Production of Saffron Apocarotenoids in Nicotiana benthamiana Plants Genome-Edited to Accumulate Zeaxanthin Precursor
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Published:2023-06-06
Issue:6
Volume:13
Page:729
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ISSN:2218-1989
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Container-title:Metabolites
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language:en
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Short-container-title:Metabolites
Author:
Demurtas Olivia Costantina1ORCID, Sulli Maria1ORCID, Ferrante Paola1, Mini Paola1, Martí Maricarmen2, Aragonés Verónica2, Daròs José-Antonio2ORCID, Giuliano Giovanni1ORCID
Affiliation:
1. Biotechnology and Agro-Industry Division, ENEA, Italian National Agency for New Technologies, Energy and Sustainable Economic Development, Casaccia Research Center, 00123 Rome, Italy 2. Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València, 46022 Valencia, Spain
Abstract
Crocins are glycosylated apocarotenoids with strong coloring power and anti-oxidant, anticancer, and neuro-protective properties. We previously dissected the saffron crocin biosynthesis pathway, and demonstrated that the CsCCD2 enzyme, catalyzing the carotenoid cleavage step, shows a strong preference for the xanthophyll zeaxanthin in vitro and in bacterio. In order to investigate substrate specificity in planta and to establish a plant-based bio-factory system for crocin production, we compared wild-type Nicotiana benthamiana plants, accumulating various xanthophylls together with α- and β-carotene, with genome-edited lines, in which all the xanthophylls normally accumulated in leaves were replaced by a single xanthophyll, zeaxanthin. These plants were used as chassis for the production in leaves of saffron apocarotenoids (crocins, picrocrocin) using two transient expression methods to overexpress CsCCD2: agroinfiltration and inoculation with a viral vector derived from tobacco etch virus (TEV). The results indicated the superior performance of the zeaxanthin-accumulating line and of the use of the viral vector to express CsCCD2. The results also suggested a relaxed substrate specificity of CsCCD2 in planta, cleaving additional carotenoid substrates.
Funder
European Commission Spanish Ministerio de Ciencia e Innovación Ministerio de Educación, Cultura y Deporte
Subject
Molecular Biology,Biochemistry,Endocrinology, Diabetes and Metabolism
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