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Many species in genebank collections are maintained vegetatively either as in vitro cultures or as plants in the field or greenhouse. In these cases, individual genotypes (cultivars) are the focus of conservation. In crop wild relatives, where the value of an accession is in the heterogeneity of segregating genotypes, seeds may be the most representative form of conservation. We used two sets of field-planted Malus sieversii apple trees that originated from seeds collected from two wild populations in Kazakhstan as a model system to determine if seeds from hand-pollinated crosses could be used to represent the allelic diversity of the parent trees in a genebank orchard. A crossing design was developed that used each of the selected trees from the two Kazakhstan collection sites as mothers and also as fathers in the form of bulked pollen pools. The sets of offspring from these crosses were genotyped using microsatellite markers. Paternity analyses revealed the percentage of offspring that was derived from each of the parent trees. We also determined the contribution of each parent to the fertilization events from the pollen pools. Although there were some unintended pollination events (fertilization by trees in unexpected pollen pools) and unequal contribution of pollen parents within pollen pools, we demonstrated that the seedlings effectively captured the allelic frequency and diversity of the parents. A seed collection strategy is the most efficient way to conserve the diversity of crop wild relatives even in collections typically conserved as individual clones.

Malus sieversii is a progenitor species of domestic apple M.×domestica. Using population "GMAL 4595” of 188 individuals derived from a cross of Royal Gala×PI 613988 (apple scab resistant, M. sieversii), 287 SSR (simple sequence repeats) loci were mapped. Of these SSRs, 80 are published anchors and 207 are newly developed EST (expressed sequence tag) contig-based SSRs, representing 1,630 Malus EST accessions in GenBank. Putative gene functions of these EST contigs are diverse, including regulating plant growth, development and response to environmental stresses. Among the 80 published SSRs, 18 are PI 613988 specific, 38 are common and 24 are Royal Gala specific. Out of the 207 newly developed EST contig-based SSRs, 79 are PI 613988 specific, 45 are common and 83 are Royal Gala specific. These results led to the construction of a M. sieversii map (1,387.0cM) of 180 SSR markers and a Royal Gala map (1,283.4cM) of 190 SSR markers. Mapping of scab resistance was independently conducted in two subsets of population "GMAL 4595” that were inoculated with Ventura inaequalis races (1) and (2), respectively. In combination with the two major resistance reactions Chl (chlorotic lesions) and SN (stellate necrosis) to each race, four subsets of resistance data, i.e., Chl/race (1), SN/race (1), Chl/race (2) and SN/race (2), were constituted and analyzed, leading to four resistance loci mapped to the linkage group 2 of PI 613988; SNR1 (stellate necrosis resistance to race (1)) and SNR2 are tightly linked in a region of known scab resistance genes, and ChlR1 (Chlorotic lesion resistance to race (1)) and ChlR2 are also linked tightly but in a region without known scab resistance genes. The utility of the two linkage maps, the new EST contig-based markers and M. sieversii as sources of apple scab resistance are discussed

There are several Central Asian Malus species and varieties in the USDA-ARS National Plant Germplasm System (NPGS) apple collection. Malus sieversii is the most comprehensively collected species native to Central Asia. Other taxa such as M. sieversii var. kirghisorum, M. sieversii var. turkmenorum, M. pumila, and M. pumila var. niedzwetzkyana have primarily been donated to the collection by other institutions and arboreta. We sought to determine if genetic and/or phenotypic differences among the individuals that make up the gene pools of these taxa in the NPGS exhibit unique characteristics. Genetic data, based on microsatellite analyses, suggested that the diversity within each taxa is significantly greater than that among taxa. Trait data also revealed very few differences among taxa, the primary characteristic being the dark red fruit coloration and tinted flesh color of the accessions assigned to M. pumila var. niedzwetzkyana resulting from a known single-gene mutation in anthocyanin production. We found that M. sieversii is a highly diverse species with a range in genetic and phenotypic trait variation that includes the characteristics of the other Central Asian taxa of interest. We conclude that the gene pools that comprise the accessions within the NPGS Central Asian Malus collection are highly overlapping with respect to both phenotypic traits and genotypic characters. The mountains of Central Asia have forest species that include Malus sieversii (Ledeb.) M. Roem., a highly diverse apple crop wild relative. The Russian scientist Vavilov ex- plored the forests of Central Asia in the 1920s and made note of the wide range of M. sieversii phenotypes, suggesting that this region, includ- ing Kazakhstan, Kyrgyzstan, Tajikistan, and western China, is a center of origin for the domesticated apple (Luby et al., 2001; Wan et al., 2011; Yan et al., 2008). Malus sieversii grows in a wide range of habitats, including areas with hot summers and short winters as well as those with long, severe winters in the Tien Shan Mountains (Dzhangaliev, 2003). Wild populations are found in montane, scrub, humid as well as dry continental forests and in diverse stream habitats in xeric areas (Forsline et al., 2003). Malus sieversii trees are phenotypically diverse, offering variation in the time of flowering, quantity and quality of fruit, ripening period, fruit biochemical composition, tree architecture, winter and

