Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/22700
標題: 浮游植物氮源利用相關基因之表現及其在海洋生態上之應用
Expression of nitrogen-utilization related genes in marine phytoplankton and its application to the study of marine ecology
作者: 康利國
Kang, Lee-Kuo
關鍵字: phytoplankton
浮游植物
nitrate transporter
nitrogen-utilization related genes
nitrogen maker gene
硝酸運輸基因
氮源利用相關基因
氮指標基因
出版社: 生命科學系所
引用: Adhitya, A., Thomas, F. I. M. & Ward, B. B. 2007. Diversity of assimilatory nitrate reductase genes from plankton and epiphytes associated with a seagrass bed. Microb. Ecol. 54:587-97. Ahmad, I. & Hellebust, J. A. 1985a. Effects of methionine sulfoximine on growth and nitrogen assimilation of the marine microalga Chlorella autotrophica. Mar. Biol. 86:85-91. Ahmad, I. & Hellebust, J. A. 1985b. Transport and assimilation of nitrogen by Stichococcus bacillaris grown in the presence of methionine sulfoximine. Plant Physiol. 79:1125-6. Ahn, S., Kulis, D. M., Erdner, D. L., Anderson, D. M. & Walt, D. R. 2006. Fiber optic microarray for simultaneous detection of multiple harmful algal bloom species. Appl. Environ. Microbiol. 72:5742-9. Allen, A. E., Ward, B. B. & Song, B. K. 2005. Characterization of diatom (Bacillariophyceae) nitrate reductase genes and their detection in marine phytoplankton communities. J. Phycol. 41:95-104. Almassy, R. J., Janson, C. A., Hamlin, R., Xuong, N. H. & Eisenberg, D. 1986. Novel subunit subunit interactions in the structure of glutamine synthetase. Nature 323:304-9. Anderson, D. M., Kulis, D. M., Keafer, B. A., Gribble, K. E., Marin, R. & Scholin, C. A. 2005. Identification and enumeration of Alexandrium spp. from the Gulf of Maine using molecular probes. Deep Sea Res. Part II Topical Stud. Oceanogr. 52:2467-90. Armbrust, E. V., Berges, J. A., Bowler, C., Green, B. R., Martinez, D., Putnam, N. H., Zhou, S., Allen, A. E., Apt, K. E., Bechner, M., Brzezinski, M. A., Chaal, B. K., Chiovitti, A., Davis, A. K., Demarest, M. S., Detter, J. C., Glavina, T., Goodstein, D., Hadi, M. Z., Hellsten, U., Hildebrand, M., Jenkins, B. D., Jurka, J., Kapitonov, V. V., Kroger, N., Lau, W. W. Y., Lane, T. W., Larimer, F. W., Lippmeier, J. C., Lucas, S., Medina, M., Montsant, A., Obornik, M., Parker, M. S., Palenik, B., Pazour, G. J., Richardson, P. M., Rynearson, T. A., Saito, M. A., Schwartz, D. C., Thamatrakoln, K., Valentin, K., Vardi, A., Wilkerson, F. P. & Rokhsar, D. S. 2004. The genome of the diatom Thalassiosira pseudonana: ecology, evolution, and metabolism. Science 306:79-86. Asai, R., Ootani, K., Nomura, Y., Nakamura, C., Ikebukuro, K., Arikawa, Y., Miyake, J. & Karube, I. 2003. PCR-based ribosomal DNA detection technique for microalga (Heterosigma carterae) causing red tide and its application to a biosensor using labeled probe. Mar. Biotechnol. 5:417-23. Barr, N. G., Tijsen, R. J. & Rees, T. A. V. 2004. Contrasting effects of methionine sulfoximine on uptake and assimilation of ammonium in Ulva intestinalis (Chlorophyceae). J. Phycol. 40:697-704. Berg, G. M., Glibert, P. M., Lomas, M. W. & Burford, M. A. 1997. Organic nitrogen uptake and growth by the chrysophyte Aureococcus anophagefferens during a brown tide event. Mar. Biol. 129:377-87. Bhattacharya, D. & Medlin, L. 1995. The phylogeny of plastids- a review based on comparisons of small subunit ribosomal RNA coding regions. J. Phycol. 31:489-98. Bhattacharya, D., Yoon, H. S. & Hackett, J. D. 2004. Photosynthetic eukaryotes unite: endosymbiosis connects the dots. Bioessays 26:50-60. Boehringer-Mannheim. 1993. The DIG System User''s Guide for Filter Hybridization. Boehringer Mannheim GmbH, Biochemica, Germany, 89 pp. Bowers, H. A., Tengs, T., Glasgow, H. B., Burkholder, J. M., Rublee, P. A. & Oldach, D. W. 2000. Development of real-time PCR assays for rapid detection of Pfiesteria piscicida and related dinoflagellates. Appl. Environ. Microbiol. 66:4641-8. Britto, D. T., Siddiqi, M. Y., Glass, A. D. M. & Kronzucker, H. J. 2001. Futile transmembrane NH4+ cycling: a cellular hypothesis to explain ammonium toxicity in plants. Proc. Natl. Acad. Sci. U. S. A. 98:4255-8. Cerezo, M., Tillard, P., Filleur, S., Munos, S., Daniel-Vedele, F. & Gojon, A. 2001. Major alterations of the regulation of root NO3- uptake are associated with the mutation of Nrt2.1 and Nrt2.2 genes in arabidopsis. Plant Physiol. 127:262-71. Chen, Q. & Silflow, C. D. 1996. Isolation and characterization of glutamine synthetase genes in Chlamydomonas reinhardtii. Plant Physiol. 112:987-96. Cheng, L. C., Hwang, S. P. L. & Chang, J. 1997. Gene sequence and expression of an analog of proliferating cell nuclear antigen (PCNA) in the alga Tetraselmis chui and detection of the encoded protein with anti-rat PCNA monoclonal antibody. Appl. Environ. Microbiol. 