연관지도를 이용한 새우난초, 금새우난초, 변이종의 화색의 유전분석 Genetic Analysis of Flower Color Traits in Calanthe discolor, C. sieboldii, and Variants Using Molecular Linkage Map원문보기
본 연구는 제주도에서 자생하는 새우난초 3개체, 금새우난초 3개체 그리고 변이종 14개체를 포함하여 총 20개체를 화색에 따라 분류하고 유전자지도를 작성하여 QTL분석을 하였다. 화색은 새우난초가 어두운 자색으로 CIE Lab값이 $40{\sim}50$ 정도였으며, 금새우난초는 황색으로 $110{\sim}130$ 정도였고, 변이종 개체들은 새우난초와 유사하거나 다소 높았다. PCR 결과 얻은 polymorphism이 인정되는 154개 marker에 대한 분리비 적합도 검정에서 51개 marker에서 5% 수준의 유의성이 인정되었으며, 유의성이 인정된 51개 marker 중에서 새우난초 type은 37개, 금새우난초 type은 14개 였다. Polymorphism이 인정된 154개 marker에 대하여 MAPL program을 이용하여 이들 marker 상호간의 연관관계를 분석한 결과는 16개의 연관군과 1개의 독립군으로 구분되었으며, 이들 연관군에 대한 분자연관지도는 전체 group의 크기가 220.4 cM (centi Morgan)이고, marker간의 평균거리는 3.3 cM이었다. 양적 형질에 대한 분자연관지도상의 QTL 분석 결과, LOD 3.0 이상인 화색과 설판색의 QTL은 각각 3개와 1개였다. 이상에서 얻어진 자료는 새우난초 속의 화색 연구에 도움이 될 것으로 사료된다.
본 연구는 제주도에서 자생하는 새우난초 3개체, 금새우난초 3개체 그리고 변이종 14개체를 포함하여 총 20개체를 화색에 따라 분류하고 유전자지도를 작성하여 QTL분석을 하였다. 화색은 새우난초가 어두운 자색으로 CIE Lab값이 $40{\sim}50$ 정도였으며, 금새우난초는 황색으로 $110{\sim}130$ 정도였고, 변이종 개체들은 새우난초와 유사하거나 다소 높았다. PCR 결과 얻은 polymorphism이 인정되는 154개 marker에 대한 분리비 적합도 검정에서 51개 marker에서 5% 수준의 유의성이 인정되었으며, 유의성이 인정된 51개 marker 중에서 새우난초 type은 37개, 금새우난초 type은 14개 였다. Polymorphism이 인정된 154개 marker에 대하여 MAPL program을 이용하여 이들 marker 상호간의 연관관계를 분석한 결과는 16개의 연관군과 1개의 독립군으로 구분되었으며, 이들 연관군에 대한 분자연관지도는 전체 group의 크기가 220.4 cM (centi Morgan)이고, marker간의 평균거리는 3.3 cM이었다. 양적 형질에 대한 분자연관지도상의 QTL 분석 결과, LOD 3.0 이상인 화색과 설판색의 QTL은 각각 3개와 1개였다. 이상에서 얻어진 자료는 새우난초 속의 화색 연구에 도움이 될 것으로 사료된다.
This study was conducted to clarify the genetic relationship between Calanthe discolor, C. sieboldii and variants, and the cause of flower color variations by using a molecular linkage map and a quantitative trait loci (QTL) analysis for flower and lip color in Calanthe species native to Korea. Twen...
This study was conducted to clarify the genetic relationship between Calanthe discolor, C. sieboldii and variants, and the cause of flower color variations by using a molecular linkage map and a quantitative trait loci (QTL) analysis for flower and lip color in Calanthe species native to Korea. Twenty plants were included in three C. discolor and three C. sieboldii, and fourteen variants were obtained from their habitat, Jeju-do in Korea. The flowers of C. discolor were brownish red, the values of Commission Internationale de I'Eclairage (CIE) Lab were between 40 and 50. The flowers of C. sieboldii were yellowish, the values of CIE Lab were between 110 and 130. The variants had various mixed colors that were thought to have originated from natural hybridization between C. discolor and C. sieboldii, and the values of CIE Lab were between 50 and 70. The colors of the lips were usually divided into white and yellow. C. discolor had a white lip, C. sieboldii had a yellow one, and the variants had a white to yellow one. The CIE Lab value of each color was 90 in white and 110 to 120 in yellow lips. A molecular linkage mapping was constructed based on the segregation of 154 RAPD markers using a MAPL program. Sixteen linkage groups containing 66 markers were established. It covered a total map distance of 220.4 cM. The distance between adjacent markers ranged from 0 to 6.6 cM, with an average distance of 3.3 cM. These markers are thought to be closely associated with flower and lip color expression. Among the 16 molecular linkage groups, 3 QTLs had flower color trait loci and 1 QTL had lip color trait loci.