The USDA-ARS National Plant Germplasm System Malus collection is maintained by the Plant Genetic Resources Unit (PGRU) in Geneva, NY. In the 1990s, a core subset of 258 trees was hand-selected to be representative of the grafted Malus collection. We used a combination of genotypic and phenotypic data to compare the diversity of the 198 diploid trees in the original core subset with that of 2114 diploid trees in the grafted field collection for which data were available. The 198 trees capture 192 of the 232 total microsatellite alleles and have 78 of the 95 phenotypic characters. An addition of 67 specific individuals increases the coverage to 100% of the allelic and phenotypic character states. Several de novo core sets that capture all the allelic and phenotypic character states in 100 individuals are also provided. Use of these proposed sets of individuals will help ensure that a broad range of Malus diversity is included in evaluations that use the core subset of grafted trees in the PGRU collection.

The U.S. Department of Agriculture, Agricultural Research Service, National Plant Germplasm System (NPGS), Plant Genetic Resources Unit apple (Malus) collection in Geneva, NY, conserves over 2500 trees as grafted clones. We have compared the genotypes of 1131 diploid Malus ×domestica cultivars with a total of 1910 wild and domesticated samples representing 41 taxonomic designations in the NPGS collection to identify those that are genetically identical based on nine simple sequence repeat (SSR) loci. We calculated the probability of identity for samples in the data set based on allelic diversity and, where possible, use fruit images to qualitatively confirm similarities. A total of 237 alleles were amplified and the nine SSRs were deemed adequate to assess duplication within the collection with the caveat that “sport families” likely would not be differentiated. A total of 238 M. ×domestica and 10 samples of other taxonomic groups shared a genotype with at least one other M. ×domestica individual. In several cases, genotypes for cultivars matched genotypes of known rootstocks and indicated that these accessions may not accurately represent the indicated named clones. Sets of individuals with identical genotypes and similar cultivar names were assigned to sport families. These 23 sport families, comprised of 104 individuals, may have mutational differences that were not identified using the nine SSR loci. Five of the selected markers (CH01h01, CH02d08, CH01f02, G12, GD147) overlap with sets of markers that have been used to fingerprint European apple collections, thus making it possible to compare and coordinate collection inventories on a worldwide scale.

Hearing-aid counseling can improve outcome but programs are often too resource-intensive to be clinically practical. Here we examined the effectiveness of single-session informational counseling with single-session performance-perceptual counseling. Design: Two forms of counseling were compared: informational counseling (IC) and performance- perceptual counseling (PPC). IC focused on discussing communication strategies and tips for hearing-aid use. PPC addressed the discrepancy between measured and perceived ability to understand speech. Outcomes were measured eight-to-ten weeks post-counseling using quantitative and qualitative measures: Hearing handicap inventory, abbreviated profi le of hearing aid benefi t, psychosocial impact of assistive devices scale (PIADS), international outcome inventory for hearing aids, and a semi-structured exit interview. Study sample: Seventy-four hearing aid-users with symmetrical sensorineural hearing loss participated. Results: Scores on the hearing questionnaires showed no change following either form of counseling. Scores on the PIADS improved for participants as a whole, and the semi-structured interview revealed increased hearing-aid use, better understanding and acceptance of hearing loss, increased use of communication strategies, and improved ability to explain hearing diffi culties to others. Conclusions: A single session of hearing-aid counseling can improve hearing-aid use and satisfaction. Open-ended interview and/or quality of life measures are more sensitive to these benefi ts than hearing questionnaires.

Potential interspecific hybrids are usually identified in natural populations by their proximity to interbreeding species or their intermediate phenotypes; hybridization can then be confirmed by comparing the genetic makeup of putative hybrids to pure species. In contrast, detecting interspecific hybridization and misclassifications in ex situ collections can be difficult because fine-scale geographic locations and species-specific phenotypic data are generally unavailable. Thus, there is little a priori information available to suggest which individuals might be hybrids. Instead, hybrids or misclassified individuals must be identified based on molecular data via population assignment and admixture detection programs. We have applied a variety of population assignment and admixture detection programs to over 400 samples of four closely related Malus species held in the US Department of Agriculture–Agricultural Research Service National Plant Germplasm System that were genotyped at 19 simple sequence repeat loci. Our findings indicate that over 10 % of the samples of the wild species Malus sieversii and Malus orientalis and nearly 20 % of the samples of the wild species Malus sylvestris may be admixed or misclassified. The percentage of admixed or misclassified samples of the domesticated species, Malus × domestica, was much lower, at