63:4010-4. Chung, C. C., Hwang, S. P. L. & Chang, J. 2003. Identification of a high-affinity phosphate transporter gene in a prasinophyte alga, Tetraselmis chui, and its expression under nutrient limitation. Appl. Environ. Microbiol. 69:754-9. Chung, C. C., Hwang, S. P. L. & Chang, J. 2005. Cooccurrence of ScDSP gene expression, cell death, and DNA fragmentation in a marine diatom, Skeletonema costatum. Appl. Environ. Microbiol. 71:8744-51. Chung, C. T., Niemela, S. L. & Miller, R. H. 1989. One-step preparation of competent Escherichia coli- transformation and storage of bacterial cells in the same solution. Proc. Natl. Acad. Sci. U. S. A. 86:2172-5. Clark, C. G. 1992. DNA purification from polysaccharide rich cells. In Lee, J. J. & Soldo, A. T. [Eds.] Protocol in protozoology. Society of Protozoology, Lawrence, KS. pp. D-31-2. Collos, Y., Descolas-Gros, C., Fontugne, M., Mortain-Bertrand, A., Chretiennot-Dinet, M. J. & Frikha, M. G. 1993. Chemical, isotopic and enzymatic monitoring of free and enclosed seawater: Implications for primary production estimates in incubation bottles. Mar. Ecol. Prog. Ser. 93:49-54. Crawford, N. M. & Glass, A. D. M. 1998. Molecular and physiological aspects of nitrate uptake in plants. Trends Plant Sci. 3:389-95. Davidson, K., Flynn, K. J. & Cunningham, A. 1992. Non-steady state ammonium-limited growth of the marine phytoflagellate, Isochrysis galbana Parke. New phytol. 122:433-8. Dolganov, N. A. M., Bhaya, D. & Grossman, A. R. 1995. Cyanobacterial protein with similarity to the chlorophyll a/b binding proteins of higher plants- evolution and regulation. Proc. Natl. Acad. Sci. U. S. A. 92:636-40. Dugdale, R. C. & Wilkerson, F. P. 1986. The use of super15N to measure nitrogen uptake in eutrophic oceans; experimental considerations. Limnol. Oceanogr. 31:673-89. Filleur, S. & Daniel-Vedele, F. 1999. Expression analysis of a high-affinity nitrate transporter isolated from Arabidopsis thaliana by differential display. Planta 207:461-9. Florencio, F. J. & Vega, J. M. 1983. Utilization of nitrate, nitrite and ammonium by Chlamydomonas reinhardii- photoproduction of ammonium. Planta 158:288-93. Fogg, G. E. & Calvariomartinez, O. 1989. Effects of bottle size in determinations of primary productivity by phytoplankton. Hydrobiologia 173:89-94. Forde, B. G. 2000. Nitrate transporters in plants: structure, function and regulation. Biochim. Biophys Acta-Biomembr. 1465:219-35. Fuller, N. J., West, N. J., Marie, D., Yallop, M., Rivlin, T., Post, A. F. & Scanlan, D. J. 2005. Dynamics of community structure and phosphate status of picocyanobacterial populations in the Gulf of Aqaba, Red Sea. Limnol. Oceanogr. 50:363-75. Galluzzi, L., Penna, A., Bertozzini, E., Vila, M., Garces, E. & Magnani, M. 2004. Development of a real-time PCR assay for rapid detection and quantification of Alexandrium minutum (a dinoflagellate). Appl. Environ. Microbiol. 70:1199-206. Galvan, A. & Fernandez, E. 2001. Eukaryotic nitrate and nitrite transporters. Cell. Mol. Life Sci. 58:225-33. Galvan, A., Rexach, J., Mariscal, V. & Fernandez, E. 2002. Nitrite transport to the chloroplast in Chlamydomonas reinhardtii: molecular evidence for a regulated process. J. Exp. Bot. 53:845-53. Gazzarrini, S., Lejay, T., Gojon, A., Ninnemann, O., Frommer, W. B. & von Wiren, N. 1999. Three functional transporters for constitutive, diurnally regulated, and starvation-induced uptake of ammonium into arabidopsis roots. Plant Cell 11:937-47. Geider, R. J. & Laroche, J. 1994. The role of iron in phytoplankton photosynthesis, and the potential for iron-limitation of primary productivity in the Sea. Photosynth. Res. 39:275-301. Geider, R. J., Laroche, J., Greene, R. M. & Olaizola, M. 1993. Response of the photosynthetic apparatus of Phaeodactylum tricornutum (Bacillariophyceae) to nitrate, phosphate, or iron starvation. J. Phycol. 29:755-66. Gerendas, J., Zhu, Z. J., Bendixen, R., Ratcliffe, R. G. & Sattelmacher, B. 1997. Physiological and biochemical processes related to ammonium toxicity in higher plants. Z. Pflanzener. Bodenk. 160:239-51. Glass, A. D. M., Brito, D. T., Kaiser, B. N., Kronzucker, H. J., Kumar, A., Okamoto, M., Rawat, S. R., Siddiqi, M. Y., Silim, S. M., Vidmar, J. J. & Zhuo, D. 2001. Nitrogen transport in plants, with an emphasis on the regulation of fluxes to match plant demand. J. Plant Nutr. Soil Sci. 164:199-207. Glass, A. D. M., Britto, D. T., Kaiser, B. N., Kinghorn, J. R., Kronzucker, H. J., Kumar, A., Okamoto, M., Rawat, S., Siddiqi, M. Y., Unkles, S. E. & Vidmar, J. J. 2002. The regulation of nitrate and ammonium transport systems in plants. J. Exp. Bot. 53:855-64. Gong, G.