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J.R. Ballington', C.M. Mainland, S.D. Rooks, A.D. Draper, and G.J. Galletta

1 Professor. 2 Blueberry Breeding Researcher. 3 Research Geneticist, retired. 4 Research Geneticist. Paper no. 12386 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC 27695 7602. We express our

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Jessica L. Gilbert, James W. Olmstead, Thomas A. Colquhoun, Laura A. Levin, David G. Clark, and Howard R. Moskowitz

pounds harvested for fresh production valued at $850.8 million U.S. in 2012 ( USDA, 2013 ). Blueberry acreage continues to increase worldwide in accordance with growing demand for these antioxidant-rich fruit. Although native to North America, breeding

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Paul M. Lyrene

were bred by recurrent selection in areas with warm winters. Breeding started with hybrids between northern highbush cultivars and low-chill wild blueberries from the southeastern United States, notably V. darrowi ( Sharpe and Darrow, 1960 ; Sharpe

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Amnon Levi, Elizabeth Ogden, and Lisa J. Rowland

Efforts are underway to develop genetic linkage maps for two interspecific blueberry populations (Vaccinium darrowi × V. elliottii and V. caesariense-derived populations). To date, 72 RAPD markers have been mapped, and another 200 markers have been identified as suitable for mapping in the V. darrowi × V. elliottii-derived population. Inheritance of 40 RAPD markers has been followed, and additional 40 RAPD markers have been identified as suitable for mapping in the V. darrowi × V. caesariense population. These two populations are comprised of individual plants that should have a wide range of chilling requirements. At a later date, plants will be classified according to their chilling requirements to identify RAPD markers that cosegregate with chilling requirements. Presently, a bulked-segregant analysis is being performed on a tetraploid breeding population (primarily V. corymbosum) to identify RAPD markers linked to chilling requirement genes.

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A.W. Stretch, M.K. Ehlenfeldt, and V. Brewster

Fifty-five highbush blueberry (Vaccinium corymbosum L.) cultivars and selections were evaluated over 2 years for their resistance to the shoot blighting phase of mummy berry disease [Monilinia vaccinii-corymbosi (Reade) Honey]. Blight incidence in 1993 ranged from 1% to 78% and differences among cultivars were significant. In 1994, infection levels were lower and ranged from 0% to 43%, again with significant differences among the entries. Several cultivars exhibited mummy berry blight resistance in both years. Ranking most resistant to less resistant were `Jersey', `Elliott', `Bluejay', `Duke', `Stanley', `Darrow', `Meader', and `Angola'. Among the cultivars consistently blightsusceptible were `Bluehaven', `Bluegold', `Northblue', `Croatan', `Northsky', `Sierra', `Harrison', `Coville', and `Murphy'. The consistent resistant reaction of certain cultivars indicates that they may be suitable as parents for introducing resistance into a breeding program. The evaluation methodology developed in these tests should be useful in screening germplasm for new sources of resistance and evaluating segregating progeny from crosses.

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A.W. Stretch and M.K. Ehlenfeldt

Sixty-eight highbush blueberry (Vaccinium corymbosum L.) cultivars and selections were evaluated over 3 years for their resistance to the fruit infection phase of mummy berry disease [Monilinia vaccinii-corymbosi (Reade) Honey]. Average incidence of fruit infection under test conditions was 34.3% in 1995, 14.4% in 1996, and 27.9% in 1997, with significant differences occurring among clones in all 3 test years. Several cultivars exhibited consistent resistance to mummy berry fruit infection across all years of testing. `Northsky', `Reka', `Northblue', `Cape Fear', `Bluegold', `Puru', and `Bluejay' were among the most resistant, and `Atlantic', `Berkeley', `Herbert', and `E-176' were among the most susceptible. The consistent resistant reaction of certain cultivars indicates that they may be suitable as parents for introducing resistance into a breeding program. No significant correlation was observed between blighting resistance and fruit infection resistance.

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Rajeev Arora, Lisa J. Rowland, Elizabeth L. Ogden, Anik L. Dhanaraj, Calin O. Marian, Mark K. Ehlenfeldt, and Bryan Vinyard

Loss of freeze tolerance, or deacclimation, is an integral part of winter survival in woody perennials because untimely mid-winter or spring thaws followed by a hard freeze can cause severe injury to dehardened tissues. This study was undertaken to investigate deacclimation kinetics, particularly the timing and speed, of five blueberry (Vaccinium L.) cultivars (`Bluecrop', `Weymouth', `Ozarkblue', `Tifblue', and `Legacy'), with different germplasm compositions and mid-winter bud hardiness levels, in response to an environmentally controlled temperature regime. Based upon bud cold hardiness evaluations in 2000 and 2001, `Tifblue', a Vaccinium ashei Reade cultivar, was one of the least hardy and the fastest to deacclimate; `Bluecrop', a predominantly V. corymbosum L. cultivar, was the most hardy and the slowest to deacclimate; and `Ozarkblue', a predominantly V. corymbosum cultivar but including southern species V. darrowi Camp. and V. ashei, was intermediate in speed of deacclimation. `Weymouth' (predominantly V. corymbosum) and `Legacy' (73.4% V. corymbosum and 25% V. darrowi) were slow to intermediate deacclimators. Deacclimation rates did not correlate strictly with mid-winter bud hardiness. Data suggest that the southern germplasm component V. ashei may be responsible for the observed faster deacclimation whereas both southern species, V. darrowi and V. ashei, may contribute genes for cold sensitivity. Strong positive correlations between stage of bud opening and bud cold hardiness existed in both years (r = 0.90 and 0.82 in 2000 and 2001 study, respectively). Previously identified major blueberry dehydrins, 65-, 60-, and 14-kDa, progressively decreased in their abundance during incremental dehardening in `Bluecrop', `Weymouth', and `Tifblue'. However, down-regulation of the 14-kDa dehydrin most closely mirrored the loss in cold hardiness during deacclimation, and, therefore, may be involved in regulation of bud dehardening. Because differences in deacclimation rate were clearly evident among the genotypes studied, rate of deacclimation of the flower buds of blueberry should be an important consideration in breeding to improve winter survival.

