Journal of Applied Botany and Food Quality 88, 197 - 201 (2015), DOI:10.5073/JABFQ.2015.088.028 1 Department of Horticulture, Faculty of Agriculture, University of Cukurova, Adana, Turkey 2Ataturk Central Horticultural Research Institute, Yalova, Turkey 3 Department of Horticulture, Faculty of Agriculture, Ataturk University, Erzurum, Turkey Genetic relatedness among quince (Cydonia oblonga Miller) accessions from Turkey using amplified fragment length polymorphisms H. Topcu1, S. Kafkas1*, A. Dogan2, M.E. Akcay2, S. Ercisli3 (Received February 8, 2015) * Corresponding author Summary Among fruit species cultivated in Turkey, quince shows a great deal of morphological variability and adaptability to the various environments. We attempted to study genetic relationships among 40 quince accessions using amplified fragment length polymor- phisms (AFLPs) for future breeding programs. The accessions were previously characterized based on their pomological and yield cha- racteristics and then the best ones were planted in a single collection in Ataturk Central Horticultural Research Institute, Yalova, Turkey. Six AFLP primer combinations generated a total of 746 bands, 493 of which were polymorphic (66.1 %). Resolving powers of the AFLP primers ranged from 48.0 to 99.6 making a total of 421.5. Unweighted Pair Group Method with Arithmetic Average (UPGMA) clustering of the accessions showed three major clusters and ‘SapancaEsme’ and ‘Esme-3’ were the closest accessions with 95 % similarity. Our study indicated that there is a high level of genetic diversity among quince accessions in Turkey and the results of this study can be used for future cultivar breeding programs in quince. Introduction The quince (Cydonia oblonga Miller) belongs the genus Cydo- nia and native to warm-temperate regions including Asia Minor (BROWICZ, 1972). The total quince production in the world is 596.532 tons, and Turkey (135.500 tons) is the leader producer country. The other important quince producer countries are China (125.000 tons), Uzbekistan (80.000 tons), Morocco (46.000 tons), Iran (36.500 tons), Argentina (27.500 tons), Azerbaijan (27.140 tons) and Spain (14.000 tons) (FAO, 2012). The quince is a deciduous tree, growing up to 5-8 m tall and 4-6 m wide. It has very close relationships with apples and pears, and has a pear or apple shaped pome fruits, which is bright golden yel- low when mature. In most quince producer countries, they are not grown in large amounts; typically several quince trees are grown in a mixed orchard with several apples and other fruit trees (WESTWOOD, 1993). The fruits of most of quince cultivars are hard, sour and astringent in maturation time and therefore a few cultivars can be eaten raw. Due to this reason, quince fruits are mainly used to make jam and jelly or they may be peeled, then roasted, baked or stewed. The flesh of the fruit turns red after cooking (SILVA et al., 2002; 2004). The quince fruit is also known as an important dietary source due to its antioxidant, antimicrobial and antiulcerative properties (SILVA et al., 2004; HAMAUZU et al., 2006; FATTOUCH et al., 2007). Quince trees have been used as a dwarf rootstock for pear for a long time. It forces scion to produce precocious fruits, and relatively more fruit-bearing branches, and of accelerating the maturity of the fruit (GULEN et al., 1999; MASSAI et al., 2008). Earlier characterization of the quince genotypes were performed pri- marily based on phenotypic traits of the plants such as color, size, shape and other agronomical characters