A DNA test for routine prediction in breeding of sweet cherry fruit color,Pav-R<Subscript>f</Subscript>-SSR

Sweet cherry fruit color is a market class-defining trait. The two main market classes in the USA are mahogany, consisting fruit with red skin and flesh, and blush, consisting clear-fleshed fruit with yellow skin and a red overcolor on less than the entire skin surface. Fruit color is a major consideration in sweet cherry breeding as resources and selection thresholds are often differentially applied to each market class. The use of DNA-based information could improve breeding efficiency and accuracy for fruit color, but a predictive DNA test is required. The objective of this study was to develop a reliable, simple DNA test for the prediction of sweet cherry color-based market classes, targeting the major locus, termed here as R _f , associated with fruit color variation. Haplotypes were developed based on 14 SNP markers from the RosBREED cherry 6K SNP array v1 that were associated with the two market classes. To convert the multiple SNP markers to a single, simple PCR-based assay, 11 PCR-based assays targeting microsatellite motifs were designed, using the peach reference genome sequence, and used to screen 20 individuals representing the most common SNP haplotypes. One assay, subsequently named Pav-R_f-SSR, was used to screen 221 phenotyped individuals of the RosBREED sweet cherry reference germplasm set and accurately differentiated individuals with mahogany and blush fruits. Pav-R_f-SSR can be used in DNA-informed breeding schemes to efficiently and accurately predict genetic potential for fruit color and is one of the first DNA tests publicly available for a sweet cherry fruit quality trait.     

Mapping quantitative trait loci associated with blush in peach [Prunus persica (L.) Batsch]

Blush is an important trait for marketing peaches. The red skin pigmentation develops through the flavonoid and anthocyanin pathways, and both genetic and environmental stimuli, and their interaction, control the regulation of these pathways. The molecular basis of blush development in peach is yet to be understood. An F₂ blush population (ZC²) derived from a cross between two peach cultivars with contrasting phenotypes for blush, “Zin Dai” (～30 % red) and “Crimson Lady” (～100 % red), was used for linkage map construction and quantitative trait loci (QTLs) mapping. The segregating population was phenotyped for blush for 4 years using a visual rating scale and quantified using a colorimeter (L*, a*, and b*) 1 year. The ZC² population was genotyped with the IPSC 9 K peach single-nucleotide polymorphism (SNP) array v1, and a high-density ZC² genetic linkage map was constructed. The map covers genetic a distance of ～452.51 cM with an average marker spacing of 2.38 cM/marker. Four QTLs were detected: one major QTL on LG3 (Blush.Pp.ZC-3.1) and three minor QTLs on LG 4 and 7 (Blush.Pp.ZC-4.1; Blush.Pp.ZC-4.2; Blush.Pp.ZC-7.1), indicating the presence of major and minor genes involved in blush development. Candidate genes involved in skin and flesh coloration of peach (PprMYB10), cherry (PavMYB10), and apple (MdMYB1/MdMYBA/MdMYB10) are located within the interval of the major QTL on LG3, suggesting the same genetic control for color development in the Rosaceae family. Marker-assisted selection (MAS) for blush is discussed.     

Comparison of the genetic determinism of two key phenological traits,flowering and maturity dates,in three Prunus species: peach,apricot and sweet cherry

The present study investigates the genetic determinism of flowering and maturity dates, two traits highly affected by global climate change. Flowering and maturity dates were evaluated on five progenies from three Prunus species, peach, apricot and sweet cherry, during 3–8 years. Quantitative trait locus (QTL) detection was performed separately for each year and also by integrating data from all years together. High heritability estimates were obtained for flowering and maturity dates. Several QTLs for flowering and maturity dates were highly stable, detected each year of evaluation, suggesting that they were not affected by climatic variations. For flowering date, major QTLs were detected on linkage groups (LG) 4 for apricot and sweet cherry and on LG6 for peach. QTLs were identified on LG2, LG3, LG4 and LG7 for the three species. For maturity date, a major QTL was detected on LG4 in the three species. Using the peach genome sequence data, candidate genes underlying the major QTLs on LG4 and LG6 were investigated and key genes were identified. Our results provide a basis for the identification of genes involved in flowering and maturity dates that could be used to develop cultivar ideotypes adapted to future climatic conditions.     