-C. 1992. Chemical hydrography of the Kuroshio front in the sea northern to Taiwan. In Institute of Oceanography. National Taiwan University, Taipei. Gonzalez-Ballester, D., Camargo, A. & Fernandez, E. 2004. Ammonium transporter genes in Chlamydomonas: the nitrate-specific regulatory gene Nit2 is involved in Amt1;1 expression. Plant Mol. Biol. 56:863-78. Gray, A., Wawrik, B., Paul, J. & Casper, E. 2003. Molecular detection and quantitation of the red tide dinoflagellate Karenia brevis in the marine environment. Appl. Environ. Microbiol. 69:5726-30. Gray, M. W. 1999. Evolution of organellar genomes. Curr. Opin. Genet. Dev. 9:678-87. Guillard, R. R. L. 1983. Culture of phytoplankton for feeding marine invertebrates. In Berg Jr, C. J. [Ed.] Culture of marine invertebrates. Seclected readings. Hutchinson Ross Publishing Co., Stroudsberg, Pensylvania, pp. 108-32. Guillard, R. R. L. & Ryther, J. H. 1962. Studies on marine planktonic diatom. I. Cyclotella nana Hustedt and Detonula confervacea (Cleve) Gran. Can. J. Microbiol. 8:229-39. Guo, F. Q., Wang, R. C., Chen, M. S. & Crawford, N. M. 2001. The Arabidopsis dual-affinity nitrate transporter gene AtNRT1.1 (CHL1) is activated and functions in nascent organ development during vegetative and reproductive growth. Plant Cell 13:1761-77. Guo, F. Q., Wang, R. C. & Crawford, N. M. 2002. The Arabidopsis dual-affinity nitrate transporter gene AtNRT1.1 (CHL1) is regulated by auxin in both shoots and roots. J. Exp. Bot. 53:835-44. Handy, S. M., Hutchins, D. A., Cary, S. C. & Coyne, K. J. 2006. Simultaneous enumeration of multiple raphidophyte species by quantitative real-time PCR: capabilities and limitations. Limnol. Oceanogr. Methods 4:193-204. Healey, F. P. & Hendzel, L. L. 1979. Fluorometric measurement of alkaline-phosphatase activity in algae. Freshw. Biol. 9:429-39. Herrero, A., Muro-Pastor, A. M. & Flores, E. 2001. Nitrogen control in cyanobacteria. J. Bacteriol. 183:411-25. Hildebrand, M. 2005. Cloning and functional characterization of ammonium transporters from the marine diatom Cylindrotheca fusiformis (Bacillariophyceae). J. Phycol. 41:105-13. Hildebrand, M. & Dahlin, K. 2000. Nitrate transporter genes from the diatom Cylindrotheca fusiformis (Bacillariophyceae): mRNA levels controlled by nitrogen source and by the cell cycle. J. Phycol. 36:702-13. Howitt, S. M. & Udvardi, M. K. 2000. Structure, function and regulation of ammonium transporters in plants. Biochim. Biophy. Acta Biomembr. 1465:152-70. Hwang, S. P. L., Wang, S. K., Wei, S. F., Cheng, L. C. & Chang, J. 1999. Identification and expression pattern of DNA polymerase alpha gene in a marine diatom, Skeletonema costatum. Mar. Biotechnol. 1:200-6. Inokuchi, R., Kuma, K., Miyata, T. & Okada, M. 2002. Nitrogen-assimilating enzymes in land plants and algae: phylogenic and physiological perspectives. Physiol. Plant. 116:1-11. Ishiyama, K., Inoue, E., Tabuchi, M., Yamaya, T. & Takahashi, H. 2004a. Biochemical background and compartmentalized functions of cytosolic glutamine synthetase for active ammonium assimilation in rice roots. Plant Cell Physiol. 45:1640-7. Ishiyama, K., Inoue, E., Tabuchi, M., Yamaya, T. & Takahashi, H. 2005. Physiological functions and regulations for metabolic pathway of glutamine synthetase in Arabidopsis and rice roots. Plant Cell Physiol. 46:S70. Ishiyama, K., Inoue, E., Takahashi-Watanbe, A., Kojima, S., Yamaya, T. & Takahashi, H. 2003. Identification of glutamine synthetase (GS) and glutamate synthase (GOGAT) responsible for assimilation of exogenous NH4+ in Arabidopsis root. Plant Cell Physiol. 44:S40. Ishiyama, K., Inoue, E., Watanabe-Takahashi, A., Obara, M., Yamaya, T. & Takahashi, H. 2004b. Kinetic properties and ammonium dependent regulation of cytosolic isoenzymes of glutamine synthetase in Arabidopsis. J. Biol. Chem. 279:16598-605. Ishiyama, K., Inoue, E., Watanabe-Takahashi, A., Obara, M., Yamaya, T. & Takahashi, H. 2004c. Nitrogen dependent regulation of cytosolic glutamine synthetase in Arabidopsis roots. Plant Cell Physiol. 45:S98. John, D. E., Patterson, S. S. & Paul, J. H. 2007. Phytoplankton group specific quantitative polymerase chain reaction assays for RuBisCO mRNA transcripts in seawater. Mar. Biotechnol. 