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- Author or Editor: Herb S. Aldwinckle x
Abstract
Apple seedlings of two progenies whose parentage was predominantly cultivated apple (Malus spp.) were forced under optimum growing conditions in the greenhouse. Without chilling, flowering started on occasional plants 16 months after germination. Manual defoliation induced synchronous bud break very efficiently. By 20 months after germination 86% and 68% of surviving seedlings in the two progenies had flowered. The phase change from juvenile to adult occurred 10-14 months after germination or possibly earlier.
Regeneration from apple (Malus × domestica Borkh.) M.26 leaf tissue was completely inhibited by (μg·ml-1) 1 geneticin, 5 kanamycin, 10 to 25 paromomycin, and 100 neomycin. nptII-transgenic M.26 had an increased tolerance to all four of the antibiotics tested, with inhibition of regeneration occurring at (μg·ml-l) 2.5 geneticin, 100 kanamycin, 375 paromomycin, and 375 neomycin. Paromomycin (100 to 250 μg·ml-l) and neomycin (250 μg·ml-1) significantly increased the amount of regeneration from nptII-transgenic M.26 apple leaf tissue. p35SGUS-INT, a plasmid with a chimeric b -glucuronidase gene containing a plant intron, was useful for studying the early events of apple transformation by eliminating GUS expression from Agrobacterium tumefaciens. It was used to determine that the optimal aminoglycoside concentrations for the selection of nptII-transgenic M.26 cells were (μg·ml-1) 2.5 to 16 kanamycin, 63 to 100 neomycin, and 25 to 63 paromomycin. Geneticin was unsuitable as a selective agent.
Abstract
Orchard production systems have experienced rapid and dramatic changes over the past 3 decades in both the techniques and geno-types used. We expect this rate of change to accelerate in the future. As production costs escalate, the demand will become ever stronger for systems providing early, heavy, reliable yield fo high-quality, easy-to-harvest fruit. These systems must, however, be reasonably priced and durable. The grower will be less able to afford tree losses, whether due to biotic, climatic, or edaphic hazard, and will need to economize on use of machinery and chemical pesticides. Growth regulators will be employed only where there are effective, economically justified, and environmentally acceptable.
The Geneva® Apple Rootstock Breeding program initiated in 1968 by Cummins and Aldwinckle of Cornell University and continued as a joint breeding program with the USDA-ARS since 1998, has released a new dwarf apple rootstock named Geneva® 41 or G.41. G.41 (a progeny from a 1975 cross of `Malling 27' × `Robusta 5') is a selection that has been tested at the N.Y. State Agricultural Experiment Station, in commercial orchards in the United States, and at research stations across the United States, Canada, and France. G.41 is a fully dwarfing rootstock with vigor similar to M.9 T337, but with less vigor than M.9 Pajam2. It is highly resistant to fire blight and Phytophthora with no tree death from these diseases in field trials or inoculated experiments. G.41 has also shown tolerance to replant disease. Its precocity and productivity have been exceptional, equaling M.9 in all trials and surpassing M.9 in some trials. It also confers excellent fruit size and induces wide crotch angles in the scion. It appears to be very winter hardy and showed no damage following the test winter of 1994 in New York. Propagation by layering in the stool bed G.41 is not consistent and may require higher layering planting densities or tissue culture mother plants to improve its rooting. G.41 also produces some side shoots in the stool bed. The nursery liners of G.41 produce a smaller tree than G.16 liners, but similar to M.9, which is very acceptable. Unlike G.16, G.41 is not sensitive to latent viruses. G.41 has similar graft union strength to M.9 and requires a trellis or individual tree stake when planted in the orchard. Suggested orchards planting densities with this rootstock are 2,000-4,000 trees/ha. This rootstock has been released for propagation and commercial sale by licensed nurseries.
The Geneva® Apple Rootstock Breeding program, which was initiated in 1968 by Dr. James Cummins and Dr. Herb Aldwinckle of Cornell University and which has been continued as a joint breeding program with the U.S. Dept. of Agriculture Agricultural Research Service (USDA-ARS) since 1998, has released a new semi-dwarfing apple rootstock which is named Geneva® 935 or G.935. G.935 (a progeny from a 1976 cross of `Ottawa 3' × `Robusta 5') is a selection that has been widely tested at the New York State Agricultural Experiment Station in Geneva, N.Y., in commercial orchards in the United States and at research stations across the United States and Canada. G.935 is a semi-dwarfing rootstock that produces a tree slightly larger than M.26. G.935 is the most precocious and productive semi-dwarf rootstock we have released. It has had similar yield efficiency to M.9 along with excellent fruit size and wide crotch angles. It showed no symptoms of winter damage during the 1994 test winter in N.Y. G.935 is resistant to fire blight and Phytophthora; however. it is susceptible to infestations by woolly apple aphids. G.935 has shown tolerance to replant disease complex in several trials. It has good propagation characteristics in the stool bed and produces a large tree in the nursery. G.935 has better graft union strength than M.9, but will require a trellis or individual tree stake in the orchard to support the large crops when the tree is young. G.935 will be a possible replacement for M.26. Suggested orchards planting densities with this rootstock are 1,500-2,500 trees/ha. It has been released for propagation and sale by licensed nurseries. Liners will be available in the near future.
