Polar Biology

, Volume 34, Issue 5, pp 637–645 | Cite as

Differential gene expression of an Antarctic Chlorella in response to temperature stress

  • Geeng-Loo Chong
  • Wan-Loy Chu
  • Rofina Yasmin Othman
  • Siew-Moi Phang
Original Paper

Abstract

Changes in gene expression are an important response of Antarctic algae to temperature stress. The objective of this study was to investigate the differential gene expression of the Antarctic alga Chlorella UMACC 234 in response to temperature stress. The RNA was extracted from the cells grown at 4, 20, and 30°C and converted to cDNA by reverse transcription. Differentially expressed genes (DEG) were isolated and identified using the GeneFishing™ DEG Kit (Seegene) with 20 arbitrary annealing control primers (ACP). The bands of interest were excised and purified from the agarose gel and then cloned and sequenced. A total of 22 DEG clones were isolated and identified, with 11 DEG detected only at 30°C and six DEG detected only at 4°C. Three DEG were detected at 4 and 20°C while two were detected at 20 and 30°C. The DEG were associated with functions such as photosynthesis, carbohydrate metabolism, electron transfer, and cell maintenance. Three DEG that showed high degree of similarity with sequences from the database were those code for Photosystem II P680 chlorophyll a apoprotein CP47 (PSII-CP47), aldose 1-epimerase, and a putative oxidoreductase. Real-time PCR analysis showed that the expression of the PSII-CP47 gene increased by threefold at 4°C while that of the aldose 1-epimerase and oxidoreductase genes increased by threefold and eightfold, respectively, at 30°C compared with 20°C (optimal growth temperature).

Keywords

Chlorella Antarctic algae Differential gene expression GeneFishing Temperature stress 