9:747-59. Kang, L. K., Hwang, S. P. L., Gong, G. C., Lini, H. J., Chen, P. C. & Chang, J. 2007. Influences of nitrogen deficiency on the transcript levels of ammonium transporter, nitrate transporter and glutamine synthetase genes in Isochrysis galbana (Isochrysidales, Haptophyta). Phycologia 46:521-33. Kawakami, S. K., Gledhill, M. & Achterberg, E. P. 2006. Effects of metal combinations on the production of phytochelatins and glutathione by the marine diatom Phaeodactylum tricornutum. Biometals 19:51-60. Keane, T. M., Creevey, C. J., Pentony, M. M., Naughton, T. J. & Mclnerney, J. O. 2006. Assessment of methods for amino acid matrix selection and their use on empirical data shows that ad hoc assumptions for choice of matrix are not justified. BMC Evol. Biol. 6:1-29. Keeling, P. 2004. A brief history of plastids and their hosts. Protist 155:3-7. Ki, J. S. & Han, M. S. 2005. Sequence based diagnostics and phylogenetic approach of uncultured freshwater dinoflagellate Peridinium (Dinophyceae) species, based on single cell sequencing of rDNA. J. Appl. Phycol. 17:147-53. Ki, J. S. & Han, M. S. 2007. Rapid molecular identification of the harmful freshwater dinoflagellate Peridinium in various life stages using genus-specific single-cell PCR. J. Appl. Phycol. 19:467-70. Ki, J. S., Jang, G. Y. & Han, M. S. 2004. Integrated method for single-cell DNA extraction, PCR amplification, and sequencing of ribosomal DNA from harmful dinoflagellates Cochlodinium polykrikoides and Alexandrium catenella. Mar. Biotechnol. 6:587-93. Kilham, P. & Hecky, R. E. 1988. Comparative ecology of marine and freshwater phytoplankton. Limnol. Oceanogr. 33:776-95. Kolber, Z., Zehr, J. & Falkowski, P. 1988. Effects of growth irradiance and nitrogen limitation on photosynthetic energy conversion in photosystem-Ii. Plant Physiol. 88:923-9. Koltermann, M., Moroni, A., Gazzarini, S., Nowara, D. & Tischner, R. 2003. Cloning, functional expression and expression studies of the nitrate transporter gene from Chlorella sorokiniana (strain 211-8k). Plant Mol. Biol. 52:855-64. La Roche, J., Boyd, P. W., McKay, R. M. L. & Geider, R. J. 1996. Flavodoxin as an in situ marker for iron stress in phytoplankton. Nature 382:802-5. La Roche, J., Geider, R. J., Graziano, L. M., Murray, H. & Lewis, K. 1993. Induction of specific proteins in eukaryotic algae grown under iron-deficient, phosphorus- deficient, or nitrogen-deficient conditions. J. Phycol. 29:767-77. La Roche, J., McKay, R. M. L. & Boyd, P. 1999. Immunological and molecular probes to detect phytoplankton responses to environmental stress in nature. Hydrobiologia 401:177-98. La Roche, J., Nuzzi, R., Waters, R., Wyman, K., Falkowski, P. G. & Wallace, D. W. R. 1997. Brown Tide blooms in Long Island''s coastal waters linked to interannual variability in groundwater flow. Glob. Change Biol. 3:397-410. Lin, S. J. & Carpenter, E. J. 1995. Growth characteristics of marine phytoplankton determined by cell cycle proteins - the cell cycle of Ethmodiscis rex (Bacillariophyceae) in the southwestern North Atlantic Ocean and Caribbean Sea. J. Phycol. 31:778-85. Lin, S. J. & Carpenter, E. J. 1998. Identification and preliminary characterization of PCNA gene in the marine phytoplankton Dunaliella tertiolecta and Isochrysis galbana. Mol. Mar. Biol. Biotechnol. 7:62-71. Lin, S. J., Chang, J. & Carpenter, E. J. 1994. Detection of proliferating cell nuclear antigen analog in 4 species of marine phytoplankton. J. Phycol. 30:449-56. Lin, S. J., Chang, J. & Carpenter, E. J. 1995. Growth characteristics of phytoplankton determined by cell cycle proteins PCNA- immunostaining of Dunaliella tertiolecta (Chlorophyceae). J. Phycol. 31:388-95. Lin, S. J. & Corstjens, P. L. A. M. 2002. Molecular cloning and expression of the proliferating cell nuclear antigen gene from the coccolithophorid Pleurochrysis carterae (Haptophyceae). J. Phycol. 38:164-73. Lindell, D., Padan, E. & Post, A. F. 1998. Regulation of ntcA expression and nitrite uptake in the marine Synechococcus sp. strain WH 7803. J. Bacteriol. 180:1878-86. Lindell, D., Penno, S., Al-Qutob, M., David, E., Rivlin, T., Lazar, B. & Post, A. F. 2005. Expression of the nitrogen stress response gene ntcA reveals nitrogen-sufficient Synechococcus populations in the oligotrophic northern Red Sea. Limnol. Oceanogr. 50:1932-44. Lindell, D. & Post, A. F. 2001. Ecological aspects of ntcA gene expression and its use as an indicator of the nitrogen status of marine Synechococcus spp. Appl. Environ. Microbiol. 67:3340-9. Liu, J. W., Jiao, N. Z., Hong, H. S., Luo, T. W. & Cai, H. Y. 2005. Proliferating cell nuclear antigen (PCNA) as a marker of cell proliferation in the marine dinoflagellate Prorocentrum donghaiense Lu and the green alga Dunaliella salina Teodoresco. J. Appl. Phycol. 