보고서 정보
주관연구기관 |
건국대학교 KonKuk University |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2006-07 |
과제시작연도 |
2005 |
주관부처 |
농림부 Ministry of Agriculture and Forestry |
등록번호 |
TRKO201400022974 |
과제고유번호 |
1380001746 |
사업명 |
농림기술개발 |
DB 구축일자 |
2014-11-10
|
초록
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○ 연구결과
제1세부과제 : 쌀 호화점도특성 다양화 계통 육성
- 쌀 식이섬유함량 다양화를 위하여 많은 잡종집단과 계통을 육성하여 최종적으로 생산력검 정시험에 공시할 12계통을 선발하였고, 초형 등 작물학적특성이 우수하고 호화점도특성이 서로 다른 찰벼 또는 메벼 51 계통을 선발하여 2006년도 육종포장에서 시험 중임.
- 잡종 개체의 쌀 알칼리붕괴도 및 쌀알의 투명도는 호화점도특성과 유전적으로 연관되어 있음이 밝혀짐.
- 쌀의 호화점도특성 관련 분자마커를 찾기 위해서 2개 여교배 조합 387개의 F8 RILs
○ 연구결과
제1세부과제 : 쌀 호화점도특성 다양화 계통 육성
- 쌀 식이섬유함량 다양화를 위하여 많은 잡종집단과 계통을 육성하여 최종적으로 생산력검 정시험에 공시할 12계통을 선발하였고, 초형 등 작물학적특성이 우수하고 호화점도특성이 서로 다른 찰벼 또는 메벼 51 계통을 선발하여 2006년도 육종포장에서 시험 중임.
- 잡종 개체의 쌀 알칼리붕괴도 및 쌀알의 투명도는 호화점도특성과 유전적으로 연관되어 있음이 밝혀짐.
- 쌀의 호화점도특성 관련 분자마커를 찾기 위해서 2개 여교배 조합 387개의 F8 RILs를 육성하였으며, 호화점도특성과 연관된 5개의 분자마커를 찾았음.
- 쌀의 호화점도특성과 연관된 RM마커의 실제 선발효과를 조사한 결과 RM587, RM412, RM230, RM42를 사용하여 나타난 밴드형태를 확인하면 호화점도특성에 대한 잡종개체 또는 계통선발을 효과적으로 수행할 수 있을 것으로 판단하였음.
- 본 연구를 통하여 육성한 품종은 쌀 호화점도특성이 특이하며 작물학적특성이 우수한 「황금찰」과 「자채찰」이며, 「황금찰」은 품종보호출원을 하였음.
- 메벼 중 최종점도, 치반점도 및 응집점도가 높은 품종의 쌀은 가래떡, 쌀 빵 및 쌀 과자용으로 맞지 않았고, 호화점도가 낮은 찰벼도 쌀 빵이나 쌀 과자 제조에 적당치 않았으며, 찰중 호화점도가 높은 품종이 찰떡용으로 적당하였음.
제2세부과제 : 쌀의 고식이섬유 및 호화점도특성 관련 DNA microarray구축과 형질전환체 모본개발
- 총 1600여개의 벼의 전분 및 cellulose 합성관련 염기서열을 확보하였고, 이 중 Blast, Clustal W등으로 염기서열 분석을 하여 131개 유전자를 증폭할 수 있는 primer 제작완료
- 벼의 Myb 전사조절 유전자 집단의 발굴과 66개 전사조절 유전자를 증폭할 수 있는 프라이머를 제작한 후 배유 및 embryogenic callus에서 추출한 RNA를 이용하여 RT-PCR을 실시하여 33개 유전자의 동정완료.
- 앞에서 발굴한 전분합성 관여 164개 유전자의 특정부분을 확보하여 보관중이고 이를 microarray chip위에 프린팅 할 예정임. 이 들 유전자와 쌀의 고식이섬유성 및 호화점도 특성관련 형질과의 연관성 검정시스템을 개발하여 starch branching enzyme과 관련된 몇 가지 형질을 발견하였으며, 이 검정 방법에 대한 특허 출원중임.
- 유전자총을 이용한 형질전환용 배지 시스템의 확보 및 벡터개발 완료. 기관분화가 잘 되는 체세포 배의 유기와 체세포 배를 이용한 전분구조 관련 유전자의 대량 검정시스템의 개발.
협동연구과제 : 쌀 고식이섬유 소재개발 및 유전연구
- 작물학학 특성이 개량된 고식이섬유 계통육성을 위하여 인공교배, F1∼F4 잡종집단 및 계통을 육성하였고, 그 중 78계통은 약배양을 거쳐 유전적 고정을 도모하였음.
