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Eastern filbert blight (EFB), caused by Anisogramma anomala, is a fungal disease threatening the european hazelnut (Corylus avellana) industry in the Willamette Valley of Oregon. The pathogen is endemic to the eastern United States where it causes little damage to the wild Corylus americana but causes severe cankers on most cultivars of the commercially important european hazelnut. The host genetic resistance in ‘Gasaway’ is conferred by a dominant allele at a single locus on linkage group 6 (LG6), and resistance from several other sources has been mapped to the same region. Some fungal isolates can overcome ‘Gasaway’ resistance, prompting a search for other sources of resistance. Resistance from other sources has been mapped to LG2 and LG7, for which additional simple sequence repeat (SSR) markers would facilitate marker-assisted selection (MAS). In this study, an in silico approach was used to develop new polymorphic SSR markers in the EFB resistance regions on LG2 and LG7. Starting with a search of 17 contigs of the ‘Jefferson’ genome sequence, 45 new polymorphic SSR markers were developed, characterized, and placed on the linkage map. The new SSR markers had an average of 10.18 alleles per locus, and average values for expected heterozygosity, observed heterozygosity, polymorphism information content, and frequency of null alleles of 0.72, 0.65, 0.68, and 0.068, respectively. Of the 42 new polymorphic SSRs segregating in the mapping population, 24 were on LG2, 12 were on LG7, and six were placed on other LGs. The new and previously developed SSR markers were used to study six new sources of EFB resistance, four from Russia and two from Crimea. Six resistant selections were crossed with susceptible selections, resulting in 7 progenies. Phenotyping for disease response revealed that segregation in progenies of the two Moscow selections (#2 and #27), one Russian selection (OSU 1187.101), and one Crimean selection (H3R12P62) fit the 1:1 segregation ratio expected for control of resistance by a dominant allele at a single locus; but in progenies of the other Russian selection (OSU 1166.123) and the other Crimean selection (H3R07P11), there was an excess of resistant seedlings. Correlation of disease scores and alleles at SSR loci indicated that resistance from three Russian selections (Moscow selections #2 and #27 and OSU 1166.123) and the Crimean selection H3R12P62 was on LG7, while resistance from Russian selection OSU 1187.101 was on LG2. Resistance from Crimean selection H3R07P11 was not correlated with markers on LG6, or LG2, or LG7. These sources and new SSR markers will be useful in MAS and the pyramiding of resistance genes in the breeding of new EFB-resistant cultivars.

The production and consumption of nuts are increasing in the world due to strong economic returns and the nutritional value of their products. With the increasing role and importance given to nuts (i.e., walnuts, hazelnut, pistachio, pecan, almond) in a balanced and healthy diet and in the prevention of various diseases, breeding of the nuts species has also been stepped up. Most recent fruit breeding programs have focused on scion genetic improvement. However, the use of locally adapted grafted rootstocks also enhanced the productivity and quality of tree fruit crops. Grafting is an ancient horticultural practice use in nut crops to manipulate scion phenotype and productivity and overcome biotic and abiotic stresses. There are complex rootstock breeding objectives and physiological and molecular aspects of rootstock–scion interactions in nut crops. In this review, we provide an overview of these, considering the mechanisms involved in nutrient and water uptake, regulation of phytohormones, and rootstock influences on the scion molecular processes, including long-distance gene silencing and trans-grafting. Understanding the mechanisms resulting from rootstock × scion × environmental interactions will contribute to developing new rootstocks with resilience in the face of climate change, but also of the multitude of diseases and pests and of the possible increase of their aggressiveness. They will also have to offer the premises of economic production, respectively yield and the quality, according to multiple destinations of nuts in the current consumption and food industry, but also the increasing exigencies of the consumer market and the profile industry.

Commercial production of hazelnut (Corylus avellana) in Oregon’s Willamette Valley is threatened by eastern filbert blight (EFB), a serious canker disease caused by the pyrenomycete Anisogramma anomala (Peck) E. Müller. The fungus also prevents the establishment of hazelnut orchards in eastern North America. Genetic resistance is considered the most effective way to control the disease. A high level of EFB resistance was first discovered in ’Gasaway’. This resistance is conferred by a dominant allele at a single locus on linkage group 6 (LG6). Resistance from several additional sources has been assigned to the same chromosomal region. In this study, new simple sequence repeat (SSR) markers were developed for the resistance region on LG6 and new sources of resistance were investigated. Forty-two new SSR markers were developed from four contigs in the genome sequence of ‘Jefferson’ hazelnut, characterized, and nine of them were placed on LG6 of the genetic map. Accessions representing 12 new sources of EFB resistance were crossed with susceptible selections resulting in 18 seedling populations. Segregation ratios in the seedling populations fit the expected 1:1 ratio for 10 sources, while one source showed an excess of resistant seedlings and another showed an excess of susceptible seedlings. Based on correlation of disease response and scores of SSR markers in the ‘Gasaway’ resistance region in the seedlings, eight resistance sources were assigned to LG6. Linkage maps were constructed for each progeny using SSR markers. The LG6 resistance sources include two selections (#23 and #26) from the Russian Research Institute of Forestry and Mechanization near Moscow, four selections from southern Russia, one selection (OSU 1185.126) from Crimea, one selection (OSU 533.129) from Michigan, Corylus heterophylla ‘Ogyoo’ from the South Korea, and the interspecific hybrid ’Estrella #1’. These new LG6 resistance sources and SSR markers should be useful in breeding new cultivars, including the pyramiding of resistance genes. For the other four resistance sources (Moscow #37, hybrid selection OSU 401.014, C. americana ‘Winkler’ and C. americana OSU 366.060), SSR marker scores on linkage groups 6, 7 and 2 were not correlated with disease response and merit further investigation.