17:323-30. Liu, K. H., Huang, C. Y. & Tsay, Y. F. 1999. CHL1 is a dual-affinity nitrate transporter of arabidopsis involved in multiple phases of nitrate uptake. Plant Cell 11:865-74. Liu, K. H. & Tsay, Y. F. 2003. Switching between the two action modes of the dual-affinity nitrate transporter CHL1 by phosphorylation. EMBO J. 22:1005-13. Livak, K. J. & Schmittgen, T. D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 25:402-8. Llorca, O., Betti, M., Gonzalez, J. M., Valencia, A., Marquez, A. J. & Valpuesta, J. M. 2006. The three-dimensional structure of an eukaryotic glutamine synthetase: Functional implications of its oligomeric structure. J. Struct. Biol. 156:469-79. Lomas, M. W., Glibert, P. M., Berg, G. M. & Burford, M. 1996. Characterization of nitrogen uptake by natural populations of Aureococcus anophagefferens (Chrysophyceae) as a function of incubation duration, substrate concentration, light, and temperature. J. Phycol. 32:907-16. Lorenz, M. C. & Heitman, J. 1998. The MEP2 ammonium permease regulates pseudohyphal differentiation in Saccharomyces cerevisiae. EMBO J. 17:1236-47. Luque, I., Flores, E. & Herrero, A. 1994. Molecular mechanism for the operation of nitrogen control in cyanobacteria. EMBO J. 13:2862-9. Marini, A. M., Boeckstaens, M., Benjelloun, F., Cherif-Zahar, B. & Andre, B. 2006. Structural involvement in substrate recognition of an essential aspartate residue conserved in Mep/Amt and Rh-type ammonium transporters. Curr. Genet. 49:364-74. Marini, A. M., SoussiBoudekou, S., Vissers, S. & Andre, B. 1997a. A family of ammonium transporters in Saccharomyces cerevisiae. Mol. Cell. Biol. 17:4282-93. Marini, A. M., Urrestarazu, A., Beauwens, R. & Andre, B. 1997b. The Rh (rhesus) blood group polypeptides are related to NH4+ transporters. Trends Biochem. Sci. 22:460-1. Marini, A. M., Vissers, S., Urrestarazu, A. & Andre, B. 1994. Cloning and expression of the Mep1 gene encoding an ammonium transporter in Saccharomyces cerevisiae. EMBO J. 13:3456-63. Matsuda, Y. & Colman, B. 1995. Characterization of sulfate transport in the green alga Chlorella ellipsoidea. Plant Cell Physiol. 36:1291-6. Maurin, C. & Le Gal, Y. 1997. Isoforms of glutamine synthetase in the marine coccolithophorid Emiliania huxleyi (Prymnesiophyceae). Comp. Biochem. Physiol. Part B Biochem. Mol. Biol. 118:903-12. McFadden, G. I. 2001. Primary and secondary endosymbiosis and the origin of plastids. J. Phycol. 37:951-9. McKay, R. M. L., Geider, R. J. & LaRoche, J. 1997. Physiological and biochemical response of the photosynthetic apparatus of two marine diatoms to Fe stress. Plant Physiol. 114:615-22. McKay, R. M. L., La Roche, J., Yakunin, A. F., Durnford, D. G. & Geider, R. J. 1999. Accumulation of ferredoxin and flavodoxin in a marine diatom in response to Fe. J. Phycol. 35:510-9. Medina, M., Collins, A. G., Silberman, J. D. & Sogin, M. L. 2001. Evaluating hypotheses of basal animal phylogeny using complete sequences of large and small subunit rRNA. Proc. Natl. Acad. Sci. U. S. A. 98:9707-12. Merchant, S. S., Prochnik, S. E., Vallon, O., Harris, E. H., Karpowicz, S. J., Witman, G. B., Terry, A., Salamov, A., Fritz-Laylin, L. K., Marechal-Drouard, L., Marshall, W. F., Qu, L. H., Nelson, D. R., Sanderfoot, A. A., Spalding, M. H., Kapitonov, V. V., Ren, Q. H., Ferris, P., Lindquist, E., Shapiro, H., Lucas, S. M., Grimwood, J., Schmutz, J., Grigoriev, I. V., Rokhsar, D. S., Grossman, A. R., Chlamydomonas Annotation & JGI Annotation Team 2007. The Chlamydomonas genome reveals the evolution of key animal and plant functions. Science 318:245-51. Miller, P. E. & Scholin, C. A. 1998. Identification and enumeration of cultured and wild Pseudonitzschia (Bacillariophyceae) using species specific LSU rRNA targeted fluorescent probes and filter based whole cell hybridization. J. Phycol. 34:371-82. Mimura, T., Reid, R. J. & Smith, F. A. 1998. Control of phosphate transport across the plasma membrane of Chara corallina. J. Exp. Bot. 49:13-9. Montesinos, M. L., Muro-Pastor, A. M., Herrero, A. & Flores, E. 1998. Ammonium/Methylammonium permeases of a cyanobacterium- Identification and analysis of three nitrogen regulated amt genes in Synechocystis sp. PCC 6803. J. Biol. Chem. 273:31463-70. Moorthi, S. D., Countway, P. D., Stauffer, B. A. & Caron, D. A. 2006. Use of quantitative real-time PCR to investigate the dynamics of the red tide dinoflagellate Lingulodinium polyedrum. Microb. Ecol. 52:136-50. Morelli, E. & Pratesi, E. 1997. Production of phytochelatins in the marine diatom Phaeodactylum tricornutum in response to copper and cadmium exposure. Bull. Environ. Contam. Toxicol. 59:657-64. Morris, A. W. & Riley, J. P. 1963. Determination of nitrate in sea water. Anal. Chim. Acta 29:272-9. Moss, T. & Stefanovsky, V. Y. 1995. Promotion and regulation of ribosomal transcription in eukaryotes by RNA polymerase I. Prog. Nucleic Acid Res. Mol. Biol. 50:25-66. Muro-Pastor, M. I., Reyes, J. C. & Florencio, F. J. 2005. Ammonium assimilation in cyanobacteria. Photosynth. Res. 83:135-50. Ninnemann, O., Jauniaux, J. C. & Frommer, W. B. 1994. Identification of a high affinity NH4+ transporter from plants. EMBO J. 13:3464-71. Okamoto, M., Vidmar, J. J. & Glass, A. D. M. 2003. Regulation of NRT1 and NRT2 gene families of Arabidopsis thaliana: Responses to nitrate provision. Plant Cell Physiol. 