of fruits in Turkey (ERCAN et al., 1992; ERCISLI et al., 1999; DUMANOGLU et al., 2009; KUDEN et al., 2009). However, information from morphological and pheno- typic characteristics are not sufficient to identify quince genotypes because of environmental plasticity on it. Thus, environmentally free genotypic traits are necessary for proper identification and estima- tion of genetic diversity among quince accessions. Molecular characterization assays provide an efficient tool for the evaluation of genetic diversity in plants (HALASZ et al., 2010; BADRI et al., 2014). Various types of molecular markers (RFLP, RAPD, SSRs, and AFLP) have been successfully used to assess the levels of genetic diversity in fruit tree species (BELAJ et al., 2003; CHEN et al., 2005; HALASZ et al., 2006; ERCISLI et al., 2008; KAFKAS et al., 2008; KAFKAS et al., 2009). AFLP markers are highly reproducible with overall error rates of less than 2 % (VOS et al., 1995), it is suit- able for high-throughput genotyping, and DNA sequence informa- tion is not a prerequisite. Although the AFLP method has been one of the most widely used marker system to identify genetic variability in many different fruit tree species, the use of this powerful and reliable method in quince is very rare. The objective of this study is to characterize 40 quince accessions of Cydonia oblonga from Turkey using AFLP markers, to determine whether AFLP markers are appropriate to discriminate quince accessions and to have a better understanding about the vari- ability within the Turkish quince germplasm. Materials and methods Plant Material Forty quince accessions (Cydonia oblonga) were used in the pre- sent study (Tab. 1). These accessions were previously selected for their better fruit and yield characteristics than others and vegeta- tively propagated, and then they were planted in the Ataturk Hor- ticultural Research Institute in the Yalova province of Turkey. The pomological characteristics of accessions were published elsewhere (BUYUKYILMAZ, 1999). DNA extraction and AFLP analysis Genomic DNA was isolated from leaf tissue by the CTAB method (DOYLE and DOYLE, 1987). DNA concentration was estimated by comparing band intensity with l DNA of known concentrations, after 0.8 % agarose gel electrophoresis and ethidium bromide stain- ing. DNA was diluted to 50 ng μL-1 for AFLP reactions. Details of AFLP assay, adaptor and primer sequences, PCR conditions for preselective and selective amplifications, and selective primer desig- nation were according to VOS et al. (1995). Genomic DNA was re- stricted with EcoRI/MseI enzyme combination and double-stranded adaptors specific to each site were ligated. Preselective amplifica- tions were done with primers complementary to the adaptors with an extra selective base on each primer (EcoRI-A/MseI-C). Selective amplifications were performed using six primer combinations with three MseI (M) and three EcoRI (E) primers (E39/M55, E39/M57, 198 H. Topcu, S. Kafkas, A. Dogan, M.E. Akcay, S. Ercisli E45/M59, E42/M59, E45/M60 and E42/M60). AFLP fragments were resolved using capillary electrophoresis on an ABI 3130xl Ge- netic Analyzer [Applied Biosystems Inc., Foster City, Calif, (ABI)] with the data collection software 3.0 (ABI). AFLP fragments were analyzed with GeneScan analysis software 4.0 (ABI) and the data were assembled in binary format. Data analysis The ability of the most informative primer pairs to differentiate the genotypes was analyzed by calculating their resolving power (Rp) according to PREVOST and WILKINSON (1999) using the formula Rp =∑ Ib, where Ib =1-(2 x | 0.5 - p | ), and p is the proportion of the 40 genotypes containing the I band. Jaccard’s similarity coefficients (JACCARD, 1908) were calculated for all pair-wise comparisons among the 40 quince genotypes. A dendrogram was generated using NTSYSpc version 2.11V (Exeter Software, Setauket, NY) (ROHLF, 2004) based on the un-weighted pair-group method of arithmetic average cluster analysis (UPGMA). Results and discussion A total of 746 fragments were amplified using six primer combina- tions, 493 (66.8 %) were polymorphic in characterizing 40 quince accessions (Tab. 2). The number of total bands produced by each primer combination ranged from 97 (E45/M60) to 156 (E42/M59) with an average of 104.1 per primer pair. The number of polymor- phic fragments per primer combination ranged from 59 to 103 with an average of 69.1. Among the six primer combinations tested, E42/ M59 and E45/M60 amplified the lowest and highest number of total as well as polymorphic fragments, respectively (Tab. 1). The percentage of polymorphic bands varied considerably among the primer combinations. The highest polymorphism ratio (72.6 %) was observed in E39/M55 primer pair and followed by E39/M57 (69.9 %), E42/M59 (66.0 %), E42/M60 (65.7 %) and E45/M59 (60.8 %) and E45/M60 (60.8 %) (Tab. 1). Resolving power (Rp) ranged from 48.0 (E45/M60) to 99.6 (E42/M59), with a total of 421.5. The average Rp value of the primers was found to be 70.3 (Tab. 1). In the present study, we obtained the highest polymorphism ratio (72.6 %) in E39/M55 primer pair and followed by E39/M57 (69.9 %) and E42/M59 (66.0 %). Screening and selection of primer combina- tions, which detect maximum genetic variation, are vital to establish dependable genetic relationships among accessions (KAFKAS et al., 2009). In this study, AFLP analysis successfully differentiated the quince accessions each other and it was confirmed that AFLP is a power- ful marker system. AFLP markers have been previously used in the analysis of different fruit species for example mulberry (SHARMA et al., 2000), tea (KAFKAS et al., 2009), walnut (KAFKAS et al., 2005), myrtle (BRUNA et al., 2007) and pomegranate (JBIR et al., 2008) and all these results confirmed the power of AFLP technique to discrimi- nate genotypes. Although quince is known self fertile, their level of polymorphism was comparable to those of previous studies. This is an indication of the high degree of polymorphism among the acces- sions tested. In this study, Resolving power (Rp) ranged from 48.0 (E45/M60) to 99.6 (E42/M59). Rp has been found to correlate strongly with its ability to distinguish between accessions. It is usually difficult to compare the value of primers used in different investigations. The use of Rp might enable direct comparison of primers both within and between studies (PREVOST and WILKINSON, 1999). A dendrogram constructed using UPGMA method of cluster analy- sis from a combined data of all primer combinations classified ac- Tab 1: The origin of quince accessions used in this study Accession Origin Accession Origin Accession Origin Altin Subasi Kocaeli Esme 8 Sakarya Esme 6 Kocaeli Altin Yalova Yalova Havan Yalova Limon 2 Sakarya Bardak Bursa Esme 14 Kocaeli Viranyadevi Yalova Demir 1 Kocaeli Tekkes Yalova Beyaz Ayva Kocaeli Ege-25 Izmir 27-1 Yalova 7-2 Yalova Ekmek Keles Bursa Gordes Yalova 6-3 Yalova Esme 2 Kocaeli Esme 10 Sakarya 10-3 Yalova Sapanca Esme Sakarya Bencikli Yalova Ekmek Yalova Yalova Esme 3 Kocaeli Ege-22 Izmir Esme Arifiye Sakarya Esme 4 