보고서 정보
주관연구기관 |
국립원예특작과학원 National Institute of Horticultural and Herbal Science |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2014-02 |
과제시작연도 |
2011 |
주관부처 |
농촌진흥청 Rural Development Administration(RDA) |
등록번호 |
TRKO201400010990 |
과제고유번호 |
1395022789 |
사업명 |
국책기술개발 |
DB 구축일자 |
2014-07-05
|
DOI |
https://doi.org/10.23000/TRKO201400010990 |
초록
Ⅳ. 연구개발결과
○ 사과 자가적과성 연관 유전자 선발하여 사과 형질전환체 획득
- MdJOINTLESS, MdIAA14
○ 배 ‘신고’ ‘황금배’ 재분화 구축과 검은별무늬병 저항성 유전자 도입
- Aldo-keto reductase, PR10
Abstract
▼
Thinning of apple fruitlets is one of the most laborious and important works for the improvement of fruit quality and for the promotion of sufficient flower bud formation to prevent alternate bearing in commercial cultivars. Lateral fruits of self-thinning apple cultivars fall naturally within 30 da
Thinning of apple fruitlets is one of the most laborious and important works for the improvement of fruit quality and for the promotion of sufficient flower bud formation to prevent alternate bearing in commercial cultivars. Lateral fruits of self-thinning apple cultivars fall naturally within 30 days after full bloom and only central fruit remains to mature. Differences of gene expression between central fruit and lateral fruit were investigated by differential display (DD) PCR. Partial cDNAs of 30 clones from the central fruit and 24 clones from the lateral fruit were selected for nucleotide sequence determination and homology searches. The levels of transcripts coding for proteins involved in pathogenesis related proteins, senescence, temperature stress, protein degradation, fruit browning, sorbitol metabolism were significantly higher in pedicels of lateral fruit than in pedicels of central fruit. On the other hand, the up-regulation of proteins involved in anthocyanin and flavanol biosynthesis and ethylene synthesis were observed in pedicels of central fruit. In Real time PCR analysis, cytochrome P450 gene was confirmed as showing a higher expression level in lateral fruit than in central fruit. The results of this study indicate that differentially expressed genes are related to self-thinning characteristics in apple tree.
Abscission is an important developmental process used to shed organs such as leaves, flowers and fruits. Despite the detailed characterization of growth dynamics and hormonal balance during the early steps of fruit development, the molecular aspects remain unclear.
Abscission of young fruit occurs by separation of cells in anatomically distinct regions between the pedicel and junction. It is regarded that the formation of abscission zone is controlled by a MADS-box gene like J OI NTLESS in tomato. Through DNA sequence comparison with the MADS-box gene family, M dJ OI NTLESS was classified into the SVP clade composed of AtSVP (Arabidopsis thaliana), P sSVP (P isum sativum), I bM AD S3 (I pomoea batatas), StM AD S16 (Solanum tuberosum), which has high sequence similarity.
For the first time we isolated M dJ OI NTLESS from young fruit pedicels in apple (M alus xdomestica) using degenerate primers based on highly conserved regions and RACE assays.
To clarify the mechanism of M dJ OI NTLESS underlying abscission zone development, we constructed a vector for transformation into tomato and apple. Overexpression of M dJ OI NTLESS in the transgenic tomato developed abscission zone-like structures on their pedicels but transformed with the antisense construct failed to develop abscission zone.