Genetic mapping of a major codominant QTL associated with β-carotene accumulation in watermelon

The common flesh color of commercially grown watermelon is red due to the accumulation of lycopene. However, natural variation in carotenoid composition that exists among heirloom and exotic accessions results in a wide spectrum of flesh colors. We previously identified a unique orange flesh watermelon accession (NY0016) that accumulates mainly β-carotene and no lycopene. We hypothesized this unique accession could serve as a viable source for increasing provitamin A content in watermelon. Here we characterize the mode of inheritance and genetic architecture of this trait. Analysis of testcrosses of NY0016 with yellow and red fruited lines indicated a codominant mode of action as F₁ fruits exhibited a combination of carotenoid profiles from both parents. We combined visual color phenotyping with genotyping-by-sequencing of an F_2:3 population from a cross of NY0016 by a yellow fruited line, to map a major locus on chromosome 1, associated with β-carotene accumulation in watermelon fruit. The QTL interval is approximately 20 cM on the genetic map and 2.4 Mb on the watermelon genome. Trait-linked marker was developed and used for validation of the QTL effect in segregating populations across different genetic backgrounds. This study is a step toward identification of a major gene involved in carotenoid biosynthesis and accumulation in watermelon. The codominant inheritance of β-carotene provides opportunities to develop, through marker-assisted breeding, β-carotene-enriched red watermelon hybrids.     

Development of a codominant CAPS marker for allelic selection between canary yellow and red watermelon based on SNP in lycopene β-cyclase (<Emphasis Type="Italic">LCYB</Emphasis>) gene

Flesh color of watermelon is an agronomically important trait that is predominantly determined by a network of the carotenoid biosynthetic pathway, which also contributes to the nutritional value of the fruit through the health-promoting function of carotenoids. We have identified a key gene, lycopene β-cyclase (LCYB) that may determine canary yellow and red flesh color of watermelon and developed a zero-distance molecular marker that identifies a critical single nucleotide polymorphism (SNP) that distinguishes different alleles of the LCYB gene. Analysis of the flesh color inheritance in segregating populations indicated that a single gene determines the color difference between canary yellow and red flesh in watermelon. The sequence comparison of full-length cDNA of LCYB, which was isolated using degenerate PCR and RACE, identified three SNPs in the coding region of LCYB between canary yellow and red. These SNPs showed perfect co-segregation with flesh color phenotypes. One of the SNPs introduces an amino acid replacement of evolutionarily conserved Phe226 to Val, which may impair the catalytic function of LCYB. This SNP was used to develop a cleaved amplified polymorphic sequence (CAPS) marker, which perfectly cosegregated with flesh color phenotype. Our results strongly suggest that LCYB may be the genetic determinant for canary yellow or red flesh color and our CAPS marker will allow breeders to economically distinguish between canary yellow and red watermelon fruit color at the seedling stage.     

A consensus linkage map identifies genomic regions controlling fruit maturity and beta-carotene-associated flesh color in melon (<Emphasis Type="Italic">Cucumis melo</Emphasis> L.)

The nutritional value and yield potential of US Western Shipping melon (USWS; Cucumis melo L.) could be improved through the introgression of genes for early fruit maturity (FM) and the enhancement of the quantity of β-carotene (QβC) in fruit mesocarp (i.e., flesh color). Therefore, a set of 116 F₃ families derived from the monoecious, early FM Chinese line ‘Q 3-2-2’ (no β-carotene, white mesocarp) and the andromonoecious, late FM USWS line ‘Top Mark’ (possessing β-carotene, orange mesocarp) were examined during 2 years in Wisconsin, USA to identify quantitative trait loci (QTL) associated with FM and QβC. A 171-point F_2–3 based map was constructed and used for QTL analysis. Three QTL associated with QβC were detected, which explained a significant portion of the observed phenotypic variation (flesh color; R ² = 4.0–50.0%). The map position of one QTL (β-carM.E.9.1) was uniformly aligned with one carotenoid-related gene (Orange gene), suggesting its likely role in QβC in this melon population and putative relationship with the melon white flesh (wf) gene. Two major (FM.6.1 and FM.11.1; R ² ≥ 20%) and one minor QTL (FM.2.1; R ² = 8%) were found to be associated with FM. This map was then merged with a previous recombinant inbred line (RIL)-based map used to identify seven QTL associated with QβC in melon fruit. This consensus map [300 molecular markers (187 co-dominant melon and 14 interspecific; 10 LG)] provides a framework for the further dissection and cloning of published QTL, which will consequently lead to more effective trait introgression in melon.     

