구두 발표
OA-01 Genetic Analysis of Shoot Fresh Weight in Soybean Using an Interspecific Cross of PI483463 and Hutcheson Krishnanand P. Kulkarni, Sovetgul Asekovaa, Minsu Kim, Cheol-Woo Park, Jae-Eun Jeong, Jong Tae Song, Jeong-Dong Lee * School of Applied Biosciences, Kyungpook National University, Daegu 41566, Korea Shoot fresh weight (SFW) is one of the parameters, used to assess the total plant biomass yield in soybean. In this study, a total of 188 F5:8 recombinant inbred line (RILs) derived from an interspecific cross of PI483463 (Glycine soja) and Hutcheson (Glycine max) were investigated for SFW variation for three consecutive years. The parental lines along with RILs were phenotyped in the field at the R6 stage by measuring total biomass in kg/plot. The phenotypic variation (PV) was exploited to identify SFW QTLs. Three QTLs qsfw6_1, qsfw15_1, and qsfw19_1 for SFW were identified on chromosome 6, 15 and 19 respectively. The QTL qsfw19_1 flanked within the markers BARC-044913-08839 - BARC-029975-06765, was the consistent QTL expressed in all the three environments. The PV explained by the QTLs across all environments ranged from 6.56 to 21.32%. The negative additive effects by the consistent QTL indicated contribution of SFW from wild parent. Additive x environment interaction effects affected the expression of SFW QTL. Screening of the RIL population with additional SSRs from the qsfw19_1 region delimited the QTL within the markers SSR19-1329 - BARC-29975-06765. QTL mapping using bin map detected two QTLs, qsfw19_1a and qsfw19_1b. The QTL qsfw19_1a mapped close to the Dt1 gene locus, which affects stem termination, plant height and floral initiation in soybean. Prospective candidate genes for SFW were pinpointed and sequence variations within their sequences were detected using high quality whole genome resequencing data. The findings in the present study could be valuable in understanding genetic and molecular basis of SFW in forage soybean. Corresponding author: Tel. 053-950-5709, E-mail: jdlee@knu.ac.kr 13
OA-02 Identification of Candidate Genes Conferring to Resistance to Foxglove Aphid in Soybean Sumin Park 1, Eunsil Kim 1, Mijung Cho 1, Kyung Hye Kim, Jin Kyo Jung 2, Jeong-Dong Lee 3, Ju Seok Lee, Sungtaeg Kang 1* 1 Department of Crop Science and Biotechnology, Dankook University, Cheonan 330-714, Korea 2 National Institute of Crop Science, RDA, 151 Seodun-dong, 441-857, Korea 3 Division of Plant Bioscience, Kyungpook National Univ, Daegu 702-701, Korea The sap-sucking insects infect a wide of plants worldwide and causes serious yield losses in soybean, especially. The objective of this study was to identify the putative genes to foxglove aphid resistance in wild soybean, PI 366121 (Glycine soja Sieb. and Zucc.). One hundred and forty-one F4:8 recombinant inbred lines developed from a cross between susceptible variety, Williams 82 and foxglove aphid resistance wild soybean, PI 366121 were used. The two type of resistance response, antibiosis and antixenosis resistance were evaluated through choice and no-choice test, graded by the degree of total plant damage and primary infestation leaf damage; a genome-wide molecular linkage map was constructed with 29,898 single-nucleotide polymorphism markers utilizing a AxiomR 180K soyasnp array. Using inclusive composite interval mapping analysis for foxglove aphid resistance, one major candidate QTL on chromosome 7 was identified. The major QTL on chromosome 7 showed both antixenosis and antibiosis resistance responses. The newly identified major QTL was consistent with previously reported QTL, Raso2, which showed around 5 times narrow down interval range with 8 candidate genes. This result could contribute to breeding for new foxglove aphid resistant soybean cultivar. Corresponding author: Tel. 010-3551-0534, E-mail: kangst@dankook.ac.kr 14
OA-03 침수처리에 따른 콩 생육특성 및 저항성 자원 선발 조미정 1, 박수민 1, 김은실 1, 김경혜 1, 문준경 2, 이주석 1, 강성택 1* 1 충청남도 천안시 단대로 119 단국대학교 생명자원과학대학 식량생명공학과 2 전북 완주군 농생명로 370 농업생명자원부 농생물게놈활용연구사업단 최근 전세계적 환경변화에 따른 장기간의 계절적 강우나 배수불량으로 인한 침수 피해는 작물의 생장과 수확량에 영향 을 미쳐 심각한 문제가 되고 있다. 현재까지 이러한 문제를 해결하기 위해서 내습성 품종개발 등에 대한 연구는 일부 있 지만 생육특성에 대한 정확한 유전자 관련 연구는 거의 없는 실정이다. 따라서 본 연구는 콩에서 침수저항성 자원을 선 발하고 선발자원의 생육특성을 규명하며 간이선발방법을 개발하여 침수피해 경감기술 개발에 도움을 주고자 실시하였 다. 콩 유전자원 약4,000종을 침수처리 및 식물체 반응을 조사 하였고, 식물체 반응 조사 결과에 따라 저항성자원을 선 발하고, 선발된 저항성자원을 정밀검정 하였다. 또한 포장처리는 명확하게 저항성자원을 선발 할 수 있으나 시간이 오래 걸리고 노력이 많이 들어 육종 등에 쉽게 이용 할 수 없기 때문에 간이 검정법 개발이 필요하게 되어 간이 검정법을 개 발했다. 약 4,000종의 콩 유전자원에서 식물체 반응 조사 결과를 바탕으로 16종의 저항성자원을 선발하였고 이 중 줄기 와 뿌리의 부패정도가 현저히 낮은 저항성자원 3점을 최종 선발하여 생육특성을 정밀검정한 결과 부정근수와 제1복엽의 엽색 변화정도가 침수처리에 따른 저항성 및 감수성자원을 구분 짓는 우수한 지표임이 확인되었다. 개발된 간이 검정법 으로 검정한 결과 부정근수가 침수처리에 따른 저항성 및 감수성자원을 구분 짓는 우수한 지표임을 확인하였다. Corresponding author: Tel. 041-550-3621, E-mail: kangst@dankook.ac.kr 15
OA-04 한국 밀 품종 육성현황 및 발전방향 강천식 1*, 정영근 1, 김경훈 2, 손재한 1, 김학신 3, 박종철 1, 오영진 1, 김경호 1, 이광원 1, 김보경 1, 박철수 4 1 전라북도 완주군 이서면 혁신로 국립식량과학원 작물육종과 2 경상남도 밀양시 점필재로 국립식량과학원 논이용작물과 3 전라북도 완주군 이서면 혁신로 국립식량과학원 작물재배생리과 4 전라북도 전주시 덕진구 백제대로 농업생명과학대학 작물생명과학과 밀은 세계 3대 작물 중 하나이며 약 126개국에서 2억 2,427만ha를 재배하여 7억 만 톤가량을 생산하고 있다. 국내에서도 약 10천ha 정도를 재배하여 4만 2천 톤을 생산하고 있다. 국내 밀 품종은 1970년 이후 답리작 적응 조숙 다수성 밀 품 종개발을 목표로 진행한 결과 현재까지 38품종이 개발되었다. 정부보급종은 2009년 금강밀 1품종, 162톤을 공급하기 시 작하여 2016년은 금강밀 등 4품종을 농가에 보급할 예정이다. 38품종 중 금강을 포함한 30품종이 중력분용으로 이용되 고 있으며, 강력분용으로 조경과 백강 2품종, 박력분으로 고소, 조아와 우리 3품종, 취반용으로 백찰, 신미찰과 신미찰1 호 3품종이 개발되었다. 육종성과 중 성숙기의 경우 1970년대는 6월 13일에서 2010년대 6월 3일로 10일 단축되었고, 수 량은 1970년대 408kg/10a에서 2010년대 532kg/10a로 124kg 증수하였다. 2010년 이후에는 용도별 다양한 밀 품종을 개발 하였는데, 과자용, 케이크용, 국수용, 빵용, 백립계 찰밀 등 용도의 다양화가 이루어졌다. 재해안정성 부분에서는 수발아 와 붉은곰팡이병 저항성인 새금강이 개발되었다. 최근 이상기후로 인하여 기존에 보고되지 않았던 생리장애와 병 등이 발생되어 생산량과 품질을 떨어뜨리고 있다. 이러한 시점에서현재까지의 답리작 적응 조숙 다수성 용도별 품종 개발을 지속적으로 진행하고, 생산안정성과 품질유지를 위한 생리장애와 병에 저항성을 증진시키기 위해 더욱더 노력하여 국산 밀 자급률 향상에 이바지해야 할 것으로 판단된다. Corresponding author: Tel. 063-238-5227, E-mail: kcs1209@korea.kr 16
OA-05 벼흰잎마름병 저항성 유전자가 집적된 자포니카 벼 계통 개발 박현수 1*, 김기영 2, 김우재 1, 정지웅 1, 이종희 2, 노태환 1, 남정권 1, 백만기 1, 신운철 1, 김정주 1, 정종민 1, 조영찬 1, 김보경 1, 안상낙 3 1 전라북도 완주군 이서면 혁신로 181 농촌진흥청 국립식량과학원 2 전라북도 전주시 완산구 농생명로 300 농촌진흥청 연구정책국 3 대전광역시 유성구 대학로 99 충남대학교 농업생명과학대학 우리나라 자포니카 벼 품종의 벼흰잎마름병 저항성 증진을 위하여 네 개의 저항성 유전자 Xa1, Xa3, xa5 및 Xa21이 집 적된 우량 계통을 개발하였다. Xa1 저항성 유전자를 보유한 일미벼 와 Xa3+xa5+Xa21이 집적된 익산575호 간 인공교 배를 하였다. 분리세대에서 저항성 유전자 연관 분자표지 선발, 벼흰잎마름병 저항성 생물검정 및 농업형질에 대한 표현 형 선발을 함께 적용하여 네 개의 저항성 유전자가 집적된 약배양 유래 7 계통과 계통육종 유래 10 계통을 각각 개발하 였다. F2 집단에서 Xa1+Xa3+xa5+Xa21 저항성 유전자가 집적된 조합이 가장 강한 저항성 반응을 나타냈다. 이 조합은 Xa3+xa5+Xa21에 비해 향상된 저항성 반응을 나타내 저항성 유전자 집적에 따른 저항성 증진 효과를 확인하였다. 네 개 의 저항성 유전자 집적 계통들은 국내 수집 벼흰잎마름병 균주 16개에 대해서 광범위 저항성을 나타냈으며, 병원성이 강한 K3a 균계 접종 시에도 수량성 감소 및 품질 저하가 관찰되지 않았다. 이들 계통들은 초형이 양호하고 진백 등 이 전의 저항성 품종들에 비해서 수량성이 증가하였다. 네 개의 저항성 유전자 집적 조합 Xa1+Xa3+xa5+Xa21은 효율적이 고 유망한 저항성 유전자 집적 조합으로 판단되었고, 개발된 저항성 유전자 집적 계통들은 자포니카 벼의 흰잎마름병 저항성 증진을 위한 유용한 육종소재로서 활용될 것으로 생각된다. Corresponding author: Tel. 063-238-5216, E-mail: mayoe@korea.kr 17
