• Molecules and Cells
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• 제25권1호
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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.

• 한국식물병리학회지
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• 제14권3호
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• pp.212-216
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• 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
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• 제3권4호
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• pp.309-315
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• 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.

• 한국작물학회지
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• 제29권3호
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• pp.227-231
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• 1984

중공의 CGMS (cytoplasmic-genetic male sterility)를 한국품종으로 이전하여 그들을 MS유지계통으로 쓸 경우 이들이 임성회복친과 교잡되어 발현될 것으로 기대되는 수중의 잡종강 세정도를 중공의 CGMS계통 및 그 유지계통들과의 잡종과 대비하여 조사검토하였다. 그 결과는 다음과 같이 요약된다. 1. 수량의 잡종강세는 heterosis(F$_1$/MP)로 표현하여 113~221.1% 전체평균 150.5%이었고 hetero-beltiosis(F$_1$/better p)로 표현하여 86.1~179.8%전체평균 125.3%이였다. 품종군별로는 중공계통들이 가장 높았는데 유지계통들은 MS계통들보다 높았다. 다음으로 높은 것은 임성회복친들이었고 한국품종으로 유지계통들은 가장 낮은 잡종강세를 보였다. 2. 수량구성요소별로 잡종강세를 heterobeltiosis로 표현해 보면 수수는 전체평균 92.7%로 감소되었고 수당입수는 120.0%로 증가했으며 임실율은 103.0%로 유의할만한 변화가 없었고 천립중은 112%로 증가하였다. 증수에 크게 기여한 수당입수와 천립중의 heterobeltiosis를 품종군별로 보면 수량의 그것과 비슷한 경향이었다. 3. 수량구성요소가 수량에 기여한 정도를 임성회복친별 교배조합군별로 검토하기 위하여 경노계수를 계산하였더니 밀양46, 수원294 및 수원287 조합 모두에서 다같이 수당입수가 0.8073~0..8649로 가장 컸고 다음은 천립중으로 0.2000~0.5032이었다. 4. 품종군별 잡종조합의 수량을 교배조합능력이라고 간주하고 이것을 전체평균에 대한 비율로 계산하여 비교했는데 임성회복친 중 밀양46과 수원294는 일반조합능력은 비슷하나 특수조합능력은 차가 있고 S. 287은 일반조합능력이 떨어진다. 유지계통별로는 중공의 유지계통이 가장 높았고 한국유지계통이 가장 낮았다. 중공의 CGMS를 한국품종으로 이전하는데 있어서의 문제점을 검토하였다.

• Molecules and Cells
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• 제21권1호
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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.

• Molecules and Cells
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• 제21권3호
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• pp.411-417
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• 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.

• 원예과학기술지
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• 제29권1호
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• pp.53-60
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• 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.

• 한국작물학회지
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• 제38권5호
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• pp.413-417
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• 1993

우리나라 japonica 벼의 일대잡종을 다양하게 육성할 목적으로 세포질 유전자적 웅성불임인 BTCMS에 대한 임성회복력이 있는 계통 AR-3을 교잡하여 그 F1을 모본으로 하고 조ㆍ중ㆍ만생의 한국 japonica형 벼 4품종씩을 교본 반복친으로 5회 backcross하여 우리나라 벼 품종 배경의 우엉불임 계통과 임성회복계통을 육성하고 이들의 주요 작물학적 특성을 각각의 반복친과 비교한 결과를 요약하면 다음과 같다. 1. 우리나라 japonica형 품종의 배경을 갖는 세포질 유전자적 웅성불임계통을 조ㆍ중ㆍ만생군으로 각각 4개씩 도합 12계통 육성하였으며, 동일한 품종의 배경을 갖는 임성회복계통친도 조ㆍ중ㆍ만생군으로 각각 4개씩 12계통 육성하였다. 2. 육성된 웅성불임계통은 조중군에서 소백벼 A, 오대벼,A, 관악벼A, 대성벼A, 중생군에서 화진벼A, 팔달A, 수원 224A, 이리 386A, 만생군에서 낙동벼A, 팔굉A, 화청벼A 및 밀양 97A 등으로 명명하였다. 3. 육성된 임성회복계통은 조생군에서 소백벼R, 오대벼R, 팔달R, 수원 224R, 이리 386R, 만생군에서 낙동벼R, 팔굉R, 화청벼R 및 밀양 97R 등으로 명명하였다. 4. 육성된 웅성불임계통들은 반복친(불임유지친)에 비하여 간장이 단축되는 경향이었으나 출수기, 수장 및 수량구성요소 등은 대체로 비슷하였다. 5. 육성된 임성회복계통들은 반복친과 비교하면 대체로 간장, 수장, 수량구성요소 및 수량의 차이가 없었다.

• Current Research on Agriculture and Life Sciences
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• 제28권
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• pp.39-46
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• 2010

칠성초에 역병 저항성을 도입한 칠복1호에 베트남 도입 풋마름병 저항성 계통을 교배하여 육성한 $F_2$ 및 $BC_1F_1$에서부터 $F_4$ 및 $BC_1F_3$까지 역병-풋마름병 복합 저항성 선발을 2009년도와 2010년도에 걸쳐 수행하였다. 매 세대 역병을 접종하여 저항성을 평가하여 선발하고 선발개체에 풋마름병을 접종하여 감염되는 개체는 도태하였다. 역병에 대한 저항성은 선발과 함께 현저히 향상되었으며, 선발계통들은 역병 저항성으로 판매되고 있는 교배종 '무한질주'와 비슷한 수준의 저항성을 나타내었다. $BC_1F_2$ 선발개체를 칠복CMS-A라인에 교배를 하여 $F_1$의 임성을 보고 화분친의 CMS-Rf유전자형을 검정하였다. 대부분Nrfrf로 고정되고, 칠복 ${\times}$ KC995, 칠복 ${\times}$ KC1009 조합의 일부 개체가 이형(heterozygote) 상태인 것으로 확인되었다.

• 한국육종학회지
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• 제41권3호
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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.