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  • Author or Editor: Shawn A. Mehlenbacher x
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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.

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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.

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Random amplified polymorphic DNA (RAPD) markers were identified for self-incompatibility (SI) alleles that will allow marker-assisted selection of desired S-alleles and assist in cloning the locus responsible for the sporophytic SI displayed in hazelnut (Corylus avellana L.). DNA was extracted from young leaves collected from field-planted parents and 27 progeny of the cross OSU 23.017 (S1 S12) × VR6-28 (S2 S26). Screening of 10-base oligonucleotide RAPD primers was performed using bulked segregant analysis. DNA samples from six trees each were pooled into four “bulks,” one for each of the following: S1 S2, S1 S26, S2 S12, and S12 S26. “Super bulks” of twelve trees each for S1, S2, S12, and S26 then were created for each allele by combining the appropriate bulks. The DNA from these four super bulks and also the parents was used as a template in the PCR assays. Amplification products were electrophoresed on 2% agarose gels and photographed under UV light after ethidium bromide staining. 200 primers were screened and one RAPD marker each was identified for alleles S2 (OPI-07700) and S1 (OPJ-141700).

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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.

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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.

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