44:304-17. Oliveira, I. C. & Coruzzi, G. M. 1999. Carbon and amino acids reciprocally modulate the expression of glutamine synthetase in arabidopsis. Plant Physiol. 121:301-9. Oliveira, I. C., Lam, H. M., Coschigano, K., MeloOliveira, R. & Coruzzi, G. 1997. Molecular genetic dissection of ammonium assimilation in Arabidopsis thaliana. Plant Physiol. Biochem. 35:185-98. Orsel, M., Eulenburg, K., Krapp, A. & Daniel-Vedele, F. 2004. Disruption of the nitrate transporter genes AtNRT2.1 and AtNRT2.2 restricts growth at low external nitrate concentration. Planta 219:714-21. Orsel, M., Filleur, S., Fraisier, V. & Daniel-Vedele, F. 2002a. Nitrate transport in plants: which gene and which control? J. Exp. Bot. 53:825-33. Orsel, M., Krapp, A. & Daniel-Vedele, F. 2002b. Analysis of the NRT2 nitrate transporter family in Arabidopsis. Structure and gene expression. Plant Physiol. 129:886-96. Pai, S. C., Tsau, Y. J. & Yang, T. I. 2001. pH and buffering capacity problems involved in the determination of ammonia in saline water using the indophenol blue spectrophotometric method. Anal. Chim. Acta 434:209-16. Palenik, B. & Wood, A. M. 1998. Molecular markers of phytoplankton physiological status and their application at the level of individula cells. In Cooksey, K. E. [Ed.] Molecular Approaches to the Study of the Ocean. Chapman & Hall, London, pp. 188-205. Pao, S. S., Paulsen, I. T. & Saier, M. H. 1998. Major facilitator superfamily. Microbiol. Mol. Biol. Rev. 62:1-34. Parkhill, J. P., Maillet, G. & Cullen, J. J. 2001. Fluorescence-based maximal quantum yield for PSII as a diagnostic of nutrient stress. J. Phycol. 37:517-29. Parsons, M. L., Scholin, C. A., Miller, P. E., Doucette, G. J., Powell, C. L., Fryxell, G. A., Dortch, Q. & Soniat, T. M. 1999. Pseudonitzschia species (Bacillariophyceae) in Louisiana coastal waters: Molecular probe field trials, genetic variability, and domoic acid analyses. J. Phycol. 35:1368-78. Partensky, F., LaRoche, J., Wyman, K. & Falkowski, P. G. 1997. The divinyl chlorophyll a/b protein complexes of two strains of the oxyphototrophic marine prokaryote Prochlorococcus- Characterization and response to changes in growth irradiance. Photosynth. Res. 51:209-22. Quesada, A. & Fernandez, E. 1994. Expression of nitrate assimilation related genes in Chlamydomonas reinhardtii. Plant Mol. Biol. 24:185-94. Quesada, A., Galvan, A. & Fernandez, E. 1994. Identification of nitrate transporter genes in Chlamydomonas reinhardtii. Plant J. 5:407-19. Quesada, A., Hidalgo, J. & Fernandez, E. 1998. Three Nrt2 genes are differentially regulated in Chlamydomonas reinhardtii. Mol. Gen. Genet. 258:373-7. Raven, J. A. 1986. Physiological consequences of extremely small size for autotrophic organism in the sea. Can. Bull. Fish. Aquat. Sci. 214:1-70. Redfield, A. C. 1958. The Biological Control of Chemical Factors in the Environment. Am. Sci. 46:205-21. Rees, T. A. V., Larson, T. R., Heldens, J. W. G. & Huning, F. G. J. 1995. In situ glutamine synthetase activity in a marine unicellular alga- development of a sensitive colorimetric assay and the effects of nitrogen status on enzyme activity. Plant Physiol. 109:1405-10. Rexach, J., Fernandez, E. & Galvan, A. 2000. The Chlamydomonas reinhardtii Nar1 gene encodes a chloroplast membrane protein involved in nitrite transport. Plant Cell 12:1441-53. Rexach, J., Llamas, A., Fernandez, E. & Galvan, A. 2002. The activity of the high affinity nitrate transport system I (NRT2;1, NAR2) is responsible for the efficient signalling of nitrate assimilation genes in Chlamydomonas reinhardtii. Planta 215:606-11. Rexach, J., Montero, B., Fernandez, E. & Galvan, A. 1999. Differential regulation of the high affinity nitrite transport systems III and IV in Chlamydomonas reinhardtii. J. Biol. Chem. 274:27801-6. Robertson, D. L. & Alberte, R. S. 1996. Isolation and characterization of glutamine synthetase from the marine diatom Skeletonema costatum. Plant Physiol. 111:1169-75. Robertson, D. L., Smith, G. J. & Alberte, R. S. 1999. Characterization of a cDNA encoding glutamine synthetase from the marine diatom Skeletonema costatum (Bacillariophyceae). J. Phycol. 35:786-97. Robertson, D. L., Smith, G. J. & Alberte, R. S. 2001. Glutamine synthetase in marine algae: new surprises from an old enzyme. J. Phycol. 