Abstract
Bitter pit is a physiological disorder of apples that develops in the latter stages of fruit development and during storage. It is characterized by localized necrotic cells that collapse and form pits in the epidermis and outer cortex of fruit. The disorder has been associated with low calcium concentrations, and poor calcium distribution within the fruit. To date, the mechanism that leads to individual cell necrosis, while surrounding cells remain healthy, is not fully understood. In order to ascertain the underlying process of bitter pit incidence in apple fruit, a large mapping population of “Braeburn” (susceptible to bitter pit) × “Cameo” (resistant to bitter pit) was used to map the trait over two growing seasons. A subset of 94 genotypes from the mapping population representing the full range of phenotypes in the same ratio as the full population were selected for genotyping and functional characterization. RNA-Seq analysis on fruit samples of three resistant and three susceptible lines at seven developmental stages (21, 42, 63, 84, 105, 126, and 147 days post fertilization) identified a number of candidate genes displaying differential gene expression. A subset of candidate genes selected based on their position within the identified QTL interval on linkage group 16 were validated by RT-qPCR, and two candidate genes displaying differential gene expression were highlighted as strong candidates for the control of bitter pit symptom expression at the Bp-2 locus.
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The work was funded by a grant from the Autonomous Province of Trento to the Dipartimento di Genomica e Biologia delle Piante da Frutto of the Fondazione Edmund Mach, Trentino, Italy.
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ESM 1
– Apple fruits at full maturity displaying different levels of bitter pit symptoms. Symptoms were scored on a five-point scale: no symptoms (0), low symptoms (1), medium symptoms (2), high symptoms (3) and very high symptoms (4). Pictures of fruits (whole and horizontal section) belonging to each scale point were reported. (PNG 1821 kb)
ESM 2
– Phenotypic distribution of bitter pit symptoms severity in the ‘B × C mapping population over the course of two growing seasons (2012 and 2014). Fruiting trees were classified on a 0–4 scale depending on the severity of the symptoms observed. The number of plants grouped into each of the five classes is shown for each year of evaluation (PNG 678 kb)
ESM 3
– Genotyping results on a subset (n = 94) of the B × C mapping population. The position of each marker across respective linkage group was given, along with its segregation type and phase. (XLSX 48 kb)
ESM 4
– Initial linkage map and QTL analysis using interval mapping for a subset of 94 progenies of B × C mapping population. Marker positions are given in centi-Morgans (cM) and accompanying graphs give LOD scores associated with bitter pit symptom expression for 2012 and 2014. (PNG 588 kb)
ESM 5
– Results of DE analysis between resistant and susceptible samples in the first and seventh developmental stages. Only results for differentially expressed genes were reported. (XLSX 1992 kb)
ESM 6
– Protein comparisons of 13 candidate genes identified at the bitter pit susceptibility locus Bp-2 in ‘Braeburn’ and ‘Cameo’, the parental genotypes of the B × C segregating population and in the ‘Golden Delicious’ v1.0 reference sequence. (DOCX 21 kb)
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Buti, M., Sargent, D.J., Bianco, L. et al. A study of gene expression changes at the Bp-2 locus associated with bitter pit symptom expression in apple (Malus pumila). Mol Breeding 38, 85 (2018). https://doi.org/10.1007/s11032-018-0840-z
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DOI: https://doi.org/10.1007/s11032-018-0840-z