Skip to main content
Log in

Cloning and differential expression analysis of geranylgeranyl diphosphate synthase gene from Dunaliella parva

  • Published:
Journal of Applied Phycology Aims and scope Submit manuscript

Abstract

Dunaliella parva can thrive and adapt to salt stress over a wide range of NaCl concentrations which also is related to carotenoid accumulation in Dunaliella. Dunaliella parva can accumulate carotenoids; however, the underlying mechanism of carotenoid accumulation needs further research. Thus, it is necessary to study biosynthesis and regulation of carotenoids for understanding carotenoid accumulation. In the present study, a gene encoding geranylgeranyl diphosphate synthase (GGPS) from the halophilic green alga D. parva has been cloned and analyzed. It is in the branch of terpene metabolism and named as DpGGPS (D. parva GGPS). The full-length complementary DNA (cDNA) of DpGGPS was 1612 bp, including an open reading frame (ORF) of 1059 bp, 189 bp 5′-untranslated region (5′-UTR) and 364 bp 3′-UTR. The 5′-flanking region was obtained by the genome walking method. Potential regulatory elements, associated with hormones, defense, and stress, were found in the 1310 bp 5′-flanking region. Functional characterization of DpGGPS in E. coli BL21(DE3) demonstrated that DpGGPS encoded a functional GGPS. Analysis of DpGGPS expression revealed a correlation between GGPS expression and the shift of NaCl concentration, which indicated that GGPS could be a salt stress responsive gene. Cloning and expression analysis of GGPS provides a foundation for studying the regulatory mechanism of carotenoid biosynthesis, adaptation mechanism to salt stress, and massive accumulation of carotenoids in D. parva.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Barbar A, Couture M, Sen SE, Béliveau C, Nisole A, Bipfubusa M, Cusson M (2013) Cloning, expression and characterization of an insect geranylgeranyl diphosphate synthase from Choristoneura fumiferana. Insect Biochem Mol Biol 43:947–958

    Article  CAS  PubMed  Google Scholar 

  • Bartley GE, Scolnik PA (1995) Plant carotenoids: pigments for photo-protection, visual attraction, and human health. Plant Cell 7:1027–1038

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ben-Amotz A, Avron M (1972) Photosynthetic activities of the halophilic alga Dunaliella parva. Plant Physiol 49:240–243

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Borowitzka MA (2013) Dunaliella: Biology, production, and markets. In: Richmond A, Hu Q (eds) Handbook of microalgal culture: applied phycology and biotechnology, 2nd edn. Wiley-Blackwell, New York, pp 359–368

  • Borowitzka MA, Siva CJ (2007) The taxonomy of the genus Dunaliella (Chlorophyta, Dunaliellales) with emphasis on the marine and halophilic species. J Appl Phycol 19:567–590

    Article  Google Scholar 

  • Borowitzka MA, Borowitzka LJ, Kessly D (1990) Effects of salinity increase on carotenoid accumulation in the green alga Dunaliella salina. J Appl Phycol 2:111–119

    Article  Google Scholar 

  • Botella-Pavia P, Rodriguez-Concepcion M (2006) Carotenoid biotechnology in plants for nutritionally improved foods. Physiol Plant 126:369–381

    Article  CAS  Google Scholar 

  • Carretero-Paulet L, Cairó A, Botella-Pavía P, Besumbes O, Campos N, Boronat A, Rodríguez-Concepción M (2006) Enhanced flux through the methylerythritol 4-phosphate pathway in Arabidopsis plants overexpressing deoxyxylulose 5-phosphate reductoisomerase. Plant Mol Biol 62:683–695

    Article  CAS  PubMed  Google Scholar 

  • Cervantes-Cervantes M, Gallagher CE, Zhu C, Wurtzel ET (2006) Maize cDNAs expressed in endosperm encode functional farnesyl diphosphate synthase with geranylgeranyl diphosphate synthase activity. Plant Physiol 141:220–231

