Characterization of cassava ORANGE proteins and their capability to increase provitamin A carotenoids accumulation
Cassava (Manihot esculenta Crantz) biofortification with provitamin A carotenoids is an ongoing process that aims to alleviate vitamin A deficiency. The moderate content of provitamin A carotenoids achieved so far limits the contribution to providing adequate dietary vitamin A levels. Strategies to...
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| creator | Jaramillo, Angélica M Sierra, Santiago Chavarriaga-Aguirre, Paul Castillo, Diana Katherine Gkanogiannis, Anestis López-Lavalle, Luis Augusto Becerra Arciniegas, Juan Pablo Sun, Tianhu Li, Li Welsch, Ralf Boy, Erick Álvarez, Daniel |
| description | Cassava (Manihot esculenta Crantz) biofortification with provitamin A carotenoids is an ongoing process that aims to alleviate vitamin A deficiency. The moderate content of provitamin A carotenoids achieved so far limits the contribution to providing adequate dietary vitamin A levels. Strategies to increase carotenoid content focused on genes from the carotenoids biosynthesis pathway. In recent years, special emphasis was given to ORANGE protein (OR), which promotes the accumulation of carotenoids and their stability in several plants. The aim of this work was to identify, characterize and investigate the role of OR in the biosynthesis and stabilization of carotenoids in cassava and its relationship with phytoene synthase (PSY), the rate-limiting enzyme of the carotenoids biosynthesis pathway. Gene and protein characterization of OR, expression levels, protein amounts and carotenoids levels were evaluated in roots of one white (60444) and two yellow cassava cultivars (GM5309-57 and GM3736-37). Four OR variants were found in yellow cassava roots. Although comparable expression was found for three variants, significantly higher OR protein amounts were observed in the yellow varieties. In contrast, cassava PSY1 expression was significantly higher in the yellow cultivars, but PSY protein amount did not vary. Furthermore, we evaluated whether expression of one of the variants, MeOR_X1, affected carotenoid accumulation in cassava Friable Embryogenic Callus (FEC). Overexpression of maize PSY1 alone resulted in carotenoids accumulation and induced crystal formation. Co-expression with MeOR_X1 led to greatly increase of carotenoids although PSY1 expression was high in the co-expressed FEC. Our data suggest that posttranslational mechanisms controlling OR and PSY protein stability contribute to higher carotenoid levels in yellow cassava. Moreover, we showed that cassava FEC can be used to study the efficiency of single and combinatorial gene expression in increasing the carotenoid content prior to its application for the generation of biofortified cassava with enhanced carotenoids levels. |
| doi_str_mv | 10.1371/journal.pone.0262412 |
| format | Article |
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The moderate content of provitamin A carotenoids achieved so far limits the contribution to providing adequate dietary vitamin A levels. Strategies to increase carotenoid content focused on genes from the carotenoids biosynthesis pathway. In recent years, special emphasis was given to ORANGE protein (OR), which promotes the accumulation of carotenoids and their stability in several plants. The aim of this work was to identify, characterize and investigate the role of OR in the biosynthesis and stabilization of carotenoids in cassava and its relationship with phytoene synthase (PSY), the rate-limiting enzyme of the carotenoids biosynthesis pathway. Gene and protein characterization of OR, expression levels, protein amounts and carotenoids levels were evaluated in roots of one white (60444) and two yellow cassava cultivars (GM5309-57 and GM3736-37). Four OR variants were found in yellow cassava roots. Although comparable expression was found for three variants, significantly higher OR protein amounts were observed in the yellow varieties. In contrast, cassava PSY1 expression was significantly higher in the yellow cultivars, but PSY protein amount did not vary. Furthermore, we evaluated whether expression of one of the variants, MeOR_X1, affected carotenoid accumulation in cassava Friable Embryogenic Callus (FEC). Overexpression of maize PSY1 alone resulted in carotenoids accumulation and induced crystal formation. Co-expression with MeOR_X1 led to greatly increase of carotenoids although PSY1 expression was high in the co-expressed FEC. Our data suggest that posttranslational mechanisms controlling OR and PSY protein stability contribute to higher carotenoid levels in yellow cassava. Moreover, we showed that cassava FEC can be used to study the efficiency of single and combinatorial gene expression in increasing the carotenoid content prior to its application for the generation of biofortified cassava with enhanced carotenoids levels.