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M.E. Garcia and Linda A. Boccuzzo

Hardiness testing of the wood of deciduous fruit trees has been conducted using a variety of techniques. In our studies, the objective was to determine an efficient method of determining freezing injury for apple (Malus × domestica Borkh.) wood. We tested 1-year old wood of two cultivars: Liberty and RedMax. The wood was tested over the course of 2 years (1998 and 1999). Collection began in the late fall and continued throughout the winter (until it was determined full hardiness had been achieved) and then again in the early spring. The wood was cut into 1-cm sections and frozen. The artificial freezing was conducted in an ethanol bath, with the temperature lowered at 5 °C/h. Samples were removed in 3-min intervals. After freezing, the wood was acclimated to 4 °C for 12 h. Three tests were conducted to determine the hardiness/injury to the tissues. The tests used were: discoloration, callus growth and vital staining (with 2,3,5-triphenyltetrazolium chloride). This was a split block design with samples collected randomly from each tree. Four replicates (12 trees) of each cultivar were tested. Results showed that the callus test predicted the same LT50 as the other two tests, discoloration and vital staining. Discoloration was not easy to differentiate and was the most time-consuming. The callus grown by the apple wood was easily formed and distinguished. The callus test does not require the tetrazolium stain; therefore, one less step was needed in comparison to the vital staining test. This reduced testing time by over 6 h.

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Carole L. Bassett, D. Michael Glenn, Philip L. Forsline, Michael E. Wisniewski and Robert E. Farrell Jr

then by humans over the silk road trade routes into Europe. The modern cultivated apple ( Malus × domestica Borkh.) is believed to have been domesticated in Turkestan, now Kazakhstan, Kyrgyzstan, Uzbekistan, Turkmenistan, and Tajikistan ( Harris et al

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Shiow Y. Wang, Miklos Faust and Michael J. Line

The effect of Indole-3-acetic acid (IAA) on apical dominance in apple (Malus domestica Borkh.) buds was examined by studying changes In proton density (free water) and membrane lipid composition in lateral buds. Decapitation induced budbreak and enhanced lateral bud growth. IAA replaced apical control of lateral bud paradormancy. Maximal inhibition was obtained when IAA was applied immediately after the apical bud was removed. Delaying this application weakens the effect of IAA. An increase in proton density in lateral buds was observable 2 days after decapitation, whereas the change in membrane lipid composition occurred 4 days later. Decapitating the terminal bud induced an increase in membrane galacto- and phospholipids. and the ratio of unsaturated to corresponding saturated fatty acids. Decapitation also induced a decrease in the ratio of free sterols to phospholipids in lateral buds. Application of IAA to the terminal end of decapitated shoots inhibited the increase of proton density and prevented changes in the membrane lipid composition of lateral buds.

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Q. Liu, S. Salih, J. Ingersoll, R. Meng, L. Owens and F. Hammerschlag

Transgenic `Royal Gala' apple (Malus × domestica Borkh.) shoots were obtained by Agrobacterium-mediated gene transfer using the plasmid binary vector pGV-osm-AC with a T-DNA encoding a chimeric gene consisting of a secretory sequence from barley-amylase joined to the modified cecropin MB39 coding sequence. Shoots were placed under the control of a wound-inducible, osmotin promoter from tobacco. The integration of the cecropin MB39 gene into apple was confirmed by Southern blot analysis. The transformation efficiency was 1.5% when internodes from etiolated shoots were used as explants and 2% when leaf explants were used. Both non- and transgenic tetraploid plants were produced by treatment of leaf explants with colchicine at 25 mg·L-1, and polyploidy was confirmed by flow cytometry. Of the diploid transgenics, three of seven were significantly more resistant to Erwinia amylovora than the non-transgenic `Royal Gala' control. Also, in one instance, a tetraploid transgenic was significantly more resistant than the diploid shoot from which it was derived.

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Edwin J. Reidel*, Brian G. Ayre, E. Robert Turgeon and Lailiang Cheng

Sorbitol (d-glucitol) is the major end product of photosynthesis in apple (Malus domestica Borkh.), as well as the predominant phloem-translocated carbohydrate. The mechanism by which sorbitol is phloem-loaded for transport to heterotrophic sink tissues is unknown. We hypothesized that a plasma membrane-bound H+/sorbitol symporter mediates apoplastic phloem-loading of sorbitol. To discover genes potentially encoding sorbitol transporters, a cDNA library was constructed from mature `Gala' apple leaves. A homologous probe was synthesized via PCR with primers were designed against the cherry fruit sorbitol transporter, PcSot1, and using library lysate as template. From an initial plating of approximately 5 × 105 clones, twelve positives were identified after three rounds of hybridization screening. Following single-pass, 5' end sequencing, the clones were sorted into four contiguous sequences. One clone was chosen from each contig for complete sequencing. The four clones, provisionally named MdSOT1-4 (Malus domesitca Sorbitol Transporter), potentially encode full-length cDNAs for sorbitol transporters: Translated-BLAST searching (blastx) revealed that the open reading frames encode the complete Pfam sugar transporter domain, and the most significant alignments are with sequences encoding known- and putative polyol and sugar transporters.

