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

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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.

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Abbreviations

ABA:

Abscisic acid

NCED :

9-cis-Epoxycarotenoid cleavage dioxygenase

PSY:

Phytoene synthase

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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.

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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

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  1. Jacobo Arango
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  2. Florian Wüst
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  3. Peter Beyer
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  4. Ralf Welsch
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Corresponding author

Correspondence to Ralf Welsch.

Electronic supplementary material

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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)

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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

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