Publication Inra prise en compte dans l'analyse bibliométrique des publications scientifiques mondiales sur les Fruits, les Légumes et la Pomme de terre. Période 2000-2012. http://prodinra.inra.fr/record/256699; International audience; 'Antonovka' has long been recognised as a major source of scab (Venturia inaequalis) resistance useful for apple breeding worldwide. Both major gene resistances in the form of the Rvi10 and Rvi17 and quantitative resistance, collectively identified as VA, have been identified in different accessions of 'Antonovka'. Most of the 'Antonovka' scab resistance used in apple-breeding programmes around the world can be traced back to Schmidt 'Antonovka' and predominantly its B VIII progenies 33,25 (PI 172623), 34,6 (PI 172633), 33,8 (PI 172612) and 34,5 (PI 172632). Using genetic profile reconstruction, we have identified "common 'Antonovka' " as the progenitor of the B VIII family, which is consistent with it having been a commercial cultivar in Poland and the single source of scab resistance used by Dr. Martin Schmidt. The major 'Antonovka' scab resistance genes mapped to date are located either very close to Rvi6, or about 20-25 cM above it, but their identities need further elucidation. The presence of the 139 bp allele of the CH-Vf1 microsatellite marker known to be associated with Rvi17 (Va1) in most of the 'Antonovka' germplasm used in breeding suggests that it plays a central role in the resistance. The nature and the genetic relationships of the scab resistance in these accessions as well as a number of apple cultivars derived from 'Antonovka', such as, 'Freedom', 'Burgundy' and 'Angold', are discussed. The parentage of 'Reglindis' is unclear, but the cultivar commercialised as 'Reglindis' was confirmed to be an Rvi6 cultivar.

Anthocyanins are flavonoid pigments imparting red, blue, or purple pigmentation to fruits, flowers and foliage. These compounds are powerful antioxidants in vitro, and are widely believed to contribute to human health. The fruit of the domestic apple (Malus x domestica) is a popular and important source of nutrients, and is considered one of the top ‘functional foods’—those foods that have inherent health-promoting benefits beyond basic nutritional value. The pigmentation of typical red apple fruits results from accumulation of anthocyanin in the skin. However, numerous genotypes of Malus are known that synthesize anthocyanin in additional fruit tissues including the core and cortex (flesh). Red-fleshed apple genotypes are an attractive starting point for development of novel varieties for consumption and nutraceutical use through traditional breeding and biotechnology. However, cultivar development is limited by lack of characterization of the diversity of genetic backgrounds showing this trait. We identified and cataloged red-fleshed apple genotypes from four Malus diversity collections representing over 3,000 accessions including domestic cultivars, wild species, and named hybrids. We found a striking range of flesh color intensity and pattern among accessions, including those carrying the MYB10R6 allele conferring ectopic expression of a key transcriptional regulator of anthocyanin biosynthesis. Although MYB10R6 was strongly associated with red-fleshed fruit among genotypes, this allele was neither sufficient nor required for this trait in all genotypes. Nearly all red-fleshed accessions tested could be traced back to ‘Niedzwetzkyana’, a presumed natural form of M. sieversii native to central Asia.

Clonal woody crop germplasm collections often originate and are grown in distinct geographical locations. Because the degree of cold-hardiness is known to be a factor in the successful use of dormant bud cryopreservation for Malus, it was suggested that material from relatively warmer climates would not respond to cryopreservation as well as material from colder environments. To test this hypothesis, the effect of growing provenance on cryosurvival of dormant buds from three Malus (apple) cultivars grown in three locations (Geneva, NY; Davis, CA; and Corvallis, OR) was tested in 3 consecutive years. Dormant winter buds were harvested at the three locations, cryopreserved, and bud viability was tested by grafting. The collective 3-year mean viability for cryopreserved dormant apple buds for the three locations ranged from 63% to 81% of the buds surviving with the highest survival from the Corvallis site; however, the Geneva twigs were exposed to the lowest preharvest temperature. These results suggest that the temperature at the growing location may not hinder application of the dormant bud cryopreservation method with Malus to the extent previously speculated.

Reduced availability of water for agricultural use has been forecast for much of the planet as a result of global warming and greater urban demand for water in large metropolitan areas. Strategic improvement of water use efficiency (WUE) and drought tolerance in perennial crops, like fruit trees, could reduce water use without compromising yield or quality. We studied water use in apple trees using ‘Royal Gala’, a relatively water use-efficient cultivar, as a standard. To examine whether genes useful for improving WUE are represented in a wild relative genetically close to M. ×domestica, we surveyed Malus sieversii for traits associated with WUE and drought resistance using material collected from xeric sites in Kazakhstan. This collection has been maintained in Geneva, NY, and surveyed for various phenotypes and has been genetically characterized using simple sequence repeats (SSRs). These data suggest that most of the diversity in this population is contained within a subpopulation of 34 individuals. Analysis of this subpopulation for morphological traits traditionally associated with WUE or drought resistance, e.g., leaf size and stomata size and arrangement, indicated that these traits were not substantially different. These results imply that some of the genetic diversity may be associated with changes in the biochemistry, uptake, and/or transport of water, carbon, or oxygen that have allowed these trees to survive in water-limited environments. Furthermore, genes responding to drought treatment were isolated from ‘Royal Gala’ and categorized according to the biological processes with which they are associated. A large fraction of upregulated genes from roots were identified as stress-responsive, whereas genes from leaves were for the most part associated with photosynthesis. We plan to examine expression of these genes in the M. sieversii population during water deficit in future studies to compare their patterns of expression with ‘Royal Gala’.

