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

• Korean Journal Plant Pathology
• /
• v.14 no.3
• /
• pp.212-216
• /
• 1998

Capsicum pepper selections fro PI163192, PI241670, PI244670, PI271322, PI308787, PI322719, and PI369994 were confirmed to be non-hypersensitively resistant to race 3 of Xanthomonas campestris pv. vesicatoria. A resistant cultivar,‘SR’, was shown to be hypersensitive. Four Koran local cultivars, a cytoplasmic male sterile line (A-line) and its maintainer (B-line) were highly susceptible. The resistant selections and cultivars were crossed with a male sterile A-line (Smsms) and fertility of their F1 hybrids was examined by observing he pollen production, testing pollen germination, and quantifying the amount of pollen produced per anther to identify the genotype interacting with the male sterile cytoplasm. The seven resistant PI selections turned out to be restorers (N(S)MsMs) and‘SR’to be a maintainer (Nmsms).

• Plant Biotechnology Reports
• /
• v.3 no.4
• /
• pp.309-315
• /
• 2009

Molecular markers developed from the flanking sequences of two cytoplasmic male sterility (CMS)-associated genes, orf456 and ${\Psi}atp6-2$, have been used for marker-assisted selection of CMS in pepper. However, in practice, the presence of orf456 and ${\Psi}atp6-2$ at substoichiometric levels even in maintainer lines hampers reliable selection of plants containing the CMS gene. In this study, we developed a novel CMS-specific molecular marker, accD-U, for reliable determination of CMS lines in pepper, and used the newly and previously developed markers to determine the cytoplasm types of pepper breeding lines and germplasms. This marker was developed from a deletion in a chloroplast-derived sequence in the mitochondrial genome of a CMS pepper line. CMS pepper lines could be unambiguously determined by presence or absence of the accD-U marker band. Application of orf456, ${\Psi}atp6-2$and accD-U to various pepper breeding lines and germplasms revealed that accD-U is the most reliable CMS selection marker. A wide distribution of orf456, but not ${\Psi}atp6-2$, in germplasms suggests that the pepper cytoplasm containing both orf456 and ${\Psi}atp6-2$ has been selected as CMS cytoplasm from cytoplasm containing only orf456. Furthermore, factors other than orf456 may be required for the regulation of male sterility in pepper.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.29 no.3
• /
• pp.227-231
• /
• 1984

Chinese CGMS (Cytoplasmic-Genetic Male Sterility) is being transfered to Korean cultivars which will be used as CGMS maintainers. These maintainer lines along with the Chinese CGMS lines and their maintainer lines were crossed with another group of Korean cultivars which will be used as the fertility restorers, and their yield heterosis were examined. The yield heterosis was calculated as high as 113.0-221.1 % in terms of heterosis (F$_1$/Midparent) with the grand mean of 150.5%, and, 86.1-179.8% in terms of heterobeltiosis (F$_1$/better parent) with the grand mean of 125.3%. Among the female parent groups, Chinese mainteiners were the highest following by Chinese MS lines and the Korean maintainers were the lowest. Heterobeltiosis of yield components were 92.7%, 120.0%, 103.0% and 112.0% for number of panicles/unit area, number of grains/panicle, fertility per cent and 1000 grain weight, respectively. Path-coefficients of yield components to the yield were calculated. The highest one was number of grains per panicle ranging 0.8073-0.8649 followed by the 1000 grain weight ranging 0.2000-0.5032. The grain yield ratios to the grand mean were calculated in terms of combining ability. It was highest with the Chinese maintainers followed by Chinese MS lines and The Korean maintainers were lowest. This facts bring cautions to the breeding efforts to transfer the Chinese CGMS in to Korean maintainers.

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

• Molecules and Cells
• /
• v.21 no.3
• /
• pp.411-417
• /
• 2006

We have developed a polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP) marker that can distinguish male-fertile (N) and male-sterile (S) cytoplasm in onions. The PCR-RFLP marker was located in a chloroplast psbA gene amplicon. Digesting the amplicons from different cytoplasm-containing varieties with the restriction enzyme MspI revealed that N-cytoplasm plants have a functional MspI site (CCGG), whereas the S-cytoplasm plants has a substitution in that site (CTGG), and thus no MspI target. The results obtained using this PCR-RFLP marker to distinguish between cytoplasmic male sterile factors in 35 onion varieties corresponded with those using a CMS-specific sequence-characterized amplified region (SCAR) marker. Moreover, the PCR-RFLP marker can identify N- ot S-cytoplasms in DNA sample mixtures in which they are in up to a 10-fold minority, indicating that use of the marker has high diagnostic precision. We also demonstrated the usefulness of the SNP detected in the psbA gene for high-throughput discrimination of CMS factors using Real-time PCR and a TaqMan probe assay.

• Horticultural Science & Technology
• /
• v.29 no.1
• /
• pp.53-60
• /
• 2011

Cytoplasmic male sterility (CMS) induced by mutant mitochondria genome, has been used for commercial seed production of $F_1$ hybrid cultivars in diverse crops. In pepper (Capsicum annuum L.), two sterile cytoplasm specific gene organization, atp6-2 and coxII were identified. An open reading frame, orf456 nearby coxII gene has been speculated to induce male sterility (MS) by mutagenic analysis. Moreover, molecular markers for atp6-2 and coxII of mitochondrial genotype (mitotype) were developed. However, the Cytoplasmic MS specific markers, atp6SCAR and coxIISCAR markers appeared in both N and S cytoplasms when polymerase chain reaction (PCR) cycles prolonged more than 40 cycles. Since the reported molecular markers were dominant markers, the presence of the faint sterile-specific band in normal cytoplasm may lead to the mis-classification of pepper breeding lines. To solve this problem, one common forward primer and two different reverse primers specific to normal coxII and sterile orf456 genes were designed after analyzing their gene organizations. By using these three primers, N and S coxII specific bands were co-amplified in male-sterile lines, but only normal coxII specific band was amplified in maintainer lines. Since the reverse primer for sterile coxII was specifically designed 275 bp downstream of orf456, relatively stable PCR amplification patterns were observed regardless of the number of PCR cycles. These primer sets easily identified different mitotypes among the divergent breeding lines, commercial cultivars and diverse germplasms.

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

• Current Research on Agriculture and Life Sciences
• /
• v.28
• /
• pp.39-46
• /
• 2010

Selection procedures for breeding lines resistant to both bacterial wilt caused by Ralstonia solanacearum and Phytophthora blight caused by P. capsici were executed to generations from $F_2$ and $BC_1F_1$ to $F_4$ and $BC_1F_3$ of crosses between a Phytophthora resistant breed, 'Chilbok No. 1' and bacterial wilt resistant accessions introduced from Vietnam during 2009 and 2010. The breeding populations or lines were tested for resistance to P. capsici and resistant plants were selected. The resistant selections were inoculated with R. solanacearum to discard susceptible plants. Resistance to P. capsici was conspicuously improved by selection from $F_2$ and $BC_1F_1$ and the final selections showed a similar level of resistance to P. capsici as a commercial Phytophthora resistant cultivar, Muhanjilju. A few $BC_1F_2$ selections were crossed to a cytoplasmic male sterile line, Chilbok-A, to identify their nuclear genotype interacting with male sterile cytoplasm. Majority of them was fixed to maintainer (Nrfrf) and only two resulted segregating into male sterile and male fertile plants indicating that the pollen parents were heterozygous in the fertility-restoring gene.

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