Physiological responses of the green alga Dunaliella parva (Volvocales, Chlorophyta) to controlled incremental changes in the N source
Mario Giordano A D , Alessandra Norici A , Daniel J. Gilmour B and John A. Raven CA Laboratorio di Fisiologia delle Alghe, Dipartimento di Scienze del Mare, Università Politecnica delle Marche, via Brecce Bianche, 60131 Ancona, Italy.
B Department of Molecular Biology and Biotechnology, University of Sheffield, S10 2TN Sheffield, UK.
C Scottish Crop Research Institute, University of Dundee at SCRI, Invergowrie, DD2 5DA Dundee, UK.
D Corresponding author. Email: m.giordano@univpm.it
Functional Plant Biology 34(10) 925-934 https://doi.org/10.1071/FP07049
Submitted: 23 February 2007 Accepted: 20 July 2007 Published: 13 September 2007
Abstract
This work is aimed at obtaining information on the acclimation processes of the green flagellate Dunaliella parva Lerche to gradual changes in the N source from NO3– to NH4+, in continuous cultures. Photosynthesis, dark respiration, and light-independent carbon fixation (LICF) rates, chlorophyll a fluorescence, RUBISCO and phosphoenolpyruvate carboxylase (PEPc) activities, plasmalemma electrical potential difference, cell volume, and absolute or relative amounts of major cell constituents were measured. Two phases characterised the response to the transition from NO3– to NH4+: (1) an initial phase in which photosynthesis and anaplerosis were stimulated and protein increased; (2) a subsequent phase in which most parameters reached new values that were close to those at the beginning of the experiment (100% NO3–). The only exceptions were PEPc activity and LICF, whose rates remained at least 2-fold higher than at 100% NO3–, when NH4+ was the sole N source. The results are indicative of a tendency to re-establish homeostasis, after an initial perturbation of the intracellular composition. The roles of different metabolic processes during acclimation are discussed.
Additional keywords: acclimation, chlorophyll fluorescence, FTIR spectroscopy, homeostasis, resource allocation.
Acknowledgements
We thank Professor Peter Horton, R. Hill Institute, Sheffield, UK, for his assistance with fluorescence measurements, Dr Giancarlo Gobbi, Dipartimento di Scienze dei Materiali, Università Politecnica delle Marche, Ancona, Italy, for his assistance with ion chromatography and Mr Matteo Palmucci for his help in algae maintenance.
Amory AM,
Vanlerberghe GC, Turpin DH
(1991) Demonstration of both a photosynthetic and a non-photosynthetic CO2 requirement for NH4
+ assimilation in the green alga Selenastrum minutum. Plant Physiology 95, 192–196.
| PubMed |
Andrade SLA,
Dickmans A,
Ficner R, Einsle O
(2005) Crystal structure of the archaeal ammonium transporter Amt-1 from Archaeoglobus fulgidus. Proceedings of the National Academy of Sciences USA 102, 14994–14999.
| Crossref | GoogleScholarGoogle Scholar |
Azov Y, Goldman JC
(1982) Free ammonia inhibition of algal photosynthesis in intensive culture. Applied and Environmental Microbiology 43, 735–739.
| PubMed |
Beardall J, Giordano M
(2002) Ecological implications of algal CO2 concentrating mechanisms and their regulation. Functional Plant Biology 29, 335–347.
| Crossref | GoogleScholarGoogle Scholar |
Berges JA
(1997) Algal nitrate reductase. European Journal of Phycology 32, 3–8.
| Crossref | GoogleScholarGoogle Scholar |
Bradford MM
(1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–254.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Braun H-P, Zabaleta E
(2007) Carbonic anhydrase subunits of the mitochondrial NADH dehydrogenase complex (complex I) in plants. Physiologia Plantarum 129, 114–122.
| Crossref | GoogleScholarGoogle Scholar |
Britto DT,
Siddiqui MY,
Glass ADM, Kronzucker HJ
(2001) Futile transmembrane NH4
+ cycling: a cellular hypothesis to explain ammonium toxicity in plants. Proceedings of the National Academy of Sciences of the United States of America 98, 4255–4258.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Britto DT,
Siddiqi MY,
Glass ADM, Kronzucker HJ
(2002) Subcellular NH4
+ flux analysis in leaf segments of wheat (Triticum aestivum). New Phytologist 155, 373–380.
