Mitochondrial-targeted Expression of orf456 Causes Male Sterility in Chinese Cabbage (Brassica rapa L.)

( Li Liu ),( Yeong Deuk Jo ),( Won Hee Kang ),( Dosun Kim ),( Byoung Cheorl Kang ) 한국육종학회 2013 Plant Breeding and Biotechnology Vol.1 No.2

Cytoplasmic male sterility (CMS) is a phenomenon specific to plant reproduction that has been extensively exploited for hybrid seed production. Orf456 was previously identified as a candidate gene mediating male sterility in pepper. To gain further insight of this candidate gene, we carried out experiments to transform Chinese cabbage (Brassica rapa L.). About 30 T1 transgenic lines were obtained and approximately 50 % of T1 transgenic Chinese cabbage lines showed male-sterility. To evaluate pollen viability, three different approaches including plasmolysis test, staining pollen and in vitro germination assay were used. Analysis of the CMS transgenic lines showed that trasgenic Chinese cabbages produced aberrant pollen development while some were unable to produce pollen. In conclusion, the mitochondrial orf456 gene could induce partial male sterility in transgenic Chinese cabbage. Nevertheless, how the orf456 gene precisely functions to induce male sterility and its biochemical function remains to be discovered.

Investigation of Cytoplasmic Male Sterility and Development of Maintainer and Restorer Lines in Rye (Secale cereale L.)

Heo, Hwa-Young,Hong, Byung-Hee,Seong, Rak-Chun,Park, Moon-Woong The Korean Society of Crop Science 2000 Korean journal of crop science Vol.45 No.5

Rye has been a major winter forage crop in Korea. Varietal improvement of rye has been practiced either by hybrid or population breeding systems. Hybrid breeding offers important advantages over population breeding since it is normally a cross-pollinated crop. The hybrid breeding in rye has been possible since cytoplasmically inherited forms of male sterility (CMS) and corresponding nuclear restorer genes were found. The objectives of this research were to develop the maintainer and restorer lines of Korean inbred lines and to estimate the effect of 'Pampa' type of CMS cytoplasm on yield and its related characteristics. For easy discrimination of male-sterile status of plants, anther scoring and the restore index system in which seed-setting and pollen quantity of viability were taken into account were established. High significant correlation between pollen quantity and pollen viability was found. For "Pampa" cytoplasm, four of 14 Korean inbred lines tested turned out to be a maintainer but no restorer was found. But for "235b" CMS cytoplasm, seven inbred lines acted as complete restorers. The Korean inbred rye lines acted mainly as maintainers in "Pampa" cytoplasm but acted mainly as restorer in "235b" cytoplasm. The 'Pampa' cytoplasm inducing male sterility reduced cohn length and plant height and increased the number of tiller, so forage yield and grain yield were enhanced. However, heading date was slightly delayed compared to the normal cytoplasm.elayed compared to the normal cytoplasm.

역병-풋마름병 복합저항성 세포질웅성불임계 고추 육성

이재무,김병수 경북대학교 농업과학기술연구소 2010 慶北大農學誌 Vol.28 No.-

칠성초에 역병 저항성을 도입한 칠복1호에 베트남 도입 풋마름병 저항성 계통을 교배하여 육성한 및 에서부터 및 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. 선발개체를 칠복CMS-A라인에 교배를 하여 의 임성을 보고 화분친의 CMS-Rf유전자형을 검정하였다. 대부분Nrfrf로 고정되고, 칠복 KC995, 칠복 KC1009 조합의 일부 개체가 이형(heterozygote) 상태인 것으로 확인되었다. 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 and to and 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 and and the final selections showed a similar level of resistance to P. capsici as a commercial Phytophthora resistant cultivar, Muhanjilju. A few 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.

Selection of Maintainer Line in Open-Pollinated Onion (Allium cepa L. cv. “Manchuhwang”) using SCAR Marker Linked to Cytoplasmic Male Sterile Factor

Kwang-Soo Cho,Su-Young Hong,Young-Seok Kwon,Jong-Gyu Woo,Ji-Young Moon,Seung-Yeol Ryu,Hyo-Guen Park 한국육종학회 2005 한국육종학회지 Vol.37 No.3

