The effects of a range of salinity (0, 100, 200 and 400 mM NaCl) on growth, ion accumulation, photosynthesis and anatomical changes of leaves were studied in the mangrove, Bruguiera parviflora of the family Rhizophoraceae under hydroponically cultured conditions. The growth rates measured in terms of plant height, fresh and dry weight and leaf area were maximal in culture treated with 100 mM NaCl and decreased at higher concentrations. A significant increase of Na + content of leaves from 46.01 mmol m -2 in the absence of NaCl to 140.55 mmol m -2 in plants treated with 400 mM NaCl was recorded. The corresponding Clcontents were 26.92 mmol m -2 and 97.89 mmol m -2 . There was no significant alteration of the endogenous level of K + and Fe 2+ in leaves. A drop of Ca 2+ and Mg 2+ content of leaves upon salt accumulation suggests increasing membrane stability and decreased chlorophyll content respectively. Total chlorophyll content decreased from 83.44 mg cm -2 in untreated plants to 46.56 mg cm -2 in plants treated with 400 mM NaCl, suggesting that NaCl has a limiting effect on photochemistry that ultimately affects photosynthesis by inhibiting chlorophyll synthesis (ca. 50% loss in chlorophyll). Light-saturated rates of photosynthesis decreased by 22% in plants treated with 400 mM NaCl compared with untreated plants. Both mesophyll and stomatal conductance by CO 2 diffusion decreased linearly in leaves with increasing salt concentration. Stomatal and mesophyll conductance decreased by 49% and 52% respectively after 45 days in 400 mM NaCl compared with conductance in the absence of NaCl. Scanning electron microscope study revealed a decreased stomatal pore area (63%) in plants treated with 400 mM NaCl compared with untreated plants, which might be responsible for decreased stomatal conductance. Epidermal and mesophyll thickness and intercellular spaces decreased significantly in leaves after treatment with 400 mM NaCl compared with untreated leaves. These changes in mesophyll anatomy might have accounted for the decreased mesophyll conductance. We conclude that high salinity reduces photosynthesis in leaves of B. parviflora, primarily by reducing diffusion of CO 2 to the chloroplast, both by stomatal closure and by changes in mesophyll structure, which decreased the conductance to CO 2 within the leaf, as well as by affecting the photochemistry of the leaves.
Exposure of two-month-old seedlings of Bruguiera parviflora to NaCl stress (0 to 400 mM) for 45 d under hydroponic culture caused notable disorganisation of the thylakoid structure of chloroplasts in NaCl-treated leaves as revealed from transmission electron microscopy. The absorption spectra of treated and control thylakoid samples were similar having a red peak at 680 nm and Soret peaks at 439 and 471 nm in the blue region of the spectrum. The spectra of treated samples differed from control samples by gradual decrease in absorbance of 100, 200, and 400 mM NaCl treated samples at 471 and 439 nm, which could be due to scattering of radiation in these samples. Thus, absorption characteristics of thylakoid membranes indicated no major alterations in the structural integrity of the photosynthetic membranes during salt stress in B. parviflora. Analysis of pigment protein complexes of thylakoids on non-denaturing gel showed that CP1 complex consisting of photosystem (PS) 1 reaction centre decreased marginally by 19 % and the CP47 constituting the core antenna of PS2 declined significantly by 30 % in 400 mM NaCl treated samples in respect to control. This decrease in structural core antenna might cause inefficient photon harvesting capacity. However, CP43 content did not alter. An increase in CP2/CP1 ratio from 3.2 in control to 4.0 in 400 mM NaCl treated samples indicated significant structural changes in the thylakoids of salt treated plants. Haem staining of thylakoids revealed significant losses in cytochrome (Cyt) f and Cyt b 6 contents by NaCl stress. However, Cyt b 559 content remained nearly constant in both control and NaCl treated samples. SDS-PAGE of thylakoid proteins showed that the intensity of many of Coomassie stained polypeptide bands ranging from 15-22 and 28-66 kDa regions decreased significantly in NaCl treated samples as compared to control. Electron transport activity of thylakoids, measured in terms of DCPIP photoreduction, was 22 % lower in 400 mM NaCl treated plants than in the control ones. Hence, NaCl induces oxidative stress in chloroplasts causing structural alterations in thylakoids. These structural alterations might be responsible for declined efficiency of photosystems and reduced electron transport activity.
Three-dimensional (3D) hydrogels incorporating a compendium of bioactive molecules can allow efficient proliferation and differentiation of cells and can thus act as successful tissue engineering scaffolds. Self-assembled peptide-based hydrogels can be worthy candidates for such applications as peptides are biocompatible, biodegradable and can be easily functionalized with desired moieties. Here, we report 3D growth and proliferation of mammalian cells (HeLa and L929) on a dipeptide hydrogel chemically functionalized with a pentapeptide containing Arg-Gly-Asp (RGD) motif. The method of functionalization is simple, direct and can be adapted to other functional moieties as well. The functionalized gel was noncytotoxic, exhibited enhanced cell growth promoting properties, and promoted 3D growth and proliferation of cells for almost 2 weeks, with simultaneous preservation of their metabolic activities. The presence of effective cell growth supporting properties in a simple and easy to functionalize dipeptide hydrogel is unique and makes it a promising candidate for tissue engineering and cell biological applications.
Two-month-old seedlings of Bruguiera parvifora were treated with varying levels of NaCl (100, 200 and 400 mm) under hydroponic culture. Total proteins were extracted from leaves of control and NaCl treated plants after 7, 14, 30 and 45 d of treatment and analysed by SDS-PAGE. As visualized from SDS-PAGE, the intensity of several protein bands of molecular weight 17, 23, 32, 33 and 34 kDa decreased as a result of NaCl treatment. The degree of decrease of these protein bands seemed to be roughly proportional to the external NaCl concentration. The most obvious change concerned a 23 kDa-polypeptide (SSP-23), which disappeared after 45 d treatment in 400 mm NaCl. Moreover, the SSP-23 protein, which disappeared in B. parviflora under salinity stress, reappeared when these salinized seedlings were desalinized. These observations suggest the possible involvement of these polypeptides for osmotic adjustment under salt stress. NaCl stress also caused an increase in the activity of both acid and alkaline protease. The increasing activity of proteases functions as a signal of salt stress in B. parviflora, which induces the reduction of protein level.
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