Skip to main content
SpringerLink
Log in

Characterization of phytoene synthases from cassava and their involvement in abiotic stress-mediated responses

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

Abstract

Abiotic stress stimuli induce the increased synthesis of abscisic acid (ABA), which is generated through the cleavage of xanthophyll precursors. To cope with the increased xanthophyll demand, maize and rice contain a third stress-induced gene copy, coding for phytoene synthase (PSY), which catalyzes the first carotenoid-specific reaction in the pathway. To investigate whether this specific response extends beyond the Poaceae, cassava was analyzed, an important tropical crop known for its drought tolerance. We also found three PSY genes in cassava, one of which (MePSY3) forms a separate branch with the stress-specific Poaceae homologs. However, MePSY3 transcripts were virtually absent in all tissues investigated and did not change upon abiotic stress treatment. In contrast, the two remaining PSY genes contributed differentially to carotenoid biosynthesis in leaves, roots, and flower organs and responded towards drought and salt-stress conditions. Detailed analyses of PSY and 9-cis-epoxycarotenoid cleavage dioxygenase (MeNCED) expression and resulting ABA levels revealed MePSY1 as the main stress-responsive paralog. In the presence of high carotenoid levels in leaves, MePSY1 appeared to support, but not to be rate-limiting for ABA formation; MeNCED represented the main driver. The inverse situation was found in roots where carotenoid levels are low. Moreover, ABA formation and the relative induction kinetics showed discrimination between drought and salt stress. Compared to rice as a drought-intolerant species, the drought response in cassava followed a different kinetic regime. The difference is thought to represent a component contributing to the large differences in the adaptation towards water supply.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

ABA:

Abscisic acid

NCED :

9-cis-Epoxycarotenoid cleavage dioxygenase

PSY:

Phytoene synthase

References

  • Arango J, Salazar B, Welsch R, Sarmiento F, Beyer P, Al-Babili S (2010) Putative storage root specific promoters from cassava and yam: cloning and evaluation in transgenic carrots as a model system. Plant Cell Rep 29:651–659

    Article  CAS  PubMed  Google Scholar 

  • Auldridge ME, McCarty DR, Klee HJ (2006) Plant carotenoid cleavage oxygenases and their apocarotenoid products. Curr Opin Plant Biol 9:315–321

    Article  CAS  PubMed  Google Scholar 

  • Bartley GE, Scolnik PA, Beyer P (1999) Two Arabidopsis thaliana carotene desaturases, phytoene desaturase and ζ-carotene desaturase, expressed in Escherichia coli, catalyze a poly-cis pathway to yield pro-lycopene. Euro J Biochem 259:396–403

    Article  CAS  Google Scholar 

  • Bouvier F, Isner J, Dogbo O, Camara B (2005) Oxidative tailoring of carotenoids: a prospect towards novel functions in plants. Trends Plant Sci 10:187–194

    Article  CAS  PubMed  Google Scholar 

  • Bouwmeester HJ, Roux C, Lopez-Raez JA, Bécard G (2007) Rhizosphere communication of plants, parasitic plants and AM fungi. Trends Plant Sci 12:224–230

    Article  CAS  PubMed  Google Scholar 

  • Bruce B (2001) The paradox of plastid transit peptides: conservation of function despite divergence in primary structure. Biochim Biophys Acta 1541:2–21

    Article  CAS  PubMed  Google Scholar 

  • Chaves MM, Flexas J, Pinheiro C (2009) Photosynthesis under drought and salt stress: regulation mechanisms from whole plant to cell. Ann Bot 103:551–560

    Article  CAS  PubMed  Google Scholar 

  • Chen Y, Li F, Wurtzel ET (2010) Isolation and characterization of the Z-ISO gene encoding a missing component of carotenoid biosynthesis in plants. Plant Physiol 153:66–79

    Article  CAS  PubMed  Google Scholar 

  • Christmann A, Weiler EW, Steudle E, Grill E (2007) A hydraulic signal in root-to-shoot signalling of water shortage. Plant J 52:167–174

    Article  CAS  PubMed  Google Scholar 

  • Demmig-Adams B, Adams WW (1992) Photoprotection and other responses of plants to high light stress. Annu Rev Plant Physiol Plant Mol Biol 43:599–626

    Article  CAS  Google Scholar 

  • Demmig-Adams B, Adams WW (2002) Antioxidants in photosynthesis and human nutrition. Science 298:2149–2153

    Article  CAS  PubMed  Google Scholar 

  • Dun EA, Brewer PB, Beveridge CA (2009) Strigolactones: discovery of the elusive shoot branching hormone. Trends Plant Sci 14:364–372