- 약배양 후대 21계통 대한 수량성 검정 결과 백미 수량이 2.17∼5.84MT/ha로 변이가 컸고, 이들의 식이섬유 함량은 1.75∼3.56%의 범위를 보여 수원464호의 56∼113% 수준이었음.
- SSR 마커 196개를 이용하여 일품벼/수원464호에서 19개(9.2%)가 일품벼에 대해 다형성을 보였고 2번, 11번 염색체 외 10개 염색체 상에서 1∼3개 마커에서 다형성 마커가 탐색됨.
Abstract
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Summary of the results
1. Development of diverse novel breeding resources related with gelatinizat-ion properties of rice grain
1) The goal of this research is development and selection of quality lines showing diverse characteristics on gelatinization viscosity of rice grain.
We have used
Summary of the results
1. Development of diverse novel breeding resources related with gelatinizat-ion properties of rice grain
1) The goal of this research is development and selection of quality lines showing diverse characteristics on gelatinization viscosity of rice grain.
We have used numbers of genetic resources to develop and carried selection based on the agricultural characteristics obtained from breeding field and analysis results regarding the properties of gelatinization viscosity of rice obtained in laboratory. Among the breeding lines selected from the year 2005's breeding field, which have shown good agricultural characteristics and didn't show any further segregation, 12 lines were selected and progressed for the year 2006's field performance trial. Beside these selected lines, 51 waxy or non-waxy rice lines showing diverse properties of gelatinization viscosity and good agricultural characteristics have been selected and currently growing in 2006's breeding field.
2) To find out the genetic relationship between the properties of gelatinization viscosity and alkali digestibility value(ADV), we screened the properties of gelatinization viscosity and ADV of individual F2 plants of a cross between near isogenic lines showing clear difference in ADV.
We classified individual F2 plants into three groups based on ADV, those are higher and lower on ADV and still segregating group and we observed that right tendency of theoretical 1:2:1 segregation ratio. The
relationship between ADV and properties of gelatinization viscosity among F2 plants were that minimum viscosity, final viscosity, setback, and consistency biased to F2 plants showing lower ADV (ADV 1-2), whereas breakdown biased to F2 plants showing higher ADV (ADV 5-6).
And these tendency was resolved as statistically significant.
Consistently, the results indicate there is a genetic relationship between ADV and the properties of gelatinization viscosity.
3) Using F3 seeds, which obtained from F2 plants of the cross between near isogenic lines showing clear difference in transparency and clearness of rice endosperm, we analyzed the relationship of transparency and clearness of rice endosperm and the properties of gelatinization viscosity. The results was that the plants with opaque endosperm showed relatively lower minimum viscosity, final viscosity, setback and consistency. Based on this observation, we found there is genetic relationship between transparency of rice endosperm and the properties of gelatinization viscosity.
4) We have developed F8 RILs of two back-crossed combinations to identify molecular marker related to the properties of gelatinization viscosity. We observed the segregation of waxy and non-waxy lines among RILs and the great variation of gelatinization viscosity properties was found both in waxy and non-waxy lines. We harvested and stored 232 lines from the cross of Jinmi/wx202-25-1-5//Jinmi and 155 lines from the cross of Jinmi/wx185-29-1-3//Jinmi. These lines will be used as highly valuable genetic resources for further researches of rice molecular genetics.
5) We have attempted to find out molecular markers related to gelatinization viscosity properties. F6 RILs and 10 different RM primers, where located on the chromosome number 6 and 8, were used for SSR analysis. We found that RM412, 42 and 230 were potentially related to the gelatinization viscosity properties in non-waxy lines of Jinmi/wx202//Jinmi. Further, RM42, 38 and 587 were identified as molecular markers which related to the gelatinization viscosity properties in non-waxy lines of Jinmi/wx185//Jinmi. A commonly occurring maker in these two crosses was RM42, which located on the chromosome number 8.
6) We studied the effect of selection when we used the RM markers identified as above, which related to the gelatinization viscosity properties, using back-crossed RILs. Obviously, we concluded that we could use RM587, 412, 230 and 42 for selection of hybrid plants and lines based on the amplified DNA band pattern obtained from DNA gel analysis.
7) We selected a breeding line showing grater difference in the gelatinization viscosity compared with the standard variety "Jinbuchal" in 2003's yield trial field. This is one of the highly generation advanced breeding lines which has been bred in the plant breeding laboratory of Konkuk University. The nomenclature of this line has been done and the name of this line is "Hwangkeumchal". The maximum viscosity, minimum viscosity, final viscosity and breakdown of this line was much higher than standard check variety, whereas setback was much lower.
