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Soon O. Park* and Kevin M. Crosby

Mature fruit size and shape are important traits of most melon types. Our objective was to identify RAPD markers associated with major QTL affecting fruit weight, length, diameter and shape by means of bulked segregant analysis in an F2 population from the ananas melon cross of Deltex (larger fruit size) × TGR1551 (smaller fruit size). Clear separations for fruit weight, length, diameter, and shape between Deltex and TGR1551 were observed. Continuous distributions for fruit weight, length, diameter and shape were found in the F2 population indicating quantitative inheritance for the fruit traits. Significant positive correlations were detected between fruit weight and shape traits (r = 0.73 to 0.80). A significant positive correlation was observed between fruit weight and glucose (r = 0.35) or fructose (r = 0.25), whereas no correlation was noted between fruit weight and sucrose or total soluble solids. Two small and large bulks for fruit weight and shape were developed from F2 plants. A total of 240 primers were used to simultaneously screen between the small and large bulks, and between Deltex and TGR1551. Twenty-six RAPD markers were polymorphic for the small and large bulks. Ten markers were found to be significantly and consistently associated with fruit size and shape traits on the basis of simple linear regression. Of the 10 markers associated, four displayed an amplified DNA fragment in the small bulk, while six showed an amplified DNA fragment in the large bulk. The associated marker OJ07.350 explained 15% to 27% of the phenotypic variation for the fruit traits. These markers associated with QTL for melon fruit size and shape are expected to be useful in melon breeding programs for modifying fruit size.

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Alexandra B. Napier, Kevin M. Crosby, and Soon O. Park

Muskmelons (Cucumis melo L.) play an important role in the American diet. Ranked as one of the top 10 most-consumed fruits by the USDA, cantaloupe melons have the highest amount of beta-carotene of all the ranked fruits. Beta-carotene, also called pro-Vitamin A, is an essential nutrient required for eye health, and may have the potential, as an antioxidant to reduce the risks associated with cancer, heart disease, and other illnesses. Breeding melons with increased levels of beta-carotene will benefit consumer health. Research has found phytonutrients are most bioavailable when consumed in their fresh form, rather than as vitamin supplements. The high level of beta-carotene found in some melons has a genotypic component, which may be exploited to breed melons high in beta-carotene. Molecular markers and marker-assisted selection (MAS) can be used to increase the efficacy of the breeding process, while lowering breeding costs. An F2 population was created using `Sunrise', the female parent, containing no beta-carotene crossed with `TAM Uvalde', a high beta-carotene variety. A field population consisting of 115 F2 individuals and a greenhouse population containing 90 F2 individuals were grown. The resulting fruit were screened phenotypically and ranked according to beta-carotene content. Chisquare values fit the previously reported model of a single dominant gene for presence of beta-carotene (orange-flesh) vs. absence (green or white flesh). A continuous distribution of beta-carotene concentrations from high to low suggested quantitative inheritance for this trait. Two eight-plant DNA bulks composed of either high or low beta-carotene F2 individuals were screened for polymorphic molecular markers using the amplified fragment-length polymorphism technique.

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Soon O. Park, Dermot P. Coyne, and James R. Steadman

Bean rust, caused by Uromyces appendiculatus, is a major disease of common bean (Phaseolus vulgaris). The objective was to identify RAPD markers linked to the gene (Ur-7) for specific resistance to rust race 59 using bulked segregant analysis in an F2 segregating population from the common bean cross GN1140 (resistant to rust) × Nebraska #1 (susceptible to rust). A single dominant gene controlling specific resistance to race 59 was found in the F2 and was confirmed in the F3. Seven RAPD markers were detected in a coupling-phase linkage with the Ur-7 gene. Coupling-phase RAPD markers OAA11.500, OAD12.550, and OAF17.900 with no recombination to the Ur-7 gene were found. Three RAPD markers were identified in a repulsion-phase linkage with the Ur-7 gene among the three markers at a distance of 8.2 cM. This is the first report on RAPD markers linked to the Ur-7 gene in common bean. The RAPD markers linked to the gene for specific rust resistance of Middle American origin detected here, along with other independent rust resistance genes from other germplasm, could be used to pyramid multiple genes into a bean cultivar for more-durable rust resistance.

