초록

Ⅳ. 연구개발결과
○ 새로운 고추 고품질 계통육성 및 품질 관련 고세대 mapping 집단의 성분분석과 우수 계통육성하였음
○ 고추 품질관련하여 유전체를 분석하고 이를 바탕으로 대량분자표지를 개발하였고 QTL 지도를 작성하였음

Abstract ▼

Next generation sequencing technologies have proven to be a rapid and cost-effective means to assemble and characterize gene content and identify molecular markers in various organisms. Pepper (Capsicum annuum L., Solanaceae), is a major staple vegetable crop, which is economically important and has

Next generation sequencing technologies have proven to be a rapid and cost-effective means to assemble and characterize gene content and identify molecular markers in various organisms. Pepper (Capsicum annuum L., Solanaceae), is a major staple vegetable crop, which is economically important and has worldwide distribution. Objectives 454 GS-FLX Titanium massive parallel pyrosequencing of polyA-selected and normalized cDNA libraries generated from a single pool of transcripts obtained from two pepper varieties, the highly pungent, Saengryeg 211, and the non-pungent, Saengryeg 213, was done for de novo transcriptome assembly, functional annotation, and in silico discovery of potential molecular markers. Results A single 454 pyrosequencing run generated 361,671 and 274,269 reads totaling 164.49 and 124.60 Mb of sequence data, which assembled into 23,821 and 17,813 isotigs and 18,147 and 15,129 singletons for both varieties, respectively. These reads were organized into 20,352 and 15,781 ‘isogroups’ for both varieties. Assembled sequences were functionally annotated based on homology to genes in multiple public databases and assigned with Gene Ontology (GO) terms. Analysis of sequence variants identified a total of 11,584 and 9,641 potential SNPs which eventually resulted in 665 and 632 genotype specific SNPs, for both the varieties, respectively, after examining SNP frequency distribution for each mapped unigenes.
High-throughput transcriptome profiling of two pepper cultivars, Mandarin and Blackcluster using 454 GS-FLX pyrosequencing yielded 279,221 and 316,357 sequenced reads with a total 120.44 and 142.54 Mb of sequence data (average read length of 431 and 450 nucleotides). These reads resulted from 17,525 and 16,341 ‘isogroups’ and were assembled into 19,388 and 18,057 isotigs, and 22,217 and 13,153 singletons for both the cultivars, respectively. Assembled sequences were annotated functionally based on homology to genes in multiple public databases. Detailed sequence variants analysis identified a total of 9,701 and 12,741 potential SNPs for both cultivars, which eventually resulted in 1,025 and 1,059 genotype specific SNPs, for both the varieties, respectively, after examining SNP frequency distribution for each mapped unigenes. These markers for pepper will be highly valuable for marker-assisted breeding and other genetic studies. The linkage map was constructed with 365 SNPs covering 844cM, with 26 linkage groups for capsaicinoids QTL analysis and 246 SNPs covering 512cM, with 21 linkage groups for carotenoids QTL analysis. Analysis of QTL revealed that two major QTL on LG 2 and 5 minor QTL on LG1, LG3, LG8 related to capsaicinoids contents, and 6 QTLs on LG1, LG6, LG9, LG11 related to carotenoids contents. These QTL linked markers would be informative to marker assisted selection for pungency level and red color level in a chili pepper breeding program. Simple and rapid diagnostic method was newly developed for the efficient analysis of pungency or total carotenoids contents. A spraying method using 0.1N NaOH and 0.2% 2,6-dichloroquinone chlorimide solutions allowed to detect on both a fruit without a part of pericarp and a copying paper blotted with fruit section by spraying with above solutions in 5 minutes. On optical observation, this method could evaluate the presence of capsaicinoids, appropriate amount, and distribution of capsaicinoids in pepper fruits. For total carotenoids contents of pepper powder, a microplate method based on a 96-wells polystyrene microplate and an ELISA reader was also developed by simplifying the color extraction and measuring steps. The method as one of alternatives of ASTA-20.1 method allowed to rapid analysis of a number of samples. For pungency, fruit color, and sugar contents, genetic analysis showed that the traits were controlled by multiple genes. After NGS sequencing each parental lines for the traits, inter or intraspecific genetic pepper maps were constructed with SNP, SSR, and CAPS markers. Composite interval mapping analysis showed that 8, 2, 6, 4 QTLs controlled for the contents of capsaicin, dihydrocapsaicin, total carotenoids, and sugar, respectively. Using a series of markers for fertility, root rot resistance, and above three traits, processing pepper varieties with high quality were developed by inter or intraspecific hybridizations and generation advancement among genetic resources of domestic inbred lines, commercila varieties, Jolokia, and jalapeno derives. For high contents of fruit color, seven domestic C and two CGMS A-B lines were developed, resulting in a commercial variety of PNB-PR4 submitted to application for protection of new varieties of plants. For high pungency, GMS/CGMS and fertile parents were finally selected through marker analysis, sib-crossing, and generation advancement. As results, two commercial varieties of PNB-HP1 and PNB-HP2 for the Eastern Asia market were submitted for application for protection of new varieties of plants. For high contents of sugar, individuals with above 10% sugar contents were selected from the progenies of commercial variety of Daedeulbo, 22 CMS with half fertility and 8 CGMS C lines were finally selected through generation advancement.

목차 Contents

• 완결과제 최종보고서 ... 1
• 제출문 ... 2
• 요약문 ... 3
• SUMMARY ... 4
• 목 차 ... 6
• 제 1 장 서 론 ... 7
• 제 2 장 국내외 기술개발 현황 ... 8
• 제 3 장 연구개발 수행 내용 및 결과 ... 12
• 1절(제1세부과제) : 고추 품질 관련 특성 연관 분자표지의 대량발굴 및 분자육종시스템 구축 ... 12
• 제 4 장 연구개발 목표 달성도 및 대외 기여도 ... 188
• 1절 : 목표대비 대외 달성도 ... 188
• 2절 : 정량적 성과 ... 189
• 제 5 장 연구개발 결과의 활용계획 ... 189
• 제 6 장 연구개발 과정에서 수집한 해외과학기술정보 ... 189
• 제 7 장 기타 중요 변동사항 ... 191
• 제 8 장 국가과학기술종합정보시스템에 등록한 연구장비 현황 ... 192
• 제 9 장 참고문헌 ... 192
• 끝페이지 ... 195