| Crossref | GoogleScholarGoogle Scholar |
Clark DR, Flynn KJ
(2002) N assimilation in the noxious flagellate Heterosigma carterae (Raphidophyceae): dependence on light, N source, and physiological state. Journal of Phycology 38, 503–512.
| Crossref | GoogleScholarGoogle Scholar |
Collins S, Bell G
(2004) Phenotypic consequences of 1000 generations of selection at elevated CO2 in a green alga. Nature 431, 566–569.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Collins S,
Sultemeyer D, Bell G
(2006) Changes in C uptake in populations of Chlamydomonas reinhardtii selected at high CO2. Plant, Cell & Environment 29, 1812–1819.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Elrifi IR, Turpin DH
(1986) Nitrate and ammonium induced photosynthetic suppression in N-limited Selenastrum minutum. Plant Physiology 81, 273–279.
| PubMed |
Flynn KJ,
Fasham MJR, Hipkin CR
(1997) Modelling the interactions between ammonium and nitrate uptake in marine phytoplankton. Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences 352, 1625–1645.
| Crossref | GoogleScholarGoogle Scholar |
Galván A, Fernández E
(2001) Eukaryotic nitrate and nitrite transporters. Cellular and Molecular Life Sciences 58, 225–233.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Garcia-Sánchez MJ,
Jaime MP,
Ramos A,
Sanders D, Fernández JA
(2000) Sodium-dependent nitrate transport at the plasma membrane of leaf cells of the marine higher plant Zostera marina L. Plant Physiology 122, 879–885.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Gilmour DJ,
Kaaden R, Gimmler H
(1985) Vanadate inhibition of ATPases of Dunaliella parva in vitro and in vivo. Journal of Plant Physiology 118, 111–126.
Gimmler H, Greenway H
(1983) Tetraphenylphosphonium (TTP+) is not suitable for the assessment of electrical potentials in Chlorella emersonii. Plant, Cell & Environment 6, 739–744.
Giordano M
(1997) Adaptation of Dunaliella salina (Volvocales, Chlorophyceae) to growth on NH4
+ as the sole nitrogen source. Phycologia 36, 345–350.
Giordano M
(2001) Interactions between C and N metabolism in Dunaliella salina cells cultured at elevated CO2 and high N concentrations. Journal of Plant Physiology 158, 577–581.
| Crossref | GoogleScholarGoogle Scholar |
Giordano M, Bowes G
(1997) Gas exchange and C allocation in Dunaliella salina cells in response to the N source and CO2 concentration used for growth. Plant Physiology 115, 1049–1056.
| PubMed |
Giordano M,
Davis JS, Bowes G
(1994) Organic carbon release by Dunaliella salina (Chlorophyta) under different growth conditions of CO2, nitrogen, and salinity. Journal of Phycology 30, 249–257.
| Crossref | GoogleScholarGoogle Scholar |
Giordano M,
Pezzoni V, Hell R
(2000) Strategies for the allocation of resources under sulfur limitation in the green alga Dunaliella salina. Plant Physiology 124, 857–864.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Giordano M,
Kansiz M,
Heraud P,
Beardall J,
Wood B, McNaughton D
(2001) Fourier transform infrared spectroscopy as a novel tool to investigate changes in intracellular macromolecular pools in the marine microalga Chaetoceros muellerii (Bacillariophyceae). Journal of Phycology 37, 271–279.
| Crossref | GoogleScholarGoogle Scholar |
Giordano M,
Norici A, Gilmour DJ
(2002) Influence of the nitrogen source and metabolites on phosphoenolpyruvate carboxylase activity in the unicellular green alga Dunaliella parva ccap 19/9. Phycologia 41, 133–137.
Giordano M,
Norici A,
Forssen M,
Eriksson M, Raven JA
(2003) An anaplerotic role for mithochondrial carbonic anhydrase in Chlamydomonas reinhardtii. Plant Physiology 132, 2126–2134.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Granum E, Myklestad SM
(1999) Effects of NH4
+ assimilation on dark carbon fixation and beta-1,3-glucan metabolism in the marine diatom Skeletonema costatum (Bacillariophyceae). Journal of Phycology 35, 1191–1199.
| Crossref | GoogleScholarGoogle Scholar |
Guo S,
Schinner K,
Sattelmacher B, Hansen U-P
(2005) Different apparent CO2 compensation points in nitrate- and ammonium-grown Phaseolus vulgaris and the relationship to non-photorespiratory CO2 evolution. Physiologia Plantarum 123, 288–301.