Facial Skin Coloration Affects Perceived Health of Human Faces

Numerous researchers have examined the effects of skin condition, including texture and color, on the perception of health, age, and attractiveness in human faces. They have focused on facial color distribution, homogeneity of pigmentation, or skin quality. We here investigate the role of overall skin color in determining perceptions of health from faces by allowing participants to manipulate the skin portions of color-calibrated Caucasian face photographs along CIELab color axes. To enhance healthy appearance, participants increased skin redness (a*), providing additional support for previous findings that skin blood color enhances the healthy appearance of faces. Participants also increased skin yellowness (b*) and lightness (L*), suggesting a role for high carotenoid and low melanin coloration in the healthy appearance of faces. The color preferences described here resemble the red and yellow color cues to health displayed by many species of nonhuman animals.     

Identification of two major QTL for yellow seed color in two crosses of resynthesized <Emphasis Type="Italic">Brassica napus</Emphasis> line No. 2127-17

Yellow seed color, which results from a thinner seed coat, is associated with improved feed quality of rapeseed (Brassica napus L.) meal and increased oil and protein content. As this trait follows various genetic models under different genetic backgrounds, a study was performed in two genetic backgrounds to gain a better understanding of the genetic mechanisms underlying yellow seed color. The quantitative trait locus (QTL) analysis was undertaken using two crosses, Quantum ×No. 2127-17 (HZ-1) and No. 2127-17 × 94,570 (HZ-2). In the HZ-1 population, three putative QTL were detected in linkage groups N18, N5, and N3, respectively. For all of them, yellow seed color arose from the No. 2127-17 alleles. Of these QTL, the one in linkage group N18 (Bnsc-18a) explained more than half of the phenotypic variation. In the HZ-2 population, three QTL were found in linkage groups N9, N18, and N8, respectively. Of these QTL, that in linkage group N9 (Bnsc-9a) explained more than half of the phenotypic variation, whereas the QTL Bnsc-18a had a low seed color value and explained only 9.03–11.72% of the phenotypic variation. Bulked segregant analysis (BSA) of the extremes of a BC1 population derived from the cross of No. 2127-17 × 94,570 (HZ-3) identified one major gene that was identical with the QTL Bnsc-9a detected in the HZ-2 population. The QTL Bnsc-18a was common in the HZ-1 and HZ-2 populations, and the others were population-specific. These results suggested that different black-seeded forms had different seed color genes.     

Qualitative inheritance of rind pattern and flesh color in watermelon

Watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai var. lanatus] is a diverse species, with fruits of different sizes, shapes, rind patterns, and flesh colors. This study measured the inheritance of novel rind phenotypes and verified the genetics of white, red, salmon yellow, and canary yellow flesh colors. For each of the 11 crosses, six generations (P(a)S1, P(b)S1, F1, F2, BC1P(a), and BC1P(b)) were produced to form 11 families. Three new genes were identified and designated as follows: Scr for the scarlet red flesh color of Dixielee and Red-N-Sweet, Yb for the yellow belly (ground spot) of Black Diamond Yellow Belly, and ins for the intermittent stripes of Navajo Sweet. The inheritance of the C gene for the canary yellow flesh color was verified as single dominant, and a new inbred type line was developed possessing that gene. Aberrations in the segregation of red, white, and salmon yellow flesh colors were recorded, raising questions on the inheritance of these traits. Finally, the spotted phenotype from Moon and Stars was combined with light green and gray rind patterns for the development of novel cultivars with distinctive rind patterns.     

Loss of autumn colors under domestication: A byproduct of selection for fruit flavor?