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Soon O. Park, Hye Y. Hwang, and Kevin M. Crosby

Our objectives were to construct a randomly amplified polymorphic DNA (RAPD) marker-based linkage map using an F2 population derived from the melon (Cucumis melo) cross of ‘Deltex’ × TGR 1551, and map quantitative trait loci (QTL) for sucrose, total soluble solids (TSS), ratio of sucrose to total sugars (RSTS), and ascorbic acid as well as the ms-3 locus for male sterility previously reported in other muskmelon crosses. Due to the dominant character of RAPD markers, we scored 192 ‘Deltex’- and 158 TGR 1551-derived markers. One hundred eighty (94%) of the 192 markers fit the expected 3:1 ratio. On the basis of the 180 markers, we constructed a ‘Deltex’ linkage map of 171 markers distributed on 12 linkage groups (LGs) with a total map distance of 1182 cM. One hundred fifty (95%) of the 158 markers were identified to be nondistorted. We developed a TGR 1551 linkage map of 138 markers distributed on 12 LGs with a total distance of 1163 cM. A combined map of 12 LGs with a total map distance of 1394 cM was made from 82 marker pairs expressing codominance. Nine LGs were integrated into those of the existing composite map by 17 anchor markers. We mapped the ms-3 locus for male sterility on LG 9, which corresponds to LG 10 of the classical map and LG VII of the composite map. Six QTL for sucrose were located on LGs 2, 3, 4, 6, and 11. Three on LGs 3, 4, and 6 and four on LGs 2, 3, 6, and 11 of the six QTL for sucrose were also noted to be QTL for TSS and RSTS, respectively. A single QTL for ascorbic acid was placed on LG 5. This map will also be used to identify QTL for fruit sweetness, quality, size, and shape traits, as well as disease resistance.

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Lianghong Chen, Ajmer S. Bhagsari, Soon O. Park, and Sarwan Dhir

This study was carried out to optimize conditions for plant regeneration of sweetpotato [Ipomoea batatas (L.) Lam] using shoot tips, petioles, and leaves of Selection 75-96-1 as explants in Murashige and Skoog (MS) with several growth regulators at different levels. Callus initiation and callus proliferation media were 9.0 μm 2,4-dichlorophenoxyacetic acid (2,4-D) and 9.0 μm 2,4-D + 1.1 μm N 6-benzyladenine (6-BA) in protocol I; 8.1 μm α-naphthaleneacetic acid (NAA) + 1.2 μm kinetin (KIN) and 5.4 μm NAA + 4.6 μm KIN in protocol II; 0.9 μm 2,4-D, and 0.9 μm 2,4-D + 1.2 μm N-isopenylamino purine (2iP) in protocol III; NAA (8.1 μm) + KIN (1.2 μm) and 2,4-D (0.9 μm) + 2ip (1.2 μm) in protocol IV, respectively. In protocol I and II, shoot tip, petiole, and leaf were used, but only petiole and leaf in protocol III and IV. In the protocol I and II, somatic embryos were obtained only from shoot tip explants; in protocol III and IV, only from petioles. The frequencies of somatic embryo development were 33.3% in protocol I, 42.1% in protocol II, 21.2% in protocol III, and 10.3% in protocol IV, respectively. The leaf explants failed to produce somatic embryos in all the experiments. In protocol I, somatic embryogenesis occurred through the well-known sequence of globular-, heart-shaped-, torpedo-, and cotyledon-type embryos. However, in protocol II, the structures resembling plumule and radicle were observed before the emergence of torpedo/cotyledon type embryo clusters. The somatic embryogenesis in protocol III and IV was similar to that in protocol I. Growth regulators influenced somatic embryo development. Further, this study showed that explant resource and growth regulators affected the frequency of plant regeneration in sweetpotato.

