Quince (Cydonia oblongaMill.) is a minor fruit crop, which is primarily used for marmalade, jam and sauce.Very few quince cultivarsare known all over the world and in many cases similar names are used for presumably different cultivars. The aim of the present study was toevaluate and characterize the genetic diversity of 36 quince cultivars and selections with SSR markers. Seven out of 8 SSR markers designed fromapple sequences could successfully yield amplification also in quince cultivars. Number of alleles per locus ranged from 2 to 3 alleles. These allelenumbers are quite low when compared to apple. It is supposed to be the consequence of a genetic bottleneck. In spite of the low allele number perlocus, the 36 quince cultivars formed 30 different genotypes. The ratio of homozygosity was low, which might be coupled with the self-(in)compatibility phenotype of quinces. SSR markers proved unable to differentiate putatively closely related cultivars (e.g. ‘Bereczki’ and‘Bereczki bôtermô’). In general, the level of polymorphism among the tested quince genotypes was much restricted due to the low allele numberdetected. However, it must be considered that the number of analysed SSR loci is not enough high to estimate the overall heterozygosity of thequince genome. Further experiments are needed and the SSR markers proved to be a reliable and useful tool for such analyses

A., Hegedűs

J., Halász

J., Nyéki

T., Szabó

V., Hoffman

Z., Szabó

University of Debrecen Electronic Archive

Introduction
The quince (Cydonia oblonga Mill.) belongs to the
Rosaceae family, Maloideae subfamily, which includes
commercially important fruits such as apples or pears. This
subfamily comprises approximately 1000 species in 30
genera and is characterized by a distinctive fruit, pome, and a
base chromosome number (x) of 17 (Rodger & Campbell,
2002). The quince was thought to have originated in Persia,
Turkistan and the Caucasus. Its tree is used as rootstock for
pear cultivars, while its fruit is used for making preserves,
jam, marmalade, sauce and juice. In Hungary, it has been
cultivated for a long time but its importance lags behind other
fruit species. Sándor Brózik performed the domestication of
foreign quince cultivars and collected native landraces and
selections. The best cultivars grown in Hungary are
‘Angersi’, ‘Bereczki bôtermô’, ‘Champion’ and ‘Mezôtúri’
(Szabó, 1998). Only few breeders or researchers work with
quince; that is why detailed descriptions of cultivars hardly
exist. Characteristics of fruit and tree are so similar that it is
very difficult to make a reliable classification. Differentiation
of cultivars based on the shape of fruit: ‘apple-shaped’
(maliformis Mill.-Schneid.) and ‘pear-shaped’ (pyriformis
Dierb.- F. Zinn) but the morphology of fruit could be
influenced by the fruit set, the number of seeds and
environmental factors (Hedrick, 1925). All over the world
approximatley 40–50 cultivars are propagated, among them
10–15 are well-known (Szabó, 1998). The use of cultivar
names is quite chaotic among different countries, for
example the Romanian ‘De Constantinapol’ is distinct from
the Hungarian ‘Konstantinápolyi’ (Bordeianu, 1964).
The DNA-based molecular markers have become
increasingly popular in the characterization and
identification of genetic resources because they are not
influenced by the environmental factors and are more
polymorphic. However, there is only one report on genetic
study of quince cultivars using SSRmarkers. Yamamoto et al.
(2004) characterized twenty quince cultivars from France,
USA, Japan, Portugal and England using 77 SSR primers
developed from the pear and apple genome. The
development of SSR markers needs preliminary sequence
