Characterization of Carotenoid-protein Complexes and Gene Expression Analysis Associated with Carotenoid Sequestration in Pigmented Cassava (Manihot Esculenta Crantz) Storage Root



Luiz Joaquim Castelo Branco Carvalho*, 1, John Lippolis2, Songbi Chen3, Claudia Regina Batista de Souza4, Eduardo A Vieira5, James V Anderson6
1 EMBRAPA Genetic Resources and Biotechnology, Brazil
2 USDA-ARS, National Animal Disease Center, Ames-AI, USA
3 CATAS Tropical Crops Genetic Resources Institute, Dangsou, China
4 Department of Genetics – UFPA, Brazil
5 EMBRAPA Cerrados, Brazil
6 USDA-ARS, Sunflower and Plant Biology Research Unit, Fargo, ND, USA


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© Carvalho et al.; Licensee Bentham Open.

open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http: //creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

* Address correspondence to this author at the EMBRAPA Genetic Re-sources and Biotechnology, Laboratory of Biochemistry and Biophysics, Caixa Postal 02372, 70 770 916 Brasilia, DF. Brazil; Tel: + 55 61 3448 4664, + 55 61 3448 4937,+ 55 61 9557 0838; Fax: + 55 61 3340 3658; E-mail: luiz.castelo@embrapa.br


Abstract

Carotenoid-protein complex (CPC) was isolated from chromoplast-enriched suspensions of cassava storage root (CSR) using size exclusion chromatography and characterized. Peptide sequences (LC_MS/MS spectrum) obtained from CPC and their corresponding proteins were obtained using publically available databases. Small Heat Shock Proteins (sHSPs) were the most abundant proteins identified in the CPC. Western blot analysis showed that Fribrillin and Or-protein were present in chromoplast-enriched suspensions of yellow root but not in the complex or white root. Results from qRT-PCR helped identify an isoform of HSP21 possessing four single point mutations in the intense yellow CSR that may be responsible for increased sequestration of b-carotene.

Keywords: Cassava root, carotenoid accumulation, carotenoid-protein complex, heat shock proteins.