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Background Many fruits, including watermelon, are proficient in carotenoid accumulation during ripening. While most genes encoding steps in the carotenoid biosynthetic pathway have been cloned, few transcriptional regulators of these genes have been defined to date.

Here we describe the identification of a set of putative carotenoid-related transcription factors resulting from fresh watermelon carotenoid and transcriptome analysis during fruit development and ripening. Our goal is to both clarify the expression profiles of carotenoid pathway genes and to identify candidate regulators and molecular targets for crop improvement. Results Total carotenoids progressively increased during fruit ripening up to ~55 μg g -1 fw in red-ripe fruits. Trans-lycopene was the carotenoid that contributed most to this increase. Many of the genes related to carotenoid metabolism displayed changing expression levels during fruit ripening generating a metabolic flux toward carotenoid synthesis.

Constitutive low expression of lycopene cyclase genes resulted in lycopene accumulation. RNA-seq expression profiling of watermelon fruit development yielded a set of transcription factors whose expression was correlated with ripening and carotenoid accumulation. Nineteen putative transcription factor genes from watermelon and homologous to tomato carotenoid-associated genes were identified. Among these, six were differentially expressed in the flesh of both species during fruit development and ripening. The isoprenoid (also known as terpenoid) pathway is one of the most important and well-studied biosynthetic pathways in plants. It involves cross-talk between the cytosolic mevalonate (MVA) and plastidial 2- C-methyl- d-erythritol 4-phosphate (MEP) pathways, to give rise to isopentenyl-diphosphate (IPP), the C5 building block required for the synthesis of a diverse group of natural products that perform numerous biochemical functions in plants. The main branch of the isoprenoid pathway leads to the synthesis and accumulation of carotenoids, C40 terpenoid compounds formed by the condensation of eight isoprene units, within plastids [ ].

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Carotenoids are important natural pigments found in all plants and algae, in many bacteria and fungi, as well as in some animals. In photosynthetic organisms, carotenoids are always present within chloroplasts associated with the light-harvesting complexes of photosystems, where they gather light energy and transfer it to the chlorophylls, in addition to contributing to protection of the photosynthetic apparatus from photooxidation [ ]. Chromoplast synthesized carotenoids accumulate during fruit ripening resulting in dramatic changes in tissue pigmentation. Carotenoids also play an essential role in attracting insects and herbivores that act as floral pollinators and seed dispersion vehicles, including the consumption of plant organs such as ripe fruits as food by humans.

When carotenoid-rich foods are ingested, carotenoids are assimilated, metabolized, transported in the plasma by lipoproteins, and stored in various tissues [ ] where they display biological activities including acting as antioxidants and free-radical scavengers, reducing the risk of certain types of cancers, and promoting immune responses. In addition, some carotenoids (e.g. Β-carotene, β-cryptoxanthin) are precursors of vitamin A, an essential nutrient for humans [ – ]. Carotenoids can undergo multiple structural modifications, namely, cyclization, hydroxylation, and epoxidation, yielding the great variety of carotenoids found in nature comprising more than 600 compounds [ ]. Increasing interest is devoted to carotenoid content and composition of food crops because of their important roles in human health [ ]. Cultivated watermelon [ Citrullus lanatus (Thunb.) Matsum. Lanatus] is important in the diets of people throughout the world.

More than 6% of the world's total area devoted to vegetable production is used for watermelon production [ ]. Throughout the Mediterranean basin, watermelon is considered an extremely important agricultural crop, with over 10 6 tons produced in 2010 at a production value of approximately $3 billion US [ ]. The ripening of watermelon fruit is a highly regulated process because color, flavor, aroma, and texture change in a coordinated manner. One of the most noticeable characteristics of watermelon ripening is the dramatic increase in carotenoids. The change in pigmentation is caused by synthesis and accumulation of lycopene within the chromoplasts in watermelon fruit resulting in their characteristic red coloration.

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