The seed pericarp color of this variety is reddish brown and anti-oxidative activity of brown rice is significantly higher than check variety and hull color turned to golden color during early grain filling stage. By this reason, we submit the plant variety protection application for saving this variety at March of 2005.
8) Another line among the highly generation advanced lines in yield trial field was named as "Jachaechal". The chararcteristics of this line is similar in the gelatinization viscosity with check variety, but it is clearly different with "Hwangkeumchal" and is shorter in stem length and showing very early stage maturity. The seed pericarp color is reddish-brown, therefore we expect this line has higher anti-oxidative activity. We are going to submit the plant variety protection application at end of 2006's field test.
9) Food processing chararcteristics in term of texture of the non-waxy rice product, "Junambyeo" which has lower adhesiveness and higher hardness is good for rice cookie. "Junambyeo" and "Saechucheongbyeo" which have higher cohesiveness and chewiness and lower setback is good for rice cake particularly for slender stick od rice cake. Ilpumbyeo and Chucheongbyeo have lower hardness and chewiness, therefore they are good for rice cake. And, Chucheongbyeo is good for rice bread.
IR50 and IR36 were not good for rice cookie, rice cake and rice bread due to their relatively higher setback and consistency.
10) Waxy rices are not good for the rice bread and rice cracker due to their higher adhesiveness. The waxy rices showing higher gelatinization viscosity such as Suwon290, Jachaechal and TaichungSen glutinous may good for extra sticky rice cake. TaichungSen glutinous, which has higher gelatinization viscosity and Hwaseonchal, which has lower gelatinization viscosity and higher adhesiveness may good for glutinous rice cake, so called "Injeolmi".
2. Development of rice edible traits related high resolution DNA microarray system and transgenic rice
1) Our research focused on developing rice edible traits related high resolution DNA microarray system and transgenic rice what has an modified metabolic pathway resulted in changes of structure of starch and starch granules of endosperm. We tried to develop high resolution DNA microarray system can be used for breeding of quality rice cultivar mainly focused on the traits related with cellulose, amylose, amylopectin contents and other ingredients those are affected on final edible quality of steamed rice. For this purpose, we identified more than 1600 gene sequences including transcription factors potentially involved in starch, sucrose and cellulose synthesis in rice. Among them, we cloned 164 genes from RT-PCR and certain part of these genes are amplified for making high resolution DNA microarray system. We have cloned 33 transcription factor genes those are expressed in developing rice endosperm and embryogenic callus.
2) Also, we set up rice transformation system in both aspects, gene delivery and tissue culture. We successfully identified and cloned embryogenic callus and observed initiation of organ differentiation, however, we failed to obtain fully grown transgenic rice plant due to a severe contamination problem occurred during 2005 in our laboratory.
We use particle bombardment system for gene delivery. These particle bombardment and embryogenic callus provide an unique opportunity to screen the function of a candidate gene, which relates to branching degree of starch found in embryogenic callus. The branching degree of starch found in embryogenic callus showing a tendency of lower branching degree overall. This gene function screening system has some advantages; we can check the function of gene in an early stage of rice transformation and can screening many genes in a short period.
In addition, we developed a method to find out relationship between a gene and a certain traits and currently we are working with patent application.
3. Development of high dietary fiber breeding lines and its genetic study
1) This experiments were carried out to develop new high dietary fiber varieties with Suweon464 by Ilpumbyeo mutation breeding using N-methyl-N-nitrosourea(MNU). The materials of this experiments were
used to sixteen crosses by Suweon464 and some varieties. The sixteen crosses were bred 494 cross seeds, F1 of thirteen crosses 226 plants, F2 of ten crosses 810 plants, F3 of four crosses 137 plants, and F4 of two crosses thirty plants.
2) Anther culture was practiced to develop rapidly a new high dietary varieties using the F1 plants of five crosses. The 78 new breeding lines were obtained from the F1 plants of five crosses by anther culture. The grain yield in observation yield trial field showed 2.17~5.84MT/ha in 21 anther culture lines. The yield indexes of the anther culture lines to Suweon464 were 51~138. Their dietary fiber in rice grain varied 1.75 to 3.56% in 21 anther culture lines and the dietary fiber indexes were 56~113 to Suweon464.
3) To study dietary fiber locus, we anchored 196 SSR markers on chromosome 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 using Ilpumbyeo and Suweon464. Among the anchored markers, 19 markers showed the polymorphism on Ilpumbyeo. The ten chromosomes were anchored and the 1~3 markers showed the polymorphism except chromosome 2, 11.
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