Kocaeli Sekergevrek Yalova Esme 1 Sakarya Limon 1 Sakarya 2-3 Yalova Esme 7 Sakarya 19-2 Yalova Esme 5 Kocaeli Esme 9 Sakarya Esme 13 Kocaeli Esme 11 Bilecik Esme 12 Bilecik Limon Yalova Yalova Tab 2: Number of total and polymorphic AFLP bands, percentage of poly- morphic bands, and resolving powers in the DNA-fingerprinting of 40 quince genotypes originating from Turkey. AFLP primer Total bands Polymorphic Polymorphism Resolving combinations (no.) bands (no.) (%) powers (Rp) E39/M55 113 82 72.6 80.9 E39/M57 123 86 69.9 57.4 E45/M59 120 73 60.8 66.5 E42/M59 156 103 66.0 99.9 E45/M60 97 59 60.8 48.0 E42/M60 137 90 65.7 69.1 Total 746 493 421.5 Mean 104.1 95 66.8 70.3 Genetic relatedness in quince by AFLP 199 cessions into three main clusters (Fig. 1). Cluster I consisted of six quince accessions. The closest accessions in this cluster were ‘Esme-7’ and ‘Esme-9’ which were surrounded by the accessions ‘Esme-12’, ‘Esme-1’, ‘Esme Arifiye’ and ‘Ekmek Yalova’ respec- tively (Fig. 1). Cluster II include only ‘10-3’ genotype. Cluster III contained 34 accessions and this cluster divided into two sub- clusters. The first subcluster included 12 accessions (‘6-3’, ‘7-2’, ‘Beyaz Ayva’, ‘Viranyadevi’, ‘Limon-2’, ‘Esme-6’, ‘Esme-11’, ‘Esme-5’, ‘2-3’, ‘Sekergevrek’, ‘Ege 22’ and ‘Bencikli’, whereas subcluster II contained 21 accessions (‘Esme-10’, ‘Gordes, 27-1’, ‘Tekkes’, ‘Esme-14’, ‘Havan’, ‘Esme-8’, ‘Limon Yalova’, ‘Esme- 13’, ’19-2’, ‘Limon-1’, ‘Esme-4’, ‘Esme-3’, ‘Sapanca Esme’, ‘Esme- 2’, ‘Ekmek Keles’, ‘Ege 25’, ‘Demir-1’, ‘Bardak’, ‘Altin Yalova’ and ‘Altin Subasi’). The accession ‘Esme-10’ took place between two subclusters. In the subcluster II, ‘Sapanca Esme’ and ‘Esme-3’ were the closest accessions in this study (95.1 % similarity), with the accessions ‘Esme’, ‘Esme-4’, and ‘Limon-1’. The pairs of ac- cessions ‘Bardak’ with ‘Demir-1’ and ‘Ekmek Keles’ with ‘Esme- 2’ were also close to each other with 92.8 % and 92.6 % similarity, respectively (Fig. 1). The UPGMA dendrogram formed from AFLP bands successfully separated the closely-related quince accessions (particularly ‘Esme’ genotypes). It is clear that there is a large clonal variability with- in ‘Esme’. This genotype was grown in different parts of Western Anatolia (Tab. 1). This fact points to cultivation since remote times in quince growing areas in Anatolia and finally the numerous ‘Esme’ types would subsequently have been generated through accumu- lation of somatic mutations during the long period of mutations (Ercisli, 2004). Local people groups would have moved over long distances in the Anatolia over time and carried with them the core of quince variation, while further ad hoc cultivation would have caused additional and location-specific variation. Moreover, previ- ous study indicating ‘Esme’ genotypes showed a certain degree of morphological and phenological variability within the population of individuals (BUYUKYILMAZ, 1999). Most fruit tree species, includ- ing quince, are vegetatively propagated to maintain agronomically valuable genotypes. However, after many propagation cycles, clones accumulate phenotypic differences in agronomic traits and clonal diversity appears (ORIVE, 2001). This diversity can then be used to select the best clones or a new improved cultivar within a given variety. A recent study of the molecular polymorphisms genera- ted along vegetative propagation (CARRIER et al., 2012) through a genome-wide comparison of spontaneous grape ‘Pinot noir’ clones showed that only a small number of SNP and indel events are at the origin