• Molecules and Cells
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• v.21 no.1
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• pp.135-140
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• 2006

Cytoplasmic male sterility (CMS) in plants, which is due to failure to produce functional pollen, is a maternally inherited trait. Specific nuclear genes that suppress CMS, termed fertility restorer (Rf) genes, have been identified in several plants. In this study, Rfl-inked molecular markers in pepper (Capsicum annuum L.) were detected by bulked segregant analysis of eight amplified fragment length polymorphisms (AFLPs). Only AFRF8 was successfully converted to a cleaved amplified polymorphic sequence (CAPS) marker. This was named AFRF8CAPS and genotype determination using it agreed with that obtained with the original AFRF8. A linkage map with a total size of 54.1 cM was constructed with AFRF8CAPS and the seven AFLP markers using the Kosambi function. The AFRF8CAPS marker was shown to be closest to Rf with a genetic distance of 1.8 cM. These markers will be useful for fast and reliable detection of restorer lines during $F_1$ hybrid seed production and breeding programs in pepper.

• Korean Journal of Breeding Science
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• v.41 no.3
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• pp.194-204
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• 2009

Cytoplasmic male sterility (CMS) and fertility restoration have been utilized as valuable tools for $F_1$-hybrid seed production in many crops despite laborious breeding processes. Molecular markers for the selection of CMS-related genes help reduce the expenses and breeding times. A previously reported genomic region containing the Ppr-B gene, which is responsible for restoration of fertility and corresponds to the Rfo locus, was used to develop gene-based or so-called "functional" markers for allelic selection of the restorer-of-fertility gene (Rfo) in $F_1$-hybrid breeding of radish (Raphanus sativus L.) Polymorphic sequences among Rfo alleles of diverse breeding lines of radish were examined by sequencing the Ppr-B alleles. However, presence of Ppr-B homolog, designated as Ppr-D, interferes on specific PCR amplification of Ppr-B in certain breeding lines. The organization of Ppr-D, resolved by genome walking, revealed extended homology with Ppr-B even in the promoter region. Interestingly, PCR amplification of Ppr-D was repeatedly unsuccessful in certain breeding lines implying the lack of Ppr-D in these radishes. Ppr-B could only be successfully amplified for analysis through designing primers based on the sequences unique to Ppr-B that exclude interference from Ppr-D gene. Four variants of Rfo alleles were identified from 20 breeding lines. A combination of three molecular markers was developed in order to genotype the Rfo locus based on polymorphisms among four different variants. These markers will be useful in facilitating $F_1$-hybrid cultivar development in radish.

• Molecules and Cells
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• v.25 no.1
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• pp.20-29
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• 2008

Cytoplasmic male sterility (CMS), one of the most important traits in crop breeding, has been used for commercial seed production by $F_1$ hybrid cultivars of pepper (Capsicum annuum L.). To develop reliable molecular markers for allelic selection of the Restorer-of-fertility (Rf) gene, which is known to be a major determinant of pollen fertility restoration in peppers, a sequence of approximately 10 kb flanking an RAPD fragment closely linked to the Rf locus was obtained by genome walking. A homology search revealed that this sequence contained an LTR retrotransposon and a non-LTR LINE-like retrotransposon. Sequencing of this Rf-linked region to search for polymorphisms between a dominant and recessive allele revealed 98% nucleotide sequence identity between them. A third polymorphic haplotype of the Rf-linked sequence, which has 94-96% nucleotide sequence identity with the two previously isolated haplotypes, was identified among a large number of breeding lines. Utilizing polymorphic sequences in the haplotypes, PCR markers were developed for selection of particular haplotypes and used to examine the distribution of the haplotypes in diverse breeding lines, cultivars, and C. annuum germplasms. Surprisingly, the third haplotype was the predominant type in C. annuum germplasms, while its frequency in $F_1$ hybrid cultivars was relatively low. Meanwhile, analysis of breeding lines whose Rf allele genotypes and male-sterility phenotypes were already known revealed that the third haplotype was mainly present in exotic breeding lines that cause unstable male-sterility when combined with sterile cytoplasms.

• Plant Resources
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• v.7 no.2
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• pp.136-140
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• 2004

Co-segregation of male fertility with DNA markers selected by RAPD analysis as being potentially linked to the restorer gene (Rf) for Cytoplasmic male sterility (CMS) was analyzed using segregating F2 population. One RAPD marker directly linked to the Rf locus was identified. Amplification of OPT-02/570 using the STS primers generated a monomorphic band of each fertile plants randomly selected F2 progenies. From these results, this specific marker would be strongly linked to be restoring gene. The use of STS marker is effective in overcoming the reliability of the RAPD phenotype and improving their utility for MAS, co-dominant STS markers are especially very useful.