OA-06 Disruption-Mediated Functional Characterization of Rice Defense-Related Genes Selected Through Genome-Wide Association Study for Core Population Feng Peng Li 1, Won-Hee Ra 1, Kyu-Won Kim 1, Yoo-Hyun Cho 2, Shin-Chul Bae 3, Duk-Ju Hwang 3, Sang-Ryeol Park 3, Yong-Jin Park 1, Il-Pyung Ahn 3* 1 Department of Plant Resources, College of Industrial Sciences, Kongju National University, Yesan 32439, Korea 2 Seedpia Corporation, Suwon 16395, Korea 3 National Institute of Agricultural Sciences, Rural Development Administration, Jeonju 54874, Korea Rice blast is one of the most notorious diseases and breeding employing resistant germplasms have been intensively performed during the last four decades, however, frequent resistance breakdown by domination of new or endemic races still causes serious problems worldwide and available germplasms are practically limited. To find resistant resources more efficiently, genome-wide association study (GWAS) was applied for comparative analysis of the relationship between nucleotide polymorphisms (NPs) and responses induced by infections of rice blast pathogen, KI197 and KJ401, in the Korean rice core set (KRICE_CORE) composed of 295 accessions. Nipponbare genome was used as a reference. Disease responses were also enumerated in the near-isogenic lines each containing single blast resistance gene to reveal effector spectrum of both races. GWAS analysis revealed significant relationships between variations of blast disease responses and NPs located on the chromosomes Ⅰ, Ⅱ, Ⅲ, Ⅳ, Ⅵ, Ⅸ, and Ⅻ. As most of the reported qualitative disease resistance genes have nucleotide-binding site-leucine rich repeat (NBS-LRR) motif, NBS-LRR genes were analyzed within 200 kb regions harboring meaningful NPs. Finally, eleven potent candidates were further found as novel blast resistance-conferring NBS-LRRs. For functional characterization of these candidates, genome editing methods such as Transcription Activator-Like Effector Nucleases (TALEN) and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 were established. A hub gene for rice defense signaling, OsNPR1/NH1, and a structural gene mediating gibberellin biosynthesis, GERANYL GERANYL PYROPHOSPHATE SYNTHASE (OsGGPPS1), were chosen for these experiments. TALEN-mediated OsNPR1/NH1 and OsGGPPS1 disruptions resulted in the genes disruption and loss of basic resistance and stunting in the transgenic plants, respectively. More accurate GWAS and haplotyping is under progress, and characterization of candidate genes will be performed. Corresponding author: Tel. 063-238-4668, E-mail: jinhyung@korea.kr 18
OA-07 Development of Cultivar-specific DNA Markers Based on Ty1-copia-like Retrotransposon-based Insertional Polymorphism in Pear (Pyrus pyrifolia) Hoy-Taek Kim, Jong-In Park, Ill-Sup Nou * Department of Horticulture, Sunchon National University, 255, Jungang-ro, Suncheon, Jeonam 57922, Korea Retrotransposons are present in all plant genomes and play important roles in genome size, genome structure remodeling, gene function, and genome evolution. Eight novel long terminal repeat retrotransposons were identified from a bacterial artificial chromosome library of Japanese pear (Pyrus pyrifolia). These newly identified retrotransposons appear to be closely related to Ty1-copia retrotransposons. Ppcrt1 8 were grouped with the copia-like retrotransposons RIRE1 and BARE-1 by phylogenetic analysis based on the amino acid sequences encoded by the gag and pol domains. Fluorescence in situ hybridization analysis showed that sequences homologous to Ppcrt4 were dispersed throughout more than half of the pear chromosomes. We also developed retrotransposon-based insertional polymorphism (RBIP) markers based on the long terminal repeat (LTR) sequences of copia-like retrotransposon Ppcrt4 and flanking genome sequences, which were derived from 454 sequencing data from Japanese pear (Pyrus pyrifolia) Hosui. Out of 40 sequences including both LTR and flanking genome regions, we developed 22 RBIP markers and used them for DNA profiling of 80 pear cultivars: 64 Japanese, 10 Chinese (Pyrus ussuriensis) and 6 European (Pyrus communis). Three RBIP markers were enough to differentiate Hosui from the other Japanese pear cultivars. The 22 RBIP markers could also distinguish 61 of the 64 Japanese pear cultivars. European pears showed almost no amplification of the 22 RBIP markers, which might suggest that retrotransposons had transposed during Asian pear evolution or reflect the genetic relationship between Asian and European pears. Sixteen of the RBIP markers could be positioned on a genetic linkage map of Hosui. The RBIP loci were distributed in 10 linkage groups, and some loci were very closely located within the same linkage group. The information obtained will be applicable to developing cultivar-specific RBIP marker sets in plants. Acknowledgements: This research was supported by Golden Seed Project (Center for Horticultural Seed Development, No. 213003-04-4-SB110) Ministry of Agriculture, Food and Rural Affairs (MAFRA), Ministry of Oceans and Fisheries (MOF), RDA and Korea Forest Service (KFS). Corresponding author: Tel. 061-750-3249, E-mail: nis@sunchon.ac.kr 19
OA-08 Breeding for Pyramiding Target-genes and Selection of F 1 Hybrids by Marker Assisted Selection in Tomato 김명권 충북 청주시 흥덕구 강내면 월곡길, 토마토생명과학연구소 토마토 육종에서 과품질과 내병성이 중요한 요소이다. 내병성, 과품질, 과색 관련 유전자를 분자마커로 계통선발과 F 1 Hybrid 선발에 활용하였다. 내병성 마커로 토마토황화잎말림바이러스(Ty1, Ty2, Ty3), 토마토모자이크바이러스(Tm2a), 토마토반점시들음바이러스(Sw5), 시들음병(I2, I3), 반신위조병(V), 근부위조병(Forl), 역병(Ph3), 잎곰팡이병(Cf9), 선충 (Mi1) 등을 검정하였고, 저장성 마커로 Rin gene, 과색 마커로 Orange, Yellow, Brown, Pink color 등을 검정하여 유묘기 에 1차로 선발하고, 정식포장에서 최종선발된 계통과 F 1 hybrid를 다시 확인하였다. Marker Assisted Selection으로 육성 한 토마토 6품종을 개발하여 농가에 보급 판매하고 있다. Corresponding author: Tel. 043-235-3368, Fax. 043-235-3369, E-mail: mkkim207@naver.com 20