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Soon O. Park, Dermot P. Coyne, Atilla Dursun, and Geunhwa Jung

Common bacterial blight (CBB), incited by Xanthomonas campestris pv. phaseoli (Xcp), is an important seed-transmitted disease of common bean (Phaseolus vulgaris L.). Tepary bean (Phaseolus acutifolius A. Gray) has high resistance to Xcp. The objective of this study was to identify RAPD markers linked to genes controlling resistance to three isolates of Xcp using bulked segregant analysis in an F2 population from the tepary bean cross CIAT-G40005 (resistant to Xcp) × Nebr.#4B (susceptible to Xcp). Twelve RAPD markers were mapped in a coupling-phase linkage with three genes for resistance to Xcp. The linkage group spanned a distance of 19.2 cM. A marker L7750 was linked to the genes for resistance to Xcp strains EK-11 and LB-2 at 8.4 cM and 2.4 cM, respectively. Markers U10400 and Y14600 were detected as flanking markers for the resistance gene to Xcp strain SC-4A at 2.4 cM and 7.2 cM, respectively. The symbols Xcp-1, Xcp-2, and Xcp-3 were assigned for the genes for resistance to Xcp strains EK-11, LB-2, and SC-4A, respectively. RAPD markers linked to the genes for resistance to Xcp could be used for transferring all of the resistance genes from P. acutifolius to a susceptible P. vulgaris cultivar.

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Soon O. Park, Dermot P. Coyne, and James R. Steadman

Bean rust, caused by Uromyces appendiculatus, is an important disease of common bean (Phaseolus vulgaris L.). The objective was to identify RAPD markers linked to the gene (Ur-6) for specific resistance to rust race 51 using bulked segregant analysis in an F2 segregating population from the common bean cross pinto `Olathe' (resistant to rust) × great northern Nebraska #1 selection 27 (susceptible to rust). A single dominant gene controlling specific resistance to race 51 was hypothesized based on F2 segregation, and then was confirmed in the F3 generation. A good fit to a 3:1 ratio for band presence to band absence for each of three markers was observed in 100 F2 plants. Three RAPD markers were detected in a coupling phase linkage with the Ur-6 gene. Coupling-phase RAPD marker OAB14.600 was the most closely linked to the Ur-6 gene at a distance of 3.5 cM among these markers. No RAPD markers were identified in a repulsion phase linkage with the Ur-6 gene. The RAPD markers linked to the gene for specific rust resistance of Middle American origin detected here, along with other independent rust resistance genes from other germplasm, could be utilized to pyramid multiple genes into a bean cultivar for more durable rust resistance.

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Soon O. Park, Dermot P. Coyne, James R. Steadman, Paul W. Skroch, and Geunhwa Jung

The objective was to detect molecular markers associated with QTL for partial physiological resistance (PPR) to two white mold (WM) isolates, partial field resistance (PFR), plant architecture (PA), and plant height (PH) in a genetic linkage map constructed using recombinant inbred lines (RILs) from the cross `PC-50' (resistant to WM) × XAN-159 (susceptible to WM). Significant correlations (+0.39 and +0.47) were noted between the WM reactions in the greenhouse and field. A significant but negative correlation (–0.33) was observed between the WM reaction and PH in the field. Six QTL affecting PPR to isolate 152 were found on LGs 4, 5, 7, and 8. Six QTL affecting PPR to isolate 279 were found on LGs 2, 3, 4, 7, and 8. Five QTL for PFR were observed on LGs 2, 5, 7, 8, and 11. Two QTL affecting PA were detected on LGs 7 and 8. Two QTL affecting PH were identified on LGs 7 and 8. On one end of LG 8 marker H19.1250 was significant for PPR to both isolates. On the other end of LG 8 the region closely linked to the C locus was significantly associated with PPR to both isolates, PFR, PA and PH. Marker J09.950 on LG 7 was significantly associated with PPR to both isolates, PFR, PH and seed weight. Marker J01.2000 on LG 2 was the most significant locus for both PPR to the isolate 279 and PFR. QTL on LG 5 were found for PPR to the isolate 152 and PFR. Overall, four of the five QTL affecting PFR were also found for PPR to one or both isolates.