Abstract
In 1973, Ross Byers at VPI's Winchester laboratory and rootstock breeder Jim Cummins and plant pathologist Herb Aldwinckle at Cornell's Geneva station initiated a cooperative project to explore Malus for resistance to the pine vole, Microtus pinetorum Leeone. The Geneva breeders furnished more than 300 clones during the next 5 years, representing 12 Malus species and including many interspecific hybrids. Byers and his associates tested the voles' responses to the many choices offered. Throughout the several years of testing, the voles consistently exhibited a strong nonpreference for a Japanese crabapple, now introduced as ‘Novole’, and later for many of its hybrids (1, 6).
To root tissue-cultured apple cultivars, shoots from proliferating cultures were first transferred to root induction medium with IBA for 1 week in the dark. Shoots were later transferred to the same medium without IBA and incubated under light for elongation of the roots. Rooted shoots were then transferred to Jiffy-7s supplemented with biological plant protectant and fertilizer, and incubated in plastic humidity trays. After 2 to 3 weeks, plants were transferred to pots and covered with plastic bags to facilitate acclimation. This technique has resulted in 70% to 100% of shoots selected in vitro producing vigorously growing, healthy plants in the greenhouse. Chemical name used: indolebutyric acid (IBA).
The overall goal of our research is to develop an efficient transformation and regeneration system for `McIntosh' apple. The first objective was to determine the optimum combination of Gelrite (G) and agar (A) to maximize regeneration and minimize vitrification. Treatments included the following combinations of agar (in g–liter–1) and Gelrite (in g–liter–1): 1) 7 and 0; 2) 5.25 and 0.625; 3) 3.5 and 1.875; 4) 1.75 and 1.875; and 5) 0 and 2.5. There were 10 replications, and a single petri plate containing six leaf pieces was the unit of replication. Both 5.25(A) and 0.62(G) and 3.5(A) and 1.25(G) provided high regeneration of healthy, nonvitrified shoots. Since modification of media affects the concentration of antibiotics used in selection due to precipitation of antibiotics, the second objective was to determine the optimal concentration of antibiotic for the selection and regeneration of transformed `McIntosh' on gelrite–agar-based media. Kanamycin was tested at 0, 10, 25, 50, 75, and 100 μg–ml–1 and paromomycin was tested at 0, 50, 100, 150, 200, and 250 μg–ml–1. Antibiotic selection will be discussed relative to optimum concentration and efficiency of selection.
Seven nptII and gus transgenic lines of the apple (Malus ×domestica Borkh.) rootstock Malling 7 (M.7) were examined by glucuronidase (GUS) histochemical testing and a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). These lines had different amounts of neomycin phosphotransferase II (NPTII). The amounts of NPTII among lines was positively correlated with the ability of the transgenic lines to regenerate in the presence of kanamycin, paromomycin, or geneticin. Regenerants derived from transgenic lines also varied greatly in GUS expression. The apical portion of regenerant stem tissues had stronger GUS staining than the basal portion of stem. All regenerated tissue of T1, a transgenic line originally classified as a uniform GUS staining line, showed non-GUS staining, while the regenerated tissues of chimeric transgenic lines showed nonstaining, chimeric staining, or uniform GUS staining, indicating the potential to select uniform GUS staining lines from chimeras. Polymerase chain reaction (PCR) indicated the gus gene was present in GUS negative (nonstaining) lines. Negative PCR results with primers derived from vir G of Agrobacterium tumefaciens, and failure to isolate A. tumefaciens from M.7 transgenics indicated that PCR and GUS staining results were not due to A. tumefaciens. A modified PCR methylation assay (MPMA) indicated that methylation of cytosines of the CCGG site in the gus gene, and in the border between the CaMV35S promoter and the gus gene, was positively correlated with complete gus gene silencing (nonstaining lines). However, the MPMA indicated that methylation was not always associated with variable GUS expression, suggesting that chimeric staining could be due to a mixture of transformed and nontransformed cells in some transgenic lines.
Bulked segregant analysis was used to identify RAPD markers that display tight linkage to the Vf gene in apple (Malus sp.) that confers resistance to five races of apple scab [Venturia inaequalis (Cke.) Wint.]. We identified several new RAPD markers linked to Vf. The most tightly linked marker in the test population, S52500, was cloned and sequenced. A linkage map of the Vf region was developed using these markers, RAPD markers previously described by other laboratories, and the isozyme locus Pgm-1. An assay was developed for Vf by multiplexing the two markers closely flanking the Vf locus. This assay has a theoretical `escape' value (discarding a resistant plant) of 3% and an error rate (selection of a susceptible plant) of 0.02%.