;3 enhanced salt/osmotic and oxidative sensitivity under high salt/osmotic and H2O2 stress. The overexpression of CrPIP2;3 in A. thaliana could enhance the survival and recovery of transgenic plants under drought stress, and the seed germination and seedling growth of the CrPIP2;3 OX (over-expression) lines showed slightly stronger tolerance to high salt/alkali than the wild-type. The transgenic plants also showed a higher response level to high-salinity and dehydration than the wild-type, mostly based on the up-regulated expression of salt/dehydration marker genes in A. thaliana plants. The reactive oxygen species (ROS) staining results indicated that the transgenic lines did not possess stronger ROS scavenging ability and stress tolerance than the wild-type under multiple stresses. The results confirmed that CrPIP2;3 is involved in the response of C. rosea to salt and drought, and primarily acts by mediating water homeostasis rather than by acting as an ROS transporter, thereby influencing physiological processes under various abiotic stresses in plants.

The effects of municipal wastewater discharge and anthropogenic sedimentation on the structure and composition of gray mangrove (Avicennia marina (Forsk.) Vierh.) communities along Tubli Bay coastlines in Bahrain were investigated. Growth and regeneration of mangrove were measured, and its community was characterized. Sediment profile was analyzed for texture, pH, and salinity. Mangrove area covered by sand depositions was measured using Google Earth Pro. ANOVA and regression tests were employed in the analysis of the data. Results indicated that mangrove overwhelmingly dominated plant community in the study area, which was zoned by a community of other salt-tolerant species. Three main habitats exist in the study area with high similarity in their floristic composition. Species richness and the number of habitats were low due to the aridity and high sediment salinity. The dilution effect of the secondary treated wastewater had a favorable effect on height and diameters of mangrove trees. However, no differences were observed in leaf area index, basal area, and density of mangrove. The long-term accumulation of anthropogenic sedimentation had a detrimental effect on the mangrove community, expressed in swath death of mangrove trees due to root burials and formation of high topography within the community boundaries.