European hazelnut (Corylus avellana L.) is an economically important edible nut producing species, which ranked sixth in world tree nut production in 2016. European hazelnut production in the United States is primarily limited to the Willamette Valley of Oregon, and currently nonexistent in the eastern United States because of the presence of a devastating endemic disease, eastern filbert blight (EFB) caused by Anisogramma anomala (Peck) E. Muller. The primary commercial means of control of EFB to date is through the development and planting of genetically resistant european hazelnut cultivars, with an R-gene introduced from the obsolete, late-shedding pollinizer ‘Gasaway’. Although the ‘Gasaway’ resistance source provides protection against EFB in the Pacific northwestern United States (PNW), recent reports have shown that it is not effective in parts of the eastern United States. This may be in part because the identification and selection of ‘Gasaway’ and ‘Gasaway’-derived cultivars occurred in an environment (PNW) with limited genetic diversity of A. anomala. The objectives of the current research were to develop a genetic linkage map using double digestion restriction site associated DNA sequencing (ddRADseq) and identify quantitative trait loci (QTL) markers associated with EFB resistance from the resistant selection Rutgers H3R07P25 from southern Russia. A mapping population composed of 119 seedling trees was evaluated in a geographic location (New Jersey) where the EFB fungus is endemic, exhibits high disease pressure, and has a high level of genetic diversity. The completed genetic linkage map included a total of 2217 markers and spanned a total genetic distance of 1383.4 cM, with an average marker spacing of 0.65 cM. A single QTL region associated with EFB resistance from H3R07P25 was located on european hazelnut linkage group (LG) 2 and was responsible for 72.8% of the phenotypic variation observed in the study. Based on its LG placement, origin, and disease response in the field, this resistance source is different from the ‘Gasaway’ source located on LG6. The current results, in combination with results from previous research, indicate that the H3R07P25 source is likely exhibiting resistance to a broader range of naturally occurring A. anomala isolates. As such, H3R07P25 will be important for the development of new european hazelnut germplasm that combines EFB resistance from multiple sources in a gene pyramiding approach. Identification of EFB resistance in high disease pressure environments representing a diversity of A. anomala populations is likely a requirement for identifying plants expressing durable EFB resistance, which is a precursor to the development of a commercially viable european hazelnut industry in the eastern United States.

Eastern filbert blight (EFB) is a hazelnut disease caused by Anisogramma anomala, a fungus endemic to North America where it can be found associated with the wild hazelnut Corylus americana. While C. americana is resistant to EFB, the commercial hazelnut C. avellana is highly susceptible and typically dies within 5 years from exposure. A hazelnut breeding program was initiated in 1996 at Rutgers University in New Brunswick, New Jersey. The early objective was to identify sources of resistance to EFB for use in breeding regionally adapted, EFB-resistant plants to support the establishment of a hazelnut industry in eastern USA. By working with Oregon State University (OSU) and international colleagues, germplasm collections were made in Russia, Ukraine, Poland, Georgia, Latvia, Lithuania, Estonia, Italy, and Turkey. Over 5,000 seedlings were grown and evaluated for response to EFB. While most plants succumbed to EFB, about three percent were found to be resistant. Interestingly, the resistant plants spanned nearly all collection locations and molecular markers have shown them to be highly diverse. Today, we have access to over 100 EFB-resistant accessions selected from more than 60 locations, which represents a significant pool of germplasm to support breeding. Recent studies have shown that most resistance seems to be controlled by only one or a few major genes; R-gene mapping projects are underway at Rutgers as well as Oregon State University. We continue to study new sources of resistance for use in a focused cultivar improvement program.

Corylus americana is native to a wide area of land in eastern North America, bounded by the Rocky Mountains in the west and the Atlantic Ocean in the east, and ranging from southern Canada to the southeastern US. The species has been shown to be highly genetically diverse and is adapted to a variety of climates and soils. It is also resistant to the disease eastern filbert blight (EFB) caused by the fungus Anisogramma anomala. Unfortunately, C. americana has thick-shelled, tiny nuts that make it unsuitable for commercial production. However, it is cross-compatible with the commercial hazelnut C. avellana and can serve as a donor for genes for disease resistance, early nut maturity, cold tolerance, etc. As part of the Hybrid Hazelnut Consortium consisting of Rutgers University, Oregon State University, the University of Nebraska-Lincoln, and the Arbor Day Foundation, we have been successfully collecting C. americana germplasm since 2009 with the help of partners, colleagues, and the interested public around the USA. Today, we have a planting established in the field at Rutgers University in New Jersey that holds 1,899 seedlings obtained from 128 individual seed lots that span 23 states and one Canadian province. These bushes are being evaluated for EFB response, other diseases and insects, flowering and vegetative bud break phenology, cold hardiness, nut yield, and nut and kernel characteristics. The genetic diversity and population structure of the new collection is also being evaluated using several approaches, including genotyping by sequencing and subsequent SNP analysis, SSR allelic variation, and chloroplast haplotype determination across the large population. Using this multi-faceted approach, we hope that superior, diverse selections can be identified for use in systematic breeding efforts to develop new cultivars adapted to the eastern USA, especially in respect to consistent cropping in regions with highly variable year-to-year climates.

The hazelnut breeding program at Oregon State University (OSU) is developing new cultivars with high nut yield, few defects, and good kernel quality. Every year, approximately 4500 seedlings are planted and evaluated for seven years. The very best of these seedlings are put through a second period of testing in replicated yield trials to determine their potential as cultivars. The fungal disease eastern filbert blight (EFB) is now widespread in the Willamette Valley, so current trials include only EFB-resistant selections. In this paper, we summarize data for three recent releases. ‘York’ (OSU 878.048) is a widely compatible mid-season pollinizer with nuts and kernels that blend well with other recent OSU releases. ‘Wepster’ (OSU 894.030) and ‘McDonald’ (OSU 880.027) were released as main crop cultivars. Trees of ‘Wepster’, ‘McDonald’, and ‘York’ have rounded, moderately upright growth habits, and mature nuts 7-14 days before ‘Barcelona’. ‘Wepster’ nuts and kernels are small, similar in size to ‘Yamhill’, and have very few defects. The percent kernel of ‘Wepster’ is only 44% on average, but total nut yields are very high. ‘McDonald’ nuts are medium to small in size and are suitable for the kernel market. Nuts and kernels of ‘York’ are of good quality, but trees yield less than what is desirable for a main cultivar. ‘York’ has incompatibility alleles S(2) and S(21), but only the uncommon S(21) is expressed in the pollen. It is an excellent mid-season pollinizer, producing large quantities of highly viable pollen over a period of several weeks, and is compatible on most other genotypes. ‘Wepster’, ‘McDonald’ and ‘York’ are fully cross-compatible and are recommended to be planted together. All three are protected with US Plant Patents.

