DOI QR코드

DOI QR Code

Identification and Isolation of Differentially Expressed Gene in Response to Cold Stress in a Green Alga, Spirogyra varians (Zygnematales)

  • Published : 2007.06.01

Abstract

The expression of genes responding to cold stress in a freshwater alga, Spirogyra varians, was studied by using differential expression gene (DEG) method. A gene strongly up-regulated in 4°C was isolated and designated as SVCR2 (Spirogyra varians cold regulated) gene. The cDNA encoding SVCR2 was cloned using λZAP cDNA library of Spirogyra varians. The deduced amino acid had a sequence similarity with trans-membrane protein in Arabidopsis thaliana (Q9M2D2, 52.7%). Northern blot analysis demonstrated that transcript level of SVCR2 increased about 10 fold under low temperature (4°C), compared with that cultured at warm (20°C) conditions. The expression of SVCR2 was also affected by light conditions. When the plants were exposed to high light (HL) (1200 μmol photon m–2 s–1), the expression of SVCR2 began within 2 hrs. This gene expression lasted for 4 hrs and decreased afterwards. Under the blue light (470 nm) condition, the expression of this gene was induced in same way as HL treatment, even under less than 100 μmol photon m–2 s–1. But red light (650 nm) and UV-A irradiation did not affect the expression of SVCR2.