Penicillium expansum and Colletotrichum acutatum cause blue mold and bitter rot of apples during storage which results in significant economic losses. Resistance to these pathogens in commercial apple cultivars has not been documented in the literature. An apple germplasm collection, from the center of origin in Kazakhstan, is maintained in Geneva, New York. This collection represents a more diverse apple gene pool than commercial cultivars which was evaluated for resistance to blue mold and bitter rot. Resistance reactions were skewed toward susceptibility for both fungi and comprised the majority of accessions examined. However, resistance to P. expansum was confirmed in select accessions over multiple years. Maturation patterns and quality indices for soluble solids and acidity, which also may affect pathogenicity, were highly variable and represent the genetic diversity of the germplasm collection. Resistance in four accessions to C. acutatum and two accessions resistant to both P. expansum and C. acutatum, are reported here for the first time. Data from this study will serve as a foundation for conventional apple breeding programs and molecular genetics investigations to provide resistance against blue mold and bitter rot in commercial apple varieties.

Malus sieversii is a progenitor species of domestic apple M. × domestica. Using population “GMAL 4595” of 188 individuals derived from a cross of Royal Gala × PI 613988 (apple scab resistant, M. sieversii), 287 SSR (simple sequence repeats) loci were mapped. Of these SSRs, 80 are published anchors and 207 are newly developed EST (expressed sequence tag) contig-based SSRs, representing 1,630 Malus EST accessions in GenBank. Putative gene functions of these EST contigs are diverse, including regulating plant growth, development and response to environmental stresses. Among the 80 published SSRs, 18 are PI 613988 specific, 38 are common and 24 are Royal Gala specific. Out of the 207 newly developed EST contig-based SSRs, 79 are PI 613988 specific, 45 are common and 83 are Royal Gala specific. These results led to the construction of a M. sieversii map (1,387.0 cM) of 180 SSR markers and a Royal Gala map (1,283.4 cM) of 190 SSR markers. Mapping of scab resistance was independently conducted in two subsets of population “GMAL 4595” that were inoculated with Ventura inaequalis races (1) and (2), respectively. In combination with the two major resistance reactions Chl (chlorotic lesions) and SN (stellate necrosis) to each race, four subsets of resistance data, i.e., Chl/race (1), SN/race (1), Chl/race (2) and SN/race (2), were constituted and analyzed, leading to four resistance loci mapped to the linkage group 2 of PI 613988; SNR1 (stellate necrosis resistance to race (1)) and SNR2 are tightly linked in a region of known scab resistance genes, and ChlR1 (Chlorotic lesion resistance to race (1)) and ChlR2 are also linked tightly but in a region without known scab resistance genes. The utility of the two linkage maps, the new EST contig-based markers and M. sieversii as sources of apple scab resistance are discussed.

The USDA-National Plant Germplasm System apple (Malus) collection has traditionally been conserved by maintaining orchards in Geneva, NY and cryopreserving dormant buds of clones. The orchard Malus collection includes hundreds of M. sieversii and M. orieutalis individuals. These wild relative species are valued primarily for their diverse alleles, rather than for their specific genotypes. We have developed core collections based on both phenotypic and genotypic (microsatellite) characters to identify a limited number of individuals that represent the diversity of the wild collections. In addition to maintaining selected individuals in orchard core collections, we propose that wild species should be conserved as seeds from strategically selected crosses. This would ensure that many diverse individuals are available for future selection and physiology programs, rather than a limited number of specific genotypes. Crosses have been performed among core collection individuals for two of the three complementary core collections proposed for the M. sieversii collection to produce seeds that can be stored for long-term storage. This conservation approach that promotes preservation of selected clones as well as diverse sets of seeds for vegetatively propagated collections has several advantages. It ensures that valuable alleles in wild collections are conserved, reduces the labor involved in cryopreserving vegetative propagules, and also maintains diverse individuals in field settings that are available for pollen and dormant budwood distribution.