OA-09 Characterization and Stress-responsive Expression Profiling of MCM genes in Brassica Ashokraj Shanmugam, Hoy-Taek Kim, Jong-In Park, Ill-Sup Nou * Department of Horticulture, Sunchon National University, 255, Jungang-ro, Suncheon, Jeonam 57922, Korea Minichromosome maintenance protein complex [MCM(2-7)] is an important replication factor which is activated at S-phase of cell cycle. This hexamer possesses helicase activity, which are regulated by various replication factor that forms the replication fork over double stranded DNA to process replication. Apart from MCM(2-7) complex, other MCM genes are also present which regulates the helicase activity of the hexamer. In yeast and mammalian cells, subunits of MCM(2-7) are regulated during replication stress, that has suggested to plants as well. Hexamer complex are also positively regulated during cold and salinity stresses in plants. In this study, we identified, characterized and modelled the tertiary structure of 12 MCM genes each from B. oleracea and B. rapa which possess two functional domains namely MCM_N and AAA (ATPases Associated) domain. Evolutionary relationship between these MCM genes are deduced using syntenic and phylogenetic analysis in B. oleracea and B. rapa. Expression profiling of MCM(2-7) complex genes and other MCM genes showed slight variation between B. oleracea and B. rapa during different stresses (replication stress, cold and salinity). Likewise, within the species, MCM gene expression exhibited differentially response when exposed to replication stress, cold and salinity stresses with variation in subunit genes of the complex. Overall study showed that MCM genes are conserved and closely related in B. oleracea and B. rapa for the proper normal DNA replication fork formation. Our expression profiling supported that MCM complex genes are significantly responding under replication stress, cold and salinity stresses. Therefore, we proposed that MCM are tolerance related genes during the respective abiotic stresses and the extension of structural, functional and molecular studies will be helpful for implication of MCM genes in stress biology in plants. Acknowledgements: This research was supported by Golden Seed Project (Center for Horticultural Seed Development, No. 213003-04-4-SB110) Ministry of Agriculture, Food and Rural Affairs (MAFRA), Ministry of Oceans and Fisheries (MOF), RDA and Korea Forest Service (KFS). Corresponding author: Tel. 061-750-3249, E-mail: nis@sunchon.ac.kr 21
OA-10 Classical and GBS-based QTL mapping downy mildew resistance loci in cucumber Khin Thanda Win 1, Chunying Zhang 1, Sanghyeob Lee 1,2* 1 Plant Genomics Laboratory, Department of Plant Biotechnology, College of Life Sciences, Sejong University, 209 Neungdong-ro, Gwanjing-gu, Seoul 143-747, Korea 2 Plant Biotechnology Research Institute, Sejong University, 209 Neungdong-ro, Gwanjing-gu, Seoul 143-747, Korea Downy mildew (DM) is a very important disease of cucumber (Cucumis sativus L.). Resistant cultivars have been deployed in production for a long time, but the genetic mechanisms of DM resistance in cucumber are not well understood. A QTL mapping study of DM resistance was conducted with 149 F2 population derived from two cucumber inbred lines R (resistant) and S (susceptible). A genetic map covering 811.7 cm in seven linkage groups was developed with 100 SSR marker loci. The 18 F2 individuals showed significant resistance and susceptible phenotype were applied GBS-based QTL analysis. Both QTL mapping analysis revealed four QTLs on chromosome 2, 4, 5 and 6. Detail information will be presented Acknowledgements: This work was supported by grants from Bio-industry Technology Development Program (111057-5) of ipet (Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry) and Korea-China Collaborative Research Program (NRF-2015K2A2A2001928) of NRF (National Research Foundation of Korea). Corresponding author: Tel. 82-2-3408-4375, E-mail: sanglee@sejong.ac.kr 22
OA-11 Genome Wide Characterization and Expression Profiling of Aquaporin Gene Family in Brassica rapa for Responsiveness to Abiotic and Biotic Stresses Md. Abdul Kayum, Hoy-Taek Kim, Jong-In Park, Ill-Sup Nou * Department of Horticulture, Sunchon National University, 255 Jungang-ro, Suncheon, Jeonnam 540-950, Korea Aquaporin (AQP) genes are those which have been so far reported as responsive for water transportation in plants. The aquaporin proteins are present in diverse forms in plants, where they function in transport of water and other nutrient molecules through living membranes, and might be essential for survival under salt and drought stresses. In this study, we have explored and identified 59 BrAQP genes from genome sequence data of B. rapa database and Br135K microarray dataset. Phylogenetic analysis of BrAQPs with AQPs from tomato (Solanum lycopersicum) and Arabidopsis thaliana revealed four distinct subfamilies, including plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs), NOD26-like intrinsic proteins (NIPs), and small basic intrinsic proteins (SIPs). Here, we have selected PIP subfamily genes for detailed expression analysis through differential expression in microarray data and to find their homology to stress related AQPs in B. rapa. All BrPIP genes showed organ-specific expression. Eventually, we have also analyzed the expression of PIP subfamily genes in response to cold, salt, drought, and water logging conditions as well as ABA (abscisic acid) treatment and Fusarium oxysporum f. sp. conglutinans infection. Twelve out of 22 BrPIP genes were differentially expressed upon cold treatment in two B. rapa inbred lines (Chiifu and Kenshin). Seven BrPIP genes showed up-regulation under drought stress and 17 BrPIP genes were responsive to salt stress. Whereas, 18 BrPIP genes were up-regulated by ABA treatment, and all BrPIP genes showed down-regulation under water logged condition stress. Four BrPIP genes were found responsive to F. oxysporum f. sp. conglutinans infection. Therefore, this study gives a comprehensive report of AQPs in B. rapa on expression profile 22 members of the BrPIP subfamily, which could be provided a basis to elucidate the stress-related biological functions of specific PIP gene. Acknowledgements: This research was supported by Golden Seed Project (Center for Horticultural Seed Development, No. 213003-04-4-SB110) Ministry of Agriculture, Food and Rural Affairs (MAFRA), Ministry of Oceans and Fisheries (MOF), RDA and Korea Forest Service (KFS). Corresponding author: Tel. 061-750-3249, E-mail: nis@sunchon.ac.kr 23