• The Plant Pathology Journal
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• v.28 no.4
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• pp.418-422
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• 2012

Eleven hot pepper accessions collected in Vietnam showed stable resistance to bacterial wilt as well-known resistance sources, MC4 and MC5, in repeated inoculation tests with different Ralstonia solanacearum isolates conducted from 2004 to 2010. Seven of these accessions (specifically KC981, KC1006, KC1021, KC1027, KC1045, KC1050, and KC1055) resulted in stable male sterile F1 plants in the crosses with a cytoplasmically male sterile (CMS) Chilseong (CMS-A, Srfrf ), and therefore, they were maintainers (CMS-B) with a genotype of Nrfrf. The rest (KC980, KC995, KC999, and KC1009) produced stable male fertile F1 plants in the crosses, and therefore, were restorers (CMS-C) with a genotype of N(S)RfRf. Therefore, the maintainer and restorer sources of resistance may be used in preference in breeding maternal (CMS and their maintainers) and paternal parents (restorers) for resistance to bacterial wilt, respectively, in the hybrid breeding system utilizing cytoplasmic male sterility.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.38 no.5
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• pp.413-417
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• 1993

This study was conducted to breed various cytoplasmic genetic male sterile (CGMS) and restorer lines with backgrounds of Korean japonica rice varieties. The CGMS line BT-CMS was crossed with the restorer line AR-3. The fertile $F_1 was emasculated and crossed with the Korean japonica rice varieties of the early maturing Sobaegbyeo, Odaebyeo, Gwanagbyeo and Daeseongbyeo, and of the medium maturing Hwajinbyeo, Paldal, Suwon 224 and Iri 386, and of the late maturing Nagdongbyeo, Palkweng, Hwacheongbyeo and Milyang 97. Each of the three way cross $F_{1S} was segregated into fertile and sterile individuals. The sterile individuals in each cross were discarded and the fertile individuals were emasculated and backcrossed with the Korean japonica rice varieties. The same process was applied from BC$_1$F$_1$ to BC$_4$F$_1$ generation. In the ${BC_4}{F_1}of each cross, the male sterile individual was crossed with the recurrent Korean japonica variety which was maintainer of male sterility. The male sterile lines of ${BC_6}{F_1}were named as Sobaegbyeo R, Odaebyeo R, Gwanagbyeo R, etc. The fertile individuals homozygous in pollen fertility were selected from the ${BC_4}{F_2}generation and named as Sobaegbyeo R, Odaebyeo R, Gwanagbyeo R, etc. Agronomic characteristics of the CGMS, restorer lines with backgrounds of Korean japonica, and the recurrent Korean japonica rice varieties grown in the field condition were compared. Culm length of the CGMS lines tended to be shorter than that of the recurrent parent, however no significant differences in heading date, panicle length and yield component were found among the CGMS, restorer and recurrent lines.

• Horticultural Science & Technology
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• v.33 no.4
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• pp.550-558
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• 2015

To analyze the linkage relationships among molecular markers recently reported to be linked to onion (Allium cepa L.) Ms, a restorer-of-fertility locus, in onion (Allium cepa L.), three single nucleotide polymorphism markers were converted into cleaved amplified polymorphic sequence (CAPS) markers based on onion transcriptome sequences and the rice genome database. Analysis of the recombinants selected from 4,273 segregating plants using CAPS and other linked markers demonstrated the jnurf13 and jnurf610 markers to perfectly co-segregate with the Ms locus. In contrast to jnurf13, the jnurf610 marker was not in perfect linkage disequilibrium with the Ms locus in diverse breeding lines. Thus, the jnurf13 marker and the marker for identification of cytoplasm types were utilized to enhance the efficiency of onion breeding through four applications. First, 89 maintainer lines containing the normal cytoplasm and homozygous recessive Ms genotypes were successfully identified from 100 breeding lines. Second, these two molecular markers were used to analyze the main sources of male-fertile contaminants frequently found in the male-sterile parental lines during F1 hybrid seed production. The majority of the contaminants contained heterozygous Ms genotypes, indicating that pollen grains harboring the dominant Ms genotype may have been introduced during propagation of the maintainer lines. Therefore, the genetic purity of the two maintainer lines was analyzed in the third application, and the results showed that both maintainer lines contained 13-21% off-types. Finally, the two markers were used to increase the seed yield potentials of two open-pollinated varieties containing sterile cytoplasms by removing the plants harboring homozygous recessive and heterozygous Ms genotypes.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.38 no.5
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• pp.418-424
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• 1993

