Complete volume with articles 1 to 32 - Cucurbit Breeding - North ...
Complete volume with articles 1 to 32 - Cucurbit Breeding - North ... Complete volume with articles 1 to 32 - Cucurbit Breeding - North ...
young fruit exterior color, and that it is dominant to light fruit color. ‘Krymka’ and ‘Eshkolit Ha’Amaqim’ bred true for exterior immature fruit color and all of the F1 plants examined had dark exterior color, closely resembling that of the dark parent, ‘Krymka’ (Table 2). The F2 segregated in accordance with a 3:1 ratio of dark to light and the backcross to ‘Eshkolit Ha’Amaqim’ segregated in accordance with a 1:1 ratio of dark to light. These results indicate that there is a single gene determining dark young fruit exterior color, and that it is dominant to light fruit color. The results of both crosses are compatible with the idea that there exists a recessive gene for light exterior immature fruit color in Cucumis melo. This gene for light fruit color apparently is not at the same locus as the heretofore reported dominant gene for light fruit color, Wi. However, it is not known what the genetic relationship is between the dominant gene for striped fruit, as in ‘Dulce’, and the dominant gene for dark fruit, as in ‘Krymka’. To determine whether these two genes are at separate loci or in an allelic series, such as occurs in Cucurbita pepo (2), a testcross for allelism will need to be conducted. Literature Cited: 1. Danin-Poleg, Y., Y. Tadmor, G. Tzuri, N. Reis, J. Hirschberg, and N. Katzir. 2002. Construction of a genetic map of melon with molecular markers and horticultural traits, and localization of genes associated with ZYMV resistance. Euphytica 125: 373–384. 2. Paris, H.S. and R.N. Brown. 2005. The genes of pumpkin and squash. HortScience 40: 1620–1630. 3. Pitrat, M. 2002. Gene list for melon. Cucurbit Genet. Coop. Rep. 25: 76–93. Table 1. Results of crossing ‘Dulce’ (striped fruit) with ‘TAM-Dew’ (light fruit). Expected Accession Total Striped rind Light rind ratio P1, Dulce 15 15 0 P2, TAM-Dew 15 0 15 F1, P2 × P1 15 15 0 F2, (P2 × P1) ⊗ 244 188 56 3:1 0.546 0.46 BC1, P2 × (P2 × P1) 88 43 45 1:1 0.045 0.83 Table 2. Results of crossing ‘Krymka’ (dark fruit) with ‘Eshkolit Ha’Amaqim’ (light fruit). Expected Accession Total Dark rind Light rind ratio χ 2 P1, Krymka 15 15 0 P2, Eshkolit Ha’Amaqim 15 0 15 F1, P2 × P1 15 15 0 F2, (P2 × P1) ⊗ 203 148 55 3:1 0.475 0.48 BC1, P2 × (P2 × P1) 45 25 20 1:1 0.556 0.45 BC1, P1 × (P2 × P1) 40 40 0 18 Cucurbit Genetics Cooperative Report 28-29: 17-18 (2005-2006) χ 2 P P
SNP Discovery and Mapping in Melon (Cucumis melo L.) Maya Lotan-Pompan, Galil Tzuri, Vitaly Portnoy, Rotem Beja and Nurit Katzir Agricultural Research Organization, Department of Genetics and Vegetable Crops, Newe Ya’ar Research Center, P. O. Box 1021, Ramat Yishay 30-095, Israel katzirn@volcani.agri.gov.il Amir Sherman, Shahar Cohen, Nir Dai and Arthur A. Schaffer Agricultural Research Organization, Dept. of Genetics and Vegetable Crops, P. O. Box 6, Bet Dagan, 50-250, Israel Jim Giovannoni Boyce Thompson Institute for Plant Research, Tower Road, Cornell University, Ithaca, NY 14853, U.S.A. Introduction: Several studies have applied various marker types (including RFLP, RAPD, AFLP and SSR) for mapping and assessing genetic variation within C. melo (1, 3, 6, 7). A partial linkage map of melon from a cross between PI 414723 (C. melo Group Acidulus) and ‘Dulce’ (C. melo Group Reticulatus) was previously described by us (2). Single nucleotide polymorphisms (SNPs) are an abundant form of genetic variation in the genome of various species that are extremely useful for gene mapping and phylogenetic studies. In recent years, SNPs have become important as genomic markers with numerous technical methods developed for their detection (4). In this study, SNP markers for genes belonging to major fruit metabolic pathways were developed. In order to link between these markers and fruit quality traits, these SNPs are currently being located on the PI 414723 × ‘Dulce’ map. Materials and Methods: Plant material. The F2 mapping population included 112 individuals derived from a cross between ‘Dulce’ and PI 414723 (2). The ‘Dulce’ fruit is aromatic, sweet, of high pH, with orangecolored flesh and a netted rind. The PI 414723 fruit is non-aromatic, non-sweet, and acidic, with salmon-colored flesh and no