Ac/Ds 삽입 변이체를 이용한 벼 유전자 기능 연구

이강섭,박성한,윤도원,안병옥,김창국,한창덕,이기환,박동수,은무영,윤웅한,Lee, Gang-Seob,Park, Sung-Han,Yun, Do-Won,Ahn, Byoung-Ohg,Kim, Chang-Kug,Han, Chang-Deok,Yi, Gi-Hwan,Park, Dong-Soo,Eun, Moo-Young,Yoon, Ung-Han 한국식물생명공학회 2010 식물생명공학회지 Vol.37 No.2

Rice is the staple food of more than 50% of the worlds population. Cultivated rice has the AA genome (diploid, 2n=24) and small genome size of only 430 megabase (haploid genome). As the sequencing of rice genome was completed by the International Rice Genome Sequencing Project (IRGSP), many researchers in the world have been working to explore the gene function on rice genome. Insertional mutagenesis has been a powerful strategy for assessing gene function. In maize, well characterized transposable elements have traditionally been used to clone genes for which only phenotypic information is available. In rice endogenous mobile elements such as MITE and Tos (Hirochika. 1997) have been used to generate gene-tagged populations. To date T-DNA and maize transposable element systems has been utilized as main insertional mutagens in rice. A main drawback of a T-DNA scheme is that Agrobacteria-mediated transformation in rice requires extensive facilities, time, and labor. In contrast, the Ac/Ds system offers the advantage of generating new mutants by secondary transposition from a single tagged gene. Revertants can be utilized to correlate phenotype with genotype. To enhance the efficiency of gene detection, advanced gene-tagging systems (i.e. activation, gene or enhancer trap) have been employed for functional genomic studies in rice. Internationally, there have been many projects to develop large scales of insertionally mutagenized populations and databases of insertion sites has been established. Ultimate goals of these projects are to supply genetic materials and informations essential for functional analysis of rice genes and for breeding using agronomically important genes. In this report, we summarize the current status of Ac/Ds-mediated gene tagging systems that has been launched by collaborative works from 2001 in Korea.

글로벌 GM작물 연구개발 및 상업화 동향

조정일,이강섭,박수철,Cho, Jung-Il,Lee, Gang-Seob,Park, Soo-Chul 한국식물생명공학회 2016 식물생명공학회지 Vol.43 No.2

Global GM crops continue to grow. They have reached 181 million hectares. A total of 28 countries have approved biotech crops for planting. More than 60 countries have approved biotech crops to be imported as food and feed since 1996, meaning that biotech crops are now commonly accepted in those countries. Although biotech crops provide key solutions for the challenge of global food security in the future due to population growth and climate change, there are still some debates on whether biotech crops should be accepted in many countries including Korea. Therefore, it is very important to make people understand that GM crops will provide benefits to both farmers and consumers. In this review, current global status of GM crop development and commercialization are summarized.

다양한 계통의 옥수수 미성숙배로부터 캘러스 유도와 식물체 재분화

홍준기,박기진,이강섭,김둘이,김주곤,이승범,서은정,이연희,Hong, Joon Ki,Park, Ki Jin,Lee, Gang-Seob,Kim, Dool Yi,Kim, Ju-Kon,Lee, Seung Bum,Suh, Eun Jung,Lee, Yeon-Hee 한국식물생명공학회 2017 식물생명공학회지 Vol.44 No.1

옥수수의 최적 조직 배양 조건을 확립하기 위하여 옥수수 국내 5 계통과 국외 11 계통 총 16 계통을 포트와 포장 재배하여 미성숙 배를 분리하여 배발생 캘러스 유도 및 식물체 재분화율을 조사하였다. MS 배지에 auxin으로 1.5 mg/L Dicamba와 0.5 mg/L 2,4-D 가 첨가된 배지에서 캘러스 형성은 본 실험에 사용된 옥수수 계통 모두에서 높은 빈도로 유도되었으며, 캘러스로부터 식물체 재분화는 5mg/L zeatin이 첨가된 재분화 배지에서 높은 재분화율을 보였다. 또한 포장에서 재배된 옥수수로부터 미성숙 배를 분리하여 사용하였을 때 캘러스 유기 및 식물체 재분화 효율이 높았던 것으로 보아 미성숙 배를 분리하기 위한 옥수수 상태 및 genotype이 중요한 영향을 준다는 것을 알 수 있었다. 본 실험을 통하여 배 발생 캘러스 형성 및 식물체 재분화 효율이 조사된 옥수수 계통들은 생명공학 기술을 활용한 신품종 개발을 위한 형질전환 시스템 개발에 유전자원으로 활용될 수 있는 정보를 제공할 것으로 사료된다. We investigated the callus induction and plant regeneration ability of 16 maize genotypes, including the Korean inbred lines, using 9 to 15 day-old immature zygotic embryos from maize grown in pots and from field cultures. Immature zygotic embryos placed on MS medium supplemented with L-proline 0.7 g/L, MES 0.5 g/L, Dicamba 1.5 mg/L, 2,4-D 0.5 mg/L, $AgNO_3$ 4 mg/L, and sucrose 20 g/L, showed the highest frequency of callus induction. The highest number of shoots regenerated when the embryogenic callus were transferred to MS medium supplemented with 5 mg/L zeatin. The root formation was observed when shoots were grown on MS medium supplemented with 0.2 mg/L indole-3-butyric acid (IBA). Additionally, under the same culture conditions, immature zygotic embryos from maize grown in the field also had a high frequency of plant regeneration. Except one genotype, 15 genotypes showed callus induction and shoot regeneration. Among the 16 genotypes tested, H99, B98, HW3, and B73 yielded the best plant regeneration. H99 showed maximum shoot formation from the primary embryogenic callus. The results suggest that genotypes and growth conditions of the maize plant plays very important roles for enhancing the embryogenesis competence of immature zygotic embryos. The successful regeneration from immature zygotic embryos of maize inbred lines provides a basis for molecular breeding of new cultivars by genetic transformation.