of clonal variation, while mobile elements of many families are involved in most polymorphisms, displaying the highest muta- tional event. In general, our results were in agreement with those of DUMANOGLU et al. (2009) who indicated a large genetic diversity within different ‘Kalecik’ quince clones in Turkey (average poly- morphism ratio with 65 %) by using SSR markers. YAMAMATO et al. (2004) used a total 118 SSR markers (developed for apple and pears) on 20 quince cultivars and they obtained relatively high discrimina- tion capacity from 39 SSR markers. YUKSEL et al. (2013) conduct- ed simple sequence repeat (SSR) analyses of 15 traditional quince (Cydonia oblonga) cultivars from Anatolian gene sources for mole- cular characterization and investigation of genetic relationships. They used eight SSR loci that previously developed from apple and pear and they found similarity ratio between 18-87 % among culti- vars. In present study, in most cases, quince accessions within the same cluster did not have similar morphological fruit characteristics. For example, in the Cluster I, the closest accessions ‘Esme-7’ and ‘Esme-9’ have different tree and fruit characteristics. ‘Esme7’ has light yellow fruit skin color at maturation and Esme9 has greenish yellow skin color. Esme7 has oval with neck fruit shape, whereas ‘Esme9’ has neck less oval fruits. The yields of these accessions are also very variable (BUYUKYILMAZ, 1999). In addition, in the Cluster I, ‘EkmekYalova’ has yellow skin color. The genotype ‘Esme-1’ has medium fiber in its fruits; however the other accessions had low fiber levels. Fruit weight of six accessions in Cluster I varied from 350 to 396 g and yield per tree varied from 62 kg (‘Esme-7’) to 110 kg (‘EsmeArifiye’) (BUYUKYILMAZ, 1999). In the Cluster II, the accessions ‘SapancaEsme’ and ‘Esme-3’ were the closest accessions with 95 % similarity ratio. However, these two accessions had different fruit characteristics. For example, ‘SapancaEsme’ had oval with neck fruit shape, however ‘Esme-3’ had oval with light neck fruit shape. The other closest pairs of ac- cessions ‘Bardak’-‘Demir-1’ and ‘EkmekKeles’-‘Esme-2’ have also different fruit characteristics: ‘Bardak’ has greenish yellow, long neck fruits while ‘Demir-1’ has light yellow oval with light neck fruits. ‘Bardak’ has more fiber characteristics than ‘Demir-1’ (BUYUKYILMAZ, 1999). The accessions from the same province were assigned in the dif- ferent cluster. For example, the selections from Esme cultivar in Kocaeli province, ‘Esme-1’ placed in Cluster I and the other ac- cessions (‘Esme-2’, ‘Esme-3’, ‘Esme-4’, ‘Esme-5’ and ‘Esme-6’) distributed in Cluster II. Likewise, the Esme accessions from Sa- karya province were also assigned in different clusters (‘Esme-7’ and ‘Esme-9’ in Cluster I and ‘Esme-8’ and ‘Esme-10’ in Cluster II (Fig. 1). The closest pair of accessions in this study, ‘SapancaEsme’ and ‘Esme-3’, were collected from the different provinces (Sakarya and Kocaeli) in Northwestern parts of Turkey. This could be at- Fig. 1: Dendrogram of 40 quince genotypes originating from Turkey result- ing from the unweighted pair-group method of arithmetic mean clus- ter analysis based on Jaccard similarity coefficient obtained from 746 AFLP markers. 