According to the coevolution hypothesis the red autumn leaves of certain tree species are a warning signal towards insects that lay their eggs on the trees. A recent study has shown that red leaves are common in wild varieties of apple (Malus pumila) but not in cultivated varieties. This suggests that autumn colors have been lost during domestication due to relaxed selection against insects. The few varieties with red leaves have small fruits, similar to their wild ancestors, which shows that they have been under less effective artificial selection. As expected by the coevolution hypothesis these red varieties are very susceptible to an insect-borne disease, fire blight. Here I report further data on the loss of autumn colors under domestication. Since red leaf color is correlated with red fruit flesh color, if red fruit flesh has more astringent taste it is possible that loss of autumn colors is not only due to relaxed selection against insect, but also to direct artificial selection against astringent taste. However even varieties with yellow flesh turn out to have astringent taste. Moreover, while red fruit flesh is common in cultivated varieties with red leaves, it is very rare in wild varieties. It is unclear, therefore, whether loss of autumn color under domestication was a byproduct of artificial selection against red fruit flesh.Key words: coevolution, autumn colors, signaling, apple, Malus pumila, domestication, artificial selection, germplasm     

High-density linkage maps constructed in sweet cherry (<Emphasis Type="Italic">Prunus avium</Emphasis> L.) using cross- and self-pollination populations reveal chromosomal homozygosity in inbred families and non-syntenic regions with the peach genome

The landrace sweet cherry (Prunus avium L.) cultivar ‘Cristobalina’ is a useful resource for sweet cherry breeding due to several important traits, including low chilling requirement, early maturity date, and self-compatibility. In this work, three families (N?=?325), derived from ‘Cristobalina’, were used to develop high-density genetic maps using the RosBREED 6K Illumina Infinium® cherry SNP array. Two of the families were derived from self-pollination, which allowed construction of the first F₂ genetic maps in the species. The other map developed was from an interspecific cross of cultivars ‘Vic’?×?‘Cristobalina’. The maps developed include 511 to 816 mapped SNPs covering 622.4 to 726.0 cM. Mapped SNP marker order and position were compared to the sweet cherry and peach genome sequences, and a high degree of synteny was observed. However, inverted and small translocated regions between peach and sweet cherry genomes were observed with the most noticeable inversion at the top of LG5. The progeny resulting from self-pollination also revealed a high level of homozygosity, as large presumably homozygous regions as well as entire homozygous LGs were observed. These maps will be used for genetic analysis of relevant traits in sweet cherry breeding by QTL analysis, and self-pollination populations will be useful for investigating inbreeding depression in a naturally outbreeding species.     

Fruit size QTL analysis of an F<Subscript>1</Subscript> population derived from a cross between a domesticated sweet cherry cultivar and a wild forest sweet cherry

Maximizing fruit size is critical for profitable sweet cherry (Prunus avium L.) production. Yet, despite its importance, little is known about the genetic control of fruit size. The objective of this study was to identify quantitative trait loci (QTLs) for fruit size and two essential components of fruit size, mesocarp cell number and size. This study utilized a double pseudo-testcross population derived from reciprocal crosses between a sweet cherry cultivar with ~8 g fruit, “Emperor Francis” (EF), and a wild forest sweet cherry selection with ~2 g fruit, “New York 54” (NY). A total of 190 F₁ progeny previously utilized for the construction of the linkage maps were evaluated in 2006 and 2007 for fruit weight, length, and diameter; mesocarp cell number and length; and pit length and diameter. In 2008, a subset of this population was again evaluated for fruit weight. Correlation analysis revealed that the three fruit size traits were highly correlated with each other, and mesocarp cell number, not cell length, was correlated with fruit size. Three QTLs were identified for each fruit size trait, and one QTL was identified for mesocarp cell number. Fruit size QTLs were found on linkage group 2 on the EF map (EF 2) and linkage groups 2 and 6 on the NY map (NY 2 and NY 6). On EF 2, the cell number QTL clustered with the fruit size QTL, suggesting that the underlying basis of the fruit size increase associated with this QTL was an increase in mesocarp cell number. On NY 6, pit length and diameter QTLs clustered with those for fruit size, suggesting that the underlying morphological basis of this fruit size QTL is the difference in pit size.     