OB-01 Role of Flowering Pathway Genes on Anthocyanin Biosynthesis and Accumulation in Brassica rapa Ujjal Kumar Nath 1, Md. Abdul Kayum 1, Rawnak Laila 1, Khadiza Khatun 1, Jong-In Park 1, Ill-Sup Nou 1* Department of Horticulture, Sunchon National University, 255 Jungang-ro, Suncheon, Jeonnam 540-950, Korea Anthocyanins are secondary metabolites in plants responsible for various colour formations in plant parts and their products. It is marked as potential source of antioxidant and plays a pivotal role in overcome stresses. Recent report on potential health benefits of anthocyanins brings attention to develop Brassica vegetables with high anthocyanins accumulations in different vegetative parts for consumption. The mechanisms of anthocyanins accumulation, deposition and its physiological functions are poorly understood in Brassica. In the purple Chinese cabbage (Brassica rapa) the anthocyanins production and accumulation remain as usual up to vegetative phase. However, the leaf pigmentation becomes fade-out when plants enter into reproductive phase with blooming of flowers. This observation creates interest to know the role of flowering pathway genes on anthocyanins biosynthesis and accumulation. To confirm this hypothesis, a Chinese cabbage inbred line Re20 (green leaf with coloured mid-rib) was crossed with another inbred line of Asia seed (designated as Asia: coloured leaf with green mid-rib). F 1 plants were confirmed and selected with the presence of coloured pigmentation both in leaf and mid-rib. Fifty eight F 2 progenies were produced by selfing from one of the best F 1 plant. RNA was extracted from all of the plant leaf blade and mid-rib separately. 7 F 2 plants were selected based on relative expression of anthocyanin biosynthesis genes and they were selfed for producing F 3 progenies. F 3 lines were grown in 5 replicates and leaf samples were collected from all the plants before put into vernalization and at first week after six weeks of vernalization. Complementary DNA (cdna) was synthesized using SuperScript III First Strand cdna kit from invitrogen. Real time quantitative PCR (qrt) was performed using 12 anthocyanins biosynthesis pathway genes including phenylpropanoid pathway(pal, C4H, 4CL), early biosynthesis pathway (CHS, CHI, F3H, F3 H), late biosynthesis pathway (DFR, ANS, UFGT, MYB, TT) and five flower regulatory genes including VRN, FRIGIDA and FLC. Relative expression was calculated following 2 -ΔΔ C T method using cq value. The correlations, multiple regressions and path-coefficient analysis results of F 2 revealed that flower integrator genes had negative effect on anthocyanin accumulation during flowering in B. rapa. This result will be confirmed by the expression profiling and estimating anthocyanins in F 3 progeny. Meanwhile, genes involved in flower pathway and anthocyanin pathway were also found co-up or co-down regulated accompanied with the accumulation of anthocyanin in leaves which indicated the possible interaction between anthocyanin and flowering genes. Acknowledgements: This research was supported by Golden Seed Project (Center for Horticultural Seed Development, No. 213003-04-4-SB110) Ministry of Agriculture, Food and Rural Affairs (MAFRA), Ministry of Oceans and Fisheries (MOF), RDA and Korea Forest Service (KFS). Corresponding author: Tel. +82 61 750 3249, Fax: +82-61-750-3208, E-mail: nis@sunchon.ac.kr 24
OB-02 Leaf Developmental and Genotypic Variations in Wax Content, Composition and Expression of Wax Biosynthetic Genes in The Inbred Lines of Brassica Oleracea Var. Capitata Rawnak Laila, Kiwoung Yang, Arif Hasan Khan Robin, Mi Chung Suh, Juyoung Kim, Jong-In Park and Ill-Sup Nou * Department of Horticulture, Sunchon National University, Suncheon, Korea Department of Bioenergy Science and Technology, Chonnam National University, Gwangju, Korea Cuticular waxes act as a protective barrier against environmental stresses. The present study was conducted to investigate the leaf developmental and genotypic variations in cuticular wax content, composition and expression level of wax biosynthetic genes in two high-wax depositing and two low-wax depositing cabbage lines. Wax deposition analyzed cuticular wax composition for the leaves of 20 days of age collected from the 10 week old plants. The waxy compounds in all cabbage lines were predominated by C 29 -alkanes, -secondary alcohols and ketones. Relative expression level of wax biosynthetic genes was measured at three different growth stages. Gene expression level at the first-formed leaf of the highest wax depositing line accounted consistently higher at three sampling points collected weekly interval but that decreased in other genotypes with increasing leaf age. In 10 weeks old plants, expression level of genes generally increased from the youngest leaf to leaves at position 3-4 at the stem axis but that greatly declined at the leaf positions 8 and 12. Overall, expression level of wax biosynthesis genes decreased at the older leaves. Principal component analysis revealed that higher level of relative expression level of LTP2 genes in the lowest wax depositing line and higher level of relative expression of CER3 gene in the highest wax depositing line probably determined comparatively lower and higher wax content in those two lines. Results of this study might be useful in modelling wax development pattern in cabbage plants and also to develop cabbage varieties with desired level wax contents. This study was supported by the Golden Seed Project (Center for Horticultural Seed Development, No. 213003-04-4-SB110) of the Ministry of Agriculture, Food and Rural Affairs in the Republic of Korea (MAFRA). Corresponding author: E-Mail: nis@sunchon.ac.kr 25
OB-03 EMS 돌연변이체 PE1905의 사포닌 특성 및 유전 연구 박철우 1, 송종태 1, 정규화 2, 이정동 1* 1 대구광역시 북구 산격동 경북대학교 농업생명과학대학 2 전라남도 여수시 대학로 50 전남대학교 여수캠퍼스 생명화학 공학부 풍산나물콩을 EMS 처리하여 얻은 돌연변이체 892라인을 TLC 분석하여 사포닌 변이개체인 PE1905를 선발하였다. PE1905와 풍산나물콩의 하배축을 LC-MS 분석한 결과 돌연변이체인 PE1905에서는 wild type인 풍산나물콩 보다 Af 사 포닌이 3배 이상 증가하였고 Ac 사포닌 또한 1.5 배 증가하였다. 반면에 Ab 사포닌은 10% 감소하였고, DDMP 사포닌 αg와 βg또한 감소하는 것을 알 수 있었다(각각 75%, 37% 감소). 또한 PE1905에서 새로운 물질이 생성되는 것을 확인 하였고 분석결과 지금까지 발견되지 않은 Af 사포닌과 Ab 사포닌의 전구체로 추정된다. 변이체의 유전특성 연구를 위해 wild type인 풍산나물콩과 교배하여 F 1 종자를 얻었고 이를 다시 파종하여 얻은 F 2 종자 를 TLC 해본 결과 wild type과 mutant type의 비가 3:1로 나타났으며 F 2 종자를 파종하여 얻은 F 2:3 계통을 수확하여 TLC 분석을 해본 결과 wild type 동형접합 개체와, 이형접합 개체, mutant type 동형접합 개체가 1:2:1 로 분리되는 것을 확인 했고 이를 통해 PE1905 돌연변이체의 표현형은 단일열성유전자에 의해 나타나는 것으로 평가되었다. 새롭게 발견 된 사포닌 변이체는 사포닌의 생합성 경로를 연구하는데 중요한 재료가 될 것으로 평가된다. Corresponding author: Tel. 053-950-5709, E-mail: jdlee@knu.ac.kr 26
