연사 발표
SYMP-01 1 1 Doubled Haploid (DH) Technology in Maize Breeding: Application and Technology for Production of DH Lines Vijaya Vardhana Reddy. Chaikam International Maize and wheat improvement center (CIMMYT), Mexico Doubled haploid technology enables rapid development of 100% homozygous inbred lines. Production of DH lines in maize involves induction of haploids using special genotypes called haploid inducers, haploid identification preferably at seed or seedling stage using anthocyanin markers, chromosomal doubling with antimitotic chemicals in haploid seedlings and self-fertilization of fertile doubled haploid plants to generate seed for DH lines. Use of DH lines in maize breeding results in increased selection gains, simplified logistics and possibly reduced costs. DH lines are also valuable in exploring and uncovering the diversity present in the landraces and open pollinated verities as fixed homozygous inbred lines. Hence, most multinational breeding companies routinely use Doubled Haploid technology in maize breeding programs. However, adoption of DH technology is lagging behind in the maize breeding programs of the developing world. Limitations include lack of adapted haploid inducers, difficulties associated with haploid identification and lack of technical knowhow on chromosomal doubling procedures. At CIMMYT, research is being conducted to address some of these limitations. CIMMYT in collaboration with University of Hohenheim developed first publicly available tropically adapted haploid inducers which were distributed to several organizations throughout the world. Further breeding for improved inducers led to development of second generation tropicalized haploid inducers with high haploid induction rates and better agronomic performance. Analysis of several tropical breeding populations and inbred lines revealed that the commonly used R1-nj phenotypic marker is completely or partially inhibited in significant proportion of tropical germplasm making it ineffective for haploid identification. To address this problem, we have developed haploid inducers expressing anthocyanin coloration in roots, which aid in haploid identification at seedling stage when R1-nj is inhibited. Also, inducers with high oil trait are also being developed which can facilitate automation of haploid identification. Different chromosomal doubling methods are being tested for improving the efficiency of doubling. In addition, the influence of source germplasm on haploid induction and chromosomal doubling has been studied which revealed a very strong influence of source germplasm. In this talk topics like advantages of using DH lines in maize breeding, different steps involved in maize DH line production, limitations at each step and new developments to address these limitations will be presented. Corresponding author: E-mail: V.Chaikam@cgiar.org 3
SYMP-02 1 Finding Mineral Element Transporters for Better and Safe Production of Rice Jian Feng Ma Institute of Plant Science and Resources, Okayama University, Kurashiki, Japan Crop productivity largely relies on mineral nutrients in soils. These mineral nutrients are taken up by the roots, translocated to the shoots and then distributed to different tissues for their growth. On the other hand, harmful mineral elements are also present in soils. Transfer of these toxic elements from soils to edible parts will affect our health through food chain. Tremendous transporters are involved in the transport of these essential and toxic mineral elements in plants, but most of them are unidentified. Therefore, identification and manipulation of these transporters will be required for better and safe production of crops. At this symposium, I will show some results on transporters of two elements (Si and Cd) in rice, which is a staple food for half population of the world. Silicon (Si) is required for high and sustainable production of rice, a high Si-accumulating species (Ma and Yamaji, 2015). We identified two transporters (Lsi1 and Lsi2) for Si uptake in rice. Lsi1 is a Si-permeable channel (Ma et al. 2006), while Lsi2 is an active efflux transporter of Si (Ma et al. 2007). Both Lsi1 and Lsi2 are localized at the exodermis and endodermis of the roots, but they show different polar localization. Lsi1 is localized at the distal side of both exodermis and endodermis, while Lsi2 is localized at the proximal side of the same cell layers. Knockout of either Lsi1 or Lsi2 results in loss of Si uptake. At next step, Si in the xylem sap is unloaded by Lsi6, a homolog of Lsi1 in rice (Yamaji et al., 2008). At reproductive stage, three transporters (Lsi2, Lsi3 