Genetic diversity of some Iranian sweet cherry (<Emphasis Type="Italic">Prunus avium</Emphasis>) cultivars using microsatellite markers and morphological traits

The aim of this study was to characterize 23 important Iranian sweet cherry (Prunus avium) cultivars collected from different provinces of Iran and 1 foreign cultivar, which was used as control, considered for breeding programs by using 21 microsatellite markers and 27 morphological traits. In sweet cherry (Prunus avium) accessions, leaf, fruit, and stone morphological characters were evaluated during two consecutive years. The study revealed a high variability in the set of evaluated sweet cherry accessions. The majority of important correlations were determined among variables representing fruit and leaf size and variables related to color. Cluster analysis distinguished sweet cherry accessions into two distinct groups. Principal component analysis (PCA) of qualitative and quantitative morphological parameters explained over 86.59% of total variability in the first seven axes. In PCA, leaf traits such as leaf length and width, and fruit traits such as length, width, and weight, and fruit flesh and juice color were predominant in the first two components, indicating that they were useful for the assessment of sweet cherry germplasm characterization. Out of 21 SSR markers, 16 were polymorphic, producing 177 alleles that varied from 4 to 16 alleles (9.35 on average) with a mean heterozygosity value of 0.82 that produced successful amplifications and revealed DNA polymorphisms. Allele size varied from 95 to 290 bp. Cluster analyses showed that the studied sweet cherry genotypes were classified into five main groups based mainly on their species characteristics and SSR data. In general, our results did not show a clear structuring of genetic variability within the Iranian diffusion area of sweet cherry, so it was not possible to draw any indications on regions of provenance delimitation. The results of this study contribute to a better understanding of sweet cherry genetic variations in Iran, thus making for more efficient programs aimed at preserving biodiversity and more rational planning of the management of reproductive material.     

SSR-based linkage map of flax (<Emphasis Type="Italic">Linum usitatissimum</Emphasis> L.) and mapping of QTLs underlying fatty acid composition traits

Flax (Linum usitatissimum L.) seeds contain nearly 50% oil which is high in linolenic acid (an omega-3 fatty acid). In this study, a genetic linkage map was constructed based on 114 expressed sequence tag-derived simple sequence repeat (SSR) markers in addition to five single nucleotide polymorphism markers, five genes (fad2A, fad2B, fad3A, fad3B and dgat1) and one phenotypic trait (seed coat color), using a doubled haploid (DH) population of 78 individuals generated from a cross between SP2047 (a yellow-seeded Solin™ line with 2–4% linolenic acid) and UGG5-5 (a brown-seeded flax line with 63–66% linolenic acid). This map consists of 24 linkage groups with 113 markers spanning ~833.8 cM. Quantitative trait locus (QTL) analysis detected two major QTLs each for linoleic acid (LIO, QLio.crc-LG7, QLio.crc-LG16), linolenic acid (LIN, QLin.crc-LG7, QLin.crc-LG16) and iodine value (IOD, QIod.crc-LG7, QIod.crc-LG16), and one major QTL for palmitic acid (PAL, QPal.crc-LG9). The mutant allele of fad3A, mapped to the chromosomal segment inherited from the parent SP2047, underlies the QTL on linkage group 7 and was positively associated with high LIO content but negatively associated with LIN and IOD. This fad3A locus accounted for approximately 34, 25 and 29% of the phenotypic variation observed in this DH population for these three traits, respectively. The QTL localized on linkage group 16 explained approximately 20, 25 and 13% of the phenotypic variation for these same traits, respectively. For palmitic acid, QPal.crc-LG9 accounted for ~42% of the phenotypic variation. This first SSR-based linkage map in flax will serve as a resource for mapping additional markers, genes and traits, in map-based cloning and in marker-assisted selection.     