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0262412</identifier><identifier>PMID: 34995328</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Accumulation ; Analysis ; Biology and Life Sciences ; Biosynthesis ; Biosynthetic Pathways ; Callus ; Carotenoids ; Carotenoids - metabolism ; Cassava ; Combinatorial analysis ; Control stability ; Cultivars ; Diet ; Food ; Friability ; Gene expression ; Gene Expression Regulation, Plant ; Genes ; Genes, Plant ; Genetic aspects ; Genetically altered foods ; Geranylgeranyl-Diphosphate Geranylgeranyltransferase - genetics ; Geranylgeranyl-Diphosphate Geranylgeranyltransferase - metabolism ; Health aspects ; Low income groups ; Manihot - genetics ; Manihot - metabolism ; Manihot esculenta ; Nutrient deficiency ; Physical Sciences ; Phytoene synthase ; Plant Proteins - genetics ; Plant Proteins - metabolism ; Plant Roots - genetics ; Plant Roots - metabolism ; Properties ; Protein biosynthesis ; Proteins ; Provitamins - metabolism ; Research and Analysis Methods ; Retinene ; Roots ; Vitamin A ; Vitamin A - metabolism</subject><ispartof>PloS one, 2022-01, Vol.17 (1), p.e0262412-e0262412</ispartof><rights>COPYRIGHT 2022 Public Library of Science</rights><rights>2022 Jaramillo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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The moderate content of provitamin A carotenoids achieved so far limits the contribution to providing adequate dietary vitamin A levels. Strategies to increase carotenoid content focused on genes from the carotenoids biosynthesis pathway. In recent years, special emphasis was given to ORANGE protein (OR), which promotes the accumulation of carotenoids and their stability in several plants. The aim of this work was to identify, characterize and investigate the role of OR in the biosynthesis and stabilization of carotenoids in cassava and its relationship with phytoene synthase (PSY), the rate-limiting enzyme of the carotenoids biosynthesis pathway. Gene and protein characterization of OR, expression levels, protein amounts and carotenoids levels were evaluated in roots of one white (60444) and two yellow cassava cultivars (GM5309-57 and GM3736-37). Four OR variants were found in yellow cassava roots. Although comparable expression was found for three variants, significantly higher OR protein amounts were observed in the yellow varieties. In contrast, cassava PSY1 expression was significantly higher in the yellow cultivars, but PSY protein amount did not vary. Furthermore, we evaluated whether expression of one of the variants, MeOR_X1, affected carotenoid accumulation in cassava Friable Embryogenic Callus (FEC). Overexpression of maize PSY1 alone resulted in carotenoids accumulation and induced crystal formation. Co-expression with MeOR_X1 led to greatly increase of carotenoids although PSY1 expression was high in the co-expressed FEC. Our data suggest that posttranslational mechanisms controlling OR and PSY protein stability contribute to higher carotenoid levels in yellow cassava. Moreover, we showed that cassava FEC can be used to study the efficiency of single and combinatorial gene expression in increasing the carotenoid content prior to its application for the generation of biofortified cassava with enhanced carotenoids levels.