The potential of anthocyanin pigments in fruits and vegetables to provide protection against oxidative damage is well known. Cyanidin-3-O-galactoside is a naturally occurring red pigment commonly found in skin of apples but also present in flesh of certain crab apple and apple genotypes. The purpose of this study was to investigate the cyanidin-3-O-galactoside concentration and antioxidant capacity of juice made from 14 genotypes of red-fleshed apples in comparison to three white-fleshed apple cultivars and three commercial apple juices. Cyanidin-3-O-galactoside was found only in the juice made from red-fleshed apple genotypes with the highest concentrations (39 mg L-1) in a crab apple Roberts Crab. The antioxidant capacity measured using the Folin-Ciocalteu, the ferric reducing antioxidant power (FRAP), and the oxygen radical absorbance capacity (ORAC) assays were the greatest in juice prepared from the red-fleshed genotypes Babine and Malus pumila Niedzwetzkyana, a red-fleshed crab apple genotype. The antioxidant capacity measures were strongly correlated with each other; however, there was no correlation between the concentration of cyanidin-3-O-galactoside and the antioxidant capacity measures. The juice quality parameters °Brix and titratable acidity values were not significantly different among the juices made from the red-fleshed apples, commercial apples and commercial apple juice products.Key words: Red-fleshed apple, Malus species, antioxidants, cyanidin-3-O-galactoside, Folin-Ciocalteu, FRAP, ORAC, functional beverage

Seeds from wild Malus orientalis trees were collected during explorations to Armenia (2001, 2002), Georgia (2004), Turkey (1999), and Russia (1998). Seedling orchards with between eight and 171 individuals from each collection location were established at the U.S. Department of Agriculture–Agricultural Research Service Plant Genetic Resources Unit (PGRU) in Geneva, NY. Genotypic (seven microsatellite markers) and disease resistance data were collected for the 776 M. orientalis trees. The genetic diversity of the 280 individuals from Armenia and Georgia was compared with data previously published for the M. orientalis individuals from Russia and Turkey. A total of 106 alleles were identified in the trees from Georgia and Armenia and the average gene diversity ranged from 0.47 to 0.85 per locus. The genetic differentiation among sampling locations was greater than that found between the two countries. Six individuals from Armenia exhibited resistance to fire blight (Erwinia amylovora), apple scab (Venturia inaequalis), and cedar apple rust (Gymnosporangium juniperi-virginianae). The allelic richness across all loci in the individuals from Armenia and Georgia was statistically the same as that across all loci in the individuals from Russia and Turkey. A core set of 27 trees was selected to capture 93% of the alleles represented by the entire PGRU collection of 776 M. orientalis trees. This core set representing all four countries was selected based on genotypic data using a modified maximization algorithm. The trees selected for the M. orientalis core collection will be added to the main field collection at the PGRU.

ADDITIONAL INDEX WORDS. simple sequence repeat, genetic diversity, Kazakhstan, genotype, phenotype ABSTRACT. We estimate the minimum core size necessary to maximally represent a portion of the U.S. Department of Agriculture's National Plant Germplasm System apple (Malus) collection. We have identified a subset of Malus sieversii individuals that complements the previously published core subsets for two collection sites within Kazakhstan. We compared the size and composition of this complementary subset with a core set composed without restrictions. Because the genetic structure of this species has been previously determined, we were able to identify the origin of individuals within this core set with respect to their geographic location and genetic lineage. In addition, this core set is structured in a way that samples all of the major genetic lineages identified in this collection. The resulting panel of genotypes captures a broad range of phenotypic and molecular variation throughout Kazakhstan. These samples will provide a manageable entry point into the larger collection and will be critical in developing a long-term strategy for ex situ wild Malus conservation.

Ma/us sieversii, a wild apple species native to Central Asia, has been recognized as the major progenitor of the domestic apple. Collection trips to Central Asia have verified that M. sieversii is very diverse and has all the qualities present in i1 x dornestica. Nearly 130,000 seeds from 950 wild M. siei'ersii trees were collected in eight distinct sites, mostl y in Kazakhstan, on four collection trips from 1989 to 1996. The investigation of 1480 seedlings from these diverse sites is summarized. All seedlings were screened as young seedlings for resistance to Veuturia iizaequalis (apple scab) and planted as own-rooted seedlings in a high-density orchard in Geneva, New York. Resistance was recorded in 41% of the seedlings (7%-66% among sites). Natural infection by Erit'inia amylovora (fire blight) was also recorded, with 45% exhibiting no infection (26%-75% among sites). Seedlings with no fire blight symptoms were grafted, and replicates were inoculated in the greenhouse and 60% of that group was resistant. Resistance to V. macqua/is was re-assessed on those grafted plants. Those with original reactions similar to the Vr gene showed 100% agreement and those similar to Vp 50% agreement. A large subset was also screened as young seedlings for resistance to G)'!nnosporangium juniJ)eri-I'irginianae (cedar apple rust), with 55% showing resistance. We are in the process of evaluating the heritability of resistance to PIn'tophtlwra, Rhizoctonia and other components of apple replant disease in these accessions. In another collaborative project, fruit from a subset of the collection are being screened for resistance to the post harvest disease, blue mold (Penjcj//jii,n expansum) with some indication of resistance. Seven of the scab resistant elite accessions were crossed with the susceptible cultivar Royal Gala. By screening >200 seedlings/population with V macqua/is, 10% to 67% of each population were observed to be resistant. Correlation of the markers with phenotype indicated some M. sieversii parents likely had known resistance genes but, in some cases, exhibited patterns suggesting they also contained novel resistance loci. Seven highly diverse microsatellite markers were selected to assess the genetic relationships among the trees in the field collections that represent the populations and the collection sites. We characterized trees and fruit for horticultural traits using multiple descriptors on 746 seedlings. Among the many descriptors measured, individual fruit sizes (17 g-158 g) were quite variable among the trees and sites. We anticipate this germplasm will offer useful genetic diversity for crop improvement.

