Determination of RNA and DNA concentrations in natural plankton samples using thiazole orange in combination with DNase and RNase digestions

Journal of Phycology

Volumn 32, Issue 6, 1996, Pages 1074-1083

  Fara, Angela ^a   Berdalet, Elisa ^a   Arin, Laura ^a  

^a INSTITUT DE CIÈNCIES DEL MAR   (Spain)

Author keywords

DNA quantification; fluorometric techniques; natural plankton; phytoplankton; RNA quantification; thiazole orange

Indexed keywords

VISCUM ALBUM;

EID: 0030447210     PISSN: 00223646     EISSN: None     Source Type: Journal    
DOI: 10.1111/j.0022-3646.1996.01074.x     Document Type: Article

References (34)

1. Berdalet, E. & Dortch, Q. 1991. New double-staining technique for RNA and DNA measurement in marine phytoplankton. Mar. Ecol Prog. Ser. 73:295-305.
2. Berdalet, E. & Estrada, M. 1993. Relationship between nucleic acid concentration and primary production in the Catalan Sea (northwestern Mediterranean). Mar. Biol 117:167-70.
3. Berdalet, E., Latasa, M. & Estrada, M. 1992. Variation in biochemical parameters of Heterocapsa sp. and Olisthodiscus luteus grown in 12:12 light:dark cycles. I. Cell cycle and nucleic acid composition. Hydrobiologia 238:139-47.
4. _ 1994. Effects of nitrogen and phosphorus starvation on nucleic acid and protein content of Heterocapsa sp. J. Plankton Res. 16:303-16.
5. Berdalet, E., Marrasé, C., Estrada, M., Arin L. & MacLean, M. L. 1996. Microbial community responses to nitrogen- and phosphorus-deficient nutrient inputs: microplankton dynamics and biochemical characterization. J. Plankton Res. 18:1627-41.
6. Berges, J. A., Fisher, A. E. & Harrison, P. J. 1993. A comparison of Lowry, Bradford and Smith protein assays using different protein standards and protein isolated from the marine diatom Thalassiosira pseudonana. Mar. Biol. 115:187-93.
7. Boehme, J., Frischer, M. E., Jiang, S. C., Kellogg, C. A., Pichard, S., Rose, J. B., Steinway, C. & Paul, J. H. 1993. Viruses, bacterioplankton, and phytoplankton in the southeastern Gulf of Mexico: distribution and contribution to oceanic DNA pools. Mar. Ecol. Prog. Ser. 97:1-10.
8. Brooks, J. E. 1987. Properties and uses of restriction endonucleases. In Berger, S. L. & Kimmel, A. R. [Eds.] Methods in Enzymology, Guide to Molecular Cloning Techniques. Academic Press, San Diego, California, pp. 113-29.
9. Burrin, D. H. 1986. Spectroscopic techniques. In Wilson, K. & Goulding, K. [Eds.] A Biologists Guide to Principles and Techniques of Practical Biochemistry. Edward Arnold LTD, London, pp. 287-333.
10. Coakley, W. T., Bater, A. J. & Lloyd, D. 1977. Disruption of microorganisms. Adv. Microb. Physiol. 16:279-341.
11. Comings, D. E. 1975. Mechanisms of chromosome banding. VIII. Hoechst 33258-DNA interaction. Chromosoma 52:229-43.
12. de Madariaga, I. & Joint, I. 1992. A comparative study of phytoplankton physiological indicators. J. Exp. Mar. Biol. Ecol. 158:149-65.
13. Dobrogosz, W. J. 1981. Enzymatic activity. In Gerhardt, P., Murray, R. G. E., Costilow, R. N., Nester, E. W, Wood, W. A., Krieg, N. R. & Phillips, G. B. [Eds.] Manual of Methods for General Microbiology. American Society for Microbiology, Washington, D. C., pp. 365-92.
14. Dortch, Q. & Packard, T. T. 1989. Differences in biomass structure between oligotrophic and eutrophic marine ecosystems. Deep-Sea Res. 36:223-40.
