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Amy F. Fulcher, Thomas G. Ranney, James D. Burton, James F. Walgenbach, and David A. Danehower

Japanese beetles (JB), Popillia japonica Newman, are destructive, highly polyphagous herbivores that show a general preference for Rosaceous plants. Choice and nochoice feeding assays were conducted to determine the level of resistance among 10 taxa of Malus spp. Mill. Under no-choice conditions, M. baccata (L.) Borkh. `Jackii', M. × `Hargozam' Harvest Gold and M. transitoria (Balatin) Schneider `Schmitcutleaf' Golden Raindrops were highly resistant, with <2 cm2 leaf area consumed in 24 hours. M. × `Radiant' was highly susceptible, with 7.6 cm2 consumed, and the remaining six cultivars were intermediate. Under choice conditions, eight taxa were resistant with <10% defoliation, M. × `Red Splendor' was intermediate with 26%, and M. × `Radiant' was susceptible with 73% defoliation. Feeding responses to eight individual phenolics were tested in artificial diets over a range from 0 to 100 mm. Phloridzin, phloretin, naringenin, and catechin were all feeding deterrents, whereas quercetin and rutin were feeding stimulants. Chlorogenic acid stimulated feeding at low concentrations and deterred feeding at higher concentrations (i.e., a peak response). Kaempferol had no effect. Analysis of endogenous foliar phenolics showed considerable variation in concentrations among taxa. Stepwise multiple regression analysis identified phloridzin as the only endogenous phenolic that was significantly related to resistance under both choice and no-choice feeding conditions.

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Nobuko Sugimoto and Randy Beaudry

The objective of the experiment was to determine developmental changes in major aroma profiles in `Jonagold' apple (Malus ×domestica Borkh.) and analyze climacteric fruit characteristics. Changes in internal ethylene production, respiration, skin color, texture, and aroma concentration were measured during maturation and ripening of `Jonagold' apple fruit. Patterns for skin color, starch, and internal ethylene content were typical for the variety. Volatile compounds and CO2 increased after a rapid increase in ethylene production. Total ester emission peak coincided with fruit softening. Hexyl acetate, 2-methylbutyl acetate, butyl acetate, and hexyl 2-methylbutanoate were found to be the major volatile compounds detected by GC/MS. Long chain esters, such as hexyl acetate and butyl acetate, contributed during the early stages of ripening and short chain esters such as n-propyl acetate and butyl propanoate increased later. Esters are formed by combining alcohol moiety with CoA derivative of fatty acid moiety by the action of alcohol acyl transferase (AAT). The alcohols butanol, 2-methylbutanol, propanol, and hexanol increased at an earlier developmental stage than the esters for which they acted as substrates.

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Minou Hemmat, Norman F. Weeden, and Susan K. Brown

We mapped DNA polymorphisms generated by 41 sets of Simple Sequence Repeat (SSR) primers, developed independently in four laboratories. All primer sets gave polymorphisms that could be located on our `White Angel' x `Rome Beauty' map for apple [Malus sylvestris (L.) Mill. Var. domestica (Borkh.) Mansf.]. The SSR primers were used to identify homologous linkage groups in `Wijcik McIntosh', NY 75441-58, `Golden Delicious', and `Liberty' cultivars for which relatively complete linkage maps have been constructed from isozyme and Random Amplified Polymorphic DNA (RAPD) markers. In several instances, two or more SSRs were syntenic, and except for an apparent translocation involving linkage group (LG) 6, these linkages were conserved throughout the six maps. Twenty-four SSR primers were consistently polymorphic, and these are recommended as standard anchor markers for apple maps. Experiments on a pear (Pyrus communis L.) population indicated that many of the apple SSRs would be useful for mapping in pear. However some of the primers produced fragments in pear significantly different in size than those in apple.