Construction of a linkage map and QTL analysis of horticultural traits for watermelon [<Emphasis Type="Italic">Citrullus lanatus</Emphasis> (THUNB.) MATSUM &; NAKAI] using RAPD,RFLP and ISSR markers

We have been constructing linkage maps for watermelon ( Citrullus lanatus) on the basis of random amplified polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP), inter-simple sequence repeats (ISSRs) and isozymes using an F(2) population derived from a crossing between a cultivated inbred line (H-7; C. lanatus) and an African wild form (SA-1; C. lanatus). A total of 120 F(2) plants was used for construction of a linkage map using 477 RAPDs, 53 RFLPs, 23 ISSRs and one isozyme markers. Linkage analysis revealed that 554 loci could be mapped to 11 linkage groups that extended for 2,384 centimorgans (cM). While a BC(1) population [(H-7 x SA-1) x H-7] consisting of 60 individuals was grown and scored for quantitative traits. Another linkage map with a total length of 1,729 cM was constructed in the BC(1) using genetic markers found to segregate in the F(2) population. A QTL analysis was applied by means of interval mapping for locating such agronomic traits as hardness of rind, Brix of flesh juice, flesh color (red and yellow) and rind color. The relative order of markers in the BC(1) map was essentially the same as that on the linkage map in the F(2). A total of five QTLs for four agronomic traits was detected. The QTL for hardness of rind was mapped on group 4. The linkage group 8 contained the QTL for sugar content of the flesh as expressed in Brix of the juice. The QTL for red flesh color was detected on groups 2 and 8. The QTL for rind color mapped on the group 3. The present map and QTL analysis may provide a useful tool for breeders by introducing valuable wild watermelon genes to cultivars.     

Cherry leaf spot resistance in cherry (Prunus) is associated with a quantitative trait locus on linkage group 4 inherited from P. canescens

Cherry leaf spot (CLS), caused by the fungal pathogen Blumeriella jaapii (Rehm) Arx (telomorph Phloeosporella padi [Lib.] Arx), is a major disease in all humid cherry-growing regions worldwide causing leaf yellowing and defoliation. The diploid Prunus species, P. canescens, had previously been identified as a source of CLS resistance. Therefore, the objective of this study was to identify quantitative trait loci (QTL) for CLS resistance derived from P. canescens in both diploid sweet cherry (P. avium) and tetraploid sour cherry (P. cerasus). Because of the simpler genetics of diploid cherry, the initial investigation was done with P. canescens-derived materials from crosses with sweet cherry, followed by validation using P. canescens-derived plant materials from sour cherry. A major QTL controlling P. canescens-derived CLS resistance, named CLSR_G4, was identified on linkage group 4 in sweet cherry and validated in sour cherry. All CLS-resistant individuals had one P. canescens-derived allele for CLSR_G4. A second QTL may be necessary for CLS resistance as one-fifth–one-third of the progeny individuals with the P. canescens-derived allele for CLSR_G4 were susceptible.     

Identification of a major quantitative trait locus (QTL) for yellow spot (<Emphasis Type="Italic">Mycovellosiella koepkei</Emphasis>) disease resistance in sugarcane

In this study we used amplified fragment length polymorphism (AFLP) and microsatellite (short sequence repeat or SSR) markers to identify a major quantitative trail locus (QTL) for yellow spot (Mycovellosiella koepkei) disease resistance in sugarcane. A bi-parental cross between a resistant variety, M 134/75, and a susceptible parent, R 570, generated a segregating population of 227 individuals. These clones were evaluated for yellow spot infection in replicated field trials in two locations across two consecutive years. A χ²-test (χ² at 98% confidence level) of the observed segregation pattern for yellow spot infection indicated a putative monogenic dominant inheritance for the trait with a 3 (resistant):1(susceptible) ratio. The AFLP and SSR markers identified 666 polymorphisms as being present in the resistant parent and absent in the susceptible one. A genetic map of M 134/75 was constructed using 557 single-dose polymorphisms, resulting in 95 linkage groups containing at least two markers based on linkages in coupling. QTL analysis using QTLCartographer v1.17d and MAPMAKER/QTL v1.1 identified a single major QTL located on LG87, flanked by an AFLP marker, actctc10, and an SSR marker, CIR12284. This major QTL, which was found to be linked at 14 cM to an AFLP marker, was detected with LOD 8.7, had an additive effect of −10.05% and explained 23.8% of the phenotypic variation of yellow spot resistance.