Leaf miner-induced morphological, physiological and molecular changes in mangrove plant Avicennia marina (Forsk.) Vierh.

Avicennia marina (Forsk.) Vierh is a widespread mangrove species along the southeast coasts of China. Recently, the outbreak of herbivorous insect, Phyllocnistis citrella Stainton, a leaf miner, have impacted on the growth of A. marina. Little is reported about the responses of A. marina to leaf miner infection at the biochemical, physiological and molecular levels. Here, we reported the responses of A. marina to leaf miner infection from the aspects of leaf structure, photosynthesis, and antioxidant system and miner responsive genes expression. A. marina leaves attacked by the leaf miner exhibited significant decreases in chlorophyll, carbon and nitrogen contents, as well as a decreased photosynthetic rate. Scanning and transmission electron microscopic observations revealed that the leaf miner only invaded the upper epidermis and destroyed the epidermal cell, which lead to the exposure of salt glands. In addition, the chloroplasts of mined leaves (ML) were swollen and the thylakoids degraded. The maximal net photosynthetic rate, stomatal conductance (Gs), carboxylation efficiency (CE), dark respiration (Rd), light respiration (Rp) and quantum yields (AQE) significantly decreased in the ML, whereas the light saturation point (Lsp), light compensation point (Lcp), water loss and CO2 compensation point (Г) increased in the ML. Moreover, chlorophyll fluorescence features also had been changed by leaf miner attacks. Interestingly, higher generation rate of O2ˉ· and lower antioxidant enzyme expression in the mined portion (MP) were found; on the contrary, higher H2O2 level and higher antioxidant enzyme expression in the non-mined portion (NMP) were revealed, implying that the NMP may be able to sense that the leaf miner attacks had happened in the MP of the A. marina leaf via H2O2 signaling. Besides, the protein expression of glutathione S-transferase (GST) and the glutathione (GSH) content were increased in the ML. In addition, insect resistance-related gene expression such as chitinase 3, RAR1, topless and PIF3 had significantly increased in the ML. Taken together, our data suggest that leaf miners could significantly affect leaf structure, photosynthesis, the antioxidant system and miner responsive gene expression in A. marina leaves.

4-Hydroxy cinnamic acid as mushroom preservation: Anti-tyrosinase activity kinetics and application.

Tyrosinase is a key enzyme in post-harvest browning of fruit and vegetable. To control and inhibit its activity is the most effective method for delaying the browning and extend the shelf life. In this paper, the inhibitory kinetics of 4-hydroxy cinnamic acid on mushroom tyrosinase was investigated using the kinetics method of substrate reaction. The results showed that the inhibition of tyrosinase by 4-hydroxy cinnamic acid was a slow, reversible reaction with fractional remaining activity. The microscopic rate constants were determined for the reaction on 4-hydroxy cinnamic acid with tyrosinase. Furthermore, the molecular docking was used to simulate 4-hydroxy cinnamic acid dock with tyrosinase. The results showed that 4-hydroxy cinnamic acid interacted with the enzyme active site mainly through the hydroxy competed with the substrate hydroxy group. The cytotoxicity study of 4-hydroxy cinnamic acid indicated that it had no effects on the proliferation of normal liver cells. Moreover, the results of effects of 4-hydroxy cinnamic acid on the preservation of mushroom showed that it could delay the mushroom browning. These results provide a comprehensive underlying the inhibitory mechanisms of 4-hydroxy cinnamic acid and its delaying post-harvest browning, that is beneficial for the application of this compound.

Inhibitory effects of cefotaxime on the activity of mushroom tyrosinase.

Tyrosinase (EC 1.14.18.1) catalyzes both the hydroxylation of tyrosine into o-diphenols and the oxidation of o-diphenols into o-quinones that form brown or black pigments. In the present paper, cefotaxime, a cephalosporin antibacterial drug, was tested as an inhibitor of tyrosinase. The results show that cefotaxime inhibits both the monophenolase and diphenolase activities of tyrosinase. For the monophenolase activity, cefotaxime increased the lag time and decreased the steady-state activity with an IC50 of 3.2 mM. For the diphenolase activity, the inhibition by cefotaxime is reversible and mix-I type with an IC50 of 0.14 mM. The inhibition constants (K(I) and K(IS)) were determined to be 0.14 and 0.36 mM, respectively. The molecular mechanism of inhibition of tyrosinase by cefotaxime was determined by fluorescence quenching and molecular docking. The results demonstrated that cefotaxime was a static quencher of tyrosinase and that cefotaxime could dock favorably in the active site of tyrosinase. This research may offer a lead for designing and synthesizing novel and effective tyrosinase inhibitors in the future.