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Nnadozie C. Oraguzie, Sue E. Gardiner, Heather C.M. Basset, Mirko Stefanati, Rod D. Ball, Vincent G.M. Bus, and Allan G. White

Four subsets of apple (Malus Mill.) germplasm representing modern and old cultivars from the repository and apple genetics population of the Horticulture and Food Research Institute of New Zealand Limited were used in this study. A total of 155 genotypes randomly chosen from the four subsets were analyzed for random amplified polymorphic DNA (RAPD) variation. Nine decamer primers generated a total of 43 fragments, 42 of which were polymorphic across the 155 genotypes. Pairwise distances were calculated between germplasm subsets using the distance metric algorithm in S-PLUS, and used to examine intra-and inter-subset variance components by analysis of molecular variation (AMOVAR). A phenogram based on unweighted pair group method with arithmetic average (UPGMA) cluster analysis was constructed from the pairwise distances and a scatter plot was generated from principal coordinate analysis. The AMOVAR showed that most of the variation in the germplasm (94.6%) was found within subsets, suggesting that there is significant variation among the germplasm. The grouping of genotypes based on the phenogram and scatter plot generally did not reflect the pedigree or provenance of the genotypes. It is possible that more RAPD markers are needed for determining genetic relationships in apple germplasm. Nevertheless, the variation observed in the study suggests that the current practice of sublining populations in the first generation to control inbreeding may not be necessary in subsequent generations. If these results are confirmed by fully informative molecular markers, germplasm managers should reassess the structure of their genetics populations. There may be a need to combine sublines in order to capture the maximum genetic diversity available and to streamline breeding efforts.

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James F. Harbage and Dennis P. Stimart

Many physiological responses in plants are influenced by pH. The present chemiosmotic hypothesis suggests that auxin uptake into plant cells is governed by pH. Since auxin is used widely to enhance rooting, the influence of pH on 1H-indole-3-butyric acid (IBA) induced adventitious root formation was examined. Roots were initiated aseptically in 5 node apical shoot cuttings of micropropagated Malus domestica 'Gala'. Initiation was induced using a four day pulse in IBA and 15 g/L sucrose at pH 5.6 and 30C in the dark. Observations showed pH rose to 7.0 or greater within 1 to 2 days from microcutting placement in unbuffered initiation medium. Root numbers from shoots in media containing 1.5 μM IBA buffered with 10 mM 2[N-morpholino] ethanesulfonic acid (MES) to pH 5.5, 6.0, 6.5 or 7.0 with KOH resulted in average root numbers of 14.2, 10.9, 8.7, and 7.1, respectively, while unbuffered medium yielded 7,6 roots per shoot. Comparison of MES buffered medium at pH 5.5, 6.25 or 7.0 in factorial combination with IBA at 0, 0.15, 1.5, 15.0, and 150.0 μM resulted in a significant pH by IBA interaction for root number. At 0, 0.15 and 1.5 μM IBA root numbers were greatest at pH 5.5. At 15.0 μM IBA, pH 6.25 was optimal and at 150.0 μM IBA all three pH levels produced equivalent root numbers. A calorimetric assay to measure IBA removal from the initiation medium by microcuttings of `Gala' and `Triple Red Delicious' showed more IBA removal at pH 5.5 than at pH 7.0. Possible reasons for the effect of pH on adventitious root formation will be discussed.

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H.S. Aldwinckle, P.L. Forsline, H.L. Gustafson, and S.C. Hokanson