Keywords

References

  1. Bischoff H. and Bold H.C. 1963. Phycological studies IV. Some soil algae from Enchanted Rock and related algal species. Univ. Tex. Publ. No. 6318, Austin
  2. Cattivelli L. and Bartels D. 1990. Molecular Cloning and Characterization of Cold-Regulated Genes in Barley. Plant Physiol. 93: 1504-1510 https://doi.org/10.1104/pp.93.4.1504
  3. Chomczynski P. and Sacchi N. 1987. Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162: 156-159
  4. Danyluk J., Houde M., Rassart E. and Sarhan F. 1994. Differential expression of a gene encoding an acidic dehydrin in chilling sensitive and freezing tolerant gramineae species. FEBS Lett. 344: 20-24 https://doi.org/10.1016/0014-5793(94)00353-X
  5. Gemel J., Golinowski W. and Kaniuga Z. 1986. Low-temperature induced changes in chloroplast ultrastructure in relation to changes of Hill reaction activity, manganese and free fatty acid levels in chloroplasts of chilling-sensitive and chilling-resistant plants. Acta Physiol. Plant. 8: 135-143
  6. Gilmour S.J., Artus N.N. and Thomashow M.F. 1992. cDNA sequence analysis and expression of two cold-regulated genes of Arabidopsis thaliana. Plant Mol. Biol. 18: 13-21 https://doi.org/10.1007/BF00018452
  7. Graham D. and Patterson B.D. 1982. Responses of plants to low, non freezing temperature: protein, metabolism, and acclimation. Annu. Rev. Plant Physiol. 33: 347-372 https://doi.org/10.1146/annurev.pp.33.060182.002023
  8. Gray G.R., Savitch L.V., Ivanov A.G., and Huner N. 1996. Photosystem II excitation pressure and development of resistance to photoinhibition. II. Adjustment of photosynthetic capacity in winter wheat and winter rye. Plant Physiol. 110: 61-71 https://doi.org/10.1104/pp.110.1.61
  9. Gray G.R., Chauvin L.P., Sarhan F., and Huner N. 1997. Cold Acclimation and Freezing Tolerance (A Complex Interaction of Light and Temperature). Plant Physiol. 114: 467-474 https://doi.org/10.1104/pp.114.2.467
  10. Guy C.L. 1990. Cold acclimation and freezing stress tolerance: Role of protein metabolism. Annu. Rev. Plant Physiol. Plant Mol. Biol. 41: 187-223 https://doi.org/10.1146/annurev.pp.41.060190.001155
  11. Guy C.L., Niemi K.J., and Brambl R 1985. Altered gene expression during cold acclimation of spinach. Proc. Natl. Acad. Sci. USA. 82: 3673-3677
  12. Hajela R.K., Horvath D.P., Gilmour. S.J. and Thomashow. M.F. 1990. Molecular Cloning and Expression of cor (Cold-Regulated) Genes in Arabidopsis thaliana. Plant Physiol. 93: 1246-1252 https://doi.org/10.1104/pp.93.3.1246
  13. Houde M., Daniel C., Lachapelle M., Allard F., Laliberte S. and Sarhan F. 1995. Immunolocalization of freezing-tolerance-associated proteins in the cytoplasm and nucleoplasm of wheat crown tissues. Plant J. 8: 583-593 https://doi.org/10.1046/j.1365-313X.1995.8040583.x
  14. Hwang I.T., Kim Y.J., Kim S.H., Kwak C.I., Gu Y.Y. and Chun J.Y. 2003. Annealing control primer system for improving specificity of PCR amplification. Biotechniques. 35: 1180-1184
  15. Kim G.H., Yoon M. and Klotchkova T.A. 2005. A moving mat: Phototaxis in the filamentous green algae Spirogyra (Chlorophyta, Zygnemataceae). J. Phycol. 41: 232-237 https://doi.org/10.1111/j.1529-8817.2005.03234.x
  16. Kim Y.J., Kwak C.I., Gu Y.Y., Hwang I.T. and Chun J.Y. 2004. Annealing control primers system for identification of differentially expressed genes on agarose gels. Biotechniques. 36: 424-426 https://doi.org/10.2144/04363ST02
  17. Kratsch H.A. and Wise. R.R. 2000. The ultrastructure of chilling stress. Plant Cell Environ. 23: 337-350 https://doi.org/10.1046/j.1365-3040.2000.00560.x
  18. Liang P., Averboukh L. and Pardee A.B. 1993. Distribution and cloning of eukaryotic mRNAs by means of differential display: refinements and optimization. Nucleic Acids Res. 21: 3269-3275 https://doi.org/10.1093/nar/21.14.3269
  19. Liang P. and Pardee A.B. 1992. Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257: 967-971 https://doi.org/10.1126/science.1354393
  20. Liang P., Zhu W., Zhang X., Guo Z., O'Connell R.P., Averboukh L., Wang F and Pardee A.B. 1994. Differential display using one-base anchored oligo-dT primers. Nucleic Acids Res. 22: 5763-5764 https://doi.org/10.1093/nar/22.25.5763
  21. Pearce R.S., C.E. Houlston, K.M. Atherton, J.E. Rixon, P. Harrison, M.A. Hughes, and M.A. Dunn. 1998. Localization of expression of three cold-induced genes, blt101, blt4.9, and b1t14 in different tissues of the crown and developing leaves of cold-acclimated cultivated barley. Plant Physiol. 117: 787-795 https://doi.org/10.1104/pp.117.3.787
  22. Thomashow M.F. 1990. Molecular genetics of cold acclimation in higher plants. Adv. Genet. 28: 99-131 https://doi.org/10.1016/S0065-2660(08)60525-8
  23. Sambrook, J. and Russell, D.W. 2001. Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Lab. Press, Plainview, NY

Cited by

  1. Cold stress regulation of a bi-functional 3-dehydroquinate dehydratase/shikimate dehydrogenase (DHQ/SDH)-like gene in the freshwater green alga Spirogyra varians vol.52, pp.2, 2009, https://doi.org/10.1515/BOT.2009.011
  2. Accumulation of galloyl derivatives in a green alga, Spirogyra varians, in response to cold stress vol.24, pp.5, 2012, https://doi.org/10.1007/s10811-011-9776-y
  3. An ELIP-like gene in the freshwater green alga, Spirogyra varians (Zygnematales), is regulated by cold stress and CO2 influx vol.25, pp.5, 2013, https://doi.org/10.1007/s10811-013-9975-9