200 H. Topcu, S. Kafkas, A. Dogan, M.E. Akcay, S. Ercisli tributed to the unrestricted movement of planting materials from region to region, high self-pollination habit and perennial nature of the crop. On the contrary, the accessions ‘Esme-11’ and ‘Esme-12’ were collected from Bilecik province in Turkey, however they were genetically far from each other. The low genetic similarity between these accessions collected from the same region could be attributed to the vast agro-ecological diversity present within each region that can contribute to the development of genetically diverse accessions through natural selection. Therefore, similarity in collection locality may not necessarily imply genetic similarity in quince, in Turkey. Dissimilarities in groupings using molecular markers or pheno- taxonomic characteristics were also reported in strawberry (GARCIA et al., 2002), mulberry (ORHAN et al., 2007) and olive plants (HAGI- DIMITRIOU et al., 2005). These discrepancies were also previously reported by ZAMANI et al. (2007), who compared data from the genetic distance matrices obtained from RAPD markers and from fruit characteristics. Their correlation coefficient, for comparison of morphological and RAPD data, was only 23 %. These results sup- port the view that morphological characteristics are not reliable for estimating genetic relationships among large and diverse groups of accessions, and should be used mainly for discrimination. Results of the present study as well as studies by YAMAMATO et al. (2004) in Japan DUMANOGLU et al. (2009) in Turkey, HALASZ et al. (2009) in Hungary and BASSIL et al. (2011) in USA indicated the presence of genetic variation both genotypic and clonal level among quince genotypes. This confirms the importance of conservation of the Turkish quince gene pool for the quince industry. Since quince is a tree and a perennial crop, it demands large area and year round management which is expensive. Moreover, morphological markers require evaluation over long periods of time and the present study indicated the inadequacy of morphological characters for character- ization of closely related accessions. Hence, genetic diversity analy- sis using DNA-based marker techniques is recommended for cost effective and efficient conservation of quince germplasm. The results of the present study may benefit breeders in selecting the most diverse accessions to begin crossing and selection programs. This may result in increased quince growing for better fruit produc- tion. This report is also demonstrates the presence of mutations of agronomical relevance within a monoclonal cultivar of Esme. References BADRI, J., YEPURI, V., GHANTA, A., SIVA, S., SIDDIQ, E.A., 2014: Develop- ment of microsatellite markers in sesame (Sesamum indicum L.). Turk. J. Agric. For. 38, 603-614. BASSIL, N.V., POSTMAN, J.D., HUMMER, K.E., MOTA, J., SUGAR, D., WILLIAMS, 2011: Quince (Cydonia oblonga) genetic relationships deter- mined using microsatellite markers. Acta Hortic. 909, 75-84. BELAJ, A., SATOVIC, Z., CIPRIANI, G., BALDONI, L., TESTOLIN, R., RALLO, L., TRUJILLO, I., 2003: Comparative study of the discriminating capacity of RAPD, AFLP and SSR markers and their effectiveness in establishing genetic relationships in olive. Theor. Appl. Genet. 107, 736-744. BROWICZ, K., 1972: Cydonia Miller. In: Davis, P.H. (ed.), Flora of Turkey and the East Aegean Islands, 157. Edinburgh University Press, Great Britain. BRUNA, S., PORTIS, E., CERVELLI, C., DE BENEDETTI, L., SCHIVA, T., MERCURI, A., 2007: AFLP-based genetic relationships in the Mediter- ranean myrtle (Myrtus communis L.). Sci. Hortic. 