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형광영상을 이용한 옥수수 반수체 종자 선발 효율성 향상
김영욱(Younguk Kim),한정헌(Jeong Heon Han),김재영(Jaeyoung Kim),김영태(Yeongtae Kim),김년희(Nyunhee Kim),이채원(Chaewon Lee),이서연(Seoyeoun Lee),김송림(Song Lim Kim),김문종(Moon Jong Kim),류시환(Si Hwan Ryu),이홍로(Hongro Lee),지현소(H 한국육종학회 2022 한국육종학회지 Vol.54 No.4
Many studies concerning breeding maize varieties are in progress in Korea and other countries. Double haploid technology is widely used for the development of commercial maize varieties worldwide, and has also been utilized in Korea since its introduction by the Maize Research Institute, Gangwondo. We performed a study to improve the efficiency of selecting haploid maize seeds using fluorescence imaging. It was verified that anthocyanin produced by the expression of R1-nj gene can be detected by fluorescence imaging, and we developed a high-throughput method for discriminating between haploid and diploid seeds. Compared with discriminating with naked eye, this method reduced the time for discriminating haploid and diploid maize by 91.7% and increased selection accuracy by 16.8% for haploid and 2.2% for diploid maize. This method enabled the acquisition of more haploid seeds and improved the efficiency of breeding research by shortening the time involved.
Delta 15 desaturase 유전자 억제에 의해 알파리놀렌산 함량이 낮은 들깨 육성
김경환(Kyung-Hwan Kim),이경렬(Kyeong-Ryeol Lee),김정봉(Jung-Bong Kim),이명희(Myoung Hee Lee),이은경(Eungyeong Lee),김년희(Nyunhee Kim),이홍석(Hongseok Lee),김송림(Song Lim Kim),백정호(JeongHo Baek),최인찬(Inchan Choi),지현소(Hyeonso 한국육종학회 2018 한국육종학회지 Vol.50 No.4
Perilla is an oilseed crop cultivated in Korea since ancient times. Due to the high α-linolenic acid content in perilla, perilla seed oil can easily become rancid. α-Linolenic acid is synthesized by two enzymes, endoplasmic reticulum-localized Δ15 desaturase (FAD3) and chloroplast-localized Δ15 desaturase (FAD7) in vivo. In order to lower the α-linolenic acid content of the seed oil without disturbing plant growth, we tried to suppress the expression of only the FAD3 gene using RNA interference, whilst maintaining the expression of the FAD7 gene. Seventeen transgenic plants with herbicide (Basta TM ) resistance were obtained by Agrobacterium-mediated transformation using hypocotyls of perilla plants. The transgenic plants were firstly confirmed by treatment with 0.3% (v/v) Basta TM herbicide, and the expression of FAD3 was measured by Northern blot analysis. The α-linolenic acid content was 10-20%, 30-40%, and 60% in two, seven, and three of the twelve T1 transgenic perilla plants which had enough seeds to be analyzed for fatty acid composition, respectively. Analysis of the fatty acid composition of T2 progeny seeds from T1 plants with the lowest α-linolenic acid content showed that the homozygous lines had 6-10% α-linolenic acid content and the heterozygous lines had 20-26% α-linolenic acid content. It is expected that the reduction in α-linolenic acid content in perilla seed oil will prevent rancidity and can be utilized for the production of high-value functional ingredients such as high γ-linolenic acid.
