Modulation of nitrate reductase activity by photosynthetic electron transport chain and nitric oxide balance in the red macroalga Gracilaria chilensis (Gracilariales, Rhodophyta)

Nitrate reductase (NR), a key enzyme in nitrogen metabolism, has been implicated in the production of nitric oxide (NO) in plants. The effect of photosynthetic electron transport chain inhibitors and NO scavengers or donors on NR activity of Gracilaria chilensis was studied under experimental laboratory conditions. Effective quantum yield (Φ _PSII) and NR activity were significantly diminished by 3-(3,4-dichlorophenyl)-1,1-dimethylurea and 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone, two photosynthetic electron flux inhibitors of photosystem (PS) II and PSI, respectively, but not by diphenyleneiodonium, a NADPH oxidase inhibitor, indicating a direct dependence of NR activity on the PSII and PSI electron flux. Nitrate reductase activity was sensitive to a decrease or increase of NO levels when NO scavenger (2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide) and NO donor (sodium nitroprusside) were added. Moreover, the addition of 8Br-cGMP, a secondary signal molecule, stimulated NR activity. These results evidence a modulation of the photosynthetic electron transport chain and NO balance on G. chilensis NR activity. This association could be linked to the crucial tight modulation of nitrogen assimilation and carbon metabolism to guarantee nitrite incorporation into organic compounds and to avoid toxicity by nitrite, reactive oxygen species, or nitric oxide in the cells. Nitric oxide showed to be an important signaling molecule regulating NR activity and cGMP could participate as secondary messenger on this regulation by phosphorylation and desphosphorylation processes.     

In vitro assay and light regulation of nitrate reductase in red alga Gracilaria chilensis

Nitrate reductase (NR) is the first enzyme in the nitrogen assimilation pathway. The in vitro NR activity of Gracilaria chilensis was assayed under different conditions to reveal its stability and biochemical characteristics, and an optimized in vitro assay is described. Maximal NR activities were observed at pH 8.0 and 15 degrees C. The apparent Km value for NADH was 8 microM and for nitrate 680 microM. Crude extracts of G. chilensis stored at 4 degrees C showed a 50% decrease of NR activity after 24 h. The highest NR activity value (253.20+/-2.60 x 10(-3) U g(-1)) was obtained when 100% von Stosch medium (500 microM NO3-) was added before extraction of apical parts. Algae under light:dark cycles of 12:12h exhibited circadian fluctuation of NR activity and photosynthesis with more than 2 times higher levels in the light phase. No evidence of endogenous diel rhythm controlling NR activity or photosynthesis was observed. Light pulses lasting 10 or 60 min during the darkness increased the NR activity by 30% and 45%, respectively. The results indicate that NR and photosynthesis are regulated mainly by light and not by a biological clock.     

Growth rate and carrageenan analyses in four strains of <Emphasis Type="Italic">Kappaphycus alvarezii</Emphasis> (Rhodophyta,Gigartinales) farmed in the subtropical waters of São Paulo State,Brazil

Growth rate, semi-refined and refined carrageenan yields, 3,6-anhydrogalactose and sulphate contents, and gel strength were investigated in four strains of Kappaphycus alvarezii (brown, green and red tetrasporophytic strains, and one strain derived from tetraspores progeny, called G11) farmed in subtropical waters of São Paulo State, Brazil. All studied strains showed higher growth rates from February to May, decreasing from July to December. The G11 strain exhibited lower growth rates, but had semi-refined and refined carrageenan yields significantly higher than the others and similar to a commercial sample. A negative correlation between growth rate and semi-refined carrageenan yield was verified for all strains. The brown strain had the lowest content of 3,6-anhydrogalactose, while G11 and green strains showed higher values. No clear pattern of variation and no relationship were observed for sulphate and gel strength. However, all strains presented gel strength values near the one obtained from commercial sample. These results show that the carrageenan extracted from brown, green, red, and G11 strains of K. alvarezii cultured in subtropical waters of São Paulo State, Brazil with commercial potentials.     

