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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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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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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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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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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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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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S. Mantha, H. Desilets, J.-A. Rioux, S. Gagne, S. Parent, and P. Moutoglis

Two experiments with Malus domestica sp. were planted in 1997 at the Laval Univ. experimental farm located south of the St. Lawrence river near Quebec City. These experiments examined the association of the mycorrhizal fungus Glomus intraradices with Malus domestica sp. The first experiment compared the vegetative growth of `McIntosh' apple trees on M.106 rootstock in presence or absence of a commercial inoculum of G. intraradices (Premier Tech, Riviere-du-Loup, Quebec) under three levels of phosphorus fertilization (P) to the soil (0%, 50%, and 100% of the usual recommandation for this crop). After two seasons, all the treatments had better growth than the control (0% P without G. intraradices). The best treatment was achieved with 100% of the P associated with mycorrhizal inoculation. The second experiment compared the vegetative growth of three apple rootstocks Bud.9, M.26, and M.106, inoculated with G. intraradices under the same three P levels as the preceding experiment. Uninoculated rootstocks receiving the usual phosphorus fertilization served as control. Two roostocks, M.26 and M.106, increased growth with G. intraradices, while the third one, Bud.9, did not respond to the presence of mycorrhizal fungus.