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cowan AK, Rose PD (1991) Abscisic acid metabolism in salt-stressed cells of Dunaliella salina: possible interrelationship with beta-carotene accumulation. Plant Physiol 97:798–803

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gimmler H, Möller EM (1981) Salinity-dependant regulation of starch and glycerol metabolism in Dunaliella parva. Plant Cell Environ 4:367–375

    Article  CAS  Google Scholar 

  • Igielska-Kalwat J, Gościańska J, Nowak I (2015) Carotenoids as natural antioxidants. Postepy Hig Med Dosw 69:418–428

    Article  Google Scholar 

  • Jin E, Polle JEW (2009) Carotenoid biosynthesis in Dunaliella (Chlorophyta). In: Ben-Amotz A, Polle JEW, Subba Rao DV (eds) The Alga Dunaliella: biodiversity, physiology, genomics and biotechnology. Science Publishers, Enfield, pp 147–171

    Chapter  Google Scholar 

  • Laskaris G, Verpoorte R (2000) Purification and partial characterisation of geranylgeranyl diphosphate synthase, from Taxus baccata cell cultures. An enzyme that regulates taxane biosynthesis. Plant Sci 153:97–105

    Article  CAS  PubMed  Google Scholar 

  • Lescot M, Déhais P, Thijs G, Marchal K, Moreau Y, Van de Peer Y, Rouzé P, Rombauts S (2002) PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Res 30:325–327

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lian T, Dong CH, Yang T, Sun J (2014) Three types of geranylgeranyl diphosphate synthases from the medicinal caterpillar fungus, Cordyceps militaris (Ascomycetes). Int J Med Mushrooms 16:115–124

    Article  CAS  PubMed  Google Scholar 

  • Lohr M, Schwender J, Polle JE (2012) Isoprenoid biosynthesis in eukaryotic phototrophs: a spotlight on algae. Plant Sci 185–186:9–22

    Article  PubMed  Google Scholar 

  • Park S, Lee Y, Jin E (2013) Comparison of the responses of two Dunaliella strains, Dunaliella salina CCAP 19/18 and Dunaliella bardawil to light intensity with special emphasis on carotenogenesis. Algae 28:203–211

    Article  Google Scholar 

  • Pateraki I, Kanellis AK (2008) Isolation and functional analysis of two Cistus creticus cDNAs encoding geranylgeranyl diphosphate synthase. Phytochemistry 69:1641–1652

    Article  CAS  PubMed  Google Scholar 

  • Raja R, Hemaiswarya S, Rengasamy R (2007) Exploitation of Dunaliella for beta-carotene production. Appl Microbiol Biotechnol 74:517–523

    Article  CAS  PubMed  Google Scholar 

  • Ruppel NJ, Kropp KN, Davis PA, Martin AE, Luesse DR, Hangarter RP (2013) Mutations in geranylgeranyl diphosphate synthase 1 affect chloroplast development in Arabidopsis thaliana (Brassicaceae). Am J Bot 100:2074–2084

    Article  CAS  PubMed  Google Scholar 

  • Sánchez-Estudillo L, Freile-Pelegrin Y, Rivera-Madrid R, Robledo D, Narváez-Zapata JA (2006) Regulation of two photosynthetic pigment-related genes during stress-induced pigment formation in the green alga, Dunaliella salina. Biotechnol Lett 28:787–791

    Article  PubMed  Google Scholar 

  • Shang CH, Shi L, Ren A, Qin L, Zhao MW (2010) Molecular cloning, characterization, and differential expression of a lanosterol synthase gene from Ganoderma lucidum. Biosci Biotech Biochem 74:974–978

    Article  CAS  Google Scholar 

  • Srinivasan R, Kumar VA, Kumar D, Ramesh N, Babu S, Gothandam KM (2015) Effect of dissolved inorganic carbon on β-carotene and fatty acid production in Dunaliella sp. Appl Biochem Biotechnol 175:2895–2906

