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Sierra, SantiagoAuthor

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February 8, 2025
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Characterization of cassava ORANGE proteins and their capability to increase provitamin A carotenoids accumulation

Publicated to:Plos One. 17 (1): e0262412- - 2022-01-07 17(1), DOI: 10.1371/journal.pone.0262412

Authors: Jaramillo, Angelica M; Sierra, Santiago; Chavarriaga-Aguirre, Paul; Katherine Castillo, Diana; Gkanogiannis, Anestis; Becerra Lopez-Lavalle, Luis Augusto; Pablo Arciniegas, Juan; Sun, Tianhu; Li, Li; Welsch, Ralf; Boy, Erick; Alvarez, Daniel

Affiliations

Alliance Biovers Int, Cali, Colombia - Author
Alliance Biovers Int, HarvestPlus, Cali, Colombia - Author
Cornell Univ, Robert W Holley Ctr Agr & Hlth, USDA ARS, Ithaca, NY USA - Author
Int Ctr Trop Agr CIAT, Cali, Colombia - Author
Int Food Policy Res Inst, HarvestPlus, Washington, DC 20036 USA - Author
Univ Freiburg, Fac Biol 2, Freiburg, Germany - Author
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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.

Keywords

Beta-caroteneBiofortificatioBiosynthetic pathwaysCarotenoidsDown-regulationDry-matterGeneGene expression regulation, plantGenes, plantGeranylgeranyl-diphosphate geranylgeranyltransferaseLow-incomeManihotPhytoene-synthasePlant proteinsPlant rootsPostharvest storageProvitaminsSweet-potatoVitamin aVitamin-a status

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal Plos One due to its progression and the good impact it has achieved in recent years, according to the agency Scopus (SJR), it has become a reference in its field. In the year of publication of the work, 2022, it was in position , thus managing to position itself as a Q1 (Primer Cuartil), in the category Multidisciplinary. Notably, the journal is positioned above the 90th percentile.

From a relative perspective, and based on the normalized impact indicator calculated from the Field Citation Ratio (FCR) of the Dimensions source, it yields a value of: 5.82, which indicates that, compared to works in the same discipline and in the same year of publication, it ranks as a work cited above average. (source consulted: Dimensions Sep 2025)

Specifically, and according to different indexing agencies, this work has accumulated citations as of 2025-09-08, the following number of citations:

  • WoS: 10
  • Europe PMC: 8

Impact and social visibility

From the perspective of influence or social adoption, and based on metrics associated with mentions and interactions provided by agencies specializing in calculating the so-called "Alternative or Social Metrics," we can highlight as of 2025-09-08:

  • The use, from an academic perspective evidenced by the Altmetric agency indicator referring to aggregations made by the personal bibliographic manager Mendeley, gives us a total of: 40.
  • The use of this contribution in bookmarks, code forks, additions to favorite lists for recurrent reading, as well as general views, indicates that someone is using the publication as a basis for their current work. This may be a notable indicator of future more formal and academic citations. This claim is supported by the result of the "Capture" indicator, which yields a total of: 40 (PlumX).

With a more dissemination-oriented intent and targeting more general audiences, we can observe other more global scores such as:

  • The Total Score from Altmetric: 11.
  • The number of mentions on the social network X (formerly Twitter): 3 (Altmetric).
  • The number of mentions in news outlets: 1 (Altmetric).

It is essential to present evidence supporting full alignment with institutional principles and guidelines on Open Science and the Conservation and Dissemination of Intellectual Heritage. A clear example of this is:

  • The work has been submitted to a journal whose editorial policy allows open Open Access publication.

Leadership analysis of institutional authors

This work has been carried out with international collaboration, specifically with researchers from: Colombia; Germany; United States of America.