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Flavor traits in citrus are the result of a blend of low molecular weight metabolites including sugars, acids, flavonoids and limonoids, these latter being mainly responsible for the characteristic bitter flavor in citrus. In this work, the genotype- and developmental stage-dependent accumulation of flavonoids and limonoids is addressed. To fulfill this goal, three models for citrus bitterness: bitter Duncan grapefruit, bittersweet Thomson orange and sweet Wase mandarin were selected from a total of eight different varieties. Compounds were annotated from LC/ESI-QqTOF-MS non-targeted metabolite profiles from albedo and pulp tissues. Results indicated that the specific blend of compounds providing the characteristic flavor trait is genotype-specific and hence under genetic control, but it is also regulated at the developmental level. Metabolite profiles in albedo mirrored those found in pulp, the edible part of the fruit, despite differences in the concentration and accumulation/depletion rates being found. This is particularly relevant for polymethoxylated flavones and glycosylated limonoids that showed a clear partitioning towards albedo and pulp tissues, respectively. Fruit ripening was characterized by a reduction in flavonoids and the accumulation of limonoid glycosides. However, bitter grapefruit showed higher levels of limonin A-ring lactone and naringin in contrast to sweeter orange and mandarin. Data indicated that the accumulation profile was compound class-specific and conserved among the studied varieties despite differing in the respective accumulation and/or depletion rate, leading to different specialized metabolite concentration at the full ripe stage, consistent with the flavor trait output.

This study reports the isolation and characterization of seven polymorphic microsatellite loci in Citrus. The loci were isolated from two libraries constructed from genomic DNA nonenriched for TC and AC repeats and enriched for AC repeats. These markers yielded four to nine alleles per locus (mean 6.14) in a survey of 32 Citrus cultivars. Average observed heterozygosity ranged from 0.43 to 0.72. The levels of polymorphism found in this study suggest that these microsatellite loci can become an important tool for genetic studies in Citrus.

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