information of the genome but it was noted that the use of
heterologous SSR primers in closely related species was an
effective approach because of the conserved flanking region
of microsatellite repeats (Moore et al., 1991; Cipriani et al.,
1999; Testolin et al., 2000; Dirlewanger et al., 2002).
Liebhard et al. (2002) established that the tested SSR
markers from apple was transferable and could make
successful cross-amplification in other genera (Cotoneaster,
Pyrus, Cydonia etc.) of the same Maloideae subfamily. In
apple, SSR markers could have been used for the assessment
of genetic diversity (Guarino et al., 2006; Pereira-Lorenzo et al.,
Characterization of quince (Cydonia oblongaMill.)
cultivars using SSR markers developed for apple
Halász, J.1, Hoffmann, V.1, Szabó, Z.2, Nyéki, J.2, Szabó, T.3 & Hegedûs, A.1
1Corvinus University of Budapest, Faculty of Horticultural Science, Department of Genetics and Plant Breeding,
1118 Budapest, 29 Villányi street, Hungary;
2Institute for Extension and Development, University of Debrecen, 138 Böszörményi street,
H-4032 Debrecen, Hungary
3Research and Extension Centre for Fruit Growing, 2 Vadastag street, H-4244 Újfehértó, Hungary
Summary:Quince (Cydonia oblongaMill.) is a minor fruit crop, which is primarily used for marmalade, jam and sauce.Very few quince cultivars
are known all over the world and in many cases similar names are used for presumably different cultivars. The aim of the present study was to
evaluate and characterize the genetic diversity of 36 quince cultivars and selections with SSR markers. Seven out of 8 SSR markers designed from
apple sequences could successfully yield amplification also in quince cultivars. Number of alleles per locus ranged from 2 to 3 alleles. These allele
numbers are quite low when compared to apple. It is supposed to be the consequence of a genetic bottleneck. In spite of the low allele number per
locus, the 36 quince cultivars formed 30 different genotypes. The ratio of homozygosity was low, which might be coupled with the self-
(in)compatibility phenotype of quinces. SSR markers proved unable to differentiate putatively closely related cultivars (e.g. ‘Bereczki’ and
‘Bereczki bôtermô’). In general, the level of polymorphism among the tested quince genotypes was much restricted due to the low allele number
detected. However, it must be considered that the number of analysed SSR loci is not enough high to estimate the overall heterozygosity of the
quince genome. Further experiments are needed and the SSR markers proved to be a reliable and useful tool for such analyses.
International Journal of Horticultural Science 2009, 15 (4): 7–10.
Agroinform Publishing House, Budapest, Printed in Hungary
ISSN 1585-0404
Key words: Cydonia oblonga, quince, genetic diversity, SSR
82007; Garkava-Gustavsson et al., 2008) and cultivar
identification (Guilford et al., 1997; Galli et al., 2005).
The aim of the present study was to evaluate and
characterize the genetic diversity of 36 quince cultivars and
selections obtained from the valuable germplasm collection
of Research and Extension Centre for Fruit Growing in
Újfehértó.
Materials and methods
Young leaves of 36 quince cultivars and selections were
collected in the germplasm collection of the Research and
Extension Centre for Fruit Growing, Újfehértó. Genomic
DNA was extracted using a DNeasy Plant Mini Kit (Qiagen,
Hilden, Germany). PCR was conducted using eight pairs of
primers flanking SSR sequences: CH01f02, CH01h01
(Gianfranceschi et al., 1998), CH02c02a, CH02c09,
CH03g04, CH04e03, CH05d11, CH05e03 (Liebhard et al.,
2002). PCR was carried out in a PTC 200 thermocycler (MJ
Research, Budapest, Hungary) using the program described
for the primers. PCR was performed in 25 µl volume