NGS 기반 밀양23호/기호벼 재조합자식 유전집단(MGRIL)을 이용한 CAPS 마커 개발 및 줄기 굵기 형질 관련 QTL 분석

이예지(Ye-Ji Lee),변은주(Eun-Ju Byeon),배선화(Seon-Hwa Bae),지현소(Hyeonso Ji),이강섭(Gang-Seob Lee),윤웅한(Ung-Han Yoon),김태호(Tae-Ho Kim) 한국육종학회 2016 한국육종학회지 Vol.48 No.3

The rice recombinant inbred lines derived from Milyang23 and Gihobyeo cross were used in genetic mapping and QTL analysis studies. In this study, we developed a new 101 CAPS markers based on the SNPs in the whole genome region between these varieties. As a result, the total genetic distance and average distances were 1,696.97 cM and 3.64 cM, respectively. In comparison to the distance of the previous genetic map constructed based on 365 DNA markers, the new genetic map was found to have a decreased distance. The map was applied for the detection of QTLs on all seven traits relevant to diameter of stem internode, length of culms, length of panicles and the number of panicles including the correlation analysis between each trait. The QTLs results were similar to the report in previous studies, whereas the distance between the markers was narrowed and accuracy increased with the addition of 101 CAPS markers. A total of 9 new QTLs were detected for stem internode traits. Among them, qI1D-6 had higher LOD of 5.1 and phenotype variation of 50.92%. In this experiment, a molecular map was constructed with CAPS markers using next generation sequencing showing high accuracy for markers and QTLs. In the future, developing more accurate QTL information on stem internode diameters with various agriculturally important traits will be possible for further rice breeding.

연구보문 : 자연과학 ; 자포니카 벼의 종자 저장단백질 유전자 구조 및 발현분석 연구 현황

윤웅한 ( Ung Han Yoon ),이정화 ( Jeong Hwa Lee ),이강섭 ( Gang Seob Lee ),김영미 ( Young Mi Kim ),한장호 ( Jang Ho Hahn ),지현소 ( Hyeon So Ji ),윤성원 ( Sung Won Yoon ),이종렬 ( Jong Yeol Lee ),김태호 ( Tae Ho Kim ) 한국국제농업개발학회 2012 韓國國際農業開發學會誌 Vol.24 No.3

Based on the outcome of the rice genome sequencing, a lot of researches for identification of genes underlying useful traits and their utilization are being carried out recently. Especially, rice seed storage proteins are used as a main nutrition source for human while they are used as the nitrogen source for plant growth during seed germination. Rice seed storage proteins are divided by solubility in solvent into weak acid and alkali soluble glutelin, alcohol soluble prolamin, and salt soluble globulin. The rice genome contains 11 glutelin genes and 33 prolamin genes. In the case of prolamin genes, 15 genes are located in 15 Mb region on the chromosome 5. Clustering of seed storage protein genes on the same chromosomal region indicates that they are evolved from the same chromosome or they share similar gene expression regulatory region. Only single globulin gene existed on the chromosome 5. As a result of tissue- specific expression pattern analysis of seed storage protein genes using microarray, glutelin and most of prolamin genes were expressed only in seed endosperm, while a few prolamin and the globulin genes are expressed in seed endosperm and germinating seed. As a result of isolating promoter regions of the seed storage protein genes and analyzing their expression pattern in seed, glutelin type C1 promoter showed expression in whole seed, and glutelin type B5 and α-globulin promoter showed strong expression. This paper reviewed on the structure and expression profile of rice seed storage protein genes, and will provide information on the recent research status to the researchers who are studying on the improvement of seed traits and production of substances.