| Crossref | GoogleScholarGoogle Scholar |
Hard BC, Gilmour DJ
(1996) The uptake of organic compounds by Dunaliella parva CCAP 19/9. European Journal of Phycology 31, 217–224.
| Crossref | GoogleScholarGoogle Scholar |
He QH,
Qiao DR,
Zhang QL,
Xu H,
Wei L,
Gu Y, Cao Y
(2004) Cloning and expression study of a putative high-affinity nitrate transporter gene from Dunaliella salina. Journal of Applied Phycology 16, 395–400.
| Crossref | GoogleScholarGoogle Scholar |
Hildebrand M
(2005) Cloning and functional characterization of ammonium transporters from the marine diatom Cylindrotheca fusiformis (Bacillariophyceae). Journal of Phycology 41, 105–113.
| Crossref | GoogleScholarGoogle Scholar |
Huppe HC, Turpin DH
(1994) Integration of carbon and nitrogen metabolism in plant and algal cells. Annual Review of Plant Physiology and Plant Molecular Biology 45, 577–607.
| Crossref | GoogleScholarGoogle Scholar |
Ishiyama K,
Inoue E,
Watanabe-Takahashi A,
Obara M,
Yamaya T, Takahashi H
(2004) Kinetic properties and ammonium-dependent regulation of cytosolic isoenzymes of glutamine synthetase in Arabidopsis. Journal of Biological Chemistry 279, 16598–16605.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Jahn TP,
Møller ALB,
Zeuthen T,
Holm LM,
Klærke DA,
Mohsin B,
Kühlbrandt W, Schjoerring JK
(2004) Aquaporin homologues in plants and mammals transport ammonia. FEBS Letters 574, 31–36.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Khademi S,
O’Connell J,
Remis J,
Robles-Colmenares Y,
Miercke LJW, Stroud RM
(2004) Mechanism of ammonia transport by Amt/Mep/Rh: structure of AmtB at 1.35 Å. Science 305, 1587–1594.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Krause GH, Weis E
(1991) Chlorophyll fluorescence and photosynthesis: the basics. Annual Review of Plant Physiology and Plant Molecular Biology 42, 313–349.
| Crossref | GoogleScholarGoogle Scholar |
Kristiansen S, Lund BA
(1989) Nitrogen cycling in the Barents Sea-I. Uptake of nitrogen in the water column. Deep-Sea Research 36, 255–268.
| Crossref | GoogleScholarGoogle Scholar |
Levasseur M,
Thompson PA, Harrison PJ
(1993) Physiological acclimation of marine phytoplankton to different nitrogen sources. Journal of Phycology 29, 587–595.
| Crossref | GoogleScholarGoogle Scholar |
Loder TC,
Ganning B, Love JA
(1996) Ammonia dynamics in coastal rockpools affected by gull guano. Journal of ExperimentalMarine Biology and Ecology 196, 113–129.
| Crossref | GoogleScholarGoogle Scholar |
Loqué D,
Ludewig U,
Yuan L, von Wirén N
(2005) Tonoplast intrinsic protein AtTIP2;1 and AtTIP2;3 facilitate NH3 transport into the vacuole. Plant Physiology 137, 671–680.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Lu C,
Torzillo G, Vonshak A
(1999) Kinetic response of photosystem II photochemistry in the cyanobacterium Spirulina platensis to high salinity is characterized by two distinct phases. Australian Journal of Plant Physiology 26, 283–292.
Ludewig U,
Wilken S,
Wu B,
Jost W, Obrdlik P , et al.
(2003) Homo- and hetero-oligomerization of ammonium transporter-1 NH4
+ uniporters. Journal of Biological Chemistry 278, 45603–45610.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Mayer M,
Schaaf G,
Mouro I,
Lopez C,
Colin Y,
Neumann P,
Cartron J-P, Ludewig U
(2006a) Different transport mechanisms in plant and human AMT/Rh-type ammonium transporters. Journal of General Physiology 127, 133–144.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Mayer M,
Dynowski M, Ludewig U
(2006b) Ammonium ion transport by the AMT/Rh homologue LeAMT1;1. The Biochemical Journal 396, 431–437.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Melo-Oliveira R,
Oliveira IC, Coruzzi GM
(1996) Arabidopsis mutant analysis and gene regulation define a non-redundant role for glutamate dehydrogenase in nitrogen assimilation. Proceedings of the National Academy of Sciences USA 93, 4718–4723.
| Crossref | GoogleScholarGoogle Scholar |
Metaxas A, Scheibling RE
(1994) Changes in phytoplankton abundance in tidepools over the period of tidal isolation. Botanica Marina 37, 301–314.