    Article  CAS  PubMed  Google Scholar 

  • Fan J, Hill L, Crooks C, Doerner P, Lamb C (2009) Abscisic acid has a key role in modulating diverse plant-pathogen interactions. Plant Physiol 150:1750–1761

    Article  CAS  PubMed  Google Scholar 

  • Fraser PD, Truesdale MR, Bird CR, Schuch W, Bramley PM (1994) Carotenoid biosynthesis during tomato fruit development (evidence for tissue-specific gene expression). Plant Physiol 105:405–413

    CAS  PubMed  Google Scholar 

  • Hieber AD, Bugos RC, Yamamoto HY (2000) Plant lipocalins: violaxanthin de-epoxidase and zeaxanthin epoxidase. Biochim Biophys Acta 1482:84–91

    CAS  PubMed  Google Scholar 

  • Isaacson T, Ronen G, Zamir D, Hirschberg J (2002) Cloning of tangerine from tomato reveals a carotenoid isomerase essential for the production of beta-carotene and xanthophylls in plants. Plant Cell 14:333–342

    Article  CAS  PubMed  Google Scholar 

  • Isaacson T, Ohad I, Beyer P, Hirschberg J (2004) Analysis in vitro of the enzyme CRTISO establishes a poly-cis-carotenoid biosynthesis pathway in plants. Plant Physiol 136:4246–4255

    Article  CAS  PubMed  Google Scholar 

  • Iuchi S, Kobayashi M, Taji T, Naramoto M, Seki M, Kato T, Tabata S, Kakubari Y, Yamaguchi-Shinozaki K, Shinozaki K (2001) Regulation of drought tolerance by gene manipulation of 9-cis-epoxycarotenoid dioxygenase, a key enzyme in abscisic acid biosynthesis in Arabidopsis. Plant J 27:325–333

    Article  CAS  PubMed  Google Scholar 

  • Jiang F, Hartung W (2008) Long-distance signalling of abscisic acid (ABA): the factors regulating the intensity of the ABA signal. J Exp Bot 59:37–43

    Article  CAS  PubMed  Google Scholar 

  • Kim J, Smith JJ, Tian L, Dellapenna D (2009) The evolution and function of carotenoid hydroxylases in Arabidopsis. Plant Cell Physiol 50:463–479

    Article  CAS  PubMed  Google Scholar 

  • Li F, Vallabhaneni R, Wurtzel ET (2008) PSY3, a new member of the phytoene synthase gene family conserved in the Poaceae and regulator of abiotic-stress-induced root carotenogenesis. Plant Physiol 146:1333–1345

    Article  CAS  PubMed  Google Scholar 

  • Lindgren LO, Stålberg KG, Höglund AS (2003) Seed-specific overexpression of an endogenous Arabidopsis phytoene synthase gene results in delayed germination and increased levels of carotenoids, chlorophyll, and abscisic acid. Plant Physiol 132:779–785

    Article  CAS  PubMed  Google Scholar 

  • Maass D, Arango J, Wüst F, Beyer P, Welsch R (2009) Carotenoid crystal formation in Arabidopsis and carrot roots caused by increased phytoene synthase protein levels. PLoS One 4:e6373

    Article  PubMed  Google Scholar 

  • Mann V, Harker M, Pecker I, Hirschberg J (2000) Metabolic engineering of astaxanthin production in tobacco flowers. Nature Biotechnol 18:888–892

    Article  CAS  Google Scholar 

  • Moehs C, Tian L, Osteryoung K, Dellapenna D (2001) Analysis of carotenoid biosynthetic gene expression during marigold petal development. Plant Mol Biol 45:281–293

    Article  CAS  PubMed  Google Scholar 

  • Nambara E, Marion-Poll A (2005) Abscisic acid biosynthesis and catabolism. Annu Rev Plant Biol 56:165–185

    Article  CAS  PubMed  Google Scholar 

  • Niyogi KK (1999) Photoprotection revisited: genetic and molecular approaches. Annu Rev Plant Physiol Plant Mol Biol 50:333–359

    Article  CAS  PubMed  Google Scholar 

  • Niyogi KK, Grossman AR, Björkman O (1998) Arabidopsis mutants define a central role for the xanthophyll cycle in the regulation of photosynthetic energy conversion. Plant Cell 10:1121–1134

    Article  CAS  PubMed  Google Scholar 

  • Ohmiya A, Kishimoto S, Aida R, Yoshioka S, Sumitomo K (2006) Carotenoid cleavage dioxygenase (CmCCD4a) contributes to white color formation in chrysanthemum petals. Plant Physiol 142:1193–1201