SNP 마커를 이용한 벼 흰잎마름병 저항성 선발 효율 증진

신운철,백소현,서춘순,강현중,김정곤,신문식,이강섭,한장호,김현순,Shin, Woon-Chul,Baek, So-Hyeon,Seo, Chun-Sun,Kang, Hyeon-Jung,Kim, Chung-Kon,Shin, Mun-Sik,Lee, Gang-Seob,Hahn, Jang-Ho,Kim, Hyun-Soon 한국식물생명공학회 2006 식물생명공학회지 Vol.33 No.4

본 연구는 흰잎마름병 K1 레이스에 감수성인 상주찰벼와 저항성인 HR13721-53-3-1-3-3-2-2를 인공교배하여 육성된 F2, F3를 재료로 하천 Kl 레이스에 대한 저항성 검정과 SNP 마커를 이용한 유전자형 분석 및 저항성과의 연관성을 분석하였다. Kl 레이스에 대한 저항성 검정 결과 $F_2,\;F_3$에서 각각 이론적 분리비인 3:1, 1:1의 분리비를 나타냈으며 SNP 마커를 이용한 유전자형 분석은 16PFXa1 primer를 이용하여 유전자를 증폭한 후 Eco RV 제한효소 처리하여 다형성을 분석하여 저항성 및 유전자형을 확인할 수 있었다. K1 레이스에 대한 저항성 검정과SNP마커를 이용한 유전자형의 연관분석 결과 저항성과 마커간에 연관성이 일치하였으며, 특히 SNP 마커를 이용한 유전자형 분석에서는 K1 레이스에 대한 저항성 검정에서 알 수 없었던 $F_2$ 개체가 동형접합체인지 이형접합체인지를 판별할 수 있어 저항성 품종 육종을 위한 선발 효율을 높일 수 있었다. Discovery of single nucleotide polymorphisms (SNPs), including small insertions and deletions, is one of the hot topics in genetic research. The most common type of sequence variant consists of single base differences or small insertions and deletions at specific nucleotide positions. Significance of SNPs in rice is increasing for genetic research, positional cloning and molecular breeding. $F_2$ 170 lines and $F_3$ 194 lines derived from Sangjuchalbyeo/HR13721-53-3-1-3-3-2-2 Were used for Searching SNP markers related to bacterial blight resistance. Sangjuchalbyeo is susceptible to bacterial blight, but HR13721-53-3-1-3-3-2-2 has Xa1 gene resistant to bacterial blight. Individual lines were inoculated with $K_1$ race of bacterial blight and resistant or susceptible was evaluated after 3 weeks from inoculation. The genotypes of population were analysed by PCR-RFLP for SNP marker developing. The segregation of $F_2\;and\;F_3$ population showed almost 3:1, 1:1 ratio, respectively. Analysis of genotype using SNP marker is capable of confirming resistance for $K_1$ race and genotype through amplifying the gene using 16PFXal primer and digested the PCR product with Eco RV. There were close relation between resistance test for $K_1$ race and SNP marker genotype. Especially, DNA analysis using SNP marker is capable of judging homozygote/heterozygote in $F_2$ population compared with resistant test for Kl race. So, it seems to improve the selection efficiency in disease resistant breeding.

일본의 식물유전체 연구현황 및 전망

윤웅한 ( Ung Han Yoon ),이정화 ( Jeong Hwa Lee ),이강섭 ( Gang Seob Lee ),김영미 ( Young Mi Kim ),지현소 ( Hyeon So Ji ),김태호 ( Tae Ho Kim ) 한국국제농업개발학회 2011 韓國國際農業開發學會誌 Vol.23 No.5

In Japan, plant genomics research is mainly leaded by the national research institutes. The various structural studies such as rice genome has allowed researchers to analyze useful traits, and to focus their commercialization. With aims to facilitate structural and functional study in plant genome, NIAS (National Institute of Agrobiological Sciences) constructed Poaceae genome DB and RIKEN (Rikagaku Kenkyusho) built DB for Arabidopsis genome and plant full-length cDNA. NIG (National Institute of Genetics) constructed a national biological resources DB (National Bio Resource Project). This compiling DB provides a variety of genome-related research materials for researchers in the field. Recently, as an effort to resolve global issues of food supply and environmental problems, New Agri-genome Project has been performed aiming to develop an innovative agricultural technologies for the quantity, disease resistance and identifying useful genes related to environmental problems. In addition, in order to improve agricultural productivity in developing countries, JIRCAS assisted technical supports for the plant genome research and developed NERICA rice, which is suitable for African area. Such these approaches are expected to contribute to solving the global issues about food, energy and environment in the world.

