보고서 정보
주관연구기관 |
충남대학교 Chungnam National University |
연구책임자 |
안상낙
|
참여연구자 |
강주원
,
이현숙
,
윤여태
,
상세티
,
김선하
,
전윤아
,
강윤주
,
심규찬
,
KoKo Maung
,
Luxin
,
Le Anh Quyhn
,
Adeva Cherly
,
Lung Ngoc HA
,
나혜린
,
안형주
,
최경호
,
김현중
,
이용규
,
조민주
,
남경윤
,
양재현
,
조현진
,
최호열
|
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2018-02 |
과제시작연도 |
2017 |
주관부처 |
농촌진흥청 Rural Development Administration(RDA) |
등록번호 |
TRKO201800043274 |
과제고유번호 |
1395049820 |
사업명 |
차세대바이오그린21 |
DB 구축일자 |
2018-12-15
|
키워드 |
야생벼.근동질계통.분자표지.수량안정성.유전자분리.rice.nearly isogenic lines.DNA markers.yield stability.gene isolation.
|
DOI |
https://doi.org/10.23000/TRKO201800043274 |
초록
▼
야생유전자원의 유용유전자를 활용하기 위하여, 야생벼의 염색체단편이 최소로 이입된 근동질계통을 육성하고 수량안정성 관련 형질연관 QTL을 탐색하였다. 화성/O. rufipogon 조합 계통을 이용하여 염색체 9번의 Ascorbate peroxidase 유전자가 출수기 등에 관여함을 밝히고, 염색체 8번의 O. rufipogon 단편에 위치하는 QTL(gw8.1) 이 화성의 배경에서 종자길이를 증가시킴을 확인하였다. 또한 저온발아성에 관여하는 신규유전자 ltg1을 탐지하고 후보유전자를 밝히기 위해 T-DNA 삽입변이체 등을 검정하고 있
야생유전자원의 유용유전자를 활용하기 위하여, 야생벼의 염색체단편이 최소로 이입된 근동질계통을 육성하고 수량안정성 관련 형질연관 QTL을 탐색하였다. 화성/O. rufipogon 조합 계통을 이용하여 염색체 9번의 Ascorbate peroxidase 유전자가 출수기 등에 관여함을 밝히고, 염색체 8번의 O. rufipogon 단편에 위치하는 QTL(gw8.1) 이 화성의 배경에서 종자길이를 증가시킴을 확인하였다. 또한 저온발아성에 관여하는 신규유전자 ltg1을 탐지하고 후보유전자를 밝히기 위해 T-DNA 삽입변이체 등을 검정하고 있다. 화성/O. minuta 후대계통을 이용하여 까락발달에 관여하는 신규유전자(awn9)를 탐지하고 유전자분리를 위한 고밀도지도를 작성하고 있다. 화성/O. grandiglumis 후대를 이용하여 GW2 (grain width 2) 유전자가 종자중 외에도 엽록소함량을 조절함을 밝혔고, GW2 유전자의 다면발현 효과를 분석 중이다. Nipponbare/Kasalath 조합근동질계통을 이용, 중배축신장에 관여하는 QTL을 탐지하고 초엽신장성과의 관계를 밝혀 직파재배 적성 품종육성을 위한 정보를 획득하였다. 이입계통 중 미질 및 저온발아성 형질이 반복친에 비해 우수한 화원8호, 화원15호를 2계통을 품종 출원하였다. 근동질계통은 다양한 표현형을 조절하는 양적형질유전자의 탐색 및 이용에 귀중한 자원이 될 것이다.
( 출처: 보고서 요약서 3p )
Abstract
▼
□ Purpose&Contents
The objectives of the present study were to develop NIL(nearly isogenic lines) from interspecific crosses (1) to identify trait-improving QTL derived from the wild and the weedy rice that are associated with improved agronomic performance in the cultivated background, (2) to de
□ Purpose&Contents
The objectives of the present study were to develop NIL(nearly isogenic lines) from interspecific crosses (1) to identify trait-improving QTL derived from the wild and the weedy rice that are associated with improved agronomic performance in the cultivated background, (2) to develop DNA markers to QTL of agronomic importance and isolate and characterize genes underlying these traits, and (3) to select promising lines from the NIL populations based on repeated backcrossing and marker-assisted selection.