OB-04 Comparative Phylogenomics to Elucidate The Effects of Whole Genome Triplication Event on glutathione transferases Superfamily Genes in Brassicaceae Harshavardhanan Vijayakumar, Senthil Kumar Thamilarasan, Jong-In Park, Ill-Sup Nou * Department of Horticulture, Sunchon National University, 255, Jungang-ro, Suncheon, Jeollanam-do 57922, Korea Glutathione S-Transferases (GSTs) are a group of multigenic enzyme family, which catalyze the addition of glutathione to the broad range of substrates for ubiquitination. Plant GSTs are mainly known for their degradation mechanism, however, they are involved in other secondary metabolite pathways which aid plant growth development and tolerance against various biotic and abiotic stresses. These superfamily enzymes are divided into 14 classes in green plants based on conserved motifs and catalytic sites (Ser/Cys), of which Tau, Phi, DHAR (dehydroascorbate reductases) and Lambda are plant specific GSTs. In this study, 431 GSTs were identified in Arabidopsis thaliana (70), Brassica oleracea (91), Brassica rapa (89) and Brassica napus (181) by a strategic computational in silico analysis. Based on catalytic sites and phylogenetic analysis, 431 GSTs were grouped into 11 classes and also evolutionary analysis showed that iota, hemerythrin and Ure2p are absent in Brassicaceae family. By macro- and micro-synteny analysis, we found that the duplicated regions were more conserved between Brassica species than Arabidopsis. Our data show that tandem duplication has been regarded as the major expansion mechanism in this superfamily. Non-synonymous substitutions per site (Ka) and synonymous substitutions per site (Ks) analyses showed that N- and C-terminal functional domains of GSTs (GST_N and GST_C) seem to have evolved under a strong purifying selection (Ka/Ks < 1). Brassica and Arabidopsis divergence occurred before 20-24 mya (million years ago), based on this B. oleracea GST genes were evolved rapidly after the Brassica split than other two Brassica species. Also, the evolution led to the formation of new genes with non-typical GST domains alone with GST domains. Our survey provides additional annotation of the GST gene family and there evolutionary divergence with Brassicaceae. Acknowledgements: This research was supported by Golden Seed Project (Center for Horticultural Seed Development, No. 213003-04-4-SB110), Ministry of Agriculture, Food and Rural Affairs (MAFRA), Ministry of Oceans and Fisheries (MOF), Rural Development Administration (RDA) and Korea Forest Service (KFS). Corresponding author: Tel. 061-750-3249, Email: nis@sunchon.ac.kr 27
OB-05 Molecular Breeding for Fruit Color in Tomato Indeok Hwang, Hee-Jeong Jung, Song-I Kang, Ill-Sup Nou * Department of Horticulture, Sunchon National University, 255 Jungangro, Suncheon, Jeonnam 57922, Korea It is well known that color changes in tomato occur according to the amount of carotenoids, flavonoid and the presence/absence of chrolophyll. Most fruit flesh color is significantly correlated with carotenoid content in tomato. In other case, brown color is caused by SlSGR1 mutation, which is related to chlorophyll degradation. Chlorophyll is not degraded in SlSGR1 mutant, which results in its remaining in the flesh of the fruit and a brown color. The peel of pink-colored tomato is transparent due to the lack of accumulation of the flavonoid naringenin chalcone during ripening. A strong correlation was found between the expression and function of SlMYB12 with naringenin chalcone accumulation. Thus, SlMYB12 is a strong candidate gene underlying the pink phenotype. Knock-out of SlPSY1 can cause pale carotenoids, resulting in yellow colored flesh. Accordingly the inheritance of factors associated with tomato pigmentation (carotenoid, flavonoid and chlorophyll) has been investigated. In this study, we have identified a new position of mutations in SlPSY1, SlMYB12 and SlSGR1 by combining traditional breeding and the analysis of DNA/molecular markers. The data reported herein can be used as a DNA based marker during breeding and development of new tomato varieties, and enable improved color selection within adapted breeding materials and varieties. Acknowledgements: This research was supported by the Golden Seed Project (Center for Horticultural Seed Development, No. 213003-04-4-SB110), Ministry of Agriculture, Food and Rural Affairs (MAFRA), Ministry of Oceans and Fisheries (MOF), Rural Development Administration (RDA) and Korea Forest Service (KFS). Corresponding author: Tel. 061-750-3249, E-mail: nis@sunchon.ac.kr 28
OB-06 곡실발효사료 조제를 위한 사료맥종별 수확시기 및 발효품질 박종호 *, 오영진, 정영근, 송태화, 박태일, 이광원, 김경호, 김양길, 박종철 전북 완주군 이서면 국립식량과학원 작물육종과 국내 배합사료 원료곡은 대부분 수입에 의존하고 있으며, 이를 국내산 곡실사료로 대체 시 외화절감 및 사료의 안정적 공급이 가능하다. 곡실사료용으로 맥류를 재배하여 이용시에는 수확시기의 분산이 가능하며, 값비싼 기계 및 인력을 더 욱 효율적으로 사용할 수 있다. 본 연구는 국내 겉보리 품종에 대해서 수확시기별로 수량 특성과 발효품질의 차이를 분 석하여 곡실발효사료에 적합한 겉보리 품종을 선정하고자 수행하였다. 2013년 ~ 2015년까지 수행하여 겉보리는 광안 등 6품종을, 쌀보리는 새쌀 등 5품종, 밀은 금강 등 5품종을 선정하여, 출수후 3일간격으로 보리는 24일부터 7회, 밀은 30 일부터 5회 수확하여, 수량 및 ph 등 발효품질을 조사하였다. 시험결과 겉보리는 36일차에 ph4.2로 곡실발효 적합하였 으며, 이후 수확에는 발효가 일어나지 않아 곡실발효에는 부적합하였다. 수량은 564kg/10a로 39일차, 42일차와도 유의적 차이가 없었으며, 사료품질은 단백질 함량은 12.7%, 조섬유 2.3%, TDN 80.2% 로 양호하였다. 쌀보리는 36일차에 ph4.5 로 곡실발효에 적합하였으며, 이후 수확에는 발효가 일어나지 않아 곡실발효에는 부적합하였다. 수량은 489kg/10a였으며 39일차, 42일차와 유의적 차이를 보이지 않으면서 사료품질은 조단백질함량 14.9%, 조섬유 2.0%, TDN 76.6%로 양호하 였다. 밀은 42일차에 ph4.3으로 곡실발효에 적합하였으며, 수량은 541kg/10a였으며, 조단백질 16.0%, 조섬유 2%, TDN 78.8% 사료품질이 양호하였으며, 42일차 이후 곡실발효 정도에 대한 추가 조사가 필요하다. Corresponding author: Tel. 063-238-5228, E-mail: ark0104@korea.kr 29