Malus sieversii (Lebed.) M. Roem. is a wild progenitor species of the domesticated apple. It is found across a mountainous region of central Asia and has been the focus of several collection expeditions by the USDA-ARS-National Plant Germplasm System. This study used microsatellite variation at seven loci to estimate diversity and differentiation within M. sieversii using several complimentary approaches. Multilocus genotypes were amplified from 949 individuals representing seedling trees from 88 half-sib families from eight M. sieversii populations collected in Kazakhstan. Apportioning of genetic variation was estimated at both the family and site level. Analyses using a hierarchical model to estimate F st showed that differentiation among individual families is more than three times greater than differentiation among sites. In addition, average gene diversity and allelic richness varied significantly among sites. A rendering of a genetic network among all sites showed that differentiation is largely congruent with geographical location. In addition, nonhierarchical Bayesian assignment methods were used to infer genetic clusters across the collection area. We detected four genetic clusters in the data set. The quality of these assignments was evaluated over multiple Markov Chain Monte Carlo runs using both posterior likelihood and stability of the assignments. The spatial pattern of genetic assignments among the eight collection sites shows two broadly distributed and two narrowly distributed clusters. These data indicate that the southwestern collection sites are more admixed and more diverse than the northern sites.

Malus sieversii (Lebed.), a wild apple species native to Central Asia, has been recognized as the major progenitor of the domestic apple. This investigation summarizes the natural infection by Erwinia amylovora (fire blight) on 2590 M. sieversu seedlings grown as own-rooted trees. At a USDA orchard in Geneva, New York (NY), 1180 seedlings (106 populations) representing germplasm from eight sites in Kazakhstan were planted in 1997 and 1998. At a University of Minnesota (MN) orchard in Excelsior, MN, 1410 seedlings representing 124 populations from two climatically diverse sites in Kazakhstan were planted in 1998. Individuals in each population are half-sibs of each other from seeds collected from the same mother tree of wild M. sieversii. One of the Kazakh sites (site 9) selected for the MN trial was a mesic site at 47° N latitude chosen for its potential for extreme cold hardiness. The other group was collected from site 6, a relatively warm, xeric site. To compare results, 32 of the populations grown in MN were also grown in NY including 10 from site 6 and 22 from site 9. Natural infection by fire blight was recorded annually using a 5-point scale (1= no infection to 5= very severe). This rating was cumulative over years. In both MN and NY, we observed a 20% greater incidence of fire blight in the trees from Kazakh site 9 than from site 6, with many infected trees dying outright. Populations grown only in NY from the other six Kazakh sites had results similar to those from site 6. Where infection was medium to severe (3-5 rating), we defined seedlings as susceptible. We determined that 535 (45%) of the seedlings grown in NY were rated 1 or 2 and potentially resistant. To determine if these genotypes were truly resistant, scions from the field trees were grafted to seedling rootstock, and shoot tips of the resulting grafted plants were inoculated in a greenhouse. Of 289 of the genotypes checked to date, nearly 60% of this group that escaped infection in the orchard were resistant.

Genetic diversity and disease resistance are described for 496 seedlings from wild populations of Malus orientalis Uglitzh. collected in southern Russia and Turkey in 1998 and 1999. Eighty-five half-sib families were genotyped using seven microsatellite markers, and disease resistance was determined for apple scab (Venturia inaequalis Cooke), cedar apple rust (Gymnosporangium juniperi-virginianae Schwein), and fire blight (Erwinia amylovora Burrill). Individuals from the two Russian Caucasus collection locations were homogeneous compared with populations from the four Turkish collection locations. Within three of the Turkish collection locations, some half-sib families were highly diverse and several of these families had unusually high levels of disease resistance. In all, twenty individuals exhibited resistance to all three diseases. Bayesian analyses of the population structure revealed six distinct clusters. Most of the individuals segregated into two clusters, one containing individuals primarily from southern Russia and the other containing individuals from both Russia and northern Turkey. Individuals in the four small clusters were specific to Turkish collection locations. These data suggest wild populations of M. orientalis from regions around the Black Sea are genetically distinguishable and show high levels of diversity.