and Lis6) localized at different cell layers of the node I are responsible for preferential distribution of Si to the panicles. Knockout of either of them resulted in increased distribution of Si to the flag leaf, but decreased distribution to the panicle (Yamaji and Ma, 2009; Yamaji et al., 2015). Furthermore, our recent mathematical modelling study showed that an apoplastic barrier and development of enlarged vascular bundles of the node are also required for the preferential distribution of Si to the panicles (Yamaji et al., 2015). On the other hand, rice is a major source of Cd intake. We have identified three transporters (OsNramp5, OsHMA2 and OsHMA3) involved in Cd accumulation in rice. OsNramp5 is a major transporter for Cd uptake in the roots (Sasaki et al., 2012). It is localized in the distal side of both exodermal and endodermal cells. OsHMA3 is localized to the tonoplast of root cells (Ueno et al., 2010), which is responsible for sequestration of Cd into the vacuoles in the roots. On the other hand, OsHMA2 is a plasma membrane-localized transporter and localized in the pericycle of the root and the phloem of enlarged and diffuse vascular bundles in the nodes (Yamaji et al., 2013). OsHMA2 is responsible for translocation of Cd from the roots to the shoots and distribution of Cd into the grain. Knockout of OsNramp5 and OsHMA2 resulted in significant reduction of Cd accumulation in the grain, but also caused decreased yield because they are also transporters of Mn and Zn, respectively. By contrast, overexpression of functional OsHMA3 selectively decreased accumulation of Cd in the grains without growth penalty and enhanced tolerance to Cd toxicity (Ueno et al., 2010; Sasaki et al., 2014). Corresponding author: E-mail: maj@rib.okayama-u.ac.jp 4
SYMP-03 1 Function of Fibrillin Protein in Photosynthetic Metabolism Hyun Uk Kim Department of Bioindustry and Bioresource Engineering, Plant Engineering Research Institute, Sejong University, Korea Fibrillins are lipid-associated proteins in plastids and are ubiquitous in plants. They accumulate in chromoplasts and sequester carotenoids during the development of flowers and fruits. However, little is known about the functions of fi brillins in leaf tissues. Here, we identified fibrillin 5 (FBN5), which is essential for plastoquinone-9 (PQ-9) biosynthesis in Arabidopsis thaliana. Homozygous fbn5-1 mutations were seedling-lethal, and XVE:FBN5-B transgenic plants expressing low levels of FBN5-B had a slower growth rate and were smaller than wild-type plants. In chloroplasts, FBN5-B specifi cally interacted with solanesyl diphosphate synthases (SPSs) 1 and 2, which biosynthesize the solanesyl moiety of PQ-9. Plants containing defective FBN5-B accumulated less PQ-9 and its cyclized product, plastochromanol-8, but the levels of tocopherols were not affected. The reduced PQ-9 content of XVE:FBN5-B transgenic plants was consistent with their lower photosynthetic performance and higher levels of hydrogen peroxide under cold stress. These results indicate that FBN5-B is required for PQ-9 biosynthesis through its interaction with SPS. Our study adds FBN5 as a structural component involved in the biosynthesis of PQ-9. FBN5 binding to the hydrophobic solanesyl moiety, which is generated by SPS1 and SPS2, in FBN5-B/SPS homodimeric complexes stimulates the enzyme activity of SPS1 and SPS2. Identification of interacting partners to fibrillins will contribute to better understanding of mechanism of photosynthetic metabolism under stress conditions. Corresponding author: E-mail: hukim64@sejong.ac.kr 5
SYMP-04 1 Revisiting Domestication to Revitalize Crop Improvement: The Florigen Revolution Zach Lippman Cold Spring Harbor Laboratory, USA A major challenge in modern agriculture is to develop new approaches that can expedite breeding and make its outcomes more predictable. The last decade of studies on crop domestication has pointed to a recurring theme: selection of genetic variation in the universal florigen flowering system, which includes the flower-promoting hormone florigen and antagonistic family members. Prominent examples include perpetual flowering in strawberries and roses, biennial flowering in sugar beet, adapted flowering in barley, and the introduction of a bushy determinate growth habit for common bean, soybean, and tomato, which enabled large-scale field production. Yet, these successes are all based on rare naturally occurring florigen pathway mutations that may not be providing optimal agronomic performance. I will discuss our recent work on translating molecular mechanisms underlying florigen-dependent flowering in tomato to develop a genetic toolkit that has allowed us to fine-tune and optimize shoot architecture, flower production, and yield. I will also present a new CRISPR-based approach that we are developing to create novel quantitative variation that has the potential to invigorate breeding in all crops. Corresponding author: E-mail: lippman@cshl.edu 6