In order to test combining ability of the cytoplasmic-genetic male sterile(CGMS) and restorer lines with backgrounds of Korean japonica rice varieties, diallel crosses were made among the early maturing CGMS lines Sobaegbyeo A, Odaebyeo A, Gwanagbyeo A and Daeseongbyeo A, and the restorer lines Sobaegbyeo R, Odaebyeo R, Gwanagbyeo Rand Daeseongbyeo R, and among the medium maturing CGMS lines Hwajinbyeo A, Paldal A, Suwon 224 A and Iri 386 A and the restorer lines Hwajinbyeo R, Paldal R, Suwon 224 Rand Iri 386 R, and among the late maturing CGMS lines Nagdongbyeo A, Palkweng A, Hwacheongbyeo A and Milyang 97 A and the restorer lines Nagdongbyeo R, Palkweng R, H wacheongbyeo Rand Milyang 97 R. The fourty eight combinations of the hybrids, their parents and the maintainers were grown in field condition in 1992. General combining ability, relative specific combining ability and relative combining ability for yield of the CGMS and restorer lines were tested. Combining ability of the CGMS lines Odaebyeo A, Iri 386 A, Nagdongbyeo A and Palkweng A, and of the restorer lines Odaebyeo R, Hwajinbyeo R, Hwacheongbyeo Rand Palkweng R was relatively high. Heterobeltiosis for grain yield over the better parent of the early maturing hybrids ranged from -17 % to 15%, and that of the medium maturing ones from -4% to 22%, and that of the late maturing ones from -46% to 30% respectively, Standard heterosis over the standard variety of the early maturing hybrids ranged from -13 % to 12 %, and that of the medium maturing ones from 0 % to 26%, and that of the late maturing ones from -38% to 26% respectively, Positive and negative heterosis for grain yield of the hybrid rices were observed depending on cross combinations, In the positive heterosis hybrids, number of panicles per hill and number of spikelets per panicle contributed mostly to grain yield, while in the negative heterosis hybrids, spikelet fertility and 1000 grain weight were closely correlated to grain yield. Amylose content of the hybrid rices, maintainer and restorer lines was mostly lower than 20 %, and alkali digestion value was higher than 6.0. Grain appearance of the hybrids tested was similar to the korean japonica rice varieties.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.25 no.2
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• pp.49-57
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• 1980

A $F_1$ hybrid out of 143 crosses of rape (Brassica napus) using cytoplasmic-genetic male sterility yielded 5.18 tons per hectare. European and native varieties used, were found out to have 78% and 58% of genes restoring fertility, respectively. The $F_1$ hybrids of three way cross using non-isogenic maintainer showed higher heteroses than those of double cross. To produce F 1 hybrids of three way cross, three stages of crossing operations are needed and seeds produced from about 60 hectares can be covered to 60, 000 hectares.

• Horticultural Science & Technology
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• v.30 no.1
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• pp.73-78
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• 2012

KC00256, KC00406, KC00462, KC00463, KC00820, and KC00821, the genetic resources that have previously been reported as moderately resistant to Phytophthora capsici, as well as the line KC01322, a new source of moderate resistance introduced from Laos, were tested against two strains (Pc003 and Pc005) of P. capsici. We also determined the nuclear restorer genotypes of these lines, in regards to their interaction with cytoplasmic male sterility, through crossing the resources with cytoplasmic male sterile Punggok-A (Srfrf) and determining the fertility of the $F_1$ hybrids. The studied lines exhibited a low level of resistance to both the strains of P. capsici compared to highly resistant CM334, but their response was fairly consistent for both P. capsici strains. KC00406, KC00462, KC00463, and KC01322 produced stable, male fertile $F_1$ plants indicating that they are restorers with genotype N(S)RfRf. KC00821 produced male sterile $F_1$ plants and was identified as a maintainer with genotype Nrfrf. The $F_1$ plants of the KC00820 cross, however, set a few male fertile flowers in the greenhouse at seedling stage, then became male sterile after being transplanted to the plastic greenhouse soil in May and remained so to the end of the growing season. Therefore, KC00820 is an unstable maintainer with genotype Nrfrf. The moderate resistance exhibited by these genetic resources may be integrated into breeding programs aimed at promoting higher levels resistance via recurrent selection or hybridization.