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James J. Polashock and Matthew Kramer

Stem diseases of blueberry (Vaccinium spp.) can cause significant crop loss as well as loss of entire bushes. Stem diseases are also more difficult to control with fungicides than foliar or fruit diseases. A screening program was initiated to test blueberry cultivars for resistance to two pathogenic fungi: botryosphaeria stem blight and phomopsis twig blight. An attached stem assay was developed to compare the host response with both fungi. The relative resistance of 50 blueberry cultivars was assessed using stem lesion lengths, analyzed on a log scale, taken at 4 weeks postinoculation. For Botryosphaeria stem blight, mean lesion length ranged from about 10 mm in resistant cultivars to about 140 mm in susceptible cultivars. The half-high cultivars Northsky, Northblue, and Chippewa, and the lowbush cultivar Putte were among the most resistant. Phomopsis twig blight lesions ranged in mean length from about 18 to 98 mm. Similar to results for Botryosphaeria stem blight, resistance was limited to half-high (`Northsky' and `Chippewa') and lowbush (`Blomidon', `Chignecto', and `Cumberland') cultivars. Individual cultivars resistant to one pathogen were not necessarily resistant to the other; although, overall, the resistances were correlated. Approximate 95% confidence intervals were established for all cultivars to predict mean performance across years. The cultivars tested varied in resistance, but the largest single factor affecting lesion length was the fungal isolate used for inoculations. These data enable us to identify cultivars resistant to both diseases that can be used for planting in problem areas, as well as selection of parental material for breeding cultivars with improved resistance.

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Nicholi Vorsa and Richard Novy

Vaccinium darrowi (D) is a wild blueberry species with low chilling requirements for budbreak, and heat and drought tolerance. Breeding efforts to incorporate these desirable traits into cultivated blueberry (V. corymbosum) (C) would be facilitated with a better understanding of the genomic homology between the two species. An interspecific tetraploid hybrid (CCDD, 2n=4x=48) was used to evaluate genome homology and interspecific recombination. Pollen mother cells examined at diakinesis and early metaphase I exhibited an average of 4.6 chain bivalents, 11.4 ring bivalents, 1.0 chain quadrivalent, and 3.0 ring quadrivalents. This data most closely fits a chromosome pairing model in which there is a greater pairing affinity between homologues than homoeologues. An analysis of the inheritance of 14 RAPD markers unique to V. darrowi in 72 backcross progeny of the V. darrowi–corymbosum hybrid also supported the pairing model: Seven of the 14 markers deviated significantly from tetrasomic inheritance ratios, expected if chromosome pairing was totally random. On the basis of the cytogenetic and RAPD analyses, the genomes of V. darrowi and V. corymbosum are divergent from one another, with preferential pairing within genomes. This outcome suggests there may be difficulty in breaking undesirable linkages when introgressing desirable traits from V. darrowi to V. corymbosum.

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Ann Marie Connor, James J. Luby, and Cindy B.S. Tong

Narrow-sense heritability and among-family and within-family variance components were estimated for antioxidant activity (AA), total phenolic content (TPH), and anthocyanin content (ACY) in blueberry (Vaccinium L. sp.) fruit. AA, TPH, and ACY were determined in the parents and in 10 offspring from each of 20 random crosses for each of 2 years at Becker, Minn. Offspring-midparent regression analysis provided combined-year heritability estimates of 0.43 ± 0.09 (P ≤ 0.0001) for AA, 0.46 ± 0.11 (P ≤ 0.0001) for TPH, and 0.56 ± 0.10 (P ≤ 0.0001) for ACY. Analyses of variance delineated variation among and within families for AA, TPH, and ACY (P ≤ 0.001). Year-to-year variation in the means for all offspring genotypes was not significant for AA or TPH, but there were changes in rank between years for families and for offspring within families for these traits. Year-to-year variation in the mean for all offspring genotypes was significant for ACY, but rank changes were observed only among offspring within families, not among families. In total, 18 of 200 offspring from 7 of the 20 crosses were transgressive segregants for AA, exceeding the higher parent of the cross by at least two sds. Estimates of variance components showed that variation among families accounted for 24% to 27% of total variance for the three traits. However, variation within families was greater than that among families, accounting for 38% to 56% of total variance for the three traits. These results suggest that increasing antioxidant activity in blueberry through breeding is feasible, and that the breeding strategies utilized should exploit the large within-family variation that exists.