SYMP-05 1 Manipulating Fruit and Vegetable Quality Traits David A. Brummell The New Zealand Institute for Plant & Food Research Limited, Private Bag 11600, Palmerston North, 4442, New Zealand Fruit and vegetable quality aspects important to the consumer include nutritional content, texture (eating quality) and the absence of harmful compounds. Metabolic engineering of plant pathways to enhance quality aspects, including the concentrations of phytochemicals with human health benefits, is a focus of many current plant biotechnology programmes. Fruit carotenoid or anthocyanin contents have been increased by genetic manipulation of enzymes or transcription factors, but not both at the same time. However, both pathways are affected by light. We achieved fruit-specific suppression of DET1, a regulatory gene that represses several signal transduction pathways controlled by light, resulting in significant increases in the contents of the carotenoids lycopene and β-carotene, as well as various flavonoids. Fruit softening and texture are controlled by a range of cell wall-modifying enzymes. Historically, polygalacturonase (PG) has been accorded a minor role in the softening process. Nevertheless, silencing the PG1 gene in apple reduced softening, and aspects of texture were altered. Interestingly, fruit water loss was also reduced, apparently due to alterations in the structure of the hypodermal layers. These observations suggest that PG may exert part of its effects on softening through altering transpirational water loss, and indicate that the function of PG in fruit ripening is more complex than previously reported. For potato tubers, storage at cold temperatures for many months is essential in order provide a constant supply of material to industry for processing. However, low temperature storage can cause the breakdown of starch to sucrose and the accumulation of the reducing sugars glucose and fructose (a process called cold-induced sweetening) due to the activity of vacuolar acid invertase. Processing these susceptible cultivars by frying or roasting results in the Maillard reaction and the generation of acrylamide, a suspected human carcinogen. We cloned two invertase inhibitors from potato tubers, and showed that overexpression of the vacuolar form reduced vacuolar invertase activity, cold-induced sweetening and the generation of acrylamide after frying. Corresponding author: E-mail: david.brummell@plantandfood.co.nz 7
SYMP-06 1 Designing Crops for Global Food Security: Digitizing Plant Phenotypes Maurice Moloney Global Institute for Food Security, Canada Information and communications technology pervades all aspects of human activity from healthcare, finance, and education to recreation. Agriculture and food production are no exceptions. Large-scale integrated data acquisition and management systems are emerging that will affect everything from plant breeding and soil science to automated farm practices. Critical technologies include genomics and bioinformatics, GIS/GPS, and high-precision satellite imaging. These are just the beginnings of a digital revolution in agriculture. Agricultural research will be profoundly affected by the development of digitized image analysis for crop phenotyping. Sophisticated imaging acquisition technologies will provide a digital signature for plant phenotypes. Advanced image analysis software utilising high-performance computation will convert the science of plant breeding from an analogue-to-digital activity to a digital-to-digital activity, allowing genotypes to be linked to phenotypes computationally. By analogy to current use of genomic bioinformatics to identify genes and functions, optimised breeding strategies will be determined by algorithms, which streamline the process of building elite germplasm. This transition will depend on advances in photonics, tomography, image recognition, and computational analytics much more than it will depend upon plant biology, but its effects on plant breeding will be profound. At a field level, the acquisition of data through imaging, sensing, and agrimetrics will result in large databases from which trends and correlations will emerge. These will be useful to individual farmers planning an optimal (and sustainable) cultivation strategy over several years. However, they will also be vital for national and international efforts to adapt crops to a changing climate. Corresponding author: E-mail: gifs.director@usask.ca 8
