Establishing marker-locus-trait associations to enable marker-assisted breeding depends on having an extensive, reliable database for phenotypic traits of interest in relevant germplasm. A reference germplasm set of 467 apple (Malus 3domestica Borkh.) cultivars, selections, and seedlings (referred to as individuals) was identified as part of the USDA-Specialty Crop Research Initiative (SCRI) project, RosBREED. The germplasm set provides efficient allelic representation of current parents in RosBREED demonstration apple breeding programs at Cornell University, Washington State University, and the University of Minnesota. Phenotyping at the three locations was conducted according to standardized protocols, focusing on fruit traits evaluated at harvest and after 10 and 20 weeks of refrigerated storage. Phenotypic data were collected for the sensory texture traits of firmness, crispness, and juiciness as well as for instrumental texture measures. In 2010 and 2011, fruit from 216 and 330 individuals, respectively, were harvested and a total of 369 individuals were evaluated over the two years. Correlations between sensory and instrumental texture measures were high in some instances. Moderate year-to-year repeatability of trait values was observed. Because each location had a largely unique set of individuals, as well as differing environmental conditions, means, ranges, and phenotypic variances differed greatly among locations for some traits. Loss of firmness and crispness during storage was more readily detected instrumentally than by the sensory evaluation. Fruit texture traits, significant to apple breeder decision-making yet unobservable until tree maturity, are ideal candidates for marker-assisted breeding (MAB) and markerassisted selection (MAS). Marker-locus-trait associations, validated in germplasm relevant to a particular breeding program, facilitate MAB (Bliss, 2010). MAB is used to select parents with favorable alleles and MAS is imposed on seedling populations to eliminate those with unfavorable allele combinations. Both MAB and MAS can reduce time and expense for new cultivar development in a tree fruit breeding program. Fruit texture is a focus of breeders because of its role in shaping consumer acceptance of new apple cultivars. Harker et al. (2003) reviewed studies that investigated consumer preferences for apple and factors influencing willingness to buy. They reported that although subsets of consumers vary in fruit quality expectations, most adults respond to texture and acidity as determinants of fruit quality. In a study of New Zealand consumers, adults preferred harder and crisper apples. Although the authors reported that consumers remember differences in apple texture for days, Harker et al. (2003) predicted that fruit quality standards will evolve as consumers’ expectations change. Speeding the breeding process through the use of molecular markers will aid apple breeders in developing higher quality fruit. A study using ‘Red Delicious’, ‘Gala’, and ‘Braeburn’ showed that in certain cultivars, firmness is of high importance to consumers, especially in combination with other fruit quality factors: firm apples, above a 53Newton threshold, can be improved on by changes in titratable acidity (TA) and soluble solids content (SSC), but soft apple acceptance cannot be improved on with changes in TA or SSC (Harker et al., 2008). These findings highlighted the use of genetic markers to select for fruit texture traits. Studies of apple texture have used both sensory panels and instrumental measures (e.g., Evans et al., 2010; Ioannides et al., 2007; McKay et al., 2011; Zdunek et al., 2011). Differences in terms used to describe texture as well as their definitions make comparing sensory panel results difficult. For instance, the meaning of the term ‘‘crispness’’ differs across studies. Fillion and Kilcast (2002), using a trained sensory panel and a consumer panel, defined the term ‘‘crunchy’’ as describing lower-pitched sounds that continue throughout chewing, whereas ‘‘crisp’’ described a higher-pitched sound resulting from the clean split of the first bite. Both crisp and crunchy designations, when applied to food, express that the material breaks in the mouth rather than buckling or deforming. By studying sounds during biting dry and wet crisp foods, Vickers and Bourne (1976) defined the crispness sensation as a characteristic sound of a range of frequencies emitted during biting. For a thorough discussion of the crispness sensation, refer to Roudaut et al. (2002). In our study, described by Evans et al. (2012), ‘‘crispness’’ refers to the intensity of the cracking noise of the first bite. ‘‘Firmness’’ is equivalent to ‘‘hardness’’ and determined while chewing. ‘‘Juiciness’’ is expressed juice on chewing. A trained sensory panel, as small as three experienced individuals, has been shown