□ Results
Heading date and plant height
A high-resolution physical map targeting a cluster of yield-related QTLs on the long arm of rice chromosome 9, a BC4F6 near isogenic lines (NILs) derived from a cross between Hwaseong and O. rufipogon were further evaluated and compared to Hwaseong for nine agronomic traits. Significant difference in plant height was observed between Hwaseong and NILs due to an increase in internodes, particularly in the 1st and 4th internode, than Hwaseong. To investigate the antioxidant activity in Hwaseong and NILs, the 3,3-diaminobenzidin (DAB) staining and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay were both conducted. The DAB staining was also performed to understand whether or not OsApx gene is important in scavenging H2O2 in rice. Based on DAB staining, dark-brown color was barely detected in NILs than Hwaseong. Also, the DPPH scavenging ability of NILs showed higher value than Hwaseong. Hence, these results suggested that the internode elongation and antioxidant activity might possibly be controlled by OsApx gene.
Grain weight and zinc content
Proteins containing a placenta-specific 8 (PLAC8) domain regulate cell growth and cancer cell development in animals, fruit weight (FW2.2) in tomato, and cell number (ZmCNRs) in maize, and as putative metal ion transporters (PCRs, plant cadmium resistance) in Arabidopsis, rice, and Indian mustard. However, it has not been demonstrated how PLAC8 domain-containing proteins fulfill such diverse functions. Here, we report that rice PCR1 regulates both the Zn contents of grains and grain weight.
OsPCR1 knockout and knockdown lines produced lighter grains, while OsPCR1 overexpression lines produced heavier grains than the wild type.
Furthermore, the Zn and Cd contents were altered in grains of the transgenic rice lines as well as in yeast cells heterologously expressing OsPCR1. Through PCR1 sequence analysis using different rice accessions collected from many countries, we found that the PCR1s in Japonica and Indica rice exhibited distinct amino acid sequences, and this difference was associated with Zn resistance and accumulation Japonica PCR1 enhanced Zn hypersensitivity in yeast, and Japonica rice contained less Zn in their grains than did Indica rice. Our functional study and genetic analysis suggests that rice PCR1 maintains metal ion homeostasis and grain weight and might have evolved during domestication. We believe that this discovery will give great interest to the readership of Plant Cell and Environment, as it provides clues as to how PLAC8 domain-containing proteins fulfill diverse functions.
Agronomic and grain quality traits
QTL associated with 12 grain quality traits were identified using 96 introgression lines (IL) of rice developed from an interspecific cross between the Korean elite O. sativa japonica cultivar 'Hwaseong' and O. rufipogon over 7 years. QTL analyses indicated that QTL qDTH6 for heading date, detected on chromosome 6 is associated with variance in grain traits. Most QTLs detected in this study clustered near the qDTH6 locus on chromosome 6, suggesting the effect of qDTH6. O. rufipogon alleles negatively affected grain quality traits except for a few QTLs, including qGCR9 for glossiness of cooked rice on chromosome 9. To characterize the effect of the O. rufipogon locus harboring qGCR9, four lines with a single but different O. rufipogon segment near qGCR9 were compared to Hwaseong. Three lines (O. rufipopgon ILs) having O. rufipogon segment between RM242 and RM245 in common showed higher glossiness of cooked rice than Hwaseong and the other line (Hwaseong IL), indicating that qGCR9 is located in the 3.4-Mb region between RM242 and RM245. Higher glossiness of cooked rice conferred by the O. rufipogon allele might be associated with protein content considering that three lines had lower protein content than Hwaseong (P < 0.1). These three O. rufipogon ILs showed higher yield than Hwaseong and Hwaseong IL due to increase in spikelets per panicle and grain weight indicating the linkage of qGCR9 and yield component QTLs.
Grain size and grain yield
A quantitative trait locus (QTL) gw8.1 was detected in the population derived from a cross between the elite japonica cultivar, ‘Hwaseong’ and Oryza rufipogon (IRGC 105491). Near isogenic lines (NILs) harboring the O. rufipogon segment on chromosome 8 showed increased grain length and weight compared to those of the recurrent parent, Hwaseong. This QTL was mapped to a 175.3-kb region containing 28 genes, of which four were considered as candidates based on the presence of mutations in their coding regions and as per the RNA expression pattern during the inflorescence stage. Leaves and panicles obtained from plants harvested 5 days after heading showed differences in gene expression between Hwaseong and gw8.1-NILs. Most genes were upregulated in O. rufipogon and gw8.1-NIL than in Hwaseong. Scanning electron microscopy analysis of the lemma inner epidermal cells indicated that cell length was higher in gw8.1 NIL than in Hwaseong, indicating that gw8.1 might regulate cell elongation. Among the candidate genes, LOC_Os08g34380 encoding a putative receptor-like kinase and LOC_Os08g34550 encoding putative RING-H2 finger protein were considered as possible candidates based on their functional similarity.