Incompatibility in European hazelnut (Corylus avellana L.) is sporophytic and under the control of a single locus on linkage group 5 between markers G05-510 and AU02-1350. In this study, two rounds of marker development and a population of 192 seedlings with known S-alleles that showed recombination between the flanking markers were used for fine mapping. Using the sequences of random amplified polymorphic DNA and simple sequence repeat (SSR) markers and bacterial artificial chromosome end sequences, 36 contigs from the genome sequence of “Jefferson” hazelnut were identified for pursuit. Di-nucleotide SSR markers in those contigs were developed, characterized, and mapped. This reduced the size to a region of 500 kb that contained the S-locus and 50 predicted genes, in which single-nucleotide polymorphism and additional SSR markers were developed. When the new markers were used in fine mapping, they fully exploited all recombination in the fine mapping population and reduced the region to 193.5 kb containing 18 genes. This 193.5-kb region most likely contains the S1 haplotype. A second region, 2 Mbp away from the first in the “Jefferson” genome (V3), is predicted to represent the S3 haplotype based on SSR marker allele sizes and the ratio of parental reads that align to them. Although they appear side-by-side in the “Jefferson” genome (V3), the mapped markers appear in both sequences in the same order. Gene annotations in the S1 and S3 haplotypes are highly similar and include five probable leucine-rich repeat receptor-like protein kinases with homology to Arabidopsis thaliana genes At1g35710 and three probable receptor-like serine/threonine protein kinases with homology to At5g15080 (PIX7).

From the genome sequence of hazelnut (Corylus avellana), 192 new polymorphic simple sequence repeat (SSR) markers were developed, characterized, and used to investigate genetic diversity in 50 accessions. Next-generation sequencing allows inexpensive sequencing of plant genomes and transcriptomes, and efficient development of polymorphic SSR markers, also known as microsatellite markers, at low cost. A search of the genome sequence of ‘Jefferson’ hazelnut identified 9094 fragments with long repeat motifs of 4, 5, or 6 base pairs (bp), from which polymorphic SSR markers were developed. The repeat regions in the ‘Jefferson’ genome were used as references to which genomic sequence reads of seven additional cultivars were aligned in silico. Visual inspection for variation in repeat number among the aligned reads identified 246 as polymorphic, for which primer pairs were designed. Polymerase chain reaction (PCR) amplification followed by agarose gel separation indicated polymorphism at 195 loci, for which fluorescent forward primers were used to amplify the DNA of 50 hazelnut accessions. Amplicons were post-PCR multiplexed for capillary electrophoresis, and allele sizes were determined for 50 accessions. After eliminating three, 192 were confirmed as polymorphic, and 169 showed only one or two alleles in each of the 50 cultivars, as expected in a diploid. At these 169 SSRs, a total of 843 alleles were found, for an average of 4.99 and a range of 2 to 17 alleles per locus. The mean observed heterozygosity, expected heterozygosity, polymorphism information content, and the frequency of null alleles were 0.51, 0.53, 0.47, and 0.03, respectively. An additional 25 primer pairs produced more than two bands in some accessions with an average of 6.8 alleles. The UPGMA dendrogram revealed a wide genetic diversity and clustered the 50 accessions according to their geographic origin. Of the new SSRs, 132 loci were placed on the linkage map. These new markers will be useful for diversity and parentage studies, cultivar fingerprinting, marker-assisted selection, and aligning the linkage map with scaffolds of the genome sequence.

The disease eastern filbert blight (EFB), caused by Anisogramma anomala, severely limits hazelnut (Corylus avellana) production in North America. In the 1970s, ‘Gasaway’, a late-flowering pollenizer, was found to be resistant to EFB in the Pacific northwestern U.S., and to transmit its resistance in a predictable Mendelian pattern. Its subsequent use in breeding at Oregon State University led to the release of a number of EFB-resistant cultivars, including Jefferson, Yamhill, Dorris, Wepster, and McDonald, which are now being widely planted across the Willamette Valley of Oregon. However, some cultivars carrying the ‘Gasaway’ R-gene develop EFB in New Jersey. Thus, the utility of this resistance source for breeding plants adapted to the eastern U.S has not been fully evaluated. In this study, we examined the EFB response of seedlings from 31 different full-sib progenies expected to segregate for the ‘Gasaway’ R-gene. Trees were exposed to the disease over at least 5 years and evaluated on a scale of 0 to 5 (0 = resistant, 5 = highly susceptible). In general, we found the number of seedlings free of EFB (class 0) to be less than expected based on prior work in Oregon, but resistant individuals were identified in each progeny, indicating the presence of heritable resistance. When disease response classes 0, 1, and 2 were combined (resistant through highly tolerant) and considered as “resistant”, the expected ratio of resistant to susceptible trees was recovered, signifying that the single dominant allele alone continues to provide a predictable level of at least tolerance. Modifying genes/factors appear to play a role in the final disease response of the individual seedling. It is thus hypothesized that some cultivars selected as resistant in Oregon that develop cankers in New Jersey may lack the necessary modifying genetic factors for full protection in the east. Until these modifying genes are elucidated, long-term field evaluation remains necessary to identify selections that carry the ‘Gasaway’ R-gene and express a high level of resistance to EFB in the eastern U.S.

Eastern filbert blight (EFB) is a serious fungal disease of european hazelnut (Corylus avellana) in North America. The causal agent is the pyrenomycete Anisogramma anomala, which is native in the eastern United States where it occasionally produces small cankers on the wild american hazelnut (C. americana). However, most commercial cultivars of european hazelnut are susceptible. Infection leads to perennial cankers, girdling of branches, and premature tree death. Cultural practices including scouting, pruning out infected branches, and fungicide applications are recommended to slow disease spread but are expensive and not completely effective. EFB resistance from ‘Gasaway’ is conferred by a dominant allele at a single locus and has been extensively used in the Oregon State University hazelnut breeding program, but there is concern that this resistance could be overcome by isolates now present in the eastern United States or that a new race of the pathogen could arise in Oregon. Segregation for EFB resistance from ‘Uebov’, a new source from Serbia, was studied in three progenies by a combination of structure exposure and greenhouse inoculation. The frequency of resistant seedlings following structure exposure was about 20% in all three progenies. The ratios failed to fit the expected 1:1 ratio but did fit a ratio of 1 resistant:3 susceptible, which would be expected if resistance were conferred by dominant alleles at two independent loci. Seedlings from a cross of susceptible selection OSU 741.105 and ‘Uebov’ were used to study correlation of disease response and presence of alleles at microsatellite marker loci. Resistance was highly correlated with the presence of alleles at marker loci on linkage group 6 (LG6), and these markers also showed segregation distortion. We conclude that EFB resistance from ‘Uebov’ maps to a single locus on LG6 in the same region as resistance from ‘Gasaway’, although only about 20% of the seedlings are resistant because of segregation distortion. ‘Uebov’ has large, well-filled, round nuts and is suitable as a parent in breeding for the in-shell market, but its low nut yields and a high frequency of shells with split sutures are the drawbacks. Its use would expand options for breeding and ‘Uebov’ resistance could be combined with other resistance alleles with an expectation of more durable EFB resistance. Durable resistance would not only sustain the hazelnut industry in Oregon but would also allow expansion of plantings to new areas.