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Mekjell Meland and Clive Kaiser

‘Summerred’ apples (Malus domestica Borkh.) are highly susceptible to biennial bearing if not properly thinned. This results in erratic yields and also affects fruit quality adversely. Between 2003 and 2005, ‘Summered’/‘M9’ trees were treated with ethephon at concentrations of 250, 375, and 500 mg·L−1 when most king flowers opened (≈20% bloom) or at concentrations of 500, 625, and 750 mg·L−1 when the average fruitlet size was 10 mm in diameter. The experiment was conducted with 2.5-m height slender spindle trees sprayed to the point of runoff with a hand applicator only when temperatures exceeded 15 °C. Within 2 weeks after the second application, fruit set was reduced linearly with increasing concentrations of ethephon to less than one fruitlet per cluster at the highest concentrations used. Most thinning treatments reduced fruit set significantly compared with unthinned trees. Fruit numbers per tree decreased significantly with increasing ethephon concentrations, and the highest concentrations of ethephon applied during bloom or when the average fruitlet size was 10 mm in diameter resulted in overthinning. Yield results confirmed the fruit set response in which yield reductions were significant at the highest concentrations of ethephon (2.1 kg/tree) compared with hand-thinned trees (7.3 kg/tree) in 2005. All thinning treatments resulted in higher percentage of fruits larger than 60 mm diameter compared with unthinned control fruit. Thinning resulted in significantly higher soluble solid contents, and this was especially so for hand-thinned trees. Other fruit quality parameters like yellow–green background color did not show a clear response to thinning. Return bloom was, however, improved on all thinned trees. It is recommended that ethephon be applied at a rate of 375 mg·L−1 when king flowers open or at a rate of 625 mg·L−1 when the average fruitlet size is 10 mm in diameter. This thins ‘Summerred’ apples to a target of approximately five fruits/cm2 per trunk cross-sectional area or 50 to 70 fruits per 100 flower clusters without impacting on fruit quality, yield, or return bloom the next year.

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Neal Mays, Curt Richard Rom, Kristofor R. Brye, Mary C. Savin and M. Elena Garcia

soil erosion ( Carter, 2002 ; Karlen et al., 1992 ; Kemper and Rosenau, 1986 ). Surface crusting and erosion may be reduced or eliminated in orchards with application of non-living groundcover mulches. Mulches useful for organic apple ( Malus

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Manfredo J. Seufferheld, Cecil Stushnoff and Philip Forsline

Cryopreservation of mature dormant vegetative buds is a useful method to preserve germplasm of a large number of cold hardy apple cultivars. However, cold tender cultivars have proven to be much more difficult to cryopreserve. Eight cultivars were harvested in September 1993 at Geneva, NY before developing cold hardiness naturally. The twigs were encapsulated with 5% alginate and treated with stepwise imbibition of 0.5 to 1.0 M sucrose. The samples were desiccated over glycerol at 0C. Half of the samples were plunged directly into liquid nitrogen (IN) and the other half were first cooled slowly to -30C. The twigs that had been exposed to prefreezing conditions showed the highest survival (20 to 100%). The samples that were plunged directly in LN survived poorly (0 to 20 %). Samples without encapsulation and no sucrose imbibition had 0% survival. We conclude that this protocol opens up the possibility to expand cryopreservation of cold tender apple cultivars, presently grown only in field gene banks, at great expense and inconvenience.

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Xuetong Fan, J.P. Mattheis, M.E. Patterson and J.K. Fellman

Several strains of Fuji apples were harvested weekly from September through October in 1990 and 1991, and evaluated for maturation and quality after 1 and 7 days at 20 °C following harvest and storage in atmospheres of 0.5%, 1.0%, 2.0% O2 and air. Results showed that Fuji apples have very low ethylene production rates and little firmness loss during maturation. A change in the postharvest respiration pattern preceded the increase ethylene synthesis. Oxygen concentration during storage directly affected apple respiration rate after removal from storage. Ethylene production rates and internal ethylene concentrations indicated that the apples were still in the preclimacteric stage after 7 to 9 months storage at 0.5%, 1.0%, or 2% O2. Fuji apples develop watercore and tend to have a particular type of corebrowing during maturation on the tree, or during and after storage. The cause is unknown.

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Jacques J. Crabbe

The flushing behavior of shoot growth and its consequences on shoot differentiation are important features in fruit tree development, with regard to flowering ability. In this respect, two different approaches were applied to young `M26' apple trees. First, poorly branched 2-year-old trees were headed back, either in the second-year or in the first-year wood, at different times from right before to 6 weeks after budbreak. Early pruning resulted in rapid and prolonged regrowth, with a final very similar shaping of the tree to that of the intact controls. Late pruning, in contrast, leads to a two-step reaction (late spring and summer flushes), sometimes stronger on 2-year-old than on 1-year-old wood. In a second experiment, buds and young shoots were sampled on pruned trees in locations where they could be supposed to remain short shoots or grow long, with one or two flushes. They were weighed, their leaves and internodes measured, and the plastochron evaluated. During budbreak and the first month afterwards, shoot differentiation appears achieved. The primary difference between long and short shoot types does not consist in faster internode elongation but, rather, in faster production (reduced plastochron) of larger leaves.