이채원(Chewon Lee),이서연(Seoyeon Lee),홍수빈(Subin Hong),김영욱(Young Uk Kim),김년희(Nyunhee Kim),백정호(Jeongho Baek),김송림(Song Lim Kim),지현소(Hyeonso Ji),조정건(Jung Gun Cho),김경환(Kyung-Hwan Kim) 한국원예학회 2021 한국원예학회 학술발표요지 Vol.2021 No.10
영상을 활용한 작물, 종자, 과실 등의 표현형 고속 대량 분석기술은 디지털 육종의 핵심기술로 각광 받고 있다. 이 기술을 이용해 사과, 배 과실의 표현형을 분석하기 위한 장치 개발 및 영상 데이터 수집 조건을 마련하기 위한 연구를 수행하였다. 기존에 과실 비대량 및 특성 조사를 위해 종경과 횡경측정에는 캘리퍼스를 사용하고, 과피색은 색차계를 이용해 측정하고 있다. 이러한 아날로그적 방법은 연구자에 따라 편차와 실수 등으로 인해 주관적인 오류가 있을 수 있다. RGB 센서를 이용한 과실 표현형 분석을 위해 사과(Red Apple, Green Apple), 배 각 10개씩 30개체를 대상으로 과실 이미지 획득 장치를 통해 RGB 영상을 촬영하고 영상분석 프로그램(ImageJ)을 활용해 과실 비대량 및 형태 관련 표현 형질을 분석하였다. 먼저 영상 데이터 획득 기준 마련을 위해서 Top, Side 두 대의 카메라를 이용해 다양한 배경지 색상(White, Blue, Black), 컬러차트 높이 조절, 조명 밝기 조절을 통해 영상획득 조건을 최적화하였다. 영상으로 분석한 형질은 면적(area), 둘레(perimeter), 장폭(Feret), 단폭(MinFeret) 등 8개 지표이며, 이 데이터를 기반으로 과실의 종경, 횡경, 부피, 밀도, 둥근 정도, 외형의 균일함 등의 표현형 특성 정보를 획득할 수 있었다. 영상분석의 신뢰도는 영상분석 데이터와 실측 데이터 간의 상관관계 분석 결과 R2값이 0.95 이상의 높은 상관관계를 나타내었다. 영상을 이용한 과실의 특성 분석 결과를 바탕으로 향후 사과, 배 외의 더 많은 품종 특성을 분석하는데 활용할 수 있을 것으로 기대되며 다양한 품질 특성을 디지털화, 객관화하여 디지털 육종의 기본 자료로 활용하고자 한다.
고추 탄저병 정밀 탐색을 위한 영상분석 기술에 대한 연구
백정호(JeongHo Baek),김년희(Nyunhee Kim),이은경(Eungyeong Lee),이홍석(Hongseok Lee),김송림(Song Lim Kim),박상렬(Sang Ryeol Park),지현소(Hyeonso Ji),최인찬(Inchan Choi),김경환(Kyung-Hwan Kim) 한국산업정보학회 2020 한국산업정보학회논문지 Vol.25 No.5
전세계적으로 널리 재배되는 고추 (Capsicum annuum)는 한국에서 소비가 많은 채소류 중 매우 중요한 작물중 하나이다. 고추는 생육기간 동안에 고추 탄저병에 심한 피해를 입어 방제나 저항성 품종에 대한 연구가 중요하다. 본 연구에서는 탄저병에 저항성이 있는 고추 4품종과 민감성을 가진 1품종에 대해 K1탄저균을 접종하였으며, 시간이 지나면서 나타나는 병 면적을 촬영하여 프로그램을 통해 정량화한 내용을 분석하였다. 고추과일 면적과 병면적의 비율을 통해 감수성 품종인 An-S는 약 40%로 약하게 나타났으며, 저항성 품종인 An-12R (23%), AR-Tan (21%), An-9R (19%)로 비교적 강하게 나타났고, PBC81는 약 11%로 탄저균에 강한 병 반응을 보였다. 이와 같은 정량적인 수치는 탄저병 품종이나 탄저균에 대한 저항성 연구를 수행하는데 비교할 수 있는 기초자료로 활용할 수 있다. One of the most important vegetables consumed in Korea, chili peppers (Capsicum annuum) are widely cultivated around the world. Chili peppers have been severely damaged by anthracnose during their growth, so it is important to study prevention and resistance varieties. K1 anthracnose was inoculated against four cultivar of chili peppers that are resistant to anthracnose and one cultivar that is sensitive. The area of the disease that appeared over time was photographed and quantified through the program was analyzed. Through the ratio of the area of chili pepper fruit and the area of the bottle, the sensitive variety An-S showed weak reactions to anthracnose with about 40%, the resistant variety An-12R (23%), An-Tan (21%), and An-9R (19%), and PBC81 showed a strong response to anthracnose with about 11%. These quantitative value can be used as a basis for comparison in conducting resistance studies for new varieties.