Inheritance of resistance to eastern filbert blight, caused by Anisogramma anomala, in European hazelnut (Corylus avellana) was evaluated in the progeny of seven cultivars crossed in 12 combinations. The progeny were subjected to inoculation with A. anomala in the greenhouse and in the field. Three disease responses were measured: disease incidence, number of cankers, and proportion of wood diseased. In both the greenhouse and the field, progeny produced by crossing VR6-28 with three susceptible cultivars segregated 1:1 for complete resistance to eastern filbert blight, confirming a previous report that VR6-28 is heterozygous for a single, dominant resistance gene. Histograms of disease responses in progeny of the remaining six parents showed continuous distributions for all crosses examined. Consequently, these parents were analyzed for general and specific combining abilities for each disease response. In the field, general and specific combining ability were both significant (P < 0.05) for all disease responses, with general combining ability having twice the magnitude of specific combining ability. These results suggest these disease responses are controlled by additive gene action in the cultivars examined, with nonadditive gene action being of some importance. Based on general combining ability values, high levels of partial resistance were transmitted by the pollen parents, Gem and Tonda di Giffoni, and the seed parent, Willamette. Heritability of disease incidence, number of cankers, and proportion of wood diseased were calculated to be 0.21, 0.39, and 0.47, respectively, for this set of nine crosses after the first exposure period in the field. This suggests that it will be possible to use partially resistant parents to breed for hazelnuts exhibiting fewer and smaller cankers.

Eleven simple sequence repeat (SSRs) markers were used to investigate population structure of 582 accessions representing hazelnut cultivars grown in different geographical regions of the world. SSR profiles for 229 accessions were assessed for the first time, and their allele sizes were combined with those of 353 accessions previously characterized and published, for an analysis of genetic diversity and population structure. The combined data for all 582 accessions was used to construct an UPGMA (unweighted pair group method of arithmetic averages) dendrogram. The UPGMA tree showed multiple groups with clustering partly based on their geographic origin. The STRUCTURE analysis indicated that the most likely number of sub-populations is three to seven. Taken together, the results showed that cultivated hazelnuts consist of multiple genetic groups and cultivars were likely selected from local wild populations rather than originating in one or two domestication centers.

European hazelnut (Corylus avellana L.) is a significant crop in Oregon, where 99% of United States hazelnuts are produced. Eastern filbert blight (EFB) caused by Anisogramma anomala (Peck) E. Müller is an important disease that infects the trees, reduces yield, and causes premature death. Managing the disease through cultural methods and fungicide applications is laborious and expensive, and genetic host resistance is considered the most viable option for control. Genetic resistance from ‘Gasaway’ has been used to develop resistant cultivars including Yamhill and Jefferson, but concern about the durability of this single resistance gene stimulated a search for additional sources of resistance. This study used three recently identified sources of EFB resistance: ‘Culplà’ from Spain, ‘Crvenje’ from Serbia, and OSU 495.072 from southern Russia. RAPD markers linked to resistance from ‘Gasaway’ were absent in all three accessions. Disease response was noted in segregating progenies following greenhouse or structure inoculation, and the resistance loci were mapped using microsatellite markers. In only four of the nine progenies did segregation for disease response fit the ratio of 1 resistant:1 susceptible expected for a single locus, a heterozygous resistant parent, and a dominant allele for resistance. Three progenies showed an excess of resistant seedlings while two showed a deficiency of resistant seedlings. The reciprocal translocations reported in several leading hazelnut cultivars may be present in the parents of the studied progenies, and affecting the segregation ratios. Microsatellite marker A614, previously mapped to linkage group (LG) 6, was closely linked to resistance from all three sources. Maps were constructed for LG6 for each resistant parent using microsatellite markers. The three resistance loci mapped to the same region on LG6 where resistance from ‘Gasaway’ and OSU 408.040 are located. The resistance alleles in all five accessions may be the same, or more likely are a cluster of different resistance genes in the same region. Markers LG628, LG610, and LG696 will be useful to breed new hazelnut cultivars with resistance from Culplà, Crvenje, and OSU 495.072.

Eastern filbert blight (EFB), caused by the pyrenomycete Anisogramma anomala, is a serious threat to the hazelnut industry in the Pacific Northwest. EFB is endemic in eastern North America where it occasionally produces small cankers on the wild American hazelnut (Corylus americana). In contrast, most cultivars of European hazelnut (Corylus avellana) are susceptible. Genetic resistance is the most promising disease control method and is an objective of the Oregon State University hazelnut breeding program. ‘Gasaway’ resistance, which is governed by a dominant allele at a single locus, has been extensively used in the program. However, ‘Gasaway’ and some of its offspring have been infected by EFB isolates from New Jersey, Minnesota, and Michigan. Efforts to create new cultivars with durable EFB resistance include identifying and studying new resistance sources. In this study, resistant accessions C. americana ‘Rush’ and interspecific hybrid selection ‘Yoder #5’ were crossed with susceptible C. avellana selections and the resulting segregating seedling populations were inoculated by either exposure of potted trees under a structure topped with diseased branches or field exposure supplemented by tying diseased branches to each tree. Disease response was scored when cankers were visible 20 months after inoculation. Resistance from both sources segregated in a 1:1 ratio, indicating control by a single locus and a dominant allele for resistance. DNA extracted from the seedlings was amplified with previously mapped microsatellite markers. Resistance from both C. americana ‘Rush’ and ‘Yoder #5’ was placed on linkage group 7 in the same position as resistance from C. avellana ‘Ratoli.’ Linked microsatellite markers B753, GB372, and B509 will be useful for marker-assisted selection and the pyramiding of genes for durable EFB resistance. Assessing response to EFB is challenging, whether the plants are inoculated under a structure topped with diseased wood or in a humidity chamber in the greenhouse, or by exposure in the field. The pathogen has a 2-year life cycle, and there is a 15-month wait between inoculation and symptom expression. A small number of escapes is commonly encountered, and resistant plants occasionally develop small cankers. Our approach of studying segregation ratios and then mapping with microsatellite markers should be a useful approach for disease resistance studies in many tree crops.