    Article  CAS  PubMed  Google Scholar 

  • Owen SJ, Abrams SR (2009) Measurement of plant hormones by liquid chromatography–mass spectrometry. Methods Mol Biol 495:39–51

    Article  CAS  PubMed  Google Scholar 

  • Paine JA, Shipton CA, Chaggar S, Howells RM, Kennedy MJ, Vernon G, Wright SY, Hinchliffe E, Adams JL, Silverstone AL, Drake R (2005) Improving the nutritional value of Golden Rice through increased pro-vitamin A content. Nature Biotechnol 23:429–430

    Article  Google Scholar 

  • Park H, Kreunen SS, Cuttriss AJ, DellaPenna D, Pogson BJ (2002) Identification of the carotenoid isomerase provides insight into carotenoid biosynthesis, prolamellar body formation, and photomorphogenesis. Plant Cell 14:321–332

    Article  CAS  PubMed  Google Scholar 

  • Parry AD, Griffiths A, Horgan R (1992) Abscisic acid biosynthesis in roots. Planta 187:192–197

    Article  CAS  Google Scholar 

  • Pilon-Smits E, Ebskamp M, Paul MJ, Jeuken M, Weisbeek PJ, Smeekens S (1995) Improved performance of transgenic fructan-accumulating tobacco under drought stress. Plant Physiol 107:125–130

    CAS  PubMed  Google Scholar 

  • Reymond P, Weber H, Damond M, Farmer EE (2000) Differential gene expression in response to mechanical wounding and insect feeding in Arabidopsis. Plant Cell 12:707–720

    Article  CAS  PubMed  Google Scholar 

  • Ronen G, Cohen M, Zamir D, Hirschberg J (1999) Regulation of carotenoid biosynthesis during tomato fruit development: expression of the gene for lycopene epsilon-cyclase is down-regulated during ripening and is elevated in the mutant Delta. Plant J 17:341–351

    Article  CAS  PubMed  Google Scholar 

  • Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing guide trees. Mol Biol Evol 4:406–425

    CAS  PubMed  Google Scholar 

  • Schaub P, Al-Babili S, Drake R, Beyer P (2005) Why is Golden Rice golden (yellow) instead of red? Plant Physiol 138:441–450

    Article  CAS  PubMed  Google Scholar 

  • Shewmaker CK, Sheehy JA, Daley M, Colburn S, Ke DY (1999) Seed-specific overexpression of phytoene synthase: increase in carotenoids and other metabolic effects. Plant J 20:401–412

    Article  CAS  PubMed  Google Scholar 

  • Sundaresan S, Sudhakaran PR (1995) Water stress-induced alterations in the proline metabolism of drought-susceptible and -tolerant cassava (Manihot esculenta) cultivars. Physiol Plant 94:635–642

    Article  CAS  Google Scholar 

  • Szabó I, Bergantino E, Giacometti GM (2005) Light and oxygenic photosynthesis: energy dissipation as a protection mechanism against photo-oxidation. EMBO Rep 6:629–634

    Article  PubMed  Google Scholar 

  • Tan B, Joseph LM, Deng W, Liu L, Li Q, Cline K, McCarty DR (2003) Molecular characterization of the Arabidopsis 9-cis epoxycarotenoid dioxygenase gene family. Plant J 35:44–56

    Article  CAS  PubMed  Google Scholar 

  • von Lintig J, Welsch R, Bonk M, Giuliano G, Batschauer A, Kleinig H (1997) Light-dependent regulation of carotenoid biosynthesis occurs at the level of phytoene synthase expression and is mediated by phytochrome in Sinapis alba and Arabidopsis thaliana seedlings. Plant J 12:625–634

    Article  Google Scholar 

  • Welsch R, Beyer P, Hugueney P, Kleinig H, von Lintig J (2000) Regulation and activation of phytoene synthase, a key enzyme in carotenoid biosynthesis, during photomorphogenesis. Planta 211:846–854

    Article  CAS  PubMed  Google Scholar 

  • Welsch R, Medina J, Giuliano G, Beyer P, von Lintig J (2003) Structural and functional characterization of the phytoene synthase promoter from Arabidopsis thaliana. Planta 216:523–534

    CAS  PubMed  Google Scholar 

  • Welsch R, Wüst F, Bär C, Al-Babili S, Beyer P (2008) A third phytoene synthase is devoted to abiotic stress-induced abscisic acid formation in rice and defines functional diversification of phytoene synthase genes. Plant Physiol 147:367–380

    Article  CAS  PubMed  Google Scholar 

  • Wilkinson S, Davies WJ (2002) ABA-based chemical signalling: the co-ordination of responses to stress in plants. Plant Cell Environ 25:195–210