This study carried out to identify the genes resistance to pear scab disease by Venturia nashicolay. To construct expressed sequence tags (ESTs) database for transcript by inoculation in pear ‘PS2’,we harvested the leaves at various times after inoculation in resistant cultivars ‘PS2’ (24, 48, 72, 96 and 120 hours) and susceptible cultivars ‘Sweat skin’ (0, 24, 48, 72, 96 and 120 hours). And then we divided into 4 groups (Group1; ‘PS2’ total mixture and ‘sweat skin’ NT, Group 2; ‘PS2’ total mixture and ‘sweat skin’ total mixture, Group3; ‘PS2’ 24 hours and ‘sweat skin’ 24 hours, Group4; ‘PS2’ 48 hours and ‘sweat skin’ 48 hours) and carried out suppression subtractive hybridization (SSH). As a result of the ESTs analysis, we found 18, 14, 10 and 7 unique sequences on ‘PS2’ specific expression in Group 1, 2, 3 and 4, respectively. And ratio of defense or stress related gene was most highly in Group 3 with 35%. In Group 3, pathogenesis-related protein 1a and Major allegen Pyr c1, they classified as pathogen-related protein (PR protein) family was specifically expressed. Group 4 expressed many resistance genes including F-Box/kelch-repeatprotein, peptidetransporter, dihydroflavonol-4-reductase, cysteinproteasethenothergroups. As a result of real-time PCR, most resistance-related genes except glutathione S-transferase(GST) were highly expressed. Especially, Mald1. 01, F-Box/kelch-repeatprotein, hypothetical protein was highly expressed in highly resistant cultivars ‘Bartlett’ and ‘PS2’, and weakly expressed in moderately susceptible cultivars ‘Gamcheonbae’ and susceptible cultivars ‘wonhwang’, and highly susceptible cultivars ‘Niitaka’ and‘ sweat skin’. This result means that these 3genes are important role in pear scab diseaseby V.nashicolay, and we need further study using transformants to know more detail mechanism of these genes In order to establish an efficient adventitious shoot regeneration conditions from leaf explants for Asian pear ‘Whangkeumbae’, the effect of concentration and kinds of plant growth regulator and carbon source was investigated. Leaf explants of cultures grown on Murashige and Skoog (MS) medium containing 8 g/L plant agar were used. When the medium contained 0.25 mg/L thidiazuron (TDZ) and 0.3 mg/L indolebutyric acid (IBA), the adventitious shoot regeneration rate (ASRR) was greater as 61.1% than others treated and higher TDZ concentrations (2.5 and 5 mg/L) treatment significantly reduced the ASRR. As the effect of IBA and indoleacetic acid (IAA) concentration on the ASRR, 0.5 mg/L TDZ plus different concentration of IAA exhibited relatively high ASRR and 0.5 mg/L TDZ plus 0.3 mg/L IAA showed the highest ASRR of 76.7%. Also the effect of sucrose and sorbitol as carbon source on regeneration was examined. The highest ASRR and the most shoots per explants averaged 94.4% and 3.49 by treatment of 30 mg/L sorbitol, respectably.
Sorbitol is considered better carbon source than sucrose for shoot regeneration of ‘Whangkeumbae’ pear.
Genetic manipulation of pear (P yrus pyrifolia Nakai) breeding is still difficult due to lack of reliable regeneration system. The aim of this research is to establish shoot regeneration system from leaf explants for pear (P . pyrifolia cv. Niitaka) using various concentrations of plant growth regulators and carbon source supplemented to medium. The highest regeneration rate of about 20% was found on a medium containing 4.4 g/L of Murashige and Skoog (MS) without vitamins, Linsmaier and Skoog (LS) vitamins were added separately. Leaf explants of pear were cultured on MS medium containing 7 g/L of Daishin agar supplemented with various concentrations of NAA (0.01, 0.05, 0.1, 0.5 mg/L) in combination with BA(3, 5, 10 mg/L) for shoot regeneration. In medium with 5 mg/L of BA and 0.01 mg/L of NAA, adventitious shoot regeneration rate was higher than others treated. The optimal results were observed using MS medium supplemented with 30 g/L sorbitol as carbon source on regeneration system. Sorbitol is considered better carbon source than sucrose for shoot regeneration of pear (P . pyrifolia cv. Niitaka). In order to increase of shoot regeneration in pear (P . pyrifolia cv. Niitaka), plant agar and Daishin agar used as gelling agents, Daishin agar is more efficient in shoot regeneration.
목차 Contents
- 표지 ... 1
- 제출문 ... 2
- 요약문 ... 3
- SUMMARY ... 6
- 목차 ... 9
- 제 1 장 서 론 ... 10
- 제 1 절 사과 자가적과성 육종소재의 경제적.산업적 중요성 및 연구개발의 필요성 ... 10
- 제 2 절 배 검은별무늬병 저항성 육종소재의 경제적.산업적 중요성 및 연구개발의 필요성 ... 11
- 제 2 장 국내외 기술개발 현황 ... 12
- 제 1 절 국외 기술개발 현황 ... 12
- 제 2 절 국내 기술개발 현황 ... 13
- 제 3 장 연구개발수행 내용 및 결과 ... 15
- 제 1 절 연구개발 수행 내용 ... 15
- 제 2 절 연구개발 수행 결과 ... 27
- 제 4 장 연구개발목표 달성도 및 대외기여도 ... 104
- 제 1 절 목표대비 달성도 ... 104
- 제 2 절 정량적 성과 ... 105
- 제 5 장 연구개발결과의 활용계획 ... 106
- 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 107
- 제 7 장 기타 중요 변동사항 ... 108
- 제 8 장 국가과학기술종합정보시스템에 등록한 연구장비 현황 ... 109
- 제 9 장 참고문헌 ... 110
- 끝페이지 ... 118
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