Blue mold of apples, incited by Penicillium expansum, causes extensive losses on stored apples worldwide. Despite the severity of this problem, apple breeders do not evaluate their crosses for resistance to this disease, because there has been little resistance to blue mold in the gene pool of the germplasm used. A new apple germplasm collection from the center of origin in Kazakhstan, maintained in Geneva, NY, and representing a much broader gene pool, was evaluated for resistance to blue mold. Apples were harvested from the Elite collection trees that were clonally propagated from budwood collected in Kazakhstan and from seedling trees originating from seeds of the same trees as the Elite budwood or from other wild seedling trees in Kazakhstan. Fruit from 83 such accessions were harvested at the preclimacteric to climacteric stage, wound-inoculated with P. expansum at 103, 104, and 105 mL−1 conidial suspension, incubated for 5 d at 24 °C, and evaluated for decay incidence and severity. Two accessions were classified as immune (no decay at 103 and 104 mL−1), four as resistant (no decay at 103 mL−1), 53 as moderately resistant (lesions less than 10 mm at 103 mL−1), and 24 as susceptible. There were positive correlations (r = 0.92, 0.86, and 0.91) between decay severity and all three inoculum levels. Our results indicate a greater genetic diversity among the Kazak apple collection than among cultivated apples as evidenced by their broad range of fruit maturity, quality, and disease resistance patterns. The immune and resistant accessions may serve as a source of resistance in breeding programs and can be useful in explaining the mechanism of resistance to blue mold in apples.

The year 2005 marked the 25th anniversary of the establishment of the U.S. Department of Agriculture (USDA) National Plant Germplasm System (NPGS), repositories devoted to clonally propagated, horticultural fruit and nut crops. During this quarter century, facilities in Hilo, Hawaii; Mayaguez, PR.; Miami, Fla.; and Riverside, Calif. were developed to preserve collections of tropical and subtropical fruit and nut crops; facilities in Brownwood, Texas; Corvallis, Ore.; Davis, Calif. and Geneva, N.Y. preserve the temperate crops. Each of these facilities now has internationally recognized, globally diverse collections of genetic resources for their assigned genera. Germplasm of unique genotypes are maintained as growing plants, evaluated for phenotypic and genotypic traits, documented in a national public germplasm database, and freely distributed as clonal propaggules to researchers and other germplasm users around the world. Seed collections represent wild populations for some crop relatives. These 8 genebanks maintain 30,000 accessions representing 1600 species of fruit and nut crops and their wild relatives. The genebanks distribute more than 15,000 accessions annually to international researchers. Although originally conceived as working collections for crop improvement, NPGS genebanks have also become invaluable in providing the raw materials for basic plant genetic research, reservoirs for rare or endangered species or vulnerable landraces, archives of historic cultivars, and field classrooms for educating the public. These collections preserve botanical treasures as well as the American horticultural heritage for now and for future generations.

Seeds and scionwood of Malus sieversii Lebed. have been collected from wild populations of apple trees in Kazakhstan. Seedlings and grafted trees were planted in the orchards at the U.S. Dept. of Agriculture Plant Genetic Resources Unit in Geneva, N.Y. We developed core collections to capture the genetic and phenotypic diversity represented in the trees from each of two of the Kazakhstan collection sites. These core collections capture more than 90% of the genetic diversity of the original populations, as determined using seven unlinked simple sequence repeat markers and 19 quantitative traits. Since phenotypic evaluations of these materials have been completed, the 35 trees within each population will be used as parents in crosses so that the genetic diversity in the orchard populations can be captured as seed for long-term ex situ conservation. This strategy of storing seeds, rather than maintaining costly field collections, could be applied to other collections of wild plant materials in the National Plant Germplasm System.

Objective: Measurement of hearing aid outcomes is necessary for demonstration of treatment efficacy, third-party payment, and cost-benefit analysis. Outcomes are usually measured with hearing-related questionnaires andlor tests of speech recognition. However, results from these two types of test often conflict. In this paper, we provide data from a new test measure, known as the Performance-Perceptual Test (PPT), in which subjective and performance aspects of hearing in noise are measured using the same test materials and procedures. A Performance Speech Reception Threshold (SRTN) and a Perceptual SRTN are measured using the Hearing In Noise Test materials and adaptive procedure. A third variable, the discrepancy between these two SRTNs, is also computed. It measures the accuracy with which subjects assess their own hearing ability and is referred to as the Performance-Perceptual Discrepancy (PPDIS). Design: One hundred seven subjects between 24 and 83 yr of age took part. Thirty-three subjects had normal hearing, while the remaining seventy-four had symmetrical sensorineural hearing loss. Of the subjects with impaired hearing, 24 wore hearing aids and 50 did not. All subjects underwent routine audiological examination and completed the PPT and the Hearing Handicap Inventory for the Elderly/Adults on two occasions, between 1 and 2 wk apart. The PPT was conducted for unaided listening with the masker level set to 50, 65, and 80 dB SPL. Results: PPT data show that the subjects with normal hearing have significantly better Performance and Perceptual SRTNs at each test level than the subjects with impaired hearing but that PPDIS values do not differ between the groups. Test-retest reliability for the PPT is excellent (r-values > 0.93 for all conditions). Stepwise multiple regression analysis showed that the Performance SRTN, the PPDIS, and age explain 40% of the variance in reported handicap (Hearing Handicap Inventory for the Elderly/Adults scores). More specifically, poorer performance, underestimation of hearing ability and younger age result in greater reported handicap, and vice versa. Conclusion: Reported handicap consists of a performance component and a (mis)perception component, as measured by the Performance SRTN and the PPDIS respectively. The PPT should thus prove to be a valuable tool for better understanding why some individuals complain of hearing difficulties but have only a mild hearing loss or conversely report few difficulties in the presence of substantial impairment. The measure would thus seem to provide both an explanation and a counseling tool for patients for whom there is a mismatch between reported and measured hearing difficulties.