Pollen-stigma incompatibility in hazelnut is sporophytic and under the control of a single locus with multiple alleles. To date, 33 alleles have been identified at the S-locus, and a designated pollen tester for each preserved in our collection. Hazelnut is diploid, and nearly all cultivars are heterozygous at the S-locus. Over the past 25 years, we have enlarged our collection by importing cultivars from several countries as scions. As the trees began to flower, we identified their S-alleles using florescence microscopy. In December, we bagged emasculated branches on trees growing in the field to protect the female inflorescences from wind-borne pollen. In January, we collected pollen from our tester trees. In February, females were collected from bagged branches and brought to the lab in petri dishes. The females were pollinated in the afternoon, one or two per tester pollen. The following day, the styles were squashed in aniline blue and observed under UV light. In compatible pollinations, pollen germination was excellent and tubes could be seen growing parallel down the style. In incompatible pollinations, germination was often reduced, pollen tubes were short and did not penetrate the stigmatic surface, and the tubes often ended in bulbs. Each pollination was scored as compatible or incompatible, and the testing of a tree required 1-3 years for completion. In this paper, we report the alleles of 170 cultivars for the first time, and summarize the alleles of 112 cultivars previously typed. Cultivars with different names often have identical microsatellite marker fingerprints and S-alleles. To reduce confusion, a list of cultivars with identical fingerprints is also included.

Pollen–stigma incompatibility in european hazelnut (Corylus avellana L.) is of the sporophytic type and under the control of a single locus with multiple alleles (haplotypes). The S-locus was previously assigned to linkage group 5 (LG5) and linked DNA markers were identified. The loci that control leaf color and style color are linked to the S-locus. We investigated segregation for leaf and style color and S-alleles in two progenies, mapped the loci, and compared the two new maps with the LG5 reference map using simple sequence repeat (SSR) markers. Segregation for color, S-alleles and SSR markers fit expectations. The color loci and the S-locus mapped to LG5 between SSR markers B028 and B774. The three maps aligned and the SSR markers were collinear. The SSR markers closest to the S-locus are KG819, KG847, and BR259. In progeny 05050, which segregated for style and leaf color, no recombination was observed between the two traits. Recombination between the S-locus and the style color locus was 5.4 cM in progeny 05050 and 10.1 cM in progeny 00064. The style color locus was placed very close to SSR marker B028 in both progenies. On the reference map, random amplified polymorphic DNA (RAPD) markers 564-500M, 345-1050dF, and 204-950dF and intersequence simple sequence repeat (ISSR) marker 815-540dF are very close to the S-locus. The identification of closely linked markers will facilitate the map-based cloning of the S-locus and color loci in hazelnut.

The european hazelnut (Corylus avellana L.) is native to most of Europe and nearby areas in Asia Minor and the Caucasus Mountains. Cross-pollination is enforced by sporophytic incompatibility under the control of a single locus with multiple alleles (haplotypes). Fluorescence microscopy is routinely used to determine if a pollination is compatible or incompatible, and use of an array of known testers allows identification of the alleles of cultivars and selections. Both alleles are expressed in the stigmas, but often only one is expressed in the pollen because of dominance. Cultivars are highly heterozygous diploids (2n = 2x = 22) and clonally propagated. Most of the world's leading cultivars were selected from local wild populations near where they are now planted on a commercial scale. Genetic improvement efforts are recent and, although tremendous genetic variability is available, such efforts have had little impact outside of Oregon and France. Studies of genetic diversity using simple sequence repeat markers have placed most cultivars in one of the four main groups: Spanish-Italian, Central European, English, or Black Sea. This study presents 17 years of data on incompatibility in hazelnut, including the discovery of six new S-alleles and determination of the dominance relationships among 105 new pairs of alleles. The total number of alleles now stands at 33. The S-alleles of 284 cultivars, 13 interspecific hybrids, and 522 selections of diverse origin are presented. The S-alleles identified in hazelnut cultivars is information that should be useful to breeders in the planning of crosses, to curators of germplasm collections, and to growers and nurseries as they choose cultivars and pollenizers when designing orchards. Differences in S-allele frequency seen in the cultivars and selections are related to geographic origin. The most common alleles of cultivars in the major geographical groups are S2 in the Spanish-Italian group, S5 in the Central European group, S3 in the English group, and S4 in the Black Sea group. Most selections belonged to the Black Sea group, and S4 was by far the most common allele. Differences in allele frequency were also observed among seed lots within a country.

Bacterial artificial chromosome (BAC) libraries are widely used in map-based cloning of plant genes. Eastern filbert blight (EFB), caused by the pyrenomycete Anisogramma anomala (Peck) E. Muller, is a devastating disease of European hazelnut (Corylus avellana L.) in the Pacific Northwest. A dominant allele at a single locus from the obsolete pollenizer “Gasaway” confers complete resistance. Our map-based cloning efforts use a BAC library for “Jefferson” hazelnut, which is heterozygous for resistance. Screening the library with primer pairs designed from RAPD markers closely linked to the EFB resistance locus identified 38 BACs. We sequenced 28 of these BACs using Illumina technology, by multiplexing with barcoded adapters. De novo sequence assembly using the programs Velvet and SOPRA and further alignment using CodonCode Aligner generated contigs whose length ranged from 393 to 108,194 bp. The number of contigs per BAC ranged from 1 to 19, and estimated coverage of assembled BACs ranged from 64 % to 100 %. Preliminary analysis of the sequences identified 779 simple sequence repeats (SSRs), from which we developed 23 markers. Of these, 17 were assigned to linkage group 6 adjacent to the disease resistance locus, five were placed on other linkage groups, and one could not be assigned to a linkage group. The BAC sequences and new SSR markers will be useful for our efforts at map-based cloning of the disease resistance gene.