Onion (Allium cepa L.) is one of the major condiment vegetables in Korea. Cytoplasmic-genic male sterility (CMS) system is used on a commercial basis for F1 hybrid development. Onion has biennial nature and strong self-depression, so breeding of inbred lines are taken a long period about 8 to 10 years. To breed a maintainer line highly adopted in long-day condition, SCAR marker and test crosses were used. We selected N-cytoplasm plants using sequence characterized amplified region (SCAR) marker from ‘Manchuhwang’ (open-pollinated cultivar), which was bred by National Institute of Highland Agriculture (NIHA). Selected N-cytoplasm plants were crossed with male sterile inbred line (W202A, Wisconsin Univ., USA). A total of 66 crosses were accomplished, and 34 crosses could be analyzed for the nuclear restore allele. Among 34 combinations, the offsprings of one combination showed all male sterility and this line was selected as the maintainer line in the ‘Manchuhwang’. This maintainer line was just available with materials for F1 hybrid seed production without fixing of characters with male sterile line.

Investigation of Cytoplasmic Male Sterility and Development of Maintainer and Restorer Lines in Rye(Secale cereale L.)

Hwa-Young Heo,Byung-Hee Hong,Rak-Chun Seong,Moon-Woong Park 韓國作物學會 2000 Korean journal of crop science Vol.45 No.5

Rye has been a major winter forage crop in Korea. Varietal improvement of rye has been practiced either by hybrid or population breeding systems. Hybrid breeding offers important advantages over population breeding since it is normally a cross-pollinated crop. The hybrid breeding in rye has been possible since cytoplasmically inherited forms of male sterility (CMS) and corresponding nuclear restorer genes were found. The objectives of this research were to develop the maintainer and restorer lines of Korean inbred lines and to estimate the effect of 'Pampa' type of CMS cytoplasm on yield and its related characteristics. For easy discrimination of male-sterile status of plants, anther scoring and the restore index system in which seed-setting and pollen quantity of viability were taken into account were established. High significant correlation between pollen quantity and pollen viability was found. For "Pampa" cytoplasm, four of 14 Korean inbred lines tested turned out to be a maintainer but no restorer was found. But for "235b" CMS cytoplasm, seven inbred lines acted as complete restorers. The Korean inbred rye lines acted mainly as maintainers in "Pampa" cytoplasm but acted mainly as restorer in "235b" cytoplasm. The 'Pampa' cytoplasm inducing male sterility reduced cohn length and plant height and increased the number of tiller, so forage yield and grain yield were enhanced. However, heading date was slightly delayed compared to the normal cytoplasm.elayed compared to the normal cytoplasm.

Evaluation of Cytoplasm types by PCR-marker in Male-Sterility of Onion (Allium cepa L.)

Cheol Woo Kim,Eul Tai Lee,Hyun Ju Park,Young Seok Jang,In Hu Choi,Jin Ki Bang,Sung Ju Ahn,Gap Chae Chung 한국육종학회 2007 한국육종학회지 Vol.39 No.2

The production of hybrid onion (Allium cepa L.) seed is economically feasible using systems of cytoplasmic-genic male sterility(CMS). The identification of cytoplasm types might be helpful to development of sterile and maintainer lines. Due to this reason, a knowledge of the cytoplasm type is essential for hybrid breeding in onion. Especially, due to the biennial generation time of onion, to distinguish of onion cytoplasm type in individual plant may be helpful to reduce the efforts in a breeding program, when new sterile lines and the corresponding maintainer lines be developed. In our study, we applied the PCR-marker (orfA-501) to identify the cytoplasmic genotypes of collected 100 accessions of bulb onion. Among accessions, S-cytoplasm was found in 57 accessions. Nineteen accessions possessed only N-cytoplasm and twenty four accessions possessed both S- and N-cytoplasm.

유채의 목포CGMS 및 유지친 개발

장영석,최인후,김철우,권병선,오용비,정병춘,방진기,이정일 한국육종학회 2002 한국육종학회지 Vol.34 No.3

Rapeseed as a partial allogamous crop(allogamy rate : 25~75%) displays depression by continuous selfing. Therefore, it is essential to produce F hybrids with high yield using cytoplasmic genetic male sterility(CGMS). This breeding system needs to develop male sterile(MS) line, MS-maintainer and restorer line in order to increase commercial value of F hybrid seeds. MS and restorer lines can be easily obtained in rapeseed, but MS-maintainer is difficult to breed artifically or to discover from germplasm. Mokpo-CGMS is originated from the cross between Tower(♀) and Isuzu(??), and its stamens are degenerated by interacting of cyto- plasmic and nuclear genes. The Mokpo-CGMS line has good composition of oil with no erucic aicd. But the MS could be maintained by the cross between Mokpo-CGMS plants and fertile sister plants of the Mokpo-CGMS and 50% of the progeny show male sterility. To develop perfect maintainer which can produce 100% male sterile plants, a cross between Altex(normal cytoplasm and FF nuclear genotype) and fertile sister line of Mokpo-CGMS(sterile cytoplasm and FS nuclear genotype) was made. It was difficult to select the line with normal cytoplasm and male sterile nuclear gene in F generation because all phenotypes including heterozygous and homozygous types were expressed in the same plant type with fertility. Test cross was made between individual F plants and Mokpo-CGMS line in order to select maintainer. After backcrossing the F plant to the Mokpo-CGMS 14 times, a perfect maintainer producing 100% male sterile plants was identified in progenies.