SYMP-07 1 CRISPR RNA-guided Genome Editing in Human Stem Cells, Animals, and Plants Jin-Soo Kim Center for Genome Engineering, Institute for Basic Science, Seoul, Korea Department of Chemistry, Seoul National University, Seoul, Korea Genome editing that allows targeted mutagenesis in higher eukaryotic cells and organisms is broadly useful in biology, biotechnology, and medicine. We have developed ZFNs, TALENs, and Cas9 RNA-guided endonucleases (RGENs), derived from the type II CRISPR-Cas prokaryotic adaptive immune system, to modify chromosomal DNA in a targeted manner. In particular, we used purified Cas9 protein and in vitro transcribed guide RNAs rather than plasmids encoding these components to correct large chromosomal inversions in the blood coagulation factor VIII gene that cause hemophilia A in patient-derived induced pluripotent stem cells (ipscs) and to modify diverse genes in large animals and plants. The resulting animals and plants contained small insertions or deletions (indels) at target sites, which are indistinguishable from naturally-occurring variations, possibly bypassing regulatory requirements associated with use of recombinant DNA. Despite broad interest in RNA-guided genome editing, RGENs are limited by off-target mutations. We developed Cas9 nuclease-digested whole genome (digenome) sequencing (Digenome-seq) to profile genome-wide specificities of Cas9 nucleases in an unbiased manner. Digenome-seq captured nuclease cleavage sites at single nucleotide resolution and identified off-target sites at which indels were induced with frequencies below 0.1%. We also showed that these off-target effects could be avoided by using modified guide RNAs that contain two extra guanine nucleotides at the 5 end. Digenome-seq is a robust, sensitive, unbiased, and cost-effective method for profiling genome-wide off-target effects of programmable nucleases including Cas9. Corresponding author: E-mail: jskim01@snu.ac.kr 9
SYMP-08 1 CRISPR Genome Editing in Outcrossing Woody Perennials Chung-Jui Tsai University of Georgia, USA The CRISPR/Cas9 technology is revolutionizing all facets of biology from medicine to agriculture, owing to its precision, efficiency, versatility and ease of adoption. We recently reported the first application of CRISPR/Cas9 in stably transformed Populus, extending the species range of this powerful technology to woody perennials. We achieved 100% mutational efficiency in two 4-coumarate:CoA ligase (4CL) genes tested all stably transformed plants contain bi-allelic DNA modifications. The primary transformants exhibited strikingly uniform phenotypes among independent transgenic lines, a consistency that is unmatched by previous gene silencing methods. With its specificity, CRISPR/Cas9 offers a less equivocal means than previous approaches for discerning functional redundancy of paralogous genes that are prevalent in plant genomes. For instance, CRISPR mutation of the Class I, lignin-associated 4CL1 resulted in reduced lignin content and S/G monolignol ratio, with a reddish-brown discoloration throughout the stem wood. Its genome duplicate 4CL5, however, was unaffected. Class II 4CL has been associated with flavonoid biosynthesis based on its expression pattern, but this function has not been confirmed by reverse genetics. CRISPR mutation of Populus 4CL2 resulted in significantly reduced levels of condensed tannins in roots without affecting stem lignin. An underappreciated obstacle in genome editing of outcrossing species is the frequent occurrence of sequence polymorphisms that can render CRISPR/Cas9 unproductive. However, current genomic resources and CRISPR grna design tools do not support exploration of sequence variations. We present experimental evidence as well as genome-wide computational analysis to demonstrate the sensitivity of CRISPR/Cas9 to allelic heterozygosity, and discuss tools and strategies that can help deal with such sequence polymorphisms. The ability to generate instant-nulls by the CRISPR/Cas9 system is highly desirable for woody perennials with long generation cycles. When used in conjunction with early flowering strategies, transgene-free progenies can be obtained in one breeding generation. Elite clones carrying targeted gene mutation(s) without foreign DNA may ultimately help increase public acceptance of bioengineered agricultural products. Corresponding author: E-mail: cjtsai@uga.edu 10
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