Resistance to apple scab (Venturia inaequalis) in apple cultivar breeding has been derived mainly from the Vf gene from Malus floribunda 821, which introgresses horticulturally unfavorable characters. M. sieversii, now thought to be the primary progenitor of M. × domestica, grows wild in many diverse habitats in Central Asia and can have fruit quality comparable to commercial cultivars. Since 1989, four major collections of M. sieversii have been made in Central Asia, where scab is endemic. Some seed collections have been made from trees with superior fruit, that were not infected with scab. Over a 6-year period, 3000 seedlings from 220 wild M. sieversii trees representing 10 diverse ecosystems in Kazakstan, Uzbekistan, Kyrgyzstan, and Tajikistan have been inoculated with conidia of five races and two wild types of V. inaequalis. Suspensions (270,000 conidia/ml) were applied to 4- to 8-leaved seedlings, which were incubated for 48 h at 19°C with constant leaf wetness. Symptoms for three resistant reactions were assessed 2 to 4 weeks after inoculation: A = chlorosis with crinkling (Vf type reaction); B = stellate necrotic lesions (Vr type reaction), and N = large necrotic areas (uncharacterized resistant reaction). Results indicated that nearly 20% of the seedlings showed one or more of the resistant reactions. The range of resistance within seedling populations from each of the 220 single-tree sources ranged from 0% to 75%. Significant differences existed among seedlings from each of the ecosystems. Most resistance reactions appeared to be similar to those observed for Vr from “Russian seedling.” Resistant selections with superior horticultural traits may constitute a genepool for increased efficiency of breeding scab-resistant cvs. This genepool may also be useful to address the breakdown of resistance to V. inaequalis race 6.

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Fuad Gasi, Silvio Simon, Naris Pojskic, Mirsad Kurtovic, Ivan Pejic, Mekjell Meland, and Clive Kaiser

pomological characterizations ( Nybom and Weising, 2010 ), we chose this approach in our study. SSR molecular markers have shown great promise as tools for managing Malus ex situ germplasm collections as well as for collection and preservation strategies of

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Jan Bizjak, Nika Weber, Maja Mikulic-Petkovsek, Ana Slatnar, Franci Stampar, Zobayer Alam, Karl Stich, Heidi Halbwirth, and Robert Veberic

mechanism, by which P-containing compounds affect the red coloration of apples. Materials and Methods Plant materials. The experiment was carried out in 2011 on 10-year-old trees of striped ‘Braeburn’ ( Malus domestica Borkh.) cultivar clone Hillwell

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Philip L. Forsline, E.E Dickson, and A.D. Djangaliev

The USDA National Plant Germplasm System (NPGS) sponsored a 1993 collection of wild Malus in Kazakhstan and Kyrgyzstan which followed a collection in 1989 from sites in Tajikistan and Uzbekistan. There is strong evidence that the domestic apple originated in the foothills of the Tian Shari mountains where Malus sieversii (Ldb.) M. Roem remains as a primary forest species. The goal of the recent expedition was to obtain additional genetic diversity of apple from some of the remote sites in that area with the assistance of the Kazakh hosts. While there, isolated pockets of other fruit in the wild (especially Vitis) were discovered and collected. Seed collections from the expedition are stored with the NPGS and seedling populations are being evaluated for valuable traits.

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Roberto Hauagge and James N. Cummins

Relationships among estimates for the length of bud dormancy over a wide range of cultivars of Malus × domestics Borkh. and related Malus spp. are reported. Flower bud dormancy state was estimated after fitting quadratic models for the number of days for 50% terminal budbreak (D50), bud development stage (BDS), and percent terminal budbreak (BB) after 21 days of forcing as a function of chilling unit (CU) accumulation. Cultivar, linear, and quadratic components of regression, as well as all interactions, were significant for D50, BDS, and BB when regressed against CU accumulation. Consequently, cultivars differ not only as to when changes in “dormancy intensity take place, but also in the patterns and rates at which these changes proceed. The number of CU needed to reach arbitrary stages of each characteristic at the end of dormancy was calculated and compared for all genotypes. Rates of CU accumulation until BB, BDS, and D50 reached 50%, 1.5 and 15 days, respectively, were well-correlated. These CU values were considered the end of bud dormancy or genotype chilling requirement (CR). Estimations of D50 were less variable and less subjective, and a wider range of data points could be used in the analysis. In addition, values for D50 can indicate the growth potential of buds when other indices do not show changes. A few sampling times during the dormant season may give a preliminary idea about the CR of a given genotype.