containing about 25 ng of genomic DNA, 1 x buffer, 2.5 mM
MgCl2, 0.2 mM dNTPs, 0.2 ìM each of primers and 1 U Taq
polymerase (Fermentas, Szeged, Hungary). To determine the
exact size of different fragments, the PCR products amplified
by fluorescently labelled forward primers (JOE or FAM)
were run in an automated sequencer ABI PRISM 3100
GeneticAnalyzer (Applied Biosystems, Budapest, Hungary).
Band scoring was analyzed using ABI Genotyper 3.7 NT
software.
Results and Discussion
Seven out of 8 SSR markers derived from apple genome
could successfully produce amplified bands in quince
cultivars. All these 7 primer pairs generated 1 or 2 discrete
reproducible fragments. In cases when only one band was
visible, its size is reported twice since the genotype is
presumed to be homozygous. The allele sizes determined by
the same markers were identical in all replications. Primer
CH05e03 could not amplify any fragment in the tested
cultivars. Since no SSR markers have been developed in
quince, it is considered that the use of transferable SSRs from
apple could be suitable for the genetic identification of
quince cultivars (Yamamoto et al., 2004). Number of alleles
per locus ranged from 2 to 3 alleles (Figure 1): 3 alleles in
CH03g07 locus (labelled as A1, A2, A3), 2 alleles in
CH04e03 locus (B1, B2), 3 alleles in CH05d11 locus (C1,
C2, C3), 3 alleles in CH02C02 locus (D1, D2, D3), 3 alleles
in CH01F02 locus (E1, E2, E3), 3 alleles in CH02C09 locus
(F1, F2, F3) and 2 alleles in CH01H01 locus (G1, G2). These
allele numbers are quite low, the same loci in apple showed
average of 9.2 allelic variants, from 6 to 13 alleles per locus
(Liebhard et al., 2002; Galli et al., 2005). It is supposed to be
the consequence of a genetic bottleneck, which might be at
least partially the explanation why only a limited number of
quince cultivars are known worldwide (Hedrick, 1925).
Using RAPD analysis the tested cultivars showed a very
similar banding pattern and it was impossible to differentiate
them (Yamamoto et al., 2004). However, an isozyme analysis
revealed a restricted genetic diversity but enabled to classify
cultivars into several types (Sanchez et al., 1988).
In spite of the low allele number per locus, the 36 quince
cultivars formed 30 different genotypes. From this point of
view these SSR markers were effective for the
characterization of quince cultivars with unknown origin.
Yamamoto et al. (2004) found 12 different genotypes among
20 cultivars. The ratio of homozygosity was low, which
seems to be very indicative considering that fruit set studies
on the self-fertility properties of quince revealed many
contradictions according to a wide range of environmental
conditions (Szabó et al., 1999; Nyéki et al., 2003). If self-
pollination is prevented by self-incompatibility, the level of
heterozygosity is high as in case of apple (Guarino et al.,
Halász, J., Hoffmann, V., Szabó, Z., Nyéki, J., Szabó, T. & Hegedûs, A.
Figure 1: Frequency of alleles observed in 36 quince cultivars at each SSR locus.
92006). As quince belongs to the Maloideae subfamily, a
functional gametophytic self-incompatible system (GSI) can
be presumed to control fruit set, nevertheless no data are
available on molecular investigations (Halász, 2008). This
would be, however, extremely important to provide more
unequivocal results required for the clarification of the above
described uncertainties. It must be mentioned that the
number of analysed SSR loci is not enough to estimate the
overall heterozygosity of the quince genome, further
experiments are needed and the SSR markers proved to be a
reliable and useful tool for such analyses.
Allelic compositions for each analysed quince cultivar