OB-07 In vitro and in vivo Antioxidant Activity and α-glucosidase Inhibitory Potential of Major Phenolic Compound Isolated from Superhongmi (Oryza sativa L.) Varieties Soo-In Jang 1, Hanna Kang 1, Suyeon Jang 1, Jung-Yun Lee 1, Tae-Ho Ham 2, Sung-Gil Hong 3, Su-Noh Ryu 2, Young-In Kwon 1* 1 Department of Food and Nutrition, Hannam University, Daejeon, 34054, Korea 2 Department of Agriculture Science, Korea National Open University, Seoul,03087, Korea 3 Erom R&D Center, Sungnam-Si, 13488, Korea In the current study, we evaluated phenolic profile and oxygen radical absorbance capacity (ORAC) of Superhongmi (Oryza sativa; SHM). The inhibitory activity of SHM extracts against porcine pancreatic α-amylase and rat intestinal α -glucosidase was also investigated in vitro and in vivo animal model. Ethyl alcohol extract of SHM bran had the highest α-glucosidase inhibitory activity, ORAC value and total phenolic content, followed by ethyl alcohol extract of whole SHM, and water extract of whole SHM. In rat fed on sucrose meals, SHM significantly reduced the postprandial blood glucose increment after sucrose loading in a dose dependent manner. The α-glucosidase inhibitory activity of the extracts was compared to selected specific phenolics detected in the extracts using HPLC. Taxifolin, a flavanonol, a type of flavonoid, was isolated from the extract of SHM along with three known compounds. Their structures were elucidated by extensive spectroscopic analysis, including 1D and 2D NMR, and FTICR-MS. Taxifolin, a major phenolic compound in SHM extract had high α-glucosidase inhibitory activity. The α-glucosidase inhibitory activity of the SHM extracts correlated to the Taxifolin content and antioxidant activity of the extracts. These results suggest that Superhongmi which has high Taxifolin content has the potential to contribute as a dietary supplement for controlling hyperglycemia and oxidative stress-linked diabetes complications. Corresponding author: Tel. 010-2238-7228, E-mail: youngk@hnu.kr 30
OB-08 식미 간접적 평가를 위한 기기활용 및 신속한 관능평가를 위한 해석 전재범 1*, 이점식 2, 윤미라 2, 이정희 3, 천아름 2, 김우재 1, 정지웅 1, 강경호 1, 김보경 1 농촌진흥청 국립식량과학원 1 작물육종과, 2 수확후이용과, 3 중부작물과 벼육종에서 신속한 식미 관능평가는 고품질품종을 육성하는데 필요한 부분이다. 그러나 관능평가는 1회에 4~5점 정도의 시료를 평가하게 되어 시간과 노력이 많이 소요된다. 그러므로 식미기기를 활용한 간접식미치로 관능평가를 추정하는 노력이 요구된다. 본 연구는 식미기기를 이용하여 어느 정도로 관능평가의 결과를 추정할 수 있는 가에 목적이 있다. 공시 재료는 2013~2014년 자포니카 19품종을 국립식량과학원 벼육종포장에서 재배 수확 후 90.4%로 도정하여 백미를 조제하였다. 각 시료에 대해 임의완전배치로 관능평가를 수행하였으며, Toyometer(MA-90R2)로 간접식미치를 측정하였 다. 그리고 아밀로스 함량과 단백질 함량을 측정하였고 식미관능평가에 대한 추정을 간접식미치인 토요윤기치를 이용하 여 잔차분석을 하였다. 그 결과 13~14년도에 수행한 관능평가와 간접식미치는 높은 상관성이 인정되었으며 결정계수는 14년도 r 2 = 0.7263 13 년도 r 2 = 0.8096로 나타났다. 잔차분석 결과 `13와 `14에 공통적으로 제외되는 품종은 대야, 미품으로 이 두 품종이 제 외된 공통 요인은 관능평가정도에 비해 간접식미치가 높게 나타났다. 특히 미품의 경우 14년도 간접식미치에서 93.76을 기록하여 가장 높은 값이었으나 관능평가에서는 중간정도에 위치하였다. 그러므로 관능평가와 간접식미치가 차이가 나 는 품종에 대해서는 세부적인 연구가 필요하다. 단백질 함량은 관능평가점수와 상관관계를 검토한 결과 알려진 대로 마 이너스의 상관을 확인할 수 있었다. 그러나 아밀로스 함량과 관능평가점수와 유의한 점을 찾기는 어려웠으며 이는 시료 로 사용된 밥쌀용 품종의 아밀로스 함량은 18~20% 내외이기 때문이다. 그러므로 간접식미치를 기반으로 관능평가의 약 70~80%로 추정이 가능하다는 점을 확인하였다. 또한 하루에 측정 가능 한 샘플 수는 관능평가와 비교하여 월등히 많으므로 계통육성과정에서 스크리닝하는 데 이용할 수 있다. Corresponding author: Tel. 063-238-5237, E-mail: jbchun01@korea.kr 31
OB-09 Inter-subspecific Indel Variations in Wild Rice Suggesting The Early Appearance of japonica-specific Alleles of The Oryza genus Joong Hyoun Chin 1*, Jonghwa Park 1, Dongryung Lee 1, Jong Seok Heo 1, Hee-Jong Koh 1* 1 Department of Plant Science, Research Institute for Agriculture and Life Sciences, and Plant Genomics and Breeding Institute, Seoul National University, Seoul 151-921, Korea The Indel (insertion/deletion) polymorphisms study using 67 selected STS markers showed that the conservation of indicaand japonica-specific alleles even in non-aa genome species of Oryza genus. Commonly Indel polymorphisms are generally irreversible and highly useful to evaluate the genetic relationship within genus Oryza. Even though the 67 SS-STS (subspecies specific-sequence Tagged Site) markers were selected using an evaluation variety panel composed of 30 indica and japonica varieties, the linked loci were highly conserved in wild relatives of rice. In this study, total of 290 accessions including 61 Asian cultivated rice (O. sativa), 27 African cultivated rice (O. glaberrima), and 202 accessions of wild Oryza species were investigated using the SS-STS markers. The average SS allele frequency of the various Oryza species, from AA-genome to BB~EE, and FF~HHKK showed an increased proportion of non-o. sativa and null alleles in the more distantly related wild species. Most of the wild species, except the more distant EE, GG, HHJJ, and HHKK genome accessions, consisted of relatively more indica than japonica alleles of SS markers. To validate the SS-STS study, PCR products of nine markers were sequenced across 24 33 accessions. Sequencing results revealed that Oryza species share indica or japonica-like conserved indel regions even across the different genomes. The presence of some japonica alleles beyond the AA genome at some SS indel loci also suggests that japonica-specific alleles occurred early in the history of the Oryza genus. Corresponding author: Tel. 02-880-4551, E-mail: joonghyoun.chin@gmail.com; heejkoh@snu.ac.kr 32
OB-10 Diachronic Analysis of Genetic Diversity in Rice Landraces Under On-Farm Conservation in Yunnan, China Di Cui 1,3, Jinmei Li 1, Cuifeng Tang 2, Jeonghwan Seo 3, Zhuo Jin 3, Yoye Yu 3, Hee-Jong Koh 3*, Longzhi Han 1* 1 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Key Laboratory of Crop Germplasm Resources and Utilization, The National Key Facility for Crop Gene Resources and Genetic Improvement, Ministry of Agriculture, No.12 Zhongguancun Nandajie, Beijing 100081, China 2 Institute of Biotech and Germplasm Resources, Yunnan Academy of Agricultural Sciences, No.9 Xueyunlu, Kunming 650205, Yunnan, China 3 Department of Plant Science, Plant Genomics and Breeding Institute of Agriculture and Life Science, Seoul National University, Seoul 151-921, Korea Rice (Oryza sativa L.) is one of the earliest domesticated crop species. Its genetic diversity has been declining as a result of natural and artificial selection. Compared with modern cultivars, traditional rice landraces that have been preserved by indigenous farmers have retained rich genetic variation. However, with the development of modern agriculture, rice landraces are no longer planted in most provinces in China, with the exception of some ethnic minority regions in areas such as Yunnan. In this study, we performed the first analysis of the levels and patterns of nucleotide variation in rice genomes under on-farm conservation in Yunnan during a 27-year period of domestication. We performed large-scale sequencing of 600 rice accessions with high diversity, which were collected in 1980 and 2007, using ten unlinked nuclear loci. Diachronic analysis showed no significant changes in the level of genetic diversity occurring over the past 