Waurich, V. | bioRxiv Science

bioRxiv2024

Differential expression ofCCD4(4B) drives natural variation in fruit carotenoidcontent in strawberry ( Fragaria spp.)

Pigments, mainly anthocyanins and carotenoids, are important contributors to fruits visual appearance and nutritional properties. In strawberry (Fragaria spp.), the genetic and molecular mechanisms regulating fruit carotenoid biosynthesis and its natural variation remain largely unexplored. In this study, we sought to identify genomic loci contributing to variation in yellow flesh pigmentation. A major QTL, qYellow Flesh-4B, accounting for 82% of the total phenotypic variation was identified on F. x ananassa chromosome 4B. Following a candidate gene approach, we determined that CCD4(4B), a carotenoid cleavage dioxygenase (CCD), was the underlying gene. Specific polymorphisms on CCD4(4B) promoter region were associated with the yellow flesh phenotype and with differential regulation of CCD4(4B) expression during fruit ripening. Furthermore, CCD4(4B) expression levels were negatively correlated with violaxanthin, lutein, zeaxanthin, {beta}-carotene and total carotenoid content. The role of CCD4(4B) in carotenoid turnover was confirmed by transient overexpression in F. x ananassa fruits, which led to a decrease in carotenoid accumulation. Notably, a -35 C>T SNP identified in CCD4(4B) promoter was found to be predictive for CCD4(4B) expression, and carotenoid content in fruits of a diverse germplasm collection, which included different octoploid Fragaria species. Taken together, these results provide important genetic insights into the natural variation of carotenoid content in strawberry. The High-Resolution Melting (HRM) DNA test here developed offers a fast and reliable method to predict high fruit carotenoid content, representing a useful tool for breeding projects aiming to enhance the nutritional value of this crop.

plant biology↗

bioRxiv2020

Allelic Variation of MYB10 is the Major Force Controlling Natural Variation of Skin and Flesh Color in Strawberry (Fragaria spp.) fruit

Anthocyanins are the principal color-producing compounds synthesized in developing fruits of strawberry (Fragaria spp.). Substantial natural variation in color have been observed in fruits of diploid and octoploid accessions, resulting from distinct accumulation and distribution of anthocyanins in fruits. Anthocyanin biosynthesis is controlled by a clade of R2R3 MYB transcription factors, among which MYB10 has been shown as the main activator in strawberry fruit. Here, we show that MYB10 mutations cause most of the anthocyanin variation observed in diploid woodland strawberry (F. vesca) and octoploid cultivated strawberry (F. xananassa). Using a mapping-by-sequencing approach, we identified a gypsy-transposon insertion in MYB10 that truncates the protein and knocks out anthocyanin biosynthesis in a white-fruited F. vesca ecotype. Two additional loss-of-function MYB10 mutations were identified among geographically diverse white-fruited F. vesca ecotypes. Genetic and transcriptomic analyses in octoploid Fragaria spp. revealed that FaMYB10-2, one of three MYB10 homoeologs identified, residing in the F. iinumae-derived subgenome, regulates the biosynthesis of anthocyanins in developing fruit. Furthermore, independent mutations in MYB10-2 are the underlying cause of natural variation in fruit skin and flesh color in octoploid strawberry. We identified a CACTA-like transposon (FaEnSpm-2) insertion in the MYB10-2 promoter of red-fleshed accessions that was associated with enhanced expression and anthocyanin accumulation. Our findings suggest that putative cis regulatory elements provided by FaEnSpm-2 are required for high and ectopic MYB10-2 expression and induction of anthocyanin biosynthesis in fruit flesh. We developed MYB10-2 (sub-genome) specific DNA markers for marker-assisted selection that accurately predicted anthocyanin phenotypes in octoploid segregating populations.

plant biology↗