보고서 정보
주관연구기관 |
국립농업과학원 National Institute of Agricultural Sciences |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2015-03 |
과제시작연도 |
2014 |
주관부처 |
농촌진흥청 Rural Development Administration(RDA) |
등록번호 |
TRKO201500010401 |
과제고유번호 |
1395035764 |
사업명 |
농업기초기반연구 |
DB 구축일자 |
2015-07-11
|
DOI |
https://doi.org/10.23000/TRKO201500010401 |
초록
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Ⅳ. 연구개발결과
후성유전적 형질인 ‘GFP-partitioned plant“의 표현형 분석하였고 형질전환체 게놈내 도입유전자(GFP-transgene)의 구조 및 배열 확인하여 2개의 유전자가 “direct repeat”의 구조를 확인 하였다. silencing 메카니즘은 RNA-directed DNA methylation(RdDM)으로 확인되었다. 배추 글루코시놀레이트 유용유전자 분석을 위해서는 결구배추, 순무, 청경채, 유채 등 다양한 아종을 포함한배추 핵심집단 100여 계통을 선발하여 ATTIRTA1 DNA 전이인자 전
Ⅳ. 연구개발결과
후성유전적 형질인 ‘GFP-partitioned plant“의 표현형 분석하였고 형질전환체 게놈내 도입유전자(GFP-transgene)의 구조 및 배열 확인하여 2개의 유전자가 “direct repeat”의 구조를 확인 하였다. silencing 메카니즘은 RNA-directed DNA methylation(RdDM)으로 확인되었다. 배추 글루코시놀레이트 유용유전자 분석을 위해서는 결구배추, 순무, 청경채, 유채 등 다양한 아종을 포함한배추 핵심집단 100여 계통을 선발하여 ATTIRTA1 DNA 전이인자 전시 결과 핵심집단의 유전적 다양성은 형태적인 특성과 거의 일치하는 3개의 소그룹으로 분류되었다. 글루코시놀레이트 profiling 에 대해서는 15종의 글루코시놀레이트 중에서 아종별 함량에 유의성 있는 차이를 보이는 8개 글루코시놀레이트 선발하였다.
배추에서 캘러스형성 및 식물체 재분화율에 연관된 QTL 탐색에 대해서는 유전력 분석(h2)에 의한 조직배양 능력의 유전자 존재를 확인하였고 캘러스 형성과 관련 2개, 재분화연관 4개(변이 43.2% 설명) 유전자좌 탐색되었다. 조직배양 능력 연관 QTL을 조절하는 후보 유전자 선발하기 위해 배추 유전체 정보로부터 QTL interval 내에 존재하는 23개 후보 유전자 선발하였고 auxin, cytokinin 등 호르몬 및 cell cycle 연관 유전자가 배추 조직배양에 주로 관여하는 것으로 확인되었다.
Abstract
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We previously reported the novel partitioning of regional GFP-silencing on leaves of 35S-GFP transgenic plants, coining the term “partitioned silencing”. We set out to delineate the mechanism of partitioned silencing. Here, we report that the partitioned plants were hemizygous for the transgene, pos
We previously reported the novel partitioning of regional GFP-silencing on leaves of 35S-GFP transgenic plants, coining the term “partitioned silencing”. We set out to delineate the mechanism of partitioned silencing. Here, we report that the partitioned plants were hemizygous for the transgene, possessing two direct-repeat copies of 35S-GFP. The detection of both siRNA expression (21 and 24 nt) and DNA methylation enrichment specifically at silenced regions indicated that both post-transcriptional (PTGS) and transcriptional gene silencing (TGS) were involved in the silencing mechanism. Using in vivo agro-infiltration of 35S-GFP/GUS and inoculation of TMV-GFP RNA, we demonstrate that PTGS, not TGS, plays a dominant role in the partitioned silencing, concluding that the underlying mechanism of partitioned silencing is analogous to RNA-directed DNA methylation (RdDM). The initial pattern of partitioned silencing was tightly maintained in a cell-autonomous manner, although partitioned-silenced regions possess a potential for systemic spread. Surprisingly, transcriptome profiling via Next-generation sequencing demonstrated that expression levels of most silencing pathway-involved genes were similar in both GFP-expressing and silenced regions although a diverse set of region-specific transcripts were detected.This suggests that partitioned silencing can be triggered and regulated by genes other than silencing pathway-involved genes.
Quantitative trait loci (QTL) controlling callus induction and plant regeneration were identified in the VCS3M-DH population of Brassica rapa. The VCS3MDH population showed wide and continuous variation in callus induction and shoot regeneration.
Significant coefficient correlations were detected between these two parameters. Broad-sense heritability (h2) for the two traits was around 0.7, indicating genetic regulation of regeneration ability in this population. In the composite interval mapping analysis, two QTLs for callus induction ability, qCi2 and qCi7, were mapped on chromosome A02 and A07, explaining 28.6 % of phenotypic variation. For plant regeneration, four QTLs, qPr6-1 qPr6-2, qPr7, and qPr9 were identified on chromosome A06, A07, and A09, which in total explained 50.1 % of phenotypic variation. Furthermore, 15 putative candidate genes were found on the interval of the six QTLs, which were related to various plant hormones, MADS-box genes, and serine/threonine related genes. These results provide important information to identify genes related to tissue culture ability in B. rapa.
A novel dominant resistance gene, TuRB07, was found to confer resistance to an isolate of TuMV strain C4 in B. rapa line VC1 and mapped on the top of chromosome A06. The inheritance of resistance to Turnip mosaic virus in Brassica rapa was investigated by crossing the resistant line, VC1 with the susceptible line, SR5, and genotyping and phenotyping diverse progenies derived from this cross. Both a doubled haploid population, VCS3M-DH, an F2 and two BC1 (F1 × VC1 and F1 × SR5) populations were created. Population tests revealed that the resistance to the TuMV C4 isolate in B. rapa is controlled by a single dominant gene. This resistance gene, TuRB07 was positioned on the top of linkage group A06 of the B. rapa genome through bulk segregation analysis and fine mapping recombinants in three doubled haploid- and one backcross population using microsatellite markers developed from BAC end sequences. Within the region between the two closely linked markers flanking TuRB07, H132A24-s1, and KS10960, in the Chiifu reference genome, two genes encoding nucleotide-binding site and leucine-rich repeat proteins with a coiled-coil motif (CC-NBS-LRR), Bra018862 and Bra018863 were identified as candidate resistance genes. The gene Bra018862 is truncated, but the gene Bra018863 has all the domains to function
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