net. SNP discovery. From the melon EST database (http://melon.bti.cornell.edu), three different methods were used for SNP detection: (i) comparative analysis of sequences of different genotypes, (ii) primers designed for the end sequences of interesting ESTs were used to amplify the genomic DNA of both parental lines (‘Dulce’ and PI 414723); this was followed by screening of the amplicons using either DHPLC (Denaturing High Performance Liquid Chromatography) or SSCP (Single Strand Conformational Polymorphisms), in order to find polymorphism between the parental amplicons. The polymorphic amplicons were re-sequenced in order to validate the SNPs, (iii) direct sequencing of parental amplicons. Genotyping. SNP genotyping was conducted using the Sequenom MassARRAY® platform, by the highthroughput genotyping assays: hME and iPLEX. These assays allowed multiple PCR reactions in a single well (5). JoinMap 3.0 (Kyazma, Holland) was used for linkage analysis and map calculations. Results: SNP Markers for ca. 100 genes belonging to major fruit metabolic pathways were developed (Table 1). These include genes encoding for key enzymatic and transport steps in carbohydrate, acid, volatile and carotenoid metabolism in melon and Cucurbit Genetics Cooperative Report 28-29: 19-21 (2005-2006) 19
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young fruit exterior color, and that it is<br />
dominant <strong>to</strong> light fruit color.<br />
‘Krymka’ and ‘Eshkolit Ha’Amaqim’ bred<br />
true for exterior immature fruit color and all<br />
of the F1 plants examined had dark exterior<br />
color, closely resembling that of the dark<br />
parent, ‘Krymka’ (Table 2). The F2<br />
segregated in accordance <strong>with</strong> a 3:1 ratio of<br />
dark <strong>to</strong> light and the backcross <strong>to</strong> ‘Eshkolit<br />
Ha’Amaqim’ segregated in accordance <strong>with</strong><br />
a 1:1 ratio of dark <strong>to</strong> light. These results<br />
indicate that there is a single gene<br />
determining dark young fruit exterior color,<br />
and that it is dominant <strong>to</strong> light fruit color.<br />
The results of both crosses are compatible<br />
<strong>with</strong> the idea that there exists a recessive<br />
gene for light exterior immature fruit color<br />
in Cucumis melo. This gene for light fruit<br />
color apparently is not at the same locus as<br />
the here<strong>to</strong>fore reported dominant gene for<br />
light fruit color, Wi. However, it is not<br />
known what the genetic relationship is<br />
between the dominant gene for striped fruit,<br />
as in ‘Dulce’, and the dominant gene for<br />
dark fruit, as in ‘Krymka’. To determine<br />
whether these two genes are at separate loci<br />
or in an allelic series, such as occurs in<br />
<strong>Cucurbit</strong>a pepo (2), a testcross for allelism<br />
will need <strong>to</strong> be conducted.<br />
Literature Cited:<br />
1. Danin-Poleg, Y., Y. Tadmor, G. Tzuri,<br />
N. Reis, J. Hirschberg, and N. Katzir.<br />
2002. Construction of a genetic map of<br />
melon <strong>with</strong> molecular markers and<br />
horticultural traits, and localization of<br />
genes associated <strong>with</strong> ZYMV resistance.<br />
Euphytica 125: 373–384.<br />
2. Paris, H.S. and R.N. Brown. 2005. The<br />
genes of pumpkin and squash.<br />
HortScience 40: 1620–1630.<br />
3. Pitrat, M. 2002. Gene list for melon.<br />
<strong>Cucurbit</strong> Genet. Coop. Rep. 25: 76–93.<br />
Table 1. Results of crossing ‘Dulce’ (striped fruit) <strong>with</strong> ‘TAM-Dew’ (light fruit).<br />
Expected<br />
Accession<br />
Total Striped rind Light rind ratio<br />
P1, Dulce 15 15 0<br />
P2, TAM-Dew 15 0 15<br />
F1, P2 × P1 15 15 0<br />
F2, (P2 × P1) ⊗ 244 188 56 3:1 0.546 0.46<br />
BC1, P2 × (P2 × P1) 88 43 45 1:1 0.045 0.83<br />
Table 2. Results of crossing ‘Krymka’ (dark fruit) <strong>with</strong> ‘Eshkolit Ha’Amaqim’ (light fruit).<br />
Expected<br />
Accession<br />
Total Dark rind Light rind ratio χ 2<br />
P1, Krymka 15 15 0<br />
P2, Eshkolit Ha’Amaqim 15 0 15<br />
F1, P2 × P1 15 15 0<br />
F2, (P2 × P1) ⊗ 203 148 55 3:1 0.475 0.48<br />
BC1, P2 × (P2 × P1) 45 25 20 1:1 0.556 0.45<br />
BC1, P1 × (P2 × P1) 40 40 0<br />
18 <strong>Cucurbit</strong> Genetics Cooperative Report 28-29: 17-18 (2005-2006)<br />
χ 2<br />
P<br />
P