이채원(Chaewon Lee),최인찬(Inchan Choi),이홍석(Hongseok Lee),김년희(Nyunhee Kim),안은숙(Eunsook An),김송림(Song Lim Kim),백정호(Jeongho Baek),지현소(Hyeonso Ji),윤인선(In-Sun Yoon),김경환(Kyung-Hwan Kim) 한국육종학회 2021 한국육종학회지 Vol.53 No.4
Fast and accurate selection is essential for breeding to cope with rapid climate changes and a steeply increasing population. Consequently, technologies for high-throughput phenotyping (HTP) are emerging. These technologies, unlike conventional phenotyping methods,enable us to evaluate agronomic traits in a fast and massive manner. Thus, the HTP facility was built to acquire and analyze crop imagesusing RGB sensors at the National Institute of Agricultural Sciences, Republic of Korea. By testing various conditions to acquire images,we determined the conditions for phenotyping using the RGB sensor as follows: exposure 30,000 ms, gamma 75, and gain 100 using LEDlights in a blue background. Based on this condition, images from 96 individual plants of rice Dongjin cultivar were obtained every weekto measure plant height and shoot area, which are directly associated with yield. The results obtained from the image analysis were comparedwith the manually collected results. The r2 value between the projected plant height obtained from image analysis and the plant height obtainedfrom manual measurement was 0.989. Furthermore, the r2 value between the projected shoot area obtained from image analysis and the shootarea obtained from manual measurement was 0.981. These results show that image analysis is highly reliable and can be used for crop phenotyping. Therefore, we expect that the new method we developed will be used for breeding in the near future.
MutMap 분석에 의한 벼 왜성 돌연변이 계통의 변이 유전자 탐색
오준(Jun Oh),천경성(Kyeong-Seong Cheon),강도유(Do-Yu Kang),김송림(Song Lim Kim),이은경(Eungyeong Lee),김년희(Nyunhee Kim),오효자(Hyoja Oh),최인찬(Inchan Choi),백정호(Jeongho Baek),윤인선(In Sun Yoon),김경환(Kyung-Hwan Kim),정남진(Nam-J 한국육종학회 2020 한국육종학회지 Vol.52 No.1
A dwarf mutant rice line was selected from an Ac/Ds insertion mutant population and named dwf1. The phenotype of F1 and F2 plants derived from a cross between dwf1 and Dongjin indicated that a single recessive gene is responsible for the mutant phenotype, and we named this gene dwf1. Resequencing of the dwf1 line and Dongjin (wild type) revealed 42,386 homozygous single nucleotide polymorphisms (SNPs) between dwf1 line and Dongjin. MutMap analysis was performed by sequencing a DNA pool prepared from 100 mutant type plants in the dwf1/Dongjin F2 population, and it was found that the dwf1 gene was located in the 23 ~ 30 Mbp region on chromosome 4. In this region, we found a non-synonymous SNP in the Os04g0469800 gene, which was reported as D11 gene encoding a cytochrome P450 family protein involved in the biosynthesis of brassinosteroids (BRs). This SNP was regarded as the causative SNP for the dwf1 phenotype, and the dwf1 gene is a novel allele of D11. We performed mapping of the dwf1 gene with five SNP markers on chromosome 4 with 190 dwf1/Dongjin F2 plants. The phenotype of F2 plants was completely co-segregated with genotypes of the J10402 marker, which was developed based on the non-synonymous SNP in the D11 gene. These results will contribute to the study of the molecular biological functions of the D11 gene and BRs.