This study was conducted to clarify the genetic relationship between Calanthe discolor, C. sieboldii and variants, and the cause of flower color variations by using a molecular linkage map and a quantitative trait loci (QTL) analysis for flower and lip color in Calanthe species native to Korea. Twenty plants were included in three C. discolor and three C. sieboldii, and fourteen variants were obtained from their habitat, Jeju-do in Korea. The flowers of C. discolor were brownish red, the values of Commission Internationale de I'Eclairage (CIE) Lab were between 40 and 50. The flowers of C. sieboldii were yellowish, the values of CIE Lab were between 110 and 130. The variants had various mixed colors that were thought to have originated from natural hybridization between C. discolor and C. sieboldii, and the values of CIE Lab were between 50 and 70. The colors of the lips were usually divided into white and yellow. C. discolor had a white lip, C. sieboldii had a yellow one, and the variants had a white to yellow one. The CIE Lab value of each color was 90 in white and 110 to 120 in yellow lips. A molecular linkage mapping was constructed based on the segregation of 154 RAPD markers using a MAPL program. Sixteen linkage groups containing 66 markers were established. It covered a total map distance of 220.4 cM. The distance between adjacent markers ranged from 0 to 6.6 cM, with an average distance of 3.3 cM. These markers are thought to be closely associated with flower and lip color expression. Among the 16 molecular linkage groups, 3 QTLs had flower color trait loci and 1 QTL had lip color trait loci.
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문제 정의
The presented color values in Commission Internationale de KEclairage (CIE) color system were effective to convert each flower colors into significant values efficient for QTL analysis. To our knowledge, this study was the first attempt in the Calanthe species to construct molecular linkage map and analyze flower color and lip color traits by using QTL analysis in order to understand genetic background. Constructing molecular linkage maps of allogamous and vegetatively reproducing species, such as forest trees [6, 17], fruit trees [5]z industrial crop trees [7, 14, 18] and ornamental plants [1, 34]/ have been constructed using a double-testcross strategy.
제안 방법
3 cM. In our studyz molecular linkage map was constructed by using C. discolor, C. sieboldii, and variantsz which are natural inter-specific crossed between C. discolor, and C. sieboldii.
We believe this study is the first to construct molecular linkage map of Calanthe species native to Korea using PCR-based molecular markers and to map the floral trait loci associated with flower color expression. Our results are important to understand the genetic basis of the traits and to apply molecular markers linked to mapped loci for map-assisted selection (MAS).
This study was conducted to clarify the genetic relationship between each species, and the cause of flower color variations by constructing molecular linkage map and quantitative trait loci (QTL) analysis for flower and lip color in Calanthe species native to Korea. These data will be useful as the basis of flower color study in Calanthe discolor, C.
DNA markers linked to the trait loci for the number of petal in the carnation[19]z the petal number and flower color in diploid rose [3] and leaf chlorosis and flower color in Rhododmdrm [4] have been identified and anthocyanin pigmentation in asiatic hybrid lily [1] have been identified, but no reports exists of the mapping of useful traits loci in other ornamental crops. We believe this study is the first to construct molecular linkage map of Calanthe species native to Korea using PCR-based molecular markers and to map the floral trait loci associated with flower color expression. Our results are important to understand the genetic basis of the traits and to apply molecular markers linked to mapped loci for map-assisted selection (MAS).
대상 데이터
Sixteen linkage groups containing 66 markers were established. All of markers were generated by one- hundred of 10-based Operon primers. It covered a total map distance of 220.
replexa and other species is impossible [10]. Molecular linkage map in this study has been constructed by using Calanthe taxa consisted of three C. discolor, three C. sieblodii and differently flowering twenty variants. The precedent studies of flower color were estimated by using Munsell color system [16]z but this measuring method has problems to translate color to value for QTL analysis.
Plant materials were selected by flower color at habitat at Jeju-do in Korea (Fig. 1). Total twenty plants included three C.
2). Sixteen linkage groups containing 66 markers were established. All of markers were generated by one- hundred of 10-based Operon primers.
이론/모형
For RAPD analysis’ genomic DNA was extracted by using modified CTAB(cetyItrimethylammonium bromide) method. And one hundred 10-based primers were purchased from Operon (Alameda, USA).
S-5, Technidyne corporation, USA). The association between markers and QTL related with this flower and lip color traits were tested using the interval mapping method with MAPL program [21]. The LOD value >3.
each measured value was used for QTL analysis. The color meter used in this study presented color value in Commission Internationale de rEclairage (CIE) color system. Presented valuesz L*a*b*, of color meter was converted into CIE L*a*b* (Table 1).
성능/효과
discolor and variants (Table 3). All of these results indicated that variants were originated from natural cross between C. discolor and C. sieboldii, and because the genes of flower color of C. discolor are major gene for color inheritance, it was considered that the flower color of variants have similar value of CIE Lab of C. discolor.
Identification of molecular markers closely linked to agronomically useful loci is the first step for marker-assisted selection (MAS). QTL analysis clarified that putative QTLs OPD195oo and OPE14620 in linkage groups of this study are closely associated with flower color regulation of Calanthe species. These QTLs are a target for MAS.
Total 305 markers amplified by 100 primers were used to construct the molecular linkage map. The segregation ratios of all markers were compared with the expected segregation ratio of 3:1 that followed Mendelian law and used the chi-square test at 5% significant level. The primer code and the amplified fragment-size indicated the marker name, The primer code starting 'OP' designated 10-base Operon primers.
1). Total twenty plants included three C. discolor, three C. sieboldii and 14 variants which have different flower color. These selected plants were used flower color analysisz molecular linkage map construction and QTL analysis.
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