Arsenic Accumulation and Translocation in Mangrove (Aegiceras corniculatum L.) Grown in Arsenic Contaminated Soils.

Mangrove wetlands serve as both a sink and source for arsenic (As), as mangrove plants are able to uptake and accumulate As. The present study used pot experiments to evaluate As accumulation and translocation in mangrove (Aegiceras corniculatum L.) seedlings grown in As contaminated soils. Results indicated that A. corniculatum seedlings grew normally under As stress with minute growth inhibition and biomass reduction at different As treatment concentrations in a range of 0-150 mg·kg(-1). As concentrations in roots, stems and leaves were increased with increasing As treatment concentrations, but As accumulated mainly in roots, with accumulation rates of 74.54%-89.26% of the total As accumulation. In particular, relatively high bioconcentration factor (BCF) in root (2.12-1.79), low BCF in stem (0.44-0.14) and leaf (0.06-0.01), and thereby a low translocation factor (TF) in stem/root (0.21-0.08) and leaf/root (0.02-0.008) were observed. These results demonstrated that A. corniculatum is an As excluder with the innate capacity to tolerate As stress and root tissues may be employed as a bio-indicator of As in polluted sediments. Additionally, A. corniculatum is a potential candidate mangrove species for As phytostabilization in tropical and subtropical estuarine wetlands.

Isolation and purification of condensed tannins from flamboyant tree and their antioxidant and antityrosinase activities.

Flamboyant tree, a kind of medicinal plant, was studied as a source of condensed tannins. The antioxidant activities of the condensed tannins from the leaf, fruit, and stem bark of flamboyant tree were screened by ABTS radical and hydroxyl radical scavenging activity methods. The results indicated that these compounds possessed potent antioxidant activity. Their structures were then characterized by high-performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS) after thiolytic degradation. The results showed that the leaf condensed tannins were composed of afzelechin/epiafzelechin, catechin/epicatechin, and gallocatechin/epigallocatechin, while the fruit and stem bark condensed tannins had afzelechin/epiafzelechin and catechin/epicatechin. In addition, the condensed tannins were evaluated for their antityrosinase ability. They were found to have significant antityrosinase activity. The IC50 values were 35 ± 2.0 and 40 ± 0.5 µg/ml for the condensed tannins of fruit and stem bark, respectively. Further, fluorescence quenching and copper interacting techniques were utilized to unravel the molecular mechanisms of the inhibition. The results showed that the hydroxyl group of the condensed tannins could chelate the dicopper center of the enzyme and interact with tryptophan residues. Our studies revealed that condensed tannins might be suitable for use in food, agriculture, cosmetic, nutraceutical, and pharmaceutical applications.

Dependence of phenanthrene dissipation in mangrove sediment on the distance to root surface of Kandelia obovata L.

We explored the relationship between the distance from roots and rhizosphere-mediated phenanthrene (PHE) dissipation in planted mangrove sediment. A compartmentalized device (rhizobox) was used that separated root surface and four consecutive rhizosphere layers that extended 0-2, 2-4, 4-6, and >6 mm away from the root surface. Kandelia obovata L. Druce seedlings were grown for 4 months in sediment spiked with 10 mgkg(-1) PHE. PHE dissipation, microbial community structure and enzymatic activities at millimeter scale in the vicinity of plant roots were analyzed. Results indicated significant differences in PHE dissipation through the various layers in the planted rhizobox. PHE dissipation exhibited the most rapid loss in the 0-2 mm near-rhizosphere layer, the lowest in far-rhizosphere (>6 mm) layer. Microbial community structure as indicated by denaturing gradient gel electrophoresis (DGGE) profiles showed that special bacterial species and larger bacterial numbers were observed in near-rhizosphere layers. Depending on the distance to root surface, near-rhizosphere bacterial species and enzymatic activities were more abundant than root compartment and far-rhizosphere, resulting in rapid dissipation of PHE.

NMR, HPLC-ESI-MS, and MALDI-TOF MS analysis of condensed tannins from Delonix regia (Bojer ex Hook.) Raf. and their bioactivities.