37:793-5. Sambrook, J. & Russell, D. W. 2001. Molecular Cloning: a laboratory manual, 3 ed. Cold Spring Harbor Laboratory Press, New York Scanlan, D. J., Silman, N. J., Donald, K. M., Wilson, W. H., Carr, N. G., Joint, I. & Mann, N. H. 1997. An immunological approach to detect phosphate stress in populations and single cells of photosynthetic picoplankton. Appl. Environ. Microbiol. 63:2411-20. Shimogawara, K. & Usuda, H. 1995. Uptake of inorganic phosphate by suspension cultured tobacco cells: Kinetics and regulation by Pi starvation. Plant Cell Physiol. 36:341-51. Slot, J. C., Hallstrom, K. N., Matheny, P. B. & Hibbett, D. S. 2007. Diversification of NRT2 and the origin of its fungal homolog. Mol. Biol. Evol. 24:1731-43. Sohlenkamp, C., Shelden, M., Howitt, S. & Udvardi, M. 2000. Characterization of Arabidopsis AtAMT2, a novel ammonium transporter in plants. FEBS Lett. 467:273-8. Sohlenkamp, C., Wood, C. C., Roeb, G. W. & Udvardi, M. K. 2002. Characterization of Arabidopsis AtAMT2, a high-affinity ammonium transporter of the plasma membrane. Plant Physiol. 130:1788-96. Solorzano, L. 1969. Determination of ammonia in natural waters by the phenol hypochlorate method. Limnol. Oceanogr. 14:799-801. Sommer, U. 1994. The impact of light intensity and daylength on silicate and nitrate competition among marine phytoplankton. Limnol. Oceanogr. 39:1680-8. Sommer, U. 2000. Scarcity of medium sized phytoplankton in the northern Red Sea explained by strong bottom up and weak top down control. Mar. Ecol. Prog. Ser. 197:19-25. Sommer, U., Stibor, H., Katechakis, A., Sommer, F. & Hansen, T. 2002. Pelagic food web configurations at different levels of nutrient richness and their implications for the ratio fish production: primary production. Hydrobiologia 484:11-20. Song, B. & Ward, B. B. 2007. Molecular cloning and characterization of high affinity nitrate transporters in marine phytoplankton. J. Phycol. 43:542-52. Song, B. K. & Ward, B. B. 2004. Molecular characterization of the assimilatory nitrate reductase gene and its expression in the marine green alga Dunaliella tertiolecta (Chlorophyceae). J. Phycol. 40:721-31. Soupene, E., He, L. H., Yan, D. L. & Kustu, S. 1998. Ammonia acquisition in enteric bacteria: Physiological role of the ammonium/methylammonium transport B (AmtB) protein. Proc. Natl. Acad. Sci. U. S. A. 95:7030-4. Stahlberg, A., Hakansson, J., Xian, X. J., Semb, H. & Kubista, M. 2004. Properties of the reverse transcription reaction in mRNA quantification. Clin. Chem. 50:509-15. Straus, N. A. 1994. Iron deprivation: physiology and gene regulation. In Bryant, D. A. [Ed.] The Molecular Biology of Cyanobacteria. Kluwer, Dordrecht, pp. 731-5. Strickland, J. D. H. & Parsons, T. R. 1972. A Practical Handbook for Analysis of Seawater. Fisheries Research Board of Canada, Ottawa, 77 pp. Tabita, F. R. 1999. Microbial ribulose 1,5-bisphosphate carboxylase/oxygenase: A different perspective. Photosynth. Res. 60:1-28. Takabayashi, M., Wilkerson, F. P. & Robertson, D. 2005. Response of glutamine synthetase gene transcription and enzyme activity to external nitrogen sources in the diatom Skeletonema costatum (Bacillariophyceae). J. Phycol. 41:84-94. Thelwell, C., Robinson, N. J. & Turner-Cavet, J. S. 1998. An SmtB-like repressor from Synechocystis PCC 6803 regulates a zinc exporter. Proc. Natl. Acad. Sci. U. S. A. 95:10728-33. Thomas, G. H., Mullins, J. G. L. & Merrick, M. 2000. Membrane topology of the Mep/Amt family of ammonium transporters. Mol. Microbiol. 37:331-44. Throndsen, J. 1978. Preservation and storage. In Sournia, A. [Ed.] Phytoplankton Manual. Unesco, Paris, pp. 75-87. Tilman, D., Kiesling, R., Sterner, R., Kilham, S. S. & Johnson, F. A. 1986. Green, bluegreen and diatom algae- Taxonomic differences in competitive ability for phosphorus, silicon and nitrogen. Arch. Hydrobiol. 106:473-85. Trueman, L. J., Onyeocha, I. & Forde, B. G. 1996. Recent advances in the molecular biology of a family of eukaryotic high affinity nitrate transporters. Plant Physiol. Biochem. 34:621-7. Tyrrell, J. V., Bergquist, P. R., Saul, D. J., MacKenzie, L. & Bergquist, P. L. 1997. Oligonucleotide probe technology as applied to the study of harmful algal blooms. N. Z. J. Mar. Freshw. Res. 31:551-60. Vassiliev, I. R., Kolber, Z., Wyman, K. D., Mauzerall, D., Shukla, V. K. & Falkowski, P. G. 1995. Effects of iron limitation on photosystem-II composition and light utilization in Dunaliella tertiolecta. Plant Physiol. 109:963-72. Vegapalas, M. A., Madueno, F., Herrero, A. & Flores, E. 1990. Identification and cloning of a regulatory gene for nitrogen assimilation in the cyanobacterium Synechococcus Sp. Strain PCC7942. J. Bacteriol. 172:643-7. Vidmar, J. J., Zhuo, D., Siddiqi, M. Y., Schjoerring, J. K., Touraine, B. & Glass, A. D. M. 2000. Regulation of high affinity nitrate transporter genes and high affinity nitrate influx by nitrogen pools in roots of barley. Plant Physiol. 123:307-18. von Wiren, N., Gazzarrini, S., Gojon, A. & Frommer, W. B. 2000. The molecular physio
