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William C. Johnson, Phil L. Forsline, Herb S. Aldwinckle, William C. Johnson, Phil L. Forsline, H. Todd Holleran, Terence L. Robinson and John J. Norelli

In 1998, the USDA-ARS and Cornell Univ. instituted a cooperative agreement that mobilized the resources for a jointly managed apple rootstock breeding and evaluation program. The program is a successor to the Cornell rootstock breeding program, formerly managed by Emeritus Professor of Horticultural Sciences James N. Cummins. The agreement broadens the scope of the program from a focus on regional concerns to address the constraints of all the U.S. apple production areas. In the future, the breeding program will continue to develop precocious and productive disease-resistant rootstock varieties with a range of vigor from fully dwarfing to near standard size, but there will be a renewed emphasis on nursery propagability, lodging resistance, tolerance to extreme temperatures, resistance to the soil pathogens of the sub-temperate regions of the U.S., and tolerance to apple replant disorder. The program draws on the expertise available at the Geneva campus through cooperation with plant pathologists, horticulturists, geneticists, biotechnologists, and the curator of the national apple germplasm repository. More than 1000 genotypes of apple rootstocks are currently under evaluation, and four fire blight- (Erwinia amylovora) resistant cultivars have been recently released from the program. As a service to U.S. apple producers, rootstock cultivars from other breeding programs will also be evaluated for productivity, size control, and tolerance to a range of biotic and abiotic stress events. The project will serve as an information source on all commercially available apple rootstock genotypes for nurseries and growers.

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Warren F. Lamboy, Jing Yu, Phil L. Forsline and Norman F. Weeden

One of the primary progenitors of the cultivated apple (Malus ×domestica) is M. sieversii, a species native to the forested regions of Central Asia. Despite the horticultural importance of M. sieversii, little is known about its genetic variation. In this study, isozyme diversity at 18 loci was determined for 259 open-pollinated offspring belonging to 31 different maternal half-sib families collected from 14 different populations in 4 regions of central Asia. Genetic diversity statistics were computed from the resulting allele and phenotype frequencies. Cluster analysis of half-sib families showed that there was some grouping based on geographic region, but 16 of the half-sib families were most closely related to half-sib families from other regions. AMOVA, the analysis of molecular variance, indicated that most of the enzyme variability (85%) was attributable to differences among half-sib families within populations, none could be assigned to populations within regions, and 15% was due to differences among regions. In addition, no alleles were found that were both fixed in a region and unique to that region. These results suggest that plants belonging to M. sieversii effectively form a single panmictic population. Thus, a thorough sampling of a few large populations will efficiently capture most of the genetic diversity present in wild M. sieversii.

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Warren F. Lamboy, Jing Yu, Phil L. Forsline and Norman F. Weeden

One of the primary progenitors of the cultivated apple is Malus sieversii L., a species native to the forested regions of central Asia. Despite the horticultural importance of M. sieversii, little is known about genetic variation in this species. In this study, allozyme diversity at 18 loci was determined for 259 seedlings belonging to 31 sib families, each consisting of the set of offspring from a different open-pollinated maternal (seed) parent. Maternal parents belonged to 14 populations from four geographic regions. Genetic diversity statistics were computed from the resulting allele and phenotype frequencies. Cluster analysis of sib families showed that there was some grouping based on geographic region, but 16 of the sib families were most closely related to sib families from other regions. Analysis of molecular variance (AMOVA) indicated that 85% of the enzyme variability was due to differences among sib families within populations and 15% was due to differences among regions. No variability could be assigned to differences among populations within regions. In addition, no alleles were found that were fixed in a region and unique to that region. These results suggest that plants belonging to M. sieversii effectively form a panmictic population. Consequently, a thorough sampling of a few large populations will efficiently capture most of the genetic diversity present in wild M. sieversii.

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Todd C. Einhorn, Cecil Stushnoff, Ann E. McSay, Phil L. Forsline, Sam Cox, Joel R.L. Ehrenkranz and Loretta Sandoval

Phlorizin is known for its role in reducing glucotoxicity and has a long history of use in diabetes research. In addition, its contribution to the pool of total phenolics adds to the overall health benefits attributed to fruit. Phlorizin is limited to Rosaceae family plants, of which apple comprises its current commercial source; however, limited information exists regarding its biodiversity among apple taxa. A subset of 22 taxa from a core collection of apple accessions representative of the global genetic diversity of apple was used to investigate the biodiversity of phlorizin present in apple shoots and in fruit relative to total phenolic content and free radical scavenging capacity. Fruit and shoots were harvested from the USDA Plant Genetic Resources Unit in Geneva, N.Y. Validation and quantification of phlorizin was conducted using a rigorous high-pressure liquid chromatography (HPLC) procedure. Total phenolics in fruit, assayed using a Folin-Ciocalteu method and expressed as gallic acid equivalents, ranged from 227 to 7181 mg·L-1

and were strongly related to 2,2' azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) antioxidant capacity for the core collection (r= 0.778). On a molar basis, phlorizin had lower antioxidant capacity than other major phenolic compounds present in apple fruit, but was more effective than ascorbic acid. Phlorizin yield in dormant apple shoots, expressed as percent weight, ranged from 0.9% to 5.5%. A rapid, 96 well micro-plate spectrophotometric assay was also developed to aid in the screening of multiple samples for selection of high phlorizin yielding apple taxa. Spectrophotometry overestimated phlorizin content as expected, but the calibration curve between HPLC and spectrophotometry was acceptable, r 2 = 0.88.

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Stan C. Hokanson, Phil L. Forsline, James R. McFerson, Warren F. Lamboy, Herb S. Aldwinckle and Aimak D. Djangaliev

Malus sieversii, the main progenitor of domesticated apple, is native to areas in Central Asia. To better represent Malus wild germplasm in the USDA–ARS germplasm collections, maintained in Geneva, N.Y., a cooperative project was initiated with the Republic if Kazakhstan to collect and assess that country's wild populations of M. sieversii and to develop more secure in situ reserves to complement ex situ holdings in the United States and Kazakhstan. To date, four exploration trips to the region have included participants from the United States, Kazakhstan, Canada, New Zealand, and South Africa. Four Kazkh scientists have toured USDA–ARS sites, exchanged information, and collected germplasm in the United States greenhouse screens of 1600 have revealed potentially new sources of resistance to apple scab, cedar apple rust, and fire blight. An isozyme analysis of maternal half-sib families from four regions suggests the populations of M. sieversii collected represent a single panmictic population, with over 85% of total genetic variation due to differences among families. The most recent collections in 1995 were directed towards more ecologically diverse regions, including a site (Tarbagatai) at the most northern limit for M. sieversii equivalent to northern Minnesota in the United States. Some trees in this region produced fruit nearly 70 mm in diameter with excellent aroma, firmness, and color. This germplasm is being systematically characterized for horticultural traits, pest and disease resistance, and molecular markers.