113, 370-375. BUYUKYILMAZ, M., 1999: Quince cultivar selection. Ataturk Horticultural Central Research Institute. Publication No:125. CARRIER, G., CUNFF, L., DEREEPER, A., LEGRAND, D., SABOT, F., BOUCHEZ, O., AUDEGUIN, L., BOURSIQUOT, J.M., THIS, P., 2012: Transposable elements are a major cause of somatic polymorphism in Vitis vinifera L. PLoS ONE 7 (3), e32973. CHEN, L., GAO, Q.K., CHEN, D.M., XU, J.C., 2005: The use of RAPD mark- ers for detecting genetic diversity, relationship and molecular identi- fication of Chinese elite tea genetic resources [Camellia sinensis (L.) O. Kuntze] preserved in tea germplasm repository. Biodivers Conserv. 14(6), 1433-1444. DOYLE, J.J., DOYLE, J.L., 1987: A rapid isolation procedure for small quan- tities of fresh leaf tissue. Phytochem. Bull. 19, 11-15. DUMANOGLU, H., TUNA GUNES, N., AYGUN, A., SAN, B., AKPINAR, A.E., BAKIR, M., 2009: Analysis of clonal variations in cultivated quince (Cy- donia oblonga ‘Kalecik’) based on fruit characteristics and SSR markers. New Zealand J. Crop and Hort. Sci. 37, 113-120. ERCAN, N., OZVARDAR, S., GONULSEN, N., BALDIRAN, E., ONAL, K., KARABIYIK, N., 1992: Determination of Quince cultivars for Aegean region. Proceedings of 1st National Horticulture Conference, 13-16 Oc- tober 1992, Izmir. Volume 1, 527-529. ERCISLI, S., GULERYUZ, M., ESITKEN, A., 1999: Fruit characteristics of Quince cultivars grown in Oltu district. Anadolu, 9 (2), 32-40. ERCISLI, S., 2004: A short review of the fruit germplasm resources of Turkey. Genet. Res. Crop Evol. 51, 419-435. ERCISLI, S., ORHAN, E., YILDIRIM, N., AGAR, G., 2008: Comparison of sea buckthorn genotypes (Hippophae rhamnoides L.) based on RAPD and FAME data. Tr. J. Agric. Forest. 32(5), 363-368. FATTOUCH, S., CABONI, P., CORONEO, V., TUBEROSO, C.I.G., ANGIONI, A., DESSI, S., MARZOUKI, N., CABRAS, P., 2007: Antimicrobial activity of Tunisian quince (Cydonia oblonga Miller) pulp and peel polyphenolic extracts. J. Agric. Food Chem. 55, 963-969. FOOD AND AGRICULTURAL ORGANIZATION OF THE UNITED NATIONS, 2012: Agriculture data [online]. Available from: http://faostat.fao.org/site/567/ DesktopDefault.aspx? PageID=567. GARCIA, M.G., ONTIVERO, M., DIAZ RICCI, R.C., CASTAGNARO, A., 2002: Morphological traits resolution RAPD markers for the identification of the main strawberry varieties cultivated in Argentina. Plant Breed. 121, 76-80. HAGIDIMITRIOU, M., KATSIOTIS, A., MENEXES, G., PONTIKIS, C., LOUKAS, M., 2005: Genetic diversity of major Greece olive cultivars using mo- lecular (AFLPs and RAPDs) markers and morphological traits. J. Amer. Soc. Hort. Sci. 130, 211-217. HALASZ, J., HEGEDUS, A., PEDRYC, A., 2006: Review of the molecular back- ground of self-compatibility in Rosaceous fruit trees. Int. J. Hortic. Sci. 12 (2), 7-18. HALASZ, J., HOFFMANN, V., SZABO, Z., NYEKI, J., SZABO, T., HEGEDUS, A., 2009: Characterization of quince (Cydonia oblonga Mill.) cultivars using SSR markers developed for apple. Inter. J. Hort. Sci. 15, 7-10. HALASZ, J., PEDRYC, A., ERCISLI, S., YILMAZ, K.U., HEGEDUS, A., 2010: S-genotyping supports the genetic relationships between Turkish and Hungarian apricot germplasm. J. Amer. Soc. Hort. Sci. 135, 410-417. HAMAUZU, Y., INNO, T., KUME, C., IRIE, M., HIRAMATSU, K., 2006: Anti- oxidant and antiulcerative properties of phenolics from Chinese quince, quince, and apple fruits. J. Agric. Food Chem. 54, 765-772. JACCARD, P., 1908: Nouvelle reserches sur la distribution florale. Bull. Soc. Vaud. Sci. Nat. 44, 223-227. JBIR, R., HASNAOUI, N., MARS, M., MARRAKCHI, M., TRIFI, M., 2008: Cha- racterization of Tunisian pomegranate (Punica granatum L.) cultivars using amplified fragment length polymorphism analysisa. Sci. Hortic. 