    Article  CAS  PubMed  Google Scholar 

  • Takaichi S (2011) Carotenoids in algae: distributions, biosyntheses and functions. Mar Drugs 9:1101–1118

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Thabet I, Guirimand G, Guihur A, Lanoue A, Courdavault V, Papon N, Bouzid S, Giglioli-Guivarc’h N, Simkin AJ, Clastre M (2012) Characterization and subcellular localization of geranylgeranyl diphosphate synthase from Catharanthus roseus. Mol Biol Rep 39:3235–3243

    Article  CAS  PubMed  Google Scholar 

  • Umeno D, Tobias AV, Arnold FH (2005) Diversifying carotenoid biosynthetic pathways by directed evolution. Microbiol Mol Biol Rev 69:51–78

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Velayos A, Papp T, Aguilar-Elena R, Fuentes-Vicente M, Eslava AP, Iturriaga EA, Alvarez MI (2003) Expression of the carG gene, encoding geranylgeranyl pyrophosphate synthase, is up-regulated by blue light in Mucor circinelloides. Curr Genet 43:112–120

    CAS  PubMed  Google Scholar 

  • Wang KC, Ohnuma S (2000) Isoprenyl diphosphate synthases. Biochim Biophys Acta 1529:33–48

    Article  CAS  PubMed  Google Scholar 

  • Wellburn AR (1994) The spectral determination of chlorophylls a and b, as well as total carotenoids, using various solvents with spectrophotometers of different resolution. J Plant Physiol 144:307–313

    Article  CAS  Google Scholar 

  • White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Shinsky JJ, White TJ (eds) PCR protocols: A guide to methods and applications. American Academic Press, Salt Lake City, pp 315–322

    Google Scholar 

  • Xu Z, Tian B, Sun Z, Lin J, Hua Y (2007) Identification and functional analysis of a phytoene desaturase gene from the extremely radioresistant bacterium Deinococcus radiodurans. Microbiology 153:1642–1652

    Article  CAS  PubMed  Google Scholar 

  • Zamir A (2009) Selected functional and molecular adaptations contributing to the exceptional stress tolerance of Dunaliella. In: Ben-Amotz A, Polle JEW, Subba Rao DV (eds) The Alga Dunaliella: biodiversity, physiology, genomics and biotechnology. Science Publishers, Enfield, pp 327–339

    Chapter  Google Scholar 

  • Zhang N, Wang F, Meng X, Luo S, Li Q, Dong H, Xu Z, Song R (2011) Molecular cloning and characterization of a trehalose-6-phosphate synthase/phosphatase from Dunaliella viridis. Mol Biol Rep 38:2241–2248

    Article  CAS  PubMed  Google Scholar 

  • Zhang S, Li XR, Xu H, Cao Y, Ma SH, Cao Y, Qiao D (2014) Molecular cloning and functional characterization of MnSOD from Dunaliella salina. J Basic Microbiol 54:438–447

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors gratefully thank Dr. Mohammad Asraful Alam for proof-reading and modification of English. The authors would also like to thank the editor in chief and four anonymous reviewers for their helpful comments and suggestions that greatly improved the manuscript. This research was supported by the National Natural Sciences Foundation of China (No. 51476177), Foundation of Key Laboratory of Renewable Energy, Chinese Academy of Sciences (No. y507j21001) and Foundation of Jiangsu Key Laboratory for Biomass-based Energy and Enzyme Technology (Huaiyin Normal University) (JSBEET1316).

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Zhenhong Yuan or Jun Xie.

Additional information

Changhua Shang and Xiaoli Xu contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Shang, C., Xu, X., Yuan, Z. et al. Cloning and differential expression analysis of geranylgeranyl diphosphate synthase gene from Dunaliella parva . J Appl Phycol 28, 2397–2405 (2016). https://doi.org/10.1007/s10811-015-0767-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10811-015-0767-2

Keywords

Navigation