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Differential expression of carotenogenic genes and associated changes in pigment profile during regeneration of Haematococcus pluvialis cysts

  • Applied Microbial and Cell Physiology
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Abstract

Haematococcus pluvialis is a green alga known to accumulate astaxanthin in extra-plastidic lipid vesicles under stress conditions. The present study revealed the influence of few cultural parameters and temperature treatments on regeneration efficiency of red cysts along with changes in pigment profile and expression of carotenogenic genes during regeneration. Regeneration efficiency has been improved by incubating less aged cyst cells in a medium containing ammonium carbonate, 16:8 light–dark cycle with a light intensity of 30 μmol m−2 s−1. During regeneration, there was a decrease in total astaxanthin, total carotenoids, and carotenoid to chlorophyll ratio, and increase in β-carotene, lutein, total chlorophyll, and chlorophyll a to b ratio. Expression analysis revealed the presence of transcripts of carotenogenic genes, phytoene synthase (PSY), phytoene desaturase (PDS), lycopene cyclase (LCY), β-carotene ketolase (BKT), and β-carotene hydroxylase (CHY) in cyst cells, and these transcripts were up regulated transiently upon transfer to favorable conditions. As the culture growth progressed, carotenogenic gene expressions were decreased and reached basal expression levels of green motile vegetative cells. In addition, this is the first report of detection of carotenogenic gene transcripts in red cysts, and their differential expression during regeneration. The present study suggests the use of red cysts as alternate inoculum for mass cultivation to combat protozoan predation.

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References

  • Boussiba S (2000) Carotenogenesis in the green alga Haematococcus pluvialis: cellular physiology and stress response. Physiol Plant 108:111–117

    Article  CAS  Google Scholar 

  • Damiani AMC, Leonardi PI, Pieroni OI, Caceres EJ (2006) Ultrastructure of the cyst wall of Haematococcus pluvialis (Chlorophyceae): wall development and behaviour during cyst germination. Phycologia 45:616–623

    Article  Google Scholar 

  • Duncan DB (1955) Multiple range and multiple F tests. Biometrics 11:1–42

    Article  Google Scholar 

  • Fabregas J, Dominguez A, Maseda A, Otero A (2003) Interactions between irradiance and nutrient availability during astaxanthin accumulation and degradation in Haematococcus pluvialis. Appl Microbiol Biotechnol 61:545–551

    Article  CAS  Google Scholar 

  • Fabregas J, Otero A, Maseda A, Dominguez A (2001) Two-stage cultures for the production of astaxanthin from Haematococcuspluvialis. J Biotechnol 89:65–71

    Article  CAS  Google Scholar 

  • Grunewald K, Eckert M, Hirschberg J, Hagen C (2000) Phytoene desaturase is localized exclusively in the chloroplast and up-regulated at the mRNA level during accumulation of secondary carotenoids in Haematococcus pluvialis (Volvocales, Chlorophyceae). Plant Physiol 122:1261–1268

    Article  CAS  Google Scholar 

  • Guerin M, Huntley ME, Olaizola M (2003) Haematococcus astaxanthin: applications for human health and nutrition. Trends Biotechnol 21:210–216

    Article  CAS  Google Scholar 

  • Hagen C, Grunewald K, Xylander M, Rothe E (2001) Effect of cultivation parameters on growth and pigment biosynthesis in flagellated cells of Haematococcus pluvialis. J Appl Phycol 13:79–87

    Article  Google Scholar 

  • Huang JC, Chen F, Sandmann G (2006) Stress-related differential expression of multiple β-carotene ketolase genes in the unicellular green alga Haematococcus pluvialis. J Biotechnol 122:176–185

    Article  CAS  Google Scholar 

  • Jin E, Lee CG, Polle JEW (2006) Secondary carotenoid accumulation in Haematococcus (Chlorophyceae): biosynthesis, regulation, and biotechnology. J Microbiol Biotechnol 16:821–831