The structures of the condensed tannins isolated from leaf, fruit, and stem bark of Delonix regia (Bojer ex Hook.) Raf. have been investigated with (13)C nuclear magnetic resonance ((13)C NMR) and high performance liquid chromatography electrospray ionization mass spectrometry (HPLC-ESI-MS) coupled with thiolysis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analyses. The results showed that these condensed tannins from D. regia possessed structural heterogeneity in monomer units and degree of polymerization. Propelargonidin (PP) and procyanidin (PC) were found in the leaf, fruit, and stem bark of D. regia, while prodelphinidin (PD) was found only in the leaves. The polymer chain lengths of condensed tannins from leaf and fruit organs were detected to be trimers to hexadecamers but from trimers to tridecamers for stem bark. B-type linkages were present in all these compounds. Condensed tannins from different parts of D. regia can be explored as tyrosinase inhibitors and food antioxidants because of their potent antityrosinase and antioxidant activities. The inhibitor concentration leading to 50% enzyme activity (IC(50)) was estimated to be 38 ± 1, 73 ± 2, and 54 ± 1.5 µg/mL for the condensed tannins of leaf, fruit, and stem bark. Condensed tannins extracted from stem bark exhibited the highest antioxidant activity; the DPPH scavenging activity (IC(50)) and the FRAP values were 90 ± 2 µg/mL and 5.42 ± 0.09 mmol AAE/g, respectively.

A new atisane-type diterpene from the bark of the mangrove plant Excoecaria agallocha.

A new atisane-type diterpene, ent-16alpha-hydroxy-atisane-3,4-lactone (4) and three known diterpenes, ent-16alpha-hydroxy-atisane-3-one (1), ent-atisane-3beta,16alpha-diol (2), ent-3,4-seco-16alpha-hydroxyatis- 4(19)-en -3-oic acid (3) were isolated from the bark of the mangrove plant Excoecaria agallocha. Their structures and relative stereochemistry were elucidated by means of extensive NMR and MS analysis. Compound 3 exhibited significant anti-microfouling activity against the adherence of Pseudomonas pseudoalcaligenes, with an EC(50) value of 0.54+/-0.01 ppm.

Contamination and potential biodegradation of polycyclic aromatic hydrocarbons in mangrove sediments of Xiamen, China.

Five stations were established in the Fenglin mangrove area of Xiamen, China to determine the concentrations of polycyclic aromatic hydrocarbons (PAHs) and the numbers of PAH-degrading bacteria in surface sediments. Assessing the biodegradation potential of indigenous microorganisms and isolating the high molecule weight (HMW)-PAH degrading bacteria was also one of the aims of this work. The results showed that the total PAH concentration of sediments was 222.59 ng g(-1) dry weight, whereas the HMW-PAH benzo(a)pyrene (BaP) had the highest concentration among 16 individual PAH compounds. The variation in the numbers of PAH-degrading bacteria was 2.62 x 10(2)-5.67 x 10(4)CFU g(-1) dry weight. The addition of PAHs showed a great influence in increasing the microbial activity in mangrove sediments. A bacterial consortium, which could utilize BaP as the sole source of carbon and energy, and which was isolated from mangrove sediments and enriched in liquid medium for nearly one year degraded 32.8% of BaP after 63 days incubation.

Changes of plasma membrane ATPase activities and proline accumulation in twigs of Casuarina under different growth conditions.

Twigs of Casuarina were collected from Dongshan and Xiamen Island, China. Crude plasma membrane preparation was obtained from Casuarina twigs by ultra-centrifugation. Sealed plasma membrane vesicles were isolated with aqueous PEG3350-Dextran T500 two phase partitioning. We studied the proline accumulation in twigs and the ATPase activities of plasma membrane of Casurina under different growth conditions. The results showed that free proline content and the PM ATPase activities from unfavorable growth conditions were obviously higher than those from favorable environment. This suggested that Casuarina responded to osmotic stress by increasing osmotic material content and the proton transportation activities of PM ATPase.

Responses of membrane protection enzyme system of tobacco leaves on Hg, Cd and Pb stresses in soil.

Pot experiment was used to study the responses of membrane protection enzyme system of tobacco leaves on Hg, Cd and Pb stresses in soil. The results showed that POD activity gradually increased with increasing concetrations of Hg, Cd and Pb. CAT and SOD activity gradually decreased under three heavy metals common existing and SOD variation curve showed unimodal curve under single or two elements existing with increase of concentration of Hg, Cd and Pb. The effects of Hg, Cd and Pb in soil: three elemets together > two elements together > single element only. The effects resulted in an imbalance--activated oxygen produce and scavenge and physiological biochemical process disorder. There was a synergistic action for the effect of Hg, Cd and Pb in soil on membrane protection enzyme system in tobacco leaves.