</description><subject>Accumulation</subject><subject>Analysis</subject><subject>Biology and Life Sciences</subject><subject>Biosynthesis</subject><subject>Biosynthetic Pathways</subject><subject>Callus</subject><subject>Carotenoids</subject><subject>Carotenoids - metabolism</subject><subject>Cassava</subject><subject>Combinatorial analysis</subject><subject>Control stability</subject><subject>Cultivars</subject><subject>Diet</subject><subject>Food</subject><subject>Friability</subject><subject>Gene expression</subject><subject>Gene Expression Regulation, Plant</subject><subject>Genes</subject><subject>Genes, Plant</subject><subject>Genetic aspects</subject><subject>Genetically altered foods</subject><subject>Geranylgeranyl-Diphosphate 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Erick</au><au>Álvarez, Daniel</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of cassava ORANGE proteins and their capability to increase provitamin A carotenoids accumulation</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2022-01-07</date><risdate>2022</risdate><volume>17</volume><issue>1</issue><spage>e0262412</spage><epage>e0262412</epage><pages>e0262412-e0262412</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Cassava (Manihot esculenta Crantz) biofortification with provitamin A carotenoids is an ongoing process that aims to alleviate vitamin A deficiency. The moderate content of provitamin A carotenoids achieved so far limits the contribution to providing adequate dietary vitamin A levels. Strategies to increase carotenoid content focused on genes from the carotenoids biosynthesis pathway. In recent years, special emphasis was given to ORANGE protein (OR), which promotes the accumulation of carotenoids and their stability in several plants. The aim of this work was to identify, characterize and investigate the role of OR in the biosynthesis and stabilization of carotenoids in cassava and its relationship with phytoene synthase (PSY), the rate-limiting enzyme of the carotenoids biosynthesis pathway. Gene and protein characterization of OR, expression levels, protein amounts and carotenoids levels were evaluated in roots of one white (60444) and two yellow cassava cultivars (GM5309-57 and GM3736-37). Four OR variants were found in yellow cassava roots. Although comparable expression was found for three variants, significantly higher OR protein amounts were observed in the yellow varieties. In contrast, cassava PSY1 expression was significantly higher in the yellow cultivars, but PSY protein amount did not vary. Furthermore, we evaluated whether expression of one of the variants, MeOR_X1, affected carotenoid accumulation in cassava Friable Embryogenic Callus (FEC). Overexpression of maize PSY1 alone resulted in carotenoids accumulation and induced crystal formation. Co-expression with MeOR_X1 led to greatly increase of carotenoids although PSY1 expression was high in the co-expressed FEC. Our data suggest that posttranslational mechanisms controlling OR and PSY protein stability contribute to higher carotenoid levels in yellow cassava. Moreover, we showed that cassava FEC can be used to study the efficiency of single and combinatorial gene expression in increasing the carotenoid content prior to its application for the generation of biofortified cassava with enhanced carotenoids levels.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>34995328</pmid><doi>10.1371/journal.pone.0262412</doi><tpages>e0262412</tpages><orcidid>https://orcid.org/0000-0002-3608-0597</orcidid><orcidid>https://orcid.org/0000-0002-5930-2295</orcidid><orcidid>https://orcid.org/0000-0002-7463-6816</orcidid><orcidid>https://orcid.org/0000-0002-0950-0074</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2022-01, Vol.17 (1), p.e0262412-e0262412 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_2617748069 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Free Full-Text Journals in Chemistry; Public Library of Science (PLoS) |
| subjects | Accumulation Analysis Biology and Life Sciences Biosynthesis Biosynthetic Pathways Callus Carotenoids Carotenoids - metabolism Cassava Combinatorial analysis Control stability Cultivars Diet Food Friability Gene expression Gene Expression Regulation, Plant Genes Genes, Plant Genetic aspects Genetically altered foods Geranylgeranyl-Diphosphate Geranylgeranyltransferase - genetics Geranylgeranyl-Diphosphate Geranylgeranyltransferase - metabolism Health aspects Low income groups Manihot - genetics Manihot - metabolism Manihot esculenta Nutrient deficiency Physical Sciences Phytoene synthase Plant Proteins - genetics Plant Proteins - metabolism Plant Roots - genetics Plant Roots - metabolism Properties Protein biosynthesis Proteins Provitamins - metabolism Research and Analysis Methods Retinene Roots Vitamin A Vitamin A - metabolism |
| title | Characterization of cassava ORANGE proteins and their capability to increase provitamin A carotenoids accumulation |
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