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Fate of free and bound phytol and tocopherols during fruit ripening of two Capsicum cultivars.

ABSTRACT: Phytol and tocopherols and their fatty acid esters (PFAE and TFAE) are isoprenoid lipid components which can be found for instance in vegetables. Their behavior during maturation of fruits and vegetables could reveal valuable information on their biosynthetic formation and biological function. As pods of the genus Capsicum contain considerable amounts of both PFAE and TFAE, two cultivars (i.e. Capsicum annuum var. Forajido and Capsicum chinense var. Habanero) were grown in a greenhouse project. The date of flowering and fruit formation of each blossom was noted and fruits were harvested in four specific periods which corresponded with different stages of ripening, i.e. unripe, semi-ripe, ripe and overripe. Quantification by means of gas chromatography mass spectrometry and creation of development profiles strongly supported the suggestion that PFAE and TFAE were formed as storage molecules during fruit ripening and parallel degradation of chlorophyll. Additionally, compound-specific carbon isotope ratios (?¹³C values (‰)) of originally in PFAE and chlorophyll bound phytol ultimately proved that PFAE, besides tocopherols, serve as sink for the cytotoxic phytol moiety released from chlorophyll degradation during fruit ripening. Furthermore, color measurements were successfully implemented to simplify the usually cumbersome separation of chili fruits into different ripening degrees.

SUBMITTER: Krauß S

PROVIDER: S-EPMC7560742 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Fate of free and bound phytol and tocopherols during fruit ripening of two Capsicum cultivars.

Krauß Stephanie S Hermann-Ene Vanessa V Vetter Walter W

Scientific reports 20201014 1

Phytol and tocopherols and their fatty acid esters (PFAE and TFAE) are isoprenoid lipid components which can be found for instance in vegetables. Their behavior during maturation of fruits and vegetables could reveal valuable information on their biosynthetic formation and biological function. As pods of the genus Capsicum contain considerable amounts of both PFAE and TFAE, two cultivars (i.e. Capsicum annuum var. Forajido and Capsicum chinense var. Habanero) were grown in a greenhouse project. ...[more]

PMID: 33057127

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Project description:Pepper (Capsicum annuum, Solanaceae) fruits are consumed worldwide and are of great economic importance. In most species ripening is characterized by important visual and metabolic changes, the latter including emission of volatile organic compounds associated with respiration, destruction of chlorophylls, synthesis of new pigments (red/yellow carotenoids plus xanthophylls and anthocyanins), formation of pectins and protein synthesis. The involvement of nitric oxide (NO) in fruit ripening has been established, but more work is needed to detail the metabolic networks involving NO and other reactive nitrogen species (RNS) in the process. It has been reported that RNS can mediate post-translational modifications of proteins, which can modulate physiological processes through mechanisms of cellular signalling. This study therefore examined the potential role of NO in nitration of tyrosine during the ripening of California sweet pepper.The NO content of green and red pepper fruit was determined spectrofluorometrically. Fruits at the breaking point between green and red coloration were incubated in the presence of NO for 1?h and then left to ripen for 3?d. Profiles of nitrated proteins were determined using an antibody against nitro-tyrosine (NO2-Tyr), and profiles of nitrosothiols were determined by confocal laser scanning microscopy. Nitrated proteins were identified by 2-D electrophoresis and MALDI-TOF/TOF analysis.Treatment with NO delayed the ripening of fruit. An enhancement of nitrosothiols and nitroproteins was observed in fruit during ripening, and this was reversed by the addition of exogenous NO gas. Six nitrated proteins were identified and were characterized as being involved in redox, protein, carbohydrate and oxidative metabolism, and in glutamate biosynthesis. Catalase was the most abundant nitrated protein found in both green and red fruit.The RNS profile reported here indicates that ripening of pepper fruit is characterized by an enhancement of S-nitrosothiols and protein tyrosine nitration. The nitrated proteins identified have important functions in photosynthesis, generation of NADPH, proteolysis, amino acid biosynthesis and oxidative metabolism. The decrease of catalase in red fruit implies a lower capacity to scavenge H2O2, which would promote lipid peroxidation, as has already been reported in ripe pepper fruit.

Project description:BACKGROUND: To gain insight into the regulation of fruit ascorbic acid (AsA) pool in tomatoes, a combination of metabolite analyses, non-labelled and radiolabelled substrate feeding experiments, enzyme activity measurements and gene expression studies were carried out in fruits of the 'low-' and 'high-AsA' tomato cultivars 'Ailsa Craig' and 'Santorini' respectively. RESULTS: The two cultivars exhibited different profiles of total AsA (totAsA, AsA + dehydroascorbate) and AsA accumulation during ripening, but both displayed a characteristic peak in concentrations at the breaker stage. Substrate feeding experiments demonstrated that the L-galactose pathway is the main AsA biosynthetic route in tomato fruits, but that substrates from alternative pathways can increase the AsA pool at specific developmental stages. In addition, we show that young fruits display a higher AsA biosynthetic capacity than mature ones, but this does not lead to higher AsA concentrations due to either enhanced rates of AsA breakdown ('Ailsa Craig') or decreased rates of AsA recycling ('Santorini'), depending on the cultivar. In the later stages of ripening, differences in fruit totAsA-AsA concentrations of the two cultivars can be explained by differences in the rate of AsA recycling activities. Analysis of the expression of AsA metabolic genes showed that only the expression of one orthologue of GDP-L-galactose phosphorylase (SlGGP1), and of two monodehydroascorbate reductases (SlMDHAR1 and SlMDHAR3) correlated with the changes in fruit totAsA-AsA concentrations during fruit ripening in 'Ailsa Craig', and that only the expression of SlGGP1 was linked to the high AsA concentrations found in red ripe 'Santorini' fruits. CONCLUSIONS: Results indicate that 'Ailsa Craig' and 'Santorini' use complementary mechanisms to maintain the fruit AsA pool. In the low-AsA cultivar ('Ailsa Craig'), alternative routes of AsA biosynthesis may supplement biosynthesis via L-galactose, while in the high-AsA cultivar ('Santorini'), enhanced AsA recycling activities appear to be responsible for AsA accumulation in the later stages of ripening. Gene expression studies indicate that expression of SlGGP1 and two orthologues of SlMDHAR are closely correlated with totAsA-AsA concentrations during ripening and are potentially good candidates for marker development for breeding and selection.

Dataset Information

Fate of free and bound phytol and tocopherols during fruit ripening of two Capsicum cultivars.

Publications

Fate of free and bound phytol and tocopherols during fruit ripening of two Capsicum cultivars.

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