Distribution of Three Cytoplasm Types in Onion (Allium cepa L.) Cultivars Bred in Korea and Japan

Sunggil Kim(김성길),Eul-Tai Lee(이을태),Cheol-Woo Kim(김철우),Moo-Kyoung Yoon(윤무경) 한국원예학회 2009 원예과학기술지 Vol.27 No.2

이전 연구에서 양파의 미토콘드리아 유전체 구조의 변이에 기초하여 양파에 존재하는 3종의 세포질(Normal, CMS-T, CMS-S)을 식별할 수 있는 분자표지를 개발하였다. CMS-T와 CMS-S는 양파의 웅성불임을 유기하는 세포질로 알려져 있으며 둘 다 F1 잡종 품종 개발에 활용되고 있다. 본 연구에서는 기 개발된 분자표지가 다양한 유전자원을 대상으로 신뢰성 있게 적용이 되는지 검정하기 위하여 116종의 한국과 일본에서 육종된 품종을 분석한 결과 성공적으로 모든 품종의 세포질 종류를 판별할 수 있었고 새로운 세포질 형태는 관찰되지 않았다. CMS-T 세포질의 빈도는 한국에서 육종된 품종에서 높게 나타났는데 CMS-T에 의해서 유기된 웅성불임은 회복유전자의 수가 적어도 3개 이상이므로 임성회복 기작이 매우 복잡하다. 반면 CMS-S 세포질의 빈도는 한국 품종에서 매우 낮게 나타났는데 CMS-S는 단 한 개의 회복유전자만 존재하기 때문에 분자육종 시스템에 적합한 세포질이라 할 수 있다. 이러한 경향은 특히 만생종에서 두드러진다. 반면 정상 세포질은 조생종에서 높은 빈도로 나타나 아직 대부분의 조생종은 고정종 품종이 주류를 이루는 것으로 추정된다. Previously, we reported a molecular marker designed on the basis of variation of mitochondrial genome organization for distinction of three onion (Allium cepa L.) cytoplasm types (Normal, CMS-T, and CMS-S). The CMS-T and CMS-S cytoplasms are involved in induction of cytoplasmic male-sterility, both of which are utilized in development of F1 hybrid cultivars. Reliable application of this molecular marker was tested for 116 onion cultivars developed in Korea and Japan. The molecular marker was successfully utilized to identify specific cytoplasm types of all tested cultivars, showing reliable application of the molecular marker and absence of another unidentified cytoplasm type in tested cultivars. The frequency of CMS-T cytoplasm was high in cultivars developed in Korea compared with those developed in Japan. CMS-T cytoplasm is responsible for CMS-T male-sterility of which inheritance pattern of fertility restoration is complex due to involvement of at least three restorer-of-fertility genes. On the other hand, a few cultivars containing CMS-S cytoplasm were identified in Korean cultivars. CMS-S male-sterility is more stable in diverse environmental conditions, and its fertility restoration is controlled by only a single gene, rendering CMS-S suitable for establishment of molecular breeding systems. This distribution pattern was the most evident in late-maturing cultivars. The frequency of normal cytoplasm was relatively high in early-maturing cultivars, implying most early-maturing cultivars are still open-pollinated varieties.

Sources of Resistance to Bacterial Wilt Found in Vietnam Collections of Pepper (Capsicum annuum) and Their Nuclear Fertility Restorer Genotypes for Cytoplasmic Male Sterility

Ngoc Hung Tran,김병수 한국식물병리학회 2012 Plant Pathology Journal Vol.28 No.4

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.

Sources of Resistance to Bacterial Wilt Found in Vietnam Collections of Pepper (Capsicum annuum) and Their Nuclear Fertility Restorer Genotypes for Cytoplasmic Male Sterility

Tran, Ngoc Hung,Kim, Byung-Soo The Korean Society of Plant Pathology 2012 Plant Pathology Journal Vol.28 No.4

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.