15. Dortch, Q., Robert, T. L., Clayton, J. R., Jr & Ahmed, S. I. 1983. RNA/DNA ratios and DNA concentration as indicators of growth rate and biomass in planktonic marine organisms. Mar. Ecol Prog. Ser. 13:67-71.
16. Guillard, R. R. L. 1975. Culture of phytoplankton for feeding marine invertebrates. In Smith, W. L. & Chanley, M. H. [Eds.] Culture of Marine Invertebrate Animals. Plenum, New York, pp. 108-32.
17. Holm-Hansen, O., Sutcliffe, W. H., Jr. & Sharp, J. 1968. Measurement of deoxyribonucleic acid in the ocean and its ecological significance. Limnol. Oceanogr. 13:507-14.
18. Iwamura, T., Kanazawa, K., Shibata, Y., Morimura, S., Ichimura, S. Maeda, O. & Tamiya, H. 1967. Preliminary studies on the feasibility of microanalytic measurement of planktonic populations. J. Oceanog. Soc. Jpn. 23:247-51.
19. Karl, D. M., Winn, C. D. & Wong, D. C. L. 1981. RNA synthesis as a measure of the microbial growth in aquatic environments. I. Evaluation, verification and optimization of methods. Mar. Biol. 64:1-12.
20. Kemp, P. F., Lee, S. & LaRoche, J. 1993. Estimating the growth rate of slowly growing marine bacteria from RNA content. Am. Soc Microbiol 59:2594-601.
21. Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. J. 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem. 193:265-80.
22. Mordy, C. W. & Carlson, D. J. 1991. An evaluation of fluorescence techniques for measuring DNA and RNA in marine microorganisms. Mar. Ecol. Prog. Ser. 73:283-93.
23. Olson, R. J., Vaulot, D. & Chisholm, S. W. 1986. Effects of environmental stresses on the cell cycle of two marine phytoplankton species. Plant Physiol. 80:918-25.
24. Paul, J. H. & Carlson, D. J. 1984. Genetic material in the marine environment: implication for bacterial DNA. Limnol. Oceanogr. 29:1091-7.
25. Paul, J. H., Jeffrey, W. H. & Deflaun, M. 1985. Particulate DNA in subtropical oceanic and estuarine planktonic environments. Mar. Biol. 90:95-101.
26. Paul, J. H. & Pichard, S. L. 1995. Extraction of DNA and RNA from aquatic environments. In Trevors, J. T. & van Elsas, J. D. [Eds.] Nucleic Acids in the Environment. Methods and Applications. Springer-Verlag, Germany, pp. 153-77.
27. Prasad, A. D., DuMouchelle E., Koniuch, D. & Oberleas, D. 1972. A simple fluorometric method for the determination of RNA and DNA in tissues. J. Lab. Clin. Med. 80:598-602.
28. Sambrook, J., Fritsch, E. F. & Maniatis, T. 1989. Molecular Cloning. A Laboratory Manual, Vol. 2, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, Appendix B.
29. Shapiro, H. M. 1995. Practical Flow Cytometry, 3rd ed. Wiley-Liss, New York, 542 pp.
30. Takahashi, M., Nagai, H., Yamaguchi, Y. & Ichimura, S. 1974. The distribution of chlorophyll a, protein, RNA and DNA in the North Pacific Ocean. J. Oceanogr. Soc. Jpn. 30:137-50.
31. Thoresen, S. S., Clayton, J. R., Jr., Dortch, Q. & Ahmed, S. I. 1983. A rapid technique for the determination of RNA and DNA in marine phytoplankton. J. Plankton Res. 5:253-61.
32. Walser, R. & Güde, H. 1994. DNA and RNA contents of bacteria-sized particles from Lake Constance. Arch. Hydrobiol 131:257-76.
33. Weisblum, B. & Haenssler, E. 1974. Fluorometric properties of the bisbenzimidazole derivative Hoechst 33258, a fluorescent probe specific for AT concentration in chromosomal DNA. Chromosoma 46:255-60.
34. Yentsch, C. S. & Menzel, D. W. 1963. A method for the determination of phytoplankton chlorophyll and phaeophytin by fluorescence. Deep-Sea Res. 10:221-31.