Eastern filbert blight (EFB) of European hazelnut (Corylus avellana L.), caused by the pyrenomycete Anisogramma anomala (Peck) E. Müller, is a major disease problem and production constraint in Oregon’s Willamette Valley. Host genetic resistance is viewed as the most economical means of controlling this disease. Marker-assisted selection has been extensively used for ‘Gasaway’ resistance in the hazelnut breeding program at Oregon State University (OSU). Concern over potential breakdown of this single resistance gene prompted a search for new sources of resistance. Selection OSU 408.040 showed no signs or symptoms of the fungus after a series of disease inoculations, and resistance was transmitted to half of its offspring, indicating control by a dominant allele at a single locus. In this study, we identified six random amplified polymorphic DNA (RAPD) and 11 simple sequence repeat (SSR) markers linked to EFB resistance from OSU 408.040. The new markers supplement the previously identified amplified fragment length polymorphism (AFLP) markers. A linkage map constructed in the progeny OSU 245.098 × OSU 408.040 spanned a distance of 19.5 cM with the resistance locus cosegregating with AFLP marker A8-150 and located between SSR markers LG675 and LG682. Using SSR markers as anchor loci, OSU 408.040 resistance was assigned to linkage group 6 (LG6). Comparison with the previously mapped ‘Gasaway’ resistance locus showed that resistance from OSU 408.040 maps to the same location.

Filbertworm, Cydia latiferreana (Walsingham) (Lepidoptera: Tortricidae), is a key insect pest of hazelnuts in North America. Susceptibility to filbertworm was determined in 25 hazelnut cultivars represented by 44 trees at the USDA National Clonal Germplasm Repository in Corvallis, OR. In addition, shell thickness of 17 of the cultivars was measured at the thinnest and thickest points of the basal scar as well as at the side of the shell wall. Filbertworm infestation, which ranged from 2% to 89% per tree, as well as shell thickness varied significantly among the cultivars. Filbertworm infestation significantly increased with decreasing shell thickness at the basal scar, where filbertworm larvae typically penetrate the nut. The cultivar Siciliana had the overall thickest shells and lowest filbertworm infestation (6% ± 2% average for the cultivar), whereas ‘Frango 2’, ‘Casina’, and ‘Daviana’ had the thinnest shells and 76%, 55%, and 82% infested nuts, respectively. Nut infestation was not correlated with the thickness of the side wall. Our data suggest that a thicker basal scar in hazelnut cultivars contributes to resistance against filbertworm infestation.

The hundred-year history of the european hazelnut (Corylus avellana L.) industry in the Pacific northwestern United States is threatened by eastern filbert blight (EFB) caused by the fungus Anisogramma anomala (Peck) E. Müller. Marker-assisted selection has been extensively used for ‘Gasaway’ resistance in the hazelnut breeding program at Oregon State University. Concern over possible breakdown of this single resistance gene provides an incentive to look for new sources of resistance. OSU 759.010, a selection from the Republic of Georgia, has remained free of EFB after inoculations over several years. Random amplified polymorphic DNA (RAPD) markers linked to resistance were identified by screening primers against three resistant seedlings, three susceptible seedlings, and the parents of a segregating seedling population. For the progeny OSU 759.010 × OSU 653.068, 13 linked markers were identified. The markers most closely linked to resistance were 695-1800 on the proximal side and H12-640, 373-700, 349-450, and F08-700 on the distal side. Four of the five markers also segregated in the progeny OSU 759.010 × OSU 665.076, whereas H12-640 was monomorphic. Segregation for disease response in the first population showed a surplus of resistant seedlings, approaching a 3:1 ratio, with closely linked RAPD markers showing similar ratios. In the second population, the observed segregation for disease response and associated markers did not deviate from the expected 1:1 ratio. Based on cosegregation with simple sequence repeat (SSR) markers, resistance from OSU 759.010 was assigned to linkage group 2. Resistance to EFB from ‘Gasaway’ and ‘Ratoli’ was previously mapped to linkage groups 6 and 7, respectively. Therefore, OSU 759.010 provides a novel source of EFB resistance and markers 695-1800, 373-700, 349-450, and F08-700 have potential for use in marker-assisted selection to pyramid EFB resistance alleles.

‘Tonda Pacifica’ is a new hazelnut (Corylus avellana L.) cultivar for the kernel market. It was released by the Oregon Agricultural Experiment Station in April 2010. Compared with ‘Barcelona’, Oregon’s leading cultivar, ‘Tonda Pacifica’ has smaller trees, higher nut yield efficiency, smaller nuts and kernels, higher kernel percentage, fewer nut and kernel defects, better suitability for the blanched kernel market, and earlier nut maturity. Kernels have a crisp texture, highly rated flavor and nutty aroma. The kernel quality is similar to that of its Italian parent ‘Tonda Gentile delle Langhe’. The small kernel size is well suited to use in chocolate products and baked goods. ‘Tonda Pacifica’ is susceptible to eastern filbert blight caused by Anisogramma anomala (Peck) E. Muller.

Eastern filbert blight (EFB), caused by the pyrenomycete Anisogramma anomala (Peck) E. Muller, is a major disease problem and production constraint in orchards of European hazelnut (Corylus avellana L.) in Oregon’s Willamette Valley. Host genetic resistance is viewed as the most economical means of controlling this disease. A dominant resistance gene from “Gasaway” has been used extensively in the hazelnut breeding program at Oregon State University, but concern about the durability of a single resistance gene stimulated a search for new sources of resistance. “Ratoli,” a minor cultivar from Spain, showed no signs or symptoms of the fungus following a series of inoculations. The objective of this study was to study segregation for disease response in two progenies from crosses of Ratoli with susceptible selections and identify linked DNA markers. About half of the seedlings were resistant, suggesting control by a dominant allele at a single locus. A total of 900 random amplified polymorphic DNA (RAPD) primers and 64 amplified fragment length polymorphism (AFLP) primer combinations were screened. Four RAPD markers and two ALFP markers were identified and a linkage map constructed. On this map, disease resistance was flanked by AFLP marker C4-255 and RAPD marker G17-800 at distances of 0.4 cM and 2.8 cM, respectively. Based on co-segregation with SSR markers, Ratoli resistance was assigned to linkage group 7 while Gasaway resistance is on linkage group 6. Ratoli provides a novel source of EFB resistance, and robust RAPD marker G17-800 is useful for marker-assisted selection.