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Soon O. Park, Dermot P. Coyne, Nedim Mutlu, Geunhwa Jung, and James R. Steadman

Common bacterial blight, incited by Xanthomonas campestris pv. phaseoli (Xcp) is a serious disease of common bean (Phaseolus vulgaris L.). Randomly amplified polymorphic DNA (RAPD) markers and flower color (V gene) previously were reported to be associated with six quantitative trait loci (QTL) affecting leaf and pod resistance to Xcp. However, the markers for the QTL were not confirmed in different populations and environments to indicate their merit in breeding. The objective was to determine if the associations of RAPD markers and the V gene with QTL for leaf and pod resistance to Xcp in a recombinant inbred (RI) backcross population from the cross BC2F6 `PC-50' × XAN-159 and for leaf resistance to Xcp in an F2 population from a different cross pinto `Chase' × XAN-159 could be confirmed. One or two genes from XAN-159 controlled leaf and pod resistance to Xcp. Among six QTL previously detected, five in the RI backcross population and three in the F2 population were confirmed to be associated with resistance to Xcp. The V gene and RAPD marker BC437.1050 on linkage group 5 were most consistently associated with leaf and pod resistance to two to five Xcp strains in the RI backcross population and with leaf resistance to two Xcp strains in the F2 population. One to three QTL affecting leaf and pod resistance to Xcp accounted for 22% to 61% of the phenotypic variation. Gene number (one to two) estimations and number of QTL (one to three) detected for leaf and pod resistance to Xcp in the RI backcross population were generally in agreement. The marker BC437.1050 and V gene, along with other resistance genes from other germplasm, could be utilized to pyramid the different genes into a susceptible or partially resistant bean line or cultivar to enhance the level of resistance to Xcp.

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Soon O. Park, Kevin M. Crosby, Rongfeng Huang, and T. Erik Mirkov

Male sterility is an important trait of melon in F1 hybrid seed production. Molecular markers linked to a male-sterile gene would be useful in transferring male sterility into fertile melon cultivars and breeding lines. However, markers linked to the ms-3 gene for male sterility present in melon have not been reported. Our objectives were to identify randomly amplified polymorphic DNA (RAPD) markers linked to the ms-3 gene controlling male sterility using bulked segregant analysis in an F2 population from the melon cross of line ms-3 (male-sterile) × `TAM Dulce' (male-fertile), convert the most tightly linked RAPD marker to the ms-3 gene into a sequence characterized amplified region (SCAR) marker based on a specific forward and reverse 20-mer primer pair, and confirm the linkage of the RAPD and SCAR markers with the ms-3 gene in an F2 population from the cross of line ms-3 × `Mission' (male-fertile). A single recessive gene controlling male sterility was found in F2 individuals and confirmed in F3 families. Two RAPD markers that displayed an amplified DNA fragment in the male-sterile bulk were detected to be linked to the ms-3 gene in the F2 population from the cross of line ms-3 × `TAM Dulce'. RAPD marker OAM08.650 was closely linked to the ms-3 gene at 2.1 cM. SCAR marker SOAM08.644 was developed on the basis of the specific primer pair designed from the sequence of the RAPD marker OAM08.650. The linked RAPD and SCAR markers were confirmed in the F2 population from the cross of line ms-3 × `Mission' to be consistently linked to the ms-3 gene at 5.2 cM. These markers were also present in 22 heterozygous fertile F1 plants having the ms-3 gene. The RAPD and SCAR markers linked to the ms-3 gene identified, and confirmed here could be utilized for backcrossing of male sterility into elite melon cultivars and lines for use as parents for F1 hybrid seed production.