    Article  CAS  PubMed  Google Scholar 

  • Ye X, Al-Babili S, Klöti A, Zhang J, Lucca P, Beyer P, Potrykus I (2000) Engineering the provitamin A (β-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm. Science 287:303–305

    Article  CAS  PubMed  Google Scholar 

  • Yoshida S, Forno DA, Cook JH, Gomez KA (1976) Routine procedures for growing rice plants in culture solution. In: Yoshida S, Forno DA, Cook JH, Gomez KA (eds) Laboratory manual for physiological studies of rice. IRRI, Philippines, pp 61–66

    Google Scholar 

  • Zhu JK (2002) Salt and drought stress signal transduction in plants. Annu Rev Plant Biol 53:247–273

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the HarvestPlus research consortium and the Grand Challenges in Global Health initiative of the Bill and Melinda Gates Foundation. We are deeply grateful to Paul Chavarriaga (CIAT) for the supply of cassava plant material.

Author information

Authors and Affiliations

  1. Faculty of Biology II, Cell Biology, University of Freiburg, Schänzlestr. 1, 79104, Freiburg, Germany

    Jacobo Arango, Florian Wüst, Peter Beyer & Ralf Welsch

Authors
  1. Jacobo Arango
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Florian Wüst
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Peter Beyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ralf Welsch
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ralf Welsch.

Electronic supplementary material

Below is the link to the electronic supplementary material.

425_2010_1250_MOESM1_ESM.doc

Supplemental Fig. S1 Alignment of PSY sequences from different taxa. Identical amino acids are shown in white letters on black background, conservative positions are shown with black letters on gray background and non-similar amino acids are shown in black on white type. Sequences from the following accession numbers were used: Me, Manihot esculenta, 1: GU111719, 2, GU111720, 3, locus: cassava32745 (www.phytozome.net), At, Arabidopsis thaliana, NP_197225, Sl, Solanum lycopersicum, 1, EF534740, 2, EF534738, Os, Oryza sativa, 1, NP_001058647, 2, ABA99494, 3, ACI62767, Zm, Zea mays, 1, AAR08445, 2, AAX13807, 3, ABD17618; Dc, Daucus carota, 1, Q9SSU8, 2, ABB52068; Rc, Ricinus communis, 1, XP_002527067, 2, XP_002532975 Supplementary material 1 (DOC 75 kb)

425_2010_1250_MOESM2_ESM.ppt

Supplemental Fig. S2 Phylogenetic tree of Arabidopsis CCDs and amino acid sequences deduced from two cassava EST contigs, calculated using the neighbor-joining algorithm (Saitou and Nei 1987). AGI numbers are as follows: NCED2: At4g18350, NCED3: At3g14440, NCED5: At1g30100, NCED6: At3g24220, NCED9: At1g78390, CCD1: At3g63520, CCD4: At4g19170, CCD7: At2g44990, CCD8: At4g32810; GenBank accession numbers for the cassava EST contigs (MeCon1 and 2) are given in Materials and methods Supplementary material 2 (PPT 26 kb)

425_2010_1250_MOESM3_ESM.ppt

Supplemental Fig. S3 PA and DPA levels in detached cassava and rice leaves following drought stress treatment. Cassava (left) and rice (right) leaves were subjected to drought stress as described in Fig. 5. Phaseic acid (PA) and dihydroxy phaseic acid (DPA) were identified by LC–MS. The diagram shows the ratios of peak areas from PA and DPA, respectively, to the peak area of the internal standard used in the extraction Supplementary material 3 (PPT 96 kb)

425_2010_1250_MOESM4_ESM.doc

Supplementary Table S1 Carotenoid and chlorophyll content of detached cassava leaves following salt and drought stress treatment. Leaves from cassava plants grown in vitro were detached and incubated in 250 mM NaCl (salt) or 40% (w/v) PEG 6000 in propagation medium (drought) for the times indicated. Control plants were transferred into fresh propagation medium and harvested at the same times (control). Carotenoids were quantified by HPLC. vio/neo, violaxanthin and neoxanthin; o. xan., other xanthophylls; lut, lutein, a-caro, α-carotene; b-caro, β-carotene, chl, chlorophyll. Data are given in nmol mg−1 DW and represent the mean ± SD of two technical replicates Supplementary material 4 (DOC 49 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Arango, J., Wüst, F., Beyer, P. et al. Characterization of phytoene synthases from cassava and their involvement in abiotic stress-mediated responses. Planta 232, 1251–1262 (2010). https://doi.org/10.1007/s00425-010-1250-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-010-1250-6

Keywords

Search

Navigation