References

  1. Anastasiou R, Leverrier P, Krestas I, Rouault A, Kalantzopoulos G, Boyaval P, Tsakalidou E, Jan G (2006) Changes in protein synthesis during thermal adaptation of Propionibacterium freudenreichii subsp. shermanii. Int J Food Microbiol 108:301–314PubMedGoogle Scholar
  2. Bailey JM, Fishman PH, Pentchev PG (1967) Studies on mutarotases: I. Purification and properties of a mutatorase from higher plants. J Biochem 242:4263–4269Google Scholar
  3. D’Aniello A, D’Onofrio G, Pischetola M, D’Aniello G, Vetere A, Petrucelli L, Fisher GH (1993) Biological role of d-amino acid oxidase and d-aspartate oxidase. Effects of d-amino acids. J Biol Chem 268:2694–26949Google Scholar
  4. Fiala M, Oriol L (1990) Light-temperature interactions on the growth of Antarctic diatoms. Polar Biol 10:629–636CrossRefGoogle Scholar
  5. Guy C, Kaplan F, Kopka J, Selbig J, Hincha DK (2008) Metabolomics of temperature stress. Physiol Plant 132:220–235PubMedGoogle Scholar
  6. Han JW, Lee KP, Yoon M, Kang SH, Kim GH (2009) Cold stress regulation of a bi-functional 3-dehydroquinate dehydratase (DHQ/SDH)-like gene in the freshwater green alga Spirogyra varians. Bot Mar 52:178–185CrossRefGoogle Scholar
  7. Hu H, Li H, Xu X (2008) Alternative cold response modes in Chlorella (Chlorophyta, Trebouxiophyceae) from the Antarctic. Phycologia 47:28–34CrossRefGoogle Scholar
  8. Hwang Y, Jung G, Jin E (2008) Transcriptome analysis of acclimatory responses to thermal stress in Antarctic algae. Biochem Biophys Res Comm 367:635–641PubMedCrossRefGoogle Scholar
  9. Jones TH, Murray A, Johns M, Gill CO, McMullen LM (2006) Differential expression of proteins in cold-adapted log-phase cultures of Escherichia coli incubated at 8, 6 or 2°C. Int J Food Microbiol 107:12–19PubMedCrossRefGoogle Scholar
  10. Jung G, Lee C, Kang S, Jin E (2007) Annotation and expression profile analysis of cDNAs from the Antarctic diatom Chaetoceros neogracile. J Microbiol Biotechnol 17(8):1330–1337PubMedGoogle Scholar
  11. Kim YJ, Kwak CI, Gu YY, Hwang IT, Chun JY (2004) Annealing control primer system for identification of differentially expressed genes on agarose gels. Biotechniques 36:424–434PubMedGoogle Scholar
  12. Li H, Liu X, Wang Y, Hu H, Xu X (2009) Enhanced expression of antifreeze protein genes drives the development of freeze tolerance in an Antarctic isolate of Chlorella vulgaris. Prog Nat Sci 19:1059–1062CrossRefGoogle Scholar
  13. Liang P, Pardee AB (1992) Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257:967–971PubMedCrossRefGoogle Scholar
  14. Mock T, Hoch N (2005) Long-term temperature acclimation of photosynthesis in steady-state cultures of the polar diatom Fragilariopsis cylindrus. Photosyn Res 85:307–317PubMedCrossRefGoogle Scholar
  15. Mock T, Valentin K (2004) Photosynthesis and cold acclimation: molecular evidence from a polar diatom. J Phycol 40:732–741CrossRefGoogle Scholar
  16. Mock T, Krell A, Glockner G, Kolikisaoglu U, Valentin K (2005) Analysis of Expressed Sequence Tags (ESTS) from the polar diatom Fragilariopsis cylindrus. J Phycol 42:78–85CrossRefGoogle Scholar
  17. Morgan-Kiss RM, Priscu JC, Poccock T, Gudynaite-Savitch L, Hunter NPA (2006) Adaptation and acclimation of photosynthetic microorganisms to permanently cold environments. Microbiol Mol Biol Rev 70:222–252PubMedCrossRefGoogle Scholar
  18. Nichols HW, Bold HC (1965) Trichorsarcina polymorpha gen. et sp. nov. J Phycol 1:34–38CrossRefGoogle Scholar
  19. Otoshi A, Yoshimura K, Tamoi M, Shigeoka S (2001) Isolation of genes involved in stress tolerance by activation tagging. In: Proceedings of 12th international congress on photosynthesis, August 18–23. Brisbane Convention & Exhibition Centre, Queensland, S36-006Google Scholar
  20. Pearson WR (1991) Identifying distantly related protein sequences. Curr Opin Struct Biol 1:321–326CrossRefGoogle Scholar
  21. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, vol 1, 3rd edn. Cold Spring Harbour Laboratory Press, New YorkGoogle Scholar
  22. Sane PV, Ivanov AG, Sveshnikov D, Huner NPA, Öquist G (2002) A transient exchange of the photosystem II reaction center protein D1:1 with D1:2 during low temperature stress of Synechococcus sp. PCC 7942 in the light lowers the redox potential of QB. J Biol Chem 277:32739–32745PubMedCrossRefGoogle Scholar
  23. Schena M, Shalon D, Davis RW, Brown PO (1995) Quantitative monitoring of gene expression patterns with a complementary DNA microarray. Science 270:467–470PubMedCrossRefGoogle Scholar
  24. Seaburg KG, Parker BC, Wharton RA, Simmons GM (1981) Temperature-growth responses of algal isolates from Antarctic oases. J Phycol 17:353–360CrossRefGoogle Scholar
  25. Suga K, Honjoh K, Furuya N, Shimizu H, Nishi K, Shinohara F, Hirabaru Y, Maruyama I, Miyamoto T, Hatano S, Iio M (2002) Two low-temperature-inducible Chlorella genes for Δ12 and ω-3 fatty acid desaturase (FAD): isolation of Δ12 and ω-3 fad cDNA clones, expression of Δ12 fad in Saccharomyces cerevisiae, and expression of ω-3 fad in Nioctiana tabacum. Biosci Biotechnol Biochem 66:1314–1327PubMedCrossRefGoogle Scholar
  26. Takahashi S, Whitney S, Itoh S, Maruyama T, Badger M (2008) Heat stress causes inhibition of the de novo synthesis of antenna proteins and photobleaching in cultured Symbiodinium. Proc Nat Acad Sci 105:4203–4208PubMedCrossRefGoogle Scholar
  27. Teoh ML, Chu WL, Marchant H, Phang SM (2004) Influence of culture temperature on the growth, biochemical composition and fatty acid profiles of six Antarctic microalgae. J Appl Phycol 16:421–430CrossRefGoogle Scholar
  28. Wong CY, Chu WL, Marchant H, Phang SM (2007) Comparing the response of Antarctic, tropical and temperate microalgae to ultraviolet radiation (UVR) stress. J Appl Phycol 19:689–699CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Geeng-Loo Chong
    • 1
  • Wan-Loy Chu
    • 2
  • Rofina Yasmin Othman
    • 1
  • Siew-Moi Phang
    • 1
    • 3
  1. 1.Institute of Biological Sciences, Faculty of ScienceUniversity of MalayaKuala LumpurMalaysia
  2. 2.International Medical UniversityKuala LumpurMalaysia
  3. 3.Institute of Ocean and Earth Sciences (IOES)University of MalayaKuala LumpurMalaysia

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