Effect of ultraviolet-B radiation in laboratory on morphological and ultrastructural characteristics and physiological parameters of selected cultivar of Oryza sativa L.

Ultraviolet-B radiation (UVBR) affects plants in many important ways, including reduction of growth rate and primary productivity, and changes in ultrastructures. Rice (Oryza sativa) is one of the most cultivated cereals in the world, along with corn and wheat, representing over 50 % of agricultural production. In this study, we examined O. sativa plants exposed to ambient outdoor radiation and laboratory-controlled photosynthetically active radiation (PAR) and PAR + UVBR conditions for 2 h/day during 30 days of cultivation. The samples were studied for morphological and ultrastructural characteristics, and physiological parameters. PAR + UVBR caused changes in the ultrastructure of leaf of O. sativa and leaf morphology (leaf index, leaf area and specific leaf area, trichomes, and papillae), plant biomass (dry and fresh weight), photosynthetic pigments, phenolic compounds, and protein content. As a photoprotective acclimation strategy against PAR + UVBR damage, an increase of 66.24 % in phenolic compounds was observed. Furthermore, PAR + UVBR treatment altering the levels of chlorophylls a and b, and total chlorophyll. In addition, total carotenoid contents decreased after PAR + UVBR treatment. The results strongly suggested that PAR + UVBR negatively affects the ultrastructure, morphology, photosynthetic pigments, and growth rates of leaf of O. sativa and, in the long term, it could affect the viability of this economically important plant.     

Seasonal variation of nutritional and antioxidant properties of different Kappaphycus alvarezii strains (Rhodophyta) farmed in Brazil

Journal of Applied Phycology - Antioxidant potential, carbohydrate content, ash, minerals, proteins, and amino acids of Kappaphycus alvarezii farmed along the São Paulo coast, Brazil, were...     

Ecophysiological characteristics of Porphyra spp. (Bangiophyceae,Rhodophyta): seasonal and latitudinal variations in northern-central Chile

The red macroalga Porphyra C. Agardh is one of the most ecologically successful genera that lives in the upper intertidal zone. Biochemical, physiological, and morphological acclimation strategies allow their growth and distribution as well as a quick recuperation between tidal regimens. Studies of Porphyra are poorly developed in Chile, and management and exploitation proposals need to be supported by biological and ecophysiological approaches. This study evaluated seasonal and latitudinal physiological performances of Porphyra spp. via maximum quantum yield (F _v?/?F _m), pigments, proteins, phenolic compounds, and antioxidant activity in order to describe how algae can acclimate to their environment and to provide insights to their management and use. Sampling was done at three costal sites in Chile between 25°S and 34°S between winters 2010 and 2011. A total of four different morphotypes were identified (one in the north, one in the center, and two in the south locations) and evaluated separately. Results showed seasonal and latitudinal patterns for all ecophysiological variables studied, with a general tendency of decrease in F _v?/?F _m, pigments, and soluble proteins during spring–summer seasons accompanied by an increase in the antioxidant capacity. Latitudinal differences were observed with a tendency of higher values for ecophysiological traits in central and southern morphotypes. Phenology patterns were different between an annual population in the north location and a perennial one for central-south populations. The taxonomic clarity should be evaluated in order to better understand if there exists intraspecific (dependent on morphology) or interspecific variation.     

Laboratory germination and sea out-planting of tetraspore progeny from <Emphasis Type="Italic">Kappaphycus striatum</Emphasis> (Rhodophyta) in subtropical waters of Brazil

Tetrasporophyte green strains of Kappaphycus striatum introduced from the Philippines and cultivated in the sea at Ubatuba, São Paulo, Brazil, produced tetraspores in pilot mariculture experiments. In the laboratory, the released tetraspores showed high viability and 79% of germination, producing robust plantlets and large differences in morphology. After 5 months under laboratory culture, some of the plantlets that grew best were attached to ropes, transferred into the sea and monitored for 6 months. The results show the potential of the tetraspore progeny for strain selection and the economic feasibility for mariculture of K. striatum with fast adaptation to the sea and satisfactory growth rates of 6.4±0.4 to 4.3±1.2% day^?1.