Montechiaro F,
Hirschmugl C,
Raven JA, Giordano M
(2006) Dark acclimation processes in a cave-dwelling cyanobacterium. Plant, Cell & Environment 29, 2198–2204.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Norici A,
Dalsass A, Giordano M
(2002) Role of phosphoenolpyruvate carboxylase in anaplerosis in the green microalga Dunaliella salina cultured under different nitrogen regimes. Physiologia Plantarum 116, 186–191.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Oliveira IC,
Brears T,
Knight TJ,
Clark A, Coruzzi GM
(2002) Over-expression of cytosolic glutamine synthetase: relation to nitrogen, light, and photorespiration. Plant Physiology 129, 1170–1180.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Oren-Shamir M,
Oick U, Avron M
(1990) Plasma membrane potential of the alga Dunaliella, and its relation to osmoregulation. Plant Physiology 93, 403–408.
| PubMed |
Price NM,
Cochlan WP, Harrison PJ
(1985) Time course of uptake of inorganic and organic nitrogen by phytoplankton in the Strait of Georgia: comparison of frontal and stratified communities. Marine Ecology Progress Series 27, 39–53.
Raven JA, Smith FA
(1976) Nitrogen assimilation and transport in vascular land plants in relation to intracellular pH regulation. New Phytologist 76, 415–431.
| Crossref | GoogleScholarGoogle Scholar |
Raven JA, Farquhar GD
(1990) The influence of N-metabolism and organic-acid synthesis on the natural abundance of isotopes of carbon in plants. New Phytologist 116, 505–529.
| Crossref | GoogleScholarGoogle Scholar |
Rees D,
Lee CB,
Gilmour DJ, Horton P
(1992) Mechanisms for controlling balance between light input and utilisation in the salt tolerant alga Dunaliella C9AA. Photosynthesis Research 32, 181–191.
| Crossref | GoogleScholarGoogle Scholar |
Remis D,
Simonis W, Gimmler H
(1992) Measurements of the transmembrane electrical potential of Dunaliella acidophilia by microelectrodes. Archives of Microbiology 158, 350–355.
| Crossref | GoogleScholarGoogle Scholar |
Ritchie RJ
(1984) A critical assessment of the use of lipophilic cations as membrane potential probes. Progress in Biophysics and Molecular Biology 43, 1–32.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Ritchie RJ
(1999) Tetraphenylphosphonium cation (TPP+) is not a reliable membrane potential probe in the cyanobacterium Synechococcus R-2 PCC 7942. New Phytologist 141, 387–399.
| Crossref | GoogleScholarGoogle Scholar |
Sasaki H,
Kataoka H,
Muratami A, Kawai H
(2004) Inorganic ion composition in brown algae, with special reference to sulfuric acid ion accumulation. Hydrobiologia 512, 255–262.
| Crossref | GoogleScholarGoogle Scholar |
Sasaki H,
Kataoka H,
Kamiya M, Kawai H
(1999) Accumulation of sulfuric acid in Dictyotales (Phaeophyceae): taxonomic distribution and ion chromatography of cell extracts. Journal of Phycology 35, 732–739.
| Crossref | GoogleScholarGoogle Scholar |
Smith FA, Raven JA
(1979) Intracellular pH and its regulation. Annual Review of Plant Physiology 30, 289–311.
| Crossref | GoogleScholarGoogle Scholar |
Soupene E,
Lee H, Kustu S
(2002) Ammonium/methylammonium transport (Amt) proteins facilitate diffusion of NH3 bidirectionally. Proceedings of the National Academy of Sciences USA 99, 3926–3931.
| Crossref | GoogleScholarGoogle Scholar |
Turpin DH
(1991) Effect of inorganic N availability on algal photosynthesis and carbon metabolism. Journal of Phycology 27, 14–20.
| Crossref | GoogleScholarGoogle Scholar |
Turpin DH, Weger HG
(1988) Steady-state chlorophyll a fluorescence transients during ammonium assimilation by the N-limited green alga Selenastrum minutum. Plant Physiology 88, 97–101.
| PubMed |
Young EB, Beardall J
(2003) Rapid ammonium- and nitrate-induced perturbations to Chl a fluorescence in nitrogen-stressed Dunaliella teriolecta (Chlorophyta). Journal of Phycology 39, 332–342.