Direct seedoing: mesocotyl and coleoptile elongation
Direct-seeding cultivation by deep-seeding of seeds (drill seeding) is becoming popular due to the scarcity of land and labor. However, poor emergence and inadequate seedling establishment can lead to yield loss in direct-seeding cultivation by deep-sowing. In rice, mesocotyl and coleoptile are primarily responsible for seedling emergence from deeper levels of soil.
Quantitative trait loci (QTLs) for mesocotyl and coleoptile length at 5-cm seeding depth were detected using 98 backcross inbred lines from a cross between Kasalath and Nipponbare. Three QTLs qMel-1, qMel-3, and qMel-6 for mesocotyl length were identified on chromosomes 1, 3, and 6, respectively, in two independent replicates. At two QTLs, qMel-1 and qMel-3, the Kasalath alleles increased mesocotyl length, whereas Nipponbare allele increased at qMel-6. The Nipponbare alleles at two QTLs (qCol-3 and qCol-5) increased the coleoptile length. Further, seeds of 54 chromosome segment substitution lines (CSSLs) from the cross between Kasalath and Nipponbare sown at 5 cm soil depth showed a significant positive correlation between seedling emergence and mesocotyl elongation(r > 0.6, P < 0.0001), but not with coleoptile elongation (r = 0.05, P = 0.7). Seedling emergence of Nipponbare, Kasalath, and the 3 of the 54 CSSLs rapidly decreased with increasing sowing depth. Seedling emergence at seeding depths of 7 and 10 cm was faster in Kasalath and CSSL-5 that harbored the Kasalath alleles across the qMel-1 and qMel-3 regions than in the other two CSSLs that contained a single QTL and Nipponbare alleles.
CSSL-5 showed the longest mesocotyl among
the 3 CSSLs, but no difference in coleoptile length was observed among the 3 CSSLs at seeding depths of 7 and 10 cm. Variation of mesocotyl elongation was found to be associated with seedling emergence at the seeding depth of 5 cm. To our knowledge, this is the first study performed using CSSLs to detect QTLs for mesocotyl or coleoptile elongation and to determine the effect of mesocotyl elongation on seedling emergence in rice.Our findings provides a foundation for developing rice cultivars that show higher seedling emergence after direct seeding by introgressing QTLs for mesocotyl elongation in rice breeding.
□ Expected Contribution
o The NILs developed in the study would be used as bridging lines for breeding purposes or directly as elite cultivars.
o The NILs would be useful in widening and enriching the diversity of cultivated rice cultivars.
o The NILs would be useful to isolate and characterize genes (QTLs) of agronomic impportance.
( 출처: SUMMARY 5p )
목차 Contents
- 표지 ... 1
- 제 출 문 ... 2
- 보고서 요약서 ... 3
- 국 문 요 약 문 ... 4
- Summary ... 5
- 목차 ... 8
- 제 1 장 연구 개발 과제의 개요 ... 9
- 제1절 연구 개발 목적 ... 9
- 제2절 연구 개발의 필요성 ... 9
- 제3절 연구 개발 범위 ... 9
- 제 2 장 국내외 기술개발 현황 ... 11
- 1절 연구개발 대상 기술의 국내·외 현황 ... 11
- 2절 선행 특허·기술 동향 분석 ... 13
- 제 3 장 연구 수행 내용 및 결과 ... 16
- 제1절 제1세부과제 연구 수행 내용 및 결과 ... 16
- 제 4 장 목표달성도 및 관련분야 기여도 ... 50
- 제1절 : 목표대비 달성도 ... 50
- 제2절 : 정량적 성과(논문게재, 특허출원, 기타)를 기술 ... 50
- 제 5 장 연구 결과의 활용 계획 ... 51
- 1절 연구개발결과의 활용방안 ... 51
- 2절 추가 연구의 필요성 ... 51
- 3절 추가적인 연구 성과 ... 52
- 제 6 장 연구 과정에서 수집한 해외 과학 기술 정보 ... 53
- 제 7 장 연구 개발 결과의 보안 등급 ... 56
- 제 8 장 국가과학기술지식정보서비스에등록한연구시설·장비현황 ... 56
- 제 9 장 연구개발과제수행에따른연구실등의안전조치이행실적 ... 56
- 1절 연구실 안전조치 이행계획 ... 56
- 2절 LMO 연구시설 ... 56
- 제 10 장 연구개발과제의 대표적 연구실적 ... 57
- 제 11 장 기타사항 ... 58
- 제 12 장 참고문헌 ... 60
- 끝페이지 ... 67
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