to be reliable in a postharvest study of fruit texture (Brookfield et al., 2011). That panel was able to discern greater separation among cultivars than was achieved with instrumental measures. Although sensory panels more closely mimic consumer perception of fruit texture, they can be time-consuming and difficult to standardize. Puncture tests, performed with various mechanized penetrometers, are typically used to determine firmness and juiciness (e.g., Harker et al., 2006). Harker et al. (2002) found puncture tests superior to chewing sounds and tensile measurements in forecasting sensory panelists’ perception of texture traits. The Mohr Digi-Test (Mohr and Associates, Richland, WA) computerized penetrometer captures data that correlate well with sensory firmness and sensory crispness by collecting constant velocity measurements (Evans et al., 2010). This is especially useful, because crispness has proven difficult to measure instrumentally with other devices. Received for publication 19 Dec. 2012. Accepted for publication 22 Jan. 2013. Funded by the Specialty Crop Research Initiative Competitive Grant 2009-51181-05808 of the USDA’s National Institute of Food and Agriculture. To whom reprint requests should be addressed; e-mail lubyx001@umn.edu. 296 HORTSCIENCE VOL. 48(3) MARCH 2013 Establishing marker-locus-trait associations for texture traits depends on having an extensive, reliable phenotype database for traits of interest in breeding germplasm. Without high-quality phenotypic data, association statistics that link genomic sequences to traits cannot realize full potential (Bassil and Volk, 2010). Moreover, when standardized phenotyping protocols are used across several breeding programs, the resulting large data sets give more power to studies that detect and characterize quantitative trait loci (QTL) than would be had if each program conducted a smaller, isolated study. A reference germplasm set of 467 individual genotypes including cultivars, selections, and seedlings was identified as part of the USDA-SCRI RosBREED project. The germplasm set provides efficient allelic representation of current parents in the large, publicly funded U.S. apple breeding programs of Cornell University (CU), Washington State University (WSU), and the University of Minnesota (UMN). Extensive phenotypic data, including instrumental and sensory measures of fruit texture, were collected on these individuals at each location in the years 2010 and 2011 under three regimes: at harvest, after 10 weeks of cold storage and 1 week at room temperature, and after 20 weeks of cold storage and 1 week at room temperature. Phenotypic data were collected adhering to a standardized protocol (Evans et al., 2012). The objective in this article is to elaborate on methods used to obtain data on sensory and instrumental measures of fruit texture traits in the RosBREED apple Crop Reference Set (CRS) and describe variation and repeatability observed for these traits. We also report correlations between sensory and instrumental measures used in this study. Materials and Methods Plant material. The RosBREED apple CRS and supplementing individuals included 154 cultivars and parental selections as well as 313 seedlings of families chosen to provide efficient allelic representation of important breeding parents for a total of 467 related individuals. Subsets of the RosBREED CRS were grown at the UMN Horticultural Research Center near Chaska, MN, at the WSU Tree Fruit Research & Extension Center in Wenatchee, WA, and at the CU New York State Agricultural Experiment Station in Geneva, NY. Evaluation of a reference Fig. 1. Apple equatorial slice demonstrating MDT-1 fruit texture measures described by Evans et al. (2010) and Mohr and Mohr (2000). R1 is the outer area of the apple directly below the skin, R2 is the main edible portion of the fruit, and R3 contains the core. Red lines indicate regions in which traits are determined. Fig. 2. Proportions of variance attributable to year, individual, sampling and year 3 individual for fruit texture measures at three locations at harvest. Analysis of variance was used. Data from University of Minnesota (UMN), Washington State University (WSU), and Cornell University (CU) are shown in shades of blue, purple, and green, respectively. Abbreviations are as follows: A1, A2 = average pressure regions 1 and 2, respectively (N); C0 = creep at boundary between regions 1 and 2 (cm); Cn = crispness measurement (derived value); E2 = pressure at core boundary (N); M1, M2 = maximum pressure regions 1 and 2, respectively (N); OAH = overall average hardness (N); OMH = overall maximum hardness (N); QF = quality factor (derived value). The sensory measures of crispness, firmness, and juiciness were assessed on a 5-point scale. HORTSCIENCE VOL. 48(3) MARCH 2013 297 | BREEDING, CULTIVARS, ROOTSTOCKS, AND GERMPLASM RESOURCES