27 years domestication, which indicated genetic diversity was successfully maintained under on-farm conservation. Population structure revealed that the rice landraces could be grouped into two subpopulations, namely the indica and japonica groups. Interestingly, the alternate distribution of indica and japonica rice landraces could be found in each ecological zone. The results of AMOVA showed that on-farm conservation provides opportunities for continued differentiation and variation of landraces. Therefore, dynamic conservation measures such as on-farm conservation (which is a backup, complementary strategy to ex situ conservation) should be encouraged and enhanced, especially in crop genetic diversity centers. The results of this study offered accurate insights into short-term evolutionary processes and provided a scientific basis for on-farm management practices. Corresponding author: Tel. 02-880-4541, E-mail: heejkon@snu.ac.kr; Tel. +8610-6217-6784, E-mail: hanlongzhi@caas.cn 33
OB-11 가공용 쌀 품종개발을 위한 배유변이 육종소재 개발 및 유전분석 정지웅 *, 전재범, 조성우, 김우재, 강경호, 고재권, 김현순, 김보경 전라북도 완주군 이서면 혁신로 181 국립식량과학원 작물육종과 근래 서구화, 음식소비패턴의 다양화 등으로 우리나라에서 주식으로써 쌀이 차지하는 비중은 계속 낮아져 1인당 쌀 소 비량은 2015년 62.9 Kg까지 감소하였다. 현재 다양한 쌀 가공제품을 개발함으로써 소비를 촉진시켜 생산기반을 유지하 려는 노력이 진행 중이다. 쌀을 이용한 다양한 식품형태를 개발하여 고부가가치 상품생산으로 연계하기 위해서는 기존 의 멥쌀과 찹쌀위주로 형성된 전분의 이화학적 특성을 보다 다양화해야 할 필요성이 있다. 이에 따라 자포니카계로 조 생 초다수 멥쌀 품종인 남일 에 화학적 돌연변이원을 처리, 그 후대계통들로부터 분질, 심백, 뽀얀메, 반찰 등 다양한 배 유변이 육종소재들을 확보하였다. 작물학적 특성 및 가공적성 등의 표현형자료에 의거하여 분질계통인 수원542호 를 건 식제분용으로, 심백이 강화된 중모1042 를 양조용으로 식물특허와 신품종으로 각각 등록하였다. 유용한 전분특성을 지 배하는 신규 대립인자들에 대한 지적재산권을 확보하기 위해 분리집단의 표현형 자료와 유전자형 자료간의 병합분석 (association analysis)에 기반을 두는 목표유전자위 표지, 유전적 작용가 추정 및 정밀분자표지를 개발 등의 유전분석 연 구를 수행하였다. 육종적 활용도가 높은 주동유전자에 의해 배유특성이 지배되는 돌연변이 계통을 선별하고자 원품종 (wild type)인 남일 과의 교잡후대의 표현형 분포가 비정규분포(abnormal distribution)를 따르는지를 우선 판단하였다. 연 관지도를 이용하는 목표유전자위 표지 등의 유전분석에는 분자표지의 높은 다형성을 보장하기 위해 돌연변이계통과 통 일계 품종인 밀양23호 간의 교잡후대를 이용하였다. 소규모분리집단(N<100)을 이용하였음에도 수원542호 의 분질특성 과 Namil(SA)- dull1 의 반찰특성을 지배하는 주동유전자위를 염색체 5번 장완중단과 염색체 6번 장완말단에서 확인하 여 이들에 대한 근접분자표지를 제시하였다. 반면에 현재까지 연관분석을 통해 확보된 근접분자표지들은 돌연변이 계통 과 원연인 밀양23호 간의 다형성에 기반을 두는 것들로 원품종(남일)에 대해서는 다형성이 대부분 상실될 뿐만 아니라, 유전분석집단 내 각 개체들은 원연교잡 후대로서 육종학적 실용성이 낮다는 한계가 있다. 이러한 문제점을 해결하기 위 해 원품종과 돌연변이계통의 전장유전체정보(Re-sequencing)를 분석하여 돌연변이 고유의 SNP를 분자표지로 활용함으로 써 원품종 뿐만 아니라 국내육성 우량품종들과의 교잡후대에서도 유용한 신규 대립인자를 지닌 개체를 원활히 선별할 수 있는 연구를 진행 중이다. Corresponding author: Tel. 063-238-5236, E-mail: jrnj@korea.kr 34
OC-01 Plant Gene Targeting with Site-Specific Nucleases and Homologous Recombination Yong-Ik Kwon 1*, Woo-Nam Kim 2, Min-ji Hong 2, Hong-Kyu Kang 1, Hyo-Yeon Lee 1,2, Kiyomi Abe 3, Seiichi Toki 3 1 Subtropical Horticulture Research Institute, Jeju National University, Jeju 63243, Korea 2 Faculty of Biotechnology, Jeju National University, Jeju 63243, Korea 3 Plant Genome Engineering Research Unit, National Institute of Agrobiological Sciences, Tsukuba 305-8602, Japan Gene targeting (GT) via homologous recombination (HR) is one of the best gene editing methods available to modify specific DNA sequences in genome. GT utilizes a DNA repair pathways. During HR-mediated DNA double-strand break (DSB) repair in eukaryotes, an initial step is the creation of a 3 single-stranded DNA (ssdna) overhang via resection of a 5 end. Rad51 polymerizes on this ssdna to search for a homologous sequence, and the gapped sequence is then repaired using an undamaged homologous DNA strand as template. Recent studies in eukaryotes indicate that resection of the DSB site is promoted by the cooperative action of RecQ helicase family proteins and exonuclease 1 (Exo1). Here, we demonstrate that overexpression of rice proteins OsRecQl4 and/or OsExo1 can enhance DSB repair processing, as evaluated by recombination substrate reporter lines in rice. Furthermore, we respect that the frequency of HR was evaluated all position of DSBs in the genome. These results could be applied to strategy of gene targeting with site-specific nucleases (ZFN/TALEN/CRISPR-Cas9) in plant. Acknowledgements: This work was supported by Basic Science Research Program through the National Research Foundation (2009-0094059), and BioGreen 21 Program (PJ0112802016), Rural Development Administration, Republic of Korea. Corresponding author: Tel. 064-754-3987, E-mail: yongikk@jejunu.ac.kr 35
OC-02 Engineering Rice to Reduce As Accumulation in Grain Won-Yong Song *, Youngsook Lee Department of Integrative Bioscience and Biotechnology, Pohang University of Science and Technology, Pohang 790-784, Korea Arsenic (As) is a chronic poison that causes severe skin lesions and cancer. Rice (Oryza sativa L.) is a major dietary source of As; therefore, reducing As accumulation in the rice grain is critical to diminish the amount of As that enters the food chain. Previously it was found that tonoplast localized C-type ABC transporters, such as Saccharomyces cerevisiae yeast cadmium factor 1 (ScYCF1), Arabidopsis thaliana C-type ABC transporter 1 and 2 (AtABCC1 and 2) and Oryza sativa C-type ABC transporter 1 (OsABCC1), enhance As resistance and, especially, OsABCC1 reduces As accumulation in rice grain by facilitating the thiol-as sequestration into vacuoles in the phloem companion cells of nodes in rice. Based on these findings, we hypothesized that the enhanced vacuolar thiol-as sequestration would reduce As accumulation in rice grain. To prove the hypothesis, we developed transgenic rice plants transformed with all ScYCF1, Ecγ-ECS and OsABCC1 and analyzed As concentrations in the roots, shoots and grains of the transgenic rice. The engineered plants exhibited reduced As translocation from root to shoot and 30% of the As concentration in brown rice compared to WT. Our result suggests that As vacuolar sequestration is one of the important steps to develop safety crop from As contamination. Corresponding author: Tel. 053-279-8373, E-mail: songwy@postech.ac.kr 36