are presented in Table 1. ‘Bereczki’ and ‘Bereczki bôtermô’
cultivars were characterized by the same genotype. It was
expectable because ‘Bereczki bôtermô’ can be supposed to
be a close relative of ‘Bereczki’. Generally, sport mutants are
indistinguishable from their progenitor genotype since their
genomes differ only in a small portion, which influences a
quantitative trait, for example yield or productivity. The
chance to find this region by using SSR markers is very
small. ‘Györkönyi almaalakú’, ‘Almaalakú’ and ‘Almaalakú
mezôtúri’ carried the same SSR-alleles. Their fruits are very
similar in morphology but the fruit shape is not the best
feature to differentiate cultivars since it may vary year by
year (Hedrick, 1925; Szabó, 1998). To confirm this, the
cultivar ‘Patyi almaalakú’ was characterized by a different
genotype. SSR-genotypes of ‘Dunaföldvári’, ‘Olasz 3’ and
‘Óriás birs’ were identical. Two other cultivars,
‘Szentlôrinci’ and ‘Nyakas birs’ also possessed the same
allelic composition. All other cultivars were characterized by
different genotypes; however, it must be added that most of
them differed only in one or two loci. The level of
polymorphism was much restricted due to the low allele
number detected. Still quince remains to be a cryptic species
within the Maloideae subfamily, requiring more detailed
analyses for a satisfactory characterization.
Acknowledgements
This work was financed by the NKTH-OTKA K68921.
Júlia Halász and Attila Hegedûs are grateful for receiving a
János Bolyai Scholarship, HAS, and Attila Hegedûs for the
prize Excellence in science, Corvinus University of
Budapest.
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Characterization of quince (Cydonia oblonga Mill.) cultivars using SSR markers developed for apple
Table 1:Allelic compositions of 7 SSR loci for each analysed quince
cultivar
Cultivars SSR genotypes
1. 162 A1A3B2B2C2C3D2D3E1E1F2F2G1G2
2. Almaalakú A1A3B1B2C2C3D1D1E1E2F1F2G1G2
3. Almaalakú mezôtúri A1A3B1B2C2C3D1D1E1E2F1F2G1G2
4. Angersi A1A3B2B2C1C2D2D3E1E2F2F2G1G2
5. Aromate A1A3B1B2C1C2D2D3E2E3F2F2G1G2
6. Auri A1A3B1B2C2C2D2D3E2E3F2F3G1G2
7. Bereczki A1A3B1B2C1C2D2D3E2E3F1F2G1G2
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11. Cserszegi birs A1A2B1B2C2C3D1D1E1E2F1F2G2G2
12. De Husi A1A3B2B2C1C2D2D3E2E3F2F2G1G2
13. Deák Csokonai A1A3B1B2C2C2D2D3E2E3F1F2G1G2
14. Dunaföldvári A2A3B1B2C2C3D2D3E1E2F2F2G1G2
15. Fehérvári A1A3B1B2C2C3D1D2E1E2F1F2G2G2
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17. Györkönyi almaalakú A1A3B1B2C2C3D1D1E1E2F1F2G1G2
18. Konstantinápolyi A1A3B1B2C2C3D2D3E1E2F2F2G1G2
19. Lengyel A1A3B1B2C2C3D2D3E1E3F1F2G2G2
20. Leskovacka A1A3B2B2C1C2D1D1E3E3F2F3G1G2
21. Mezôkövesdi A1A3B1B1C1C2D2D3E1E3F2F3G1G2
22. Mezôtúri A2A3B1B2C2C3D2D3E2E3F2F3G1G2
23. Moldova nesti A2A2B1B2C2C2D2D3E2E3F2F2G1G2
24. Nyakas birs A1A3B1B2C2C3D2D3E1E2F2F3G1G2
25. Olasz 3 A2A3B1B2C2C3D2D3E1E2F2F2G1G2
26. Óriás birs A2A3B1B2C2C3D2D3E1E2F2F2G1G2
27. Ôrsi birs A1A3B2B2C3C3D1D1E2E3F1F2G1G2
28. Patyi almaalakú A1A3B1B2C2C3D2D3E1E2F1F2G1G2
29. Pomázi A1A3B1B2C2C3D2D3E1E2F1F2G2G2
30. Radónia A1A3B1B2C2C2D1D1E2EeF2F2G1G2
31. Román portugál A1A3B2B2C2C2D1D1E2E2F2F2G1G2
32. Szentlôrinci A1A3B1B2C2C3D2D3E1E2F2F3G1G2
33. Trimoacium A1A3B2B2C2C3D2D3E1E2F1F2G2G2
34. Triumph A1A3B1B2C1C2D2D3E1E2F2F2G2G2
35. Váli birs A2A3B1B2C2C3D1D1E2E3F1F1G1G2
36. Vranja A1A3B1B2C1C2D1D2E2E3F2F3G1G2
10
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Characterization of quince (Cydonia oblonga Mill.) cultivars using SSR markers developed for apple

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Characterization of quince (Cydonia oblonga Mill.) cultivars using SSR markers developed for apple

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