摘要: 浮游植物是水域生態系中最主要的初級生產者,而在海洋環境中它們的生產力常受到含氮營養鹽的限制,為了深入暸解各種海洋浮游植物受限制的程度,必須發展分子指標以直接評估浮游植物的生理狀態。在本研究中,首先挑選在氮吸收及同化途徑中重要的基因,包括硝酸運輸蛋白 (Nrt2)、銨運輸蛋白 (Amt),硝酸還原酵素 (Nr)、及glutamine synthetase (glnII) 等做為具有潛力之氮指標基因,其次在純化馴養的藻種中,包括矽藻綱之Skeletonema costatum及Thalassiosira weissflogii、著鞭毛藻綱之Isochrysis galbana、及綠鞭藻綱之Tetraselmis chui進行基因之選殖。之後以I. galbana為實驗室之模式藻種,利用定量反轉錄聚合酶連鎖反應偵測不同氮生長條件下之基因表現情形,結果顯示其中以Nrt2基因的表現對於氮源之種類及濃度最為敏感。在缺氮時呈現飢餓反應並且大量表現,當以硝酸為氮源時則維持中等之表現量,但是在以銨為氮源時則明顯降低表現量,而且在缺氮時的表現量是銨培養下的160倍。顧及此種Nrt2基因表現範圍可能隨物種而變化,於是要發展相對定量法來正確判讀海洋中所測到的Nrt2表現量。在使用I. galbana和Thalassiosira pseudonana為實驗材料的測試中,發現添加銨進行培養可獲得Nrt2基因的最低表現量,而以缺氮培養可獲得Nrt2的最高表現量,藉此定出海洋中浮游植物Nrt2基因之表現範圍做為判斷氮鹽利用情形的依據。另一方面,本研究廣泛搜尋和定序已知矽藻種類之Nrt2序列,以瞭解物種間Nrt2基因在序列上的差異,同時在東海運用退化性引子定序海洋樣本中之Nrt2同源基因,將海洋中獲得的序列與已知序列比對,發現在馬祖海域Nrt2序列有96% 和Skeletonema屬類似;而在湧升流海域中除了有60% 與Skeletonema屬類似外,還有10% 與Chaetoceros屬類似。這些結果有助於針對特定海域設計出具專一性之引子,配合上定量反轉錄聚合酶連鎖反應便可以偵測到海洋樣本中特定種類之Nrt2基因表現。這種使用指標基因表現量的方法,可以方便的以單一技術同時研究不同浮游植物如何受到含氮營養鹽的影響,而且可以擴充到其他的指標基因來探討浮游植物在海洋中的各種生理狀態。
Phytoplankton serve as the primary producer in marine food chain, but their productivities are often limited by the availability of nitrogen in the ocean. For a better understanding of nitrogen limitation among phytoplankton species, it is necessary to develop suitable molecular probes to directly evaluate their physiological states. In this study, 4 genes involved in nitrogen uptake and assimilation, including nitrate transporter genes (Nrt2), ammonium transporter genes (Amt), nitrate reductase genes (Nr) and glutamine synthetase genes (glnII), were selected as candidates for a nitrogen-stress maker. In the first stage, 4 sets of degenerate primers were designed to clone fragments of these genes in cultured marine phytoplankton species, including Skeletonema costatum (Bacillariophyceae), Thalassiosira weissflogii (Bacillariophyceae), Isochrysis galbana (Haptophyceae) and Tetraselmis chui (Prasinophyceae). Next, transcript levels of Nrt2, Amt, and glnII genes in I. galbana grown under various forms and concentrations of nitrogenous nutrients were monitored by quantitative real-time PCR. Our results demonstrated that the mRNA levels of Nrt2 responded sensitively to ambient nitrogen sources with a moderate expression under nitrate-sufficient condition and a high expression under nitrogen-deprived condition. In contrast, the transcript level was severely decreased in the presence of ammonium, which was 160-fold lower than that under nitrogen deprivation. Therefore, Nrt2 genes were chosen to be the first candidate gene for evaluating the nitrogen status of phytoplankton in the ocean. Furthermore, for the proper interpretation of measured Nrt2 transcript abundances in the ocean, a relative expression assay was proposed and tested in I. galbana and Thalassiosira pseudonana. Results from both species indicated that the minimal and the maximal Nrt2 mRNA abundances could be achieved by ammonium addition and nitrogen deprivation, respectively. Considering the range of Nrt2 expression may vary from species to species, the relative expression assay will allow a more precise evaluation of nitrogen deficiency in untested species. On the other hand, to investigate sequence diversity of Nrt2 genes in phytoplankton, data banks were searched and Nrt2 gene fragments of several cultured diatoms were cloned. In addition, Nrt2 fragments were amplified with degenerate primers from samples collected in the East China Sea. Sequence alignments revealed that 96% of Nrt2 sequences at the Matsu station clustered with Skeletonema Nrt2 sequences. In the upwelling area, 60% of sequences clustered with Skeletonema and 10% clustered with Chaetoceros Nrt2. According to the sequences alignments, specific primers targeted to particular phytoplankton Nrt2 were designed. Using these primers with quantitative real-time PCR, our preliminary work demonstrated that the Nrt2 mRNA abundances of specific phytoplankton could be detected in the ocean. By detecting the expression levels of maker genes, not only the nitrogen-utilization status of various phytoplankton can be evaluated using a unified and simple procedure, the same algorithm can also be expanded to study other physiological activities of phytoplankton in the ocean.
URI: http://hdl.handle.net/11455/22700
其他識別: U0005-1907200802020000
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