Eastern filbert blight (EFB), caused by the pyrenomycete Anisogramma anomala (Peck) E. Müller, is a devastating disease of European hazelnut (Corylus avellana L.) in the Pacific Northwest. Host genetic resistance from ‘Gasaway’ has been used extensively for breeding hazelnuts at Oregon State University. Concern over the durability of this single-gene resistance prompted a search for new sources of resistance. In this study, 86 accessions from 11 countries were evaluated for their response to greenhouse inoculation with the pathogen. Nine accessions showed complete resistance, including one from Chile (‘Amarillo Tardio’), two from Serbia (‘Crvenje’ and ‘Uebov’), one from southern Russia (OSU 495.072) and five from Moscow, Russia. These new sources of EFB resistance have geographically diverse origins and will broaden the genetic base of EFB-resistant hazelnut germplasm. The previously reported resistance of ‘Grand Traverse’ from Michigan and the susceptibility of ‘Closca Molla’ from Spain were confirmed.

Microsatellite-containing sequences for the Betulaceae (Betula, Corylus, and Alnus) were retrieved from GenBank and used to develop twelve new microsatellite marker primer pairs that amplified and were polymorphic in european hazelnut (Corylus avellana). The primer pairs were characterized using 50 european hazelnut accessions. Nine of these microsatellites that segregated in a mapping population were assigned to linkage groups. The 12 new primer pairs will be useful in genetic studies in Corylus and Betula. To investigate transferability of microsatellite primer pairs in the family Betulaceae, we assessed the ability of 129 simple sequence repeat (SSR) primer pairs (75 from Corylus, 52 from Betula, and two from Alnus) to amplify DNA of 69 accessions representing diverse taxa. Microsatellite primer pairs from Betula amplified 92% of Betula, 51% of Alnus, 41% of Corylus, 37% of Carpinus, 35% of Ostryopsis, and 34% of Ostrya accessions. In the 69 accessions, microsatellite primer pairs from Corylus amplified 81% of Corylus, 55% of Carpinus, 53% of Ostrya, 51% of Ostryopsis, 41% of Alnus, and 39% of Betula accessions. An additional 147 SSR primer pairs developed from Corylus, used to amplify a subset of 32 accessions, gave similar values: 92% in Corylus, 33% in Carpinus, 33% in Ostrya, 44% in Ostryopsis, 35% in Alnus, and 54% in Betula. The high transferability (>39%) of microsatellite primer pairs between Betula and Corylus will allow comparative studies of the two genera with the greatest economic importance.

Simple sequence repeat (SSR) or microsatellite markers have many applications in breeding and genetic studies of plants, including fingerprinting of cultivars and investigations of genetic diversity, and therefore provide information for better management of germplasm collections. They are repeatable, co-dominant, highly polymorphic, and technically simple to use. We developed more than 150 SSR markers for hazelnut (Corylus avellana) and characterized them using a diverse set of 50 accessions. Some hazelnut SSR loci amplify other genera in the Betulaceae. In this study we identify two sets of a dozen markers each that are useful for fingerprinting hazelnut accessions. Loci were chosen based on high heterozygosity, PIC values, low frequency of null alleles, coverage of the hazelnut genome, and ease of scoring. The number of alleles per locus for these 24 primers ranged from 5-16. Polymorphic information content (PIC) values ranged from 0.50 to 0.87. Most loci segregated 1:1, 1:2:1 or 1:1:1:1 in our mapping population, allowing us to assign them to a linkage group.

A diverse collection of 58 hazelnut accessions, including Corylus avellana L. and interspecific hybrids, were evaluated for their response to the eastern filbert blight pathogen Anisogramma anomala (Peck) E. Müller after greenhouse inoculation. Evaluations were made using enzyme-linked immunosorbent assay and visual inspection. Forty-five of these became infected, 12 remained free of infection, and one gave inconclusive results. The 12 accessions showing complete resistance were European hazelnuts ‘Culpla’ from Spain and CCOR 187 from Finland; C. americana × C. avellana hybrids ‘G081S’, CCOR 506, and Weschcke selections TP1, TP2 and TP3; C. colurna × C. avellana hybrids Chinese Trazels Gellatly #6 and #11; Turkish Trazel Gellatly #3 and backcross hybrid ‘Lisa’; and C. heterophylla var. sutchuensis × C. avellana hybrid ‘Estrella #1’. In a second test, exposure of potted trees under structures topped with diseased wood confirmed the complete resistance of ‘Santiam’, four pollinizers, and ‘Ratoli’. However, a few small cankers were observed on ‘Closca Molla’ from Spain and OSU 729.012, with resistance from C. californica (A.DC.) Rose, in contrast to the results of earlier greenhouse inoculations.

Six hundred five hazelnut (Corylus avellana L.) seedlings from a diverse germplasm collection made in the Russian Federation and the Crimean peninsula of the Ukraine were inoculated with the eastern filbert blight (EFB) pathogen Anisogramma anomala (Peck) E. Müller and their responses evaluated. Responses were rated on a scale of 0 to 5, in which 0 represents no sign of EFB and 5 represents all branches exhibiting cankers. At final evaluation, eight seedlings showed no signs of the pathogen or symptoms of the disease. Five additional seedlings expressed only very minor signs of the pathogen (rating = 1). The remainder ranged in disease expression from moderately to severely infected to dead with 89.7% (470 of 524) of the surviving seedlings rating 4 or 5. Of the 13 apparently resistant seedlings (rating 0 or 1), seven originated from nuts purchased from roadside vendors near Simferopol, Crimea, Ukraine; five from nuts purchased at an outdoor market near Krasnodar, Russia; and one from nuts obtained from the hazelnut breeding program of the Nikita Botanical Gardens, Yalta, Crimea, Ukraine. Random amplified polymorphic DNA (RAPD) markers generated by the primers UBC 152800 and OP AA12850, which are tightly linked to the single dominant resistance gene ‘Gasaway’, were not present in all 13 resistant seedlings, providing support, along with their geographic origins, that they represent novel sources of genetic resistance to EFB.