OC-03 Development of Co-Dominant Female (F ) Locus Diagnosis Marker in Cucumber (Cucumis sativus L.) Chunying Zhang 1, Khin Thanda Win 1, Sanghyeob Lee 1,2* 1 Plant Genomics Laboratory, Department of Plant Biotechnology, College of Life Sciences, Sejong University, 209 Neungdong-ro, Gwanjing-gu, Seoul 143-747, Korea 2 Plant Biotechnology Research Institute, Sejong University, 209 Neungdong-ro, Gwanjing-gu, Seoul 143-747, Korea Cucumber is a typical monoecious having separate male and female flowers on the same plant and sex expression in cucumber is mainly determined by three major genes: F/f, M/m and A/a. Gynoecy plays an important role in cucumber hybrid breeding and the use of gynoecious lines as maternal parents ensures high productivity. This study aimed to identify a co-dominant marker linked to Female (F) locus to distinguish homozygous and heterozygous gynoecious lines for efficient selection of gynoecy in cucumber breeding. Acknowledgements: This work was supported by grants from Bio-industry Technology Development Program (111057-5) of ipet (Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry) and Korea-China Collaborative Research Program (NRF-2015K2A2A2001928) of NRF (National Research Foundation of Korea). Corresponding author: Tel. 82-2-3408-4375, E-mail: sanglee@sejong.ac.kr 37
OC-04 Drought-Tolerant Qtls from Samgang Lead to Tiller Formation at The Vegetative Stage in Rice Dongjin Shin *, Tae-Heon Kim, Sang-Ik Han, Jun-HyunCho, You-Chun Song, Yeong-Up Kwon Department of Southern Area Crop Science, Paddy Crop Research Division, National Institute of Crop Science, RDA, Miryang, 50424, Korea During the past 50 years, rice varieties which are sensitive to drought stress were developed with irrigation system and the rice production was increased at two fold and more. However, recently drought caused by global climate change is seriously giving a threat to world rice production and food security in rice field. Herein, we screened 218 rice varieties to identify 32 drought-tolerant cultivars. Strongly drought-tolerant cultivar Samgang shows stable yield in rain-fed conditions and was selected for further study. To pinpoint QTLs for drought tolerance, we examined visual drought tolerance (VDT) and relative water content (RWC) phenotypes in a double haploid (DH) population of 101 individuals derived from a Samgang Nagdong cross of a drought-tolerant and drought-sensitive variety, respectively. Three drought-tolerant QTLs for VDT derived from Samgang were identified. The phenotypic variance explained (PVE) by these QTLs was 41.8%. In particular, qvdt11 was found to contribute 20.3% of the PVE for RWC. To determine QTL effects on drought tolerance, 7 DH lines were selected according to numbers of QTLs and tillers under non-irrigated conditions. Of the drought tolerance-associated QTLs, qvdt2 and qvdt6 did not affect tiller formation, but qvdt11 increased tillers under rain-fed paddy conditions. Interestingly, tiller formation was most stable when qvdt2 and qvdt11 were combined. Together, these results suggest that qvdt11 is important for drought tolerance and stable tiller formation under rain-fed paddy conditions. 38
OC-05 Genetic Analysis of Seedling Traits Regulated by Light in Weedy Rice Hyun-Sook Lee 1, Yun-Joo Kang 1, Ju-Won Kang 1,2, Sang-Nag Ahn 1* 1 Department of Agronomy, Chungnam National University, Daejeon 34134, Korea 2 Department of Southern Area Crop Science, National Institute of Crop Science, NICS, RDA, Milyang 50424, Korea The Korean weedy rice, PBR (photoblastic rice) was reported to have light-inducible seed germination (photoblastism) and longer mesocotyl than cultivar rice. In previous studies, two QTL for photoblastism were mapped on chromosomes 1 and 12 using 124 F4 lines from a cross between Ilpumbyeo and PBR. Using 120 F8 lines advanced from the F4, we identified two QTL controlling mesocotyl length on chromosomes 1 and 3. The QTL for photoblastism on chromosome 1 was co-localized with the QTL for mesocotyl length near the SSR markers RM8260-RM246. To know whether these two genes regulated by light are functionally related, 110 F3 lines were developed from a cross between Ilpumbyeo and CR7124. The line, CR7124 having photoblastism and long mesocotyl was selected from 120 F8 lines. The germination rate in the dark condition and the mesocotyl length in the 110 F3 were in the range between two parents. The mesocotyl length in F3 lines showed significant correlation with germination rate in dark condition (r = 0.7, P < 0.0001). These results suggest the linkage of two QTL for two traits regulated by light in weedy rice. Fine mapping of the two QTL is underway to analyze their functional relationship. Acknowledgements: This work was supported by a grant from the Next-Generation BioGreen 21 Program (No. PJ011048012016), Rural Development Administration, Republic of Korea. Corresponding author: Tel. 031-821-7038, E-mail: ahnsn@cnu.ac.kr 39
OC-06 QTL Mapping of Cold Tolerance at The Seedling Stage in Rice and Development of Advanced Backcross Lines Using Marker-Assisted Selection Suk-Man Kim 1,2*, Kyung-Ho Kang 3, Bo-Kyung Kim 3, Jong-Min Jeong 3, Russell Reinke 1,2, Kshirod Jena 1 1 Plant Breeding, Division, International Rice Research Institute, Metro Manila, Philippines 2 C/o IRRI-Korea Office, National Institute of Crop Science, Rural Development Administration, Iseo-myeon, Wanju-gun, Jeollabuk-do 565-851, Korea 3 Crop breeding Research Division, National Institute of Crop Science, Rural Development Administration, Iseo-myeon, Wanju-gun, Jeollabuk-do 565-851, Korea Low temperature is a major abiotic stress for rice cultivation, causing serious yield loss in many countries. To identify QTL controlling low temperature an F 7 recombinant inbred line (RIL) population of 123 individuals derived from a cross of the cold-tolerant japonica cultivar Jinbu and the cold-susceptible indica cultivar BR29 for QTL mapping was used in this study. A total of six QTLs including two on chromosome 1 and one each on chromosomes 2, 4, 10, and 11 for seedling cold tolerance was identified with phenotypic variation (R 2 ) ranging from 6.1 to 16.5 %. Out of them two QTLs (qsct1 and qsct11) on chromosomes 1 and 11, respectively, were fine mapped. To improve seedling cold tolerance of indica cultivar, BR29, Jinbu was back crossed with BR29 by MAS. Two advanced backcross population (BC 2 F 4 and BC 3 F 3 ) were developed and used for confirming QTL effects against cold stress by comparison of QTLs distribution on the lines (qsct1, qsct11 and qsct1+qsct11). In the 18 DAT the average cold damage score of lines with one or two SCT QTL were included in the degree of moderated tolerance (5.63-6.83) based on the SES (standard evaluation system for seedling cold) and the score of lines without SCT showed susceptible reaction to cold stress along with BR29. Corresponding author: Tel. 063-238-5497, E-mail: s.kim@irri.org 40