115, 231-237. KAFKAS, S., OZGEN, M., DOGAN, Y., OZCAN, B., ERCISLI, S., SERCE, S., 2008: Molecular characterization of mulberry accessions in Turkey by AFLP Markers. J. Am. Soc. Hortic. Sci. 133, 593-597. KAFKAS, S., OZKAN, H., SUTYEMEZ, M., 2005: DNA polymorphism and as- sessment of genetic relationships in walnut genotypes based on AFLP and SAMPL markers. J. Amer. Soc. Hort. Sci. 130, 585-590. KAFKAS, S., ERCISLI, S., DOGAN, Y., ERTURK, Y., HAZNEDAR, A., SEKBAN, R., 2009: Polymorphism and genetic relationships among tea genotypes from Turkey revealed by AFLP markers. J. Amer. Soc. Hort. Sci. 134(3), 1-5. KUDEN, A., TUMER, M.A., GUNGOR, M.K., IMRAK, B., 2009: Pomological traits of some selected Quince types. Acta Hort. 818, 73-76. Genetic relatedness in quince by AFLP 201 MASSAI, R., LORETI, F., FEI, C., 2008: Growth and yield of ‘Conference’ pears grafted on quince and pear rootstocks. Acta Hort. 800, 617-624. ORHAN, E., ERCISLI, S., YILDIRIM, N., AGAR, G., 2007: Genetic variations among mulberry genotypes (Morus alba) as revealed by random ampli- fied polymorphic DNA (RAPD) markers. Plant Syst. Evol. 265, 251- 258. ORIVE, M.E., 2001: Somatic mutations in organisms with complex life histo- ries. Theor. Popul. Biol. 59, 235-249. PREVOST, A., WILKINSON, M.J., 1999: A new system of comparing PCR primers applied to ISSR fingerprinting of potato cultivars. Theor. Appl. Genet. 98, 107-112. ROHLF, F.J., 2004: NTSYS-pc numerical taxonomy and multivariate analysis system. Version 2.11V. Exeter software, Setauket, New York. SHARMA, A., SHARMA, R., MACHII, H., 2000: Assessment of genetic diver- sity in a Morus germplasm collection using fluorescence-based AFLP markers. Theor. Appl. Genet. 101, 1049-1055. SILVA, B.M., ANDRADE, P.B., MENDES, G.C., SEABRA, R.M., FERREIRA, M.A., 2002: Study of the organic acids composition of quince (Cydonia oblonga Miller) fruit and jam. J. Agric. Food Chem. 50, 2313-2317. SILVA, B.M., ANDRADE, P.B., VALENTAO, P., FERRERES, F., SEABRA, R.M., FERREIRA, M.A., 2004: Quince (Cydonia oblonga Miller) fruit (pulp, peel, and seed) and jam: antioxidant activity. J. Agric. Food Chem. 52, 4705-4712. WESTWOOD, M.N., 1993: Temperate-Zone Pomology Physiology and Cul- ture. 3rd ed. Portland, Oregon, Timber Press Inc. VOS, P., HOGERS, L., BLEEKER, M., VAN DE LEE, T., HORNES, M., FRIJTERS, © The Author(s) 2015. This is an Open Access article distributed under the terms of the Creative Commons Attribution Share-Alike License (http://creative- commons.org/licenses/by-sa/4.0/). A., POT, J., PELEMAN, J., KUIPER, M., ZABEAU, M., 1995: AFLP: A new technique for DNA fingerprinting. Nucleic Acids Res. 23, 4407-4414. YAMAMOTO, T., KIMURA, T.J., SOEJIMA, T., SANADA, T., BAN, Y., HAYASHI, T., 2004: Identification of quince varieties using SSR marker developed from pear and apple. Breed. Sci. 54, 239-244. YUKSEL, C., MUTAF, F., DEMIRTAS, I., OZTURK, G., PEKTAS, M., ERGUL, A., 2013: Characterization of Anatolian traditional quince cultivars, based on microsatellite markers. Genet. Mol. Res. 12 (4), 5880-5888. ZAMANI, Z., SARKHOSH, A., FATAHI, R., EBADI, A., 2007: Genetic relation- ships among pomegranate genotypes studied by fruit characteristics and RAPD markers. J. Hort. Sci. Biotechnol. 82, 11-18. Addresses of the authors: H. Topcu, S. Kafkas, Department of Horticulture, Faculty of Agriculture, University of Cukurova, 01330, Adana, Turkey E-mail corresponding author: skafkas@cu.edu.tr A. Dogan, M.E. Akcay, Ataturk Central Horticultural Research Institute, Yalova, 77102, Turkey S. Ercisli, Department of Horticulture, Faculty of Agriculture, Ataturk Uni- versity, 25240, Erzurum, Turkey