벼 microarray를 이용한 유전자발현 profiling 연구동향

윤웅한,김연기,김창국,한장호,이태호,김동헌,이강섭,박수철,남백희,Yoon, Ung-Han,Kim, Yeon-Ki,Kim, Chang-Kug,Hahn, Jang-Ho,Kim, Dong-Hern,Lee, Tae-Ho,Lee, Gang-Seob,Park, Soo-Chul,Nahm, Baek-Hie 한국식물생명공학회 2010 식물생명공학회지 Vol.37 No.2

As the International Rice Genome Sequencing Project (IRGSP) was completed in 2005 and opened to the public, many countries are making a lot of investments in researches on the utilization of sequence information along with system development. Also, the necessity of the functional genomics researches using microarray is increased currently to secure unique genes related with major agricultural traits and analyze metabolic pathways. Microrarray enables efficient analysis of large scale gene expression and related transcription regulation. This review aims to introduce available microarrays made based on rice genome information and current status of gene expression analysis using these microarrays integrated with the databases available to the public. Also, we introduce the researches on the large scale functional analysis of genes related with useful traits and genetic networks. Understanding of the mechanism related with mutual interaction between proteins with co-expression among rice genes can be utilized in the researches for improving major agricultural traits. The direct and indirect interactions of various genes would provide new functionality of rice. The recent results of the various expression profiling analysis in rice will promote functional genomic researches in plants including rice and provide the scientists involved in applications researches with wide variety of expression informations.

단백질 함량 증가 고품질 특수미 ‘드리미5호’ 육성

박재령(Jae-Ryoung Park),김은경(Eun-Gyeong Kim),장윤희(Yoon-Hee Jang),이강섭(Gang-Seob Lee),김경민(Kyung-Min Kim) 한국육종학회 2022 한국육종학회지 Vol.54 No.4

Rice is a staple food in most countries, and thus, diversifying its value is necessary. Recently, the focus on good health has increased, and rice cultivars with improved properties should be bred based on consumer demands. At the Plant Molecular Breeding Laboratory of Kyungpook National University, “Hwayeong” was tissue-cultured to derive mutant populations. In the mutant lines, “P35” with high protein content and excellent agricultural traits, was backcrossed with “Ilmi” three times, and the final selected line was called “Drimi5ho.” The heading date of “Drimi5ho” was August 11 (107 d after sowing), which was 5 d later than that of “Ilmi” (August 5, 101 d after sowing). The culm length of “Drimi5ho” was 71 cm, the panicle length was 18 cm, the number of panicles per hill was 13, the number of spikelets per panicle was 113, the ratio of ripened grain was 90.5%, and the 1,000-grain weight was 22.6 g. When “Drimi5ho” was planted ordinary plantation, the yield in the southern mountainous area was 526 kg/10a. Lodging was not observed when “Drimi5ho” was grown in the field with standard fertilization (N-P2O5-K2O = 9-4.5-5.7 kg/10a). This rice cultivar was resistant to leaf/neck blast, bacterial blight (K1, K2, and K3), rice stripe virus, rice black-streaked dwarf virus, and rice dwarf virus. The milled grain of “Drimi5ho” was clear and transparent without any chalkiness; moreover, the grain shape was round and short. Although it is a specialty rice with increased protein content, the taste of the rice and the grain quality were excellent (Registration No. 6829).

GM작물 연구개발 및 상용화 동향

조정일(Jung-Il Cho),박성한(Sung-Han Park),이강섭(Gang-Seob Lee),김수민(Su-Min Kim),임수미(Su-Mi Lim),김연식(Youn-Shic Kim),박수철(Soo-Chul Park) 한국육종학회 2020 한국육종학회지 Vol.52 No.S

The global area of GM Crops continued to grow in 2018 and reached 191.7 million hectares. Twenty-six countries approved biotech crops for planting and an additional 44 countries (18 + 26 EU countries) officially imported biotech crops for food, feed, and processing, meaning that biotech crops are now commonly accepted in those countries. First-generation GM seed is being commercialized by global agricultural companies in advanced countries such as the United States and parts of Europe. The fact that more than 90% of first-generation GM seeds, which have been commercialized for 20 years, are both insect resistant and herbicide resistant proves that they continue to have an effect on improving agricultural productivity and increasing farmers incomes. As the effectiveness of GM crops has been proven and technology has been developed, the GM crop development trend has recently changed. In other words, it has moved from being producer-oriented to benefiting both farmers and consumers. In Korea, the National Program for GM Crops (NCGC), one of the Next-Generation BioGreen 21 Programs organized by Rural Development Administration (RDA), was established in 2011 to develop biotech crops that will be used in the future to solve our agricultural problems. To accomplish this mission, the NCGC carried out the exploration of useful functional genes, the development of qualified events, and the safety assessment of developed events. Here, we introduce the current status of GM crop development and commercialization in the world and in Korea.