The United States National Plant Germplasm System has sponsored several explorations to collect a diversity of apple germplasm from Asia and Europe, in order to conserve it for posterity, and after evaluation, to make it available to breeders and geneticists worldwide. Germplasm was acquired from Sichuan Province, China (Malus hupehensis, M. kansuensis, M. prattii, M. sieboldii, M. toringoides, M. transitoria, M. yunnanensis, and M. zhaojiaoensis); Russian Caucasus (M. orientalis); Turkey (M. orientalis); and Germany (M. sylvestris). Samples of germplasm were evaluated as seedlings for resistance to fire blight, by inoculating potted plants in the greenhouse with Erwinia amylovora strain Ea273. Plants with < 20 % of shoot length blighted were characterized as resistant. Considerable variability in resistance was observed in seedlings from almost all collections of all species from individual sites. Seedlings from small-fruited Sichuanese species were also evaluated by inoculation of 2nd leaf plants in the field. Greenhouse and field evaluations of the Sichuanese species were in close agreement. Proportion of resistant seedlings ranged, in the different sites collected, from 0 - 3 % of M. yunnanensis to 0 - 100 % of M. hupehensis. Most populations of M. hupehensis, M. sieboldii, M. toringoides, and M. transitoria had a high frequency of resistant seedlings. Seedlings from different populations of trees with ~ 30 mm fruit size (M. orientalis) from Russian Caucasus were 50 - 93 % resistant, whereas M. orientalis seedlings from Turkey were only 6 - 50 % resistant. Seedlings from small-fruited M. sylvestris from Germany were almost all susceptible. The resistant seedlings from the Sichuanese species may have breeding potential especially for rootstocks and ornamentals. However, Russian M. orientalis may convey resistance to fire blight accompanied by less sensitivity to latent viruses, than small-fruited Malus species. This would be especially useful for breeding of fire blight resistant rootstocks.

Malus sieversii is the main ancestor of the cultivated apple (M. domestica) and is mainly endangered in its center of diversity in Kazakstan. Seeds from 35 wild Malus sieversii across 7 locations (1089 seedlings in total) were selected and germinated for evaluation at the fruit genebank at Dresden-Pillnitz. The seeds were supplied by the USDA-ARS Plant Genetic Resources Unit, Geneva, NY, USA, which obtained them during expeditions to Central Asia. The assessment of the genetic diversity of these seedlings should give assistance in preserving (in situ) the original habitats and for further collecting strategies. The main practical objective of this research is to find new sources of disease resistances and improved horticultural qualities for breeding new apple varieties. The broad range of variability within the plant material can already be observed after 2 to 3 years of evaluation (e.g. start and length of the vegetation period, plant height). Evaluations of apple scab and powdery mildew symptoms, caused by natural infections, showed significant differences within and between offsprings. A total of 64 individuals showed complete absence of both diseases. Therefore, new sources of resistance for use in breeding programs are expected to be identified. With regard to apple scab, this assumption was supported by initial PCR experiments employing closely linked SCAR and RAPD markers for the Vf -gene. Neither of 21 preselected M. sieversii individuals from 6 different sites displayed the respective marker bands, although being scored resistant to the disease. In PCR analyses aimed at determining the genetic diversity of 26 M. sieversii individuals from the different populations, a very good correlation between genetic similarity and the origin of the different individuals could be observed. The evaluation of this material and its characterization by molecular markers is planned to be intensified in future years.

The center of diversity of the genus Malus is situated in East Asia. The diversity of wild and cultivated apples, as a whole represent a great pool of traits for multiple use. This irretrievable diversity is to be preserved now and in the future by genebanks. This is to be done by: 1) keeping the trees in situ at the natural site (wild species) or on farm; 2) ex situ as grafted trees (cultivars), or as seedlings, and seed lots in the genebanks. For decades, classical collections of wild and cultivated apples have existed. Mostly, the material was collected a long time ago, exchanged between different arboreta, often as open-pollinated seeds. Usually, the original passport data is entirely incomplete. Additionally in collections, many representative Malus species are restricted to only a few accessions. Therefore they are neither representative for the extent of variability within a species nor for usable traits. As a result, in recent years collecting of wild specimens has been revived. For multiple use of genebank material, evaluation and characterization are necessary. The most important traits to be defined are for resistances to biotic and abiotic factors so that they may be used in breeding new cultivars.