    CAS  Google Scholar 

  • Kobayashi M, Kurimura Y, Sakamoto Y, Tsuji Y (1997) Selective extraction of astaxanthin and chlorophyll from the green alga Haematococcus pluvialis. Biotechnol Tech 11:657–660

    Article  CAS  Google Scholar 

  • Lee YK, Ding SY (1994) Cell cycle and accumulation of astaxanthin in Haematococcus lacustris (Chlorophyta). J Phycol 30:445–449

    Article  CAS  Google Scholar 

  • Lichtenthaler HK (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Methods Enzymol 148:350–382

    Article  CAS  Google Scholar 

  • Margalith PZ (1999) Production of ketocarotenoids by microalgae. Appl Microbiol Biotechnol 51:431–438

    Article  CAS  Google Scholar 

  • Montsant A, Zarka A, Boussiba S (2001) Presence of a nonhydrolyzable biopolymer in the cell wall of vegetative cells and astaxanthin-rich cysts of Haematococcus pluvialis (Chlorophyceae). Mar Biotechnol (NY) 3:515–521

    Article  CAS  Google Scholar 

  • Sarada R, Vidhyavathi R, Usha D, Ravishankar GA (2006) An efficient method for extraction of astaxanthin from green alga Haematococcus pluvialis. J Agric Food Chem 54:7585–7588

    Article  CAS  Google Scholar 

  • Steinbrenner J, Linden H (2001) Regulation of two carotenoid biosynthesis genes coding for phytoene synthase and carotenoid hydroxylase during stress-induced astaxanthin formation in the green alga Haematococcus pluvialis. Plant Physiol 125:810–817

    Article  CAS  Google Scholar 

  • Steinbrenner J, Linden H (2003) Light induction of carotenoid biosynthesis genes in the green alga Haematococcuspluvialis: regulation by photosynthetic redox control. Plant Mol Biol 52:343–356

    Article  CAS  Google Scholar 

  • Sun Z, Cunningham FX, Gantt E (1998) Differential expression of two isopentenyl pyrophosphate isomerases and enhanced carotenoid accumulation in a unicellular chlorophyte. Proc Natl Acad Sci USA 95:11482–11488

    Article  CAS  Google Scholar 

  • Tan S, Cunningham FX, Youmans M, Grabowski B, Sun Z, Gantt E (1995) Cytochrome f loss in astaxanthin-accumulating red cells of Haematococcus pluvialis (Chlorophyceae): comparison of photosynthetic activity, photosynthetic enzymes, and thylakoid membrane polypeptides in red and green cells. J Phycol 31:897–905

    Article  CAS  Google Scholar 

  • Triki A, Maillard P, Gudin C (1997) Gametogenesis in Haematococcuspluvialis Flowtow (Volvocales, Chlorophyta). Phycologia 36:190–194

    Article  Google Scholar 

  • Usha T, Sarada R, Rao SR, Ravishankar GA (1999) Production of astaxanthin in Haematococcus pluvialis cultured in various media. Bioresour Technol 68:197–199

    Article  Google Scholar 

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Acknowledgment

The authors thank the Department of Biotechnology, Government of India, for the financial support. The authors R.V., L.V., and B.S.K. acknowledge the Council of Scientific and Industrial Research, India, for the research fellowships. Encouragement by Dr. V. Prakash, Director, CFTRI for research activities, is gratefully acknowledged. The authors are extremely grateful to Dr. Vinod Kumar, CFTRI, for assisting in conducting the experiments.

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Correspondence to Ravi Sarada.

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Vidhyavathi, R., Venkatachalam, L., Kamath, B.S. et al. Differential expression of carotenogenic genes and associated changes in pigment profile during regeneration of Haematococcus pluvialis cysts. Appl Microbiol Biotechnol 75, 879–887 (2007). https://doi.org/10.1007/s00253-007-0876-1

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  • DOI: https://doi.org/10.1007/s00253-007-0876-1

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