Eastern filbert blight (EFB), caused by the fungus Anisogramma anomala (Peck) E. Müller, is an important disease of european hazelnut (Corylus avellana L.) in the Pacific northwestern United States. In 1989, a chance seedling free of EFB was discovered adjacent to a severely diseased orchard near Troutdale, Ore. This selection, subsequently named `Zimmerman', was crossed with three susceptible selections. Based on morphological characters and incompatibility alleles, we speculated that `Zimmerman' (S1 S3) was a hybrid between `Barcelona' (S1 S2) and `Gasaway' (S3 S26). The three seedling populations were inoculated with spores of the pathogen in a greenhouse test and assayed by indirect enzyme-linked immunosorbent assay (ELISA) and by observation of canker incidence. The observed segregation fit a 3 resistant : 1 susceptible ratio in all three progenies, in contrast to the 1 : 1 ratio found when the resistant pollinizer `Gasaway' was crossed to susceptible genotypes. Random amplified polymorphic DNA (RAPD) marker UBC 152800 linked to the resistance gene in `Gasaway' co-segregated with the resistant phenotype in all three populations with 2%, 4%, and 6% recombination, respectively. Seed germination and transplanting records did not provide evidence of selection in favor of resistant seedlings. Pollen germination was 71% in `Gasaway', 29% in `Zimmerman', and 18% in `Barcelona', indicating possible selection at the gametophytic level. Subsequently 16 resistant seedlings of `Zimmerman' were crossed with the highly susceptible selection OSU 313.078. Segregation fit a 3 : 1 ratio in 14 of the 16 progenies, and showed a surplus of resistant seedlings in the other two. None showed a 1 : 1 segregation. Resistance co-segregated with two RAPD markers that flank the `Gasaway' resistance allele. To test allelism of resistance from `Gasaway' and `Zimmerman', VR 6-28 with resistance from `Gasaway' was crossed with `Zimmerman'. Eight resistant selections from this progeny were crossed with OSU 313.078. Five of the eight progenies segregated 3 : 1, two progenies segregated 1 : 1, and OSU 313.078 × OSU 720.056 gave only resistant offspring. The ratios indicate that OSU 720.056 is homozygous resistant and that `Zimmerman' and `Gasaway' share a common resistance allele. Reciprocal translocations have been reported in hazelnut cultivars, including `Barcelona', the leading cultivar in Oregon. `Zimmerman' appears to be a hybrid of `Barcelona' and `Gasaway', but because of cytogenetic abnormalities, `Zimmerman' may have inherited two copies of the chromosome region that contain the resistance locus and flanking RAPD markers. If the region containing the resistance were attached to two independent centromeres, a 3 : 1 segregation ratio for disease response and flanking markers would be expected, and we propose this as the most likely explanation. Resistance from `Gasaway' and `Zimmerman' has been called “immunity” or “complete resistance.” However, we noted a few seedlings with small cankers, nearly all of which lacked sporulating stromata. Flanking RAPD markers indicate that the resistance allele is present in these seedlings. Although not “immune” or “completely resistant,” `Gasaway' and `Zimmerman' transmit a very high level of resistance.

Three microsatellite-enriched libraries of the european hazelnut (Corylus avellana L.) were constructed: library A for CA repeats, library B for GA repeats, and library C for GAA repeats. Twenty-five primer pairs amplified easy-to-score single loci and were used to investigate polymorphism among 20 C. avellana genotypes and to evaluate cross-species amplification in seven Corylus L. species. Microsatellite alleles were estimated by fluorescent capillary electrophoresis fragment sizing. The number of alleles per locus ranged from 2 to 12 (average = 7.16) in C. avellana and from 5 to 22 overall (average = 13.32). With the exception of CAC-B110, di-nucleotide SSRs were characterized by a relatively large number of alleles per locus (≥5), high average observed and expected heterozygosity (Ho and He > 0.6), and a high mean polymorphic information content (PIC ≥ 0.6) in C. avellana. In contrast, tri-nucleotide microsatellites were more homozygous (Ho = 0.4 on average) and less informative than di-nucleotide simple sequence repeats (SSRs) as indicated by a lower mean number of alleles per locus (4.5), He (0.59), and PIC (0.54). Cross-species amplification in Corylus was demonstrated. These microsatellite markers were highly heterozygous and polymorphic and differentiated among genotypes of C. avellana irrespective of geographical origin. They will aid in fingerprinting genotypes of the european hazelnut and other Corylus species, genome mapping, and genetic diversity assessments.

ADDITIONAL INDEX WORDS. Corylus avellana, fi lbert, Anisogramma anomala, marker-assisted selection ABSTRACT. Eastern fi lbert blight (EFB), caused by Anisogramma anomala (Peck) E. Muller, is a devastating disease to european hazelnut (Corylus avellana L.) orchards in the Willamette Valley of Oregon. Selection OSU 408.040 showed no symptoms or signs of the fungus following greenhouse inoculations, and enzyme-linked immunosorbant assays (ELI- SAs) were negative. Segregation ratios in three progenies indicate that a single dominant gene controls the resistance. A total of 64 amplifi ed fragment length polymorphism (AFLP) primer combinations were screened using three resistant and three susceptible individuals as well as the parents of the cross OSU 245.098 × OSU 408.040. Primer combinations that showed no more than one recombinant in these six seedlings were investigated in 30 additional seedlings. Markers that showed

and subtropical fruit and nut species holds 7,685 accessions made up of cultivars, selections, and unique forms (Table 1). The collections include peach and nectarine [Prunus persica (L.) Batsch.], apricot (Prunus armeniaca L.), European plum (Prunus domestica L.), cherry-plum (Prunus cerasifera Ehrh.), sour cherry (Prunus cerasus Ehrh.), apple (Malus xdomestica Borkh.), pear (Pyrus communis L.), quince (Cydonia oblonga Mill.), almond [Prunus dulcis (Mill.) D. A. Webb.], walnut (Juglans regia L.), hazelnut (Corylus avellana L.), pomegranate (Punica granatum L.), fi g (Ficus carica L.), persimmon (Diospyros spp.), Chinese date (Zyzyphus jujuba Mill.), hardy kiwifruit (Actinidia spp.), pawpaw [Asimina triloba (L.) Dunal], lemon [Citrus limon (L.) Burm.], and feijoa or pineapple guava (Feijoa sellowiana Berg.). There are nine departments in the Gardens: fl ora and vegetation, dendrology and fl oriculture, subtropical fruit culture, essential oil-bearing and medicinal plants, agroecology Table 1. Number of accessions of temperate and subtropical fruit and nut species held in collections at the Nikita Botanical Gardens.