벼 종간잡종 유래 근동질 유전자계통 이용 종자중 관여 유전자 분석

강주원(Ju-Won Kang),양바오로(Paul Yang),윤여태(Yeo-Tae Yun),안상낙(Sang-Nag Ahn) 한국육종학회 2011 한국육종학회지 Vol.43 No.4

1. 선행 연구에서 염색체 3번의 RM60~RM231 부근에서 종자중, 수당립수에 관여하는 QTL이 탐지되었고, 이를 확인하기 위하여 이 지역에 크기와 위치가 다른 O. glaberrima 단편이 이입된 5 계통을 선발하여 표현형을 조사하였다. 실험 결과 계통별로 출수기, 임실률, 수당립수, 종자중을 제외한 형태적 특성들이 밀양23호와 비슷한 양상을 보였다. 이는 대부분의 염색체 지역이 밀양23호로 회복되었기 때문이라고 보여진다. 2. 유전자의 위치를 자세히 알기 위해 RM60과 RM22 부위에 위치하는 SSR 마커를 이용하여 5개 계통의 유전자형을 검정하였다. 종자중과 수당립수에서 차이를 보이는 4계통, IL3, IL26, IL25와 IL51을 비교한 결과 종자중과 수당 립수를 조절하는 유전자는 RM60-RM523 사이의 재조환 지점과 단완 끝 부분 사이에 위치하는 것으로 판단되며 그 거리는 약 1.2-Mb이다. 3. 본 연구에서 탐지된 연관된 종자중과 수당립수 QTL은 재 배벼의 수량성 증진에 효과적으로 이용될 수 있을 것으로 판단된다. In previous studies, we reported QTLs for grain weight (GW), qGW3 and for spikelets per panicle (SPP), qSPP3 linked to RM60 on chromosome 3 using advanced backcross lines derived from a cross between Oryza sativa ssp. Indica cv. Milyang 23 and O. glaberrima. The O. glaberrima alleles at this locus increased GW and spikelets per panicle in the Milyang 23 background. To further confirm and narrow down the position of the QTLs on chromosome 3, substitution mapping was performed using five lines containing the target O. glaberrima segment on chromosome 3. The size and position of the O. glaberrima segment on chromosome 3 were different in each line. These lines possessed 3-10 non-target O. glaberrima introgressions in the Milyang 23 background. These five lines were evaluated for seven agronomic traits including 1,000 grain weight and spikelets per panicle and also genotyped with seven SSR markers. Four lines were informative in delimiting the position of QTLs, qGW3 and qSPP3. Two lines with the O. glaberrima segment flanked by SSR markers, RM60 and RM523 displayed significantly higher values than Milyang 23 in GW and SPP whereas two lines without that O. glaberrima segment displayed no difference in GW and SPP compared to Milyang 23. The result indicates that two QTL, qGW3 and qSPP3 are located in the interval between RM60 and RM523 which are 1.2-Mb apart. Introgression lines having QTLs, qGW3 and qSPP3 would be useful materials not only to indentify the relationship between these two yield QTLs, but also to develop high yielding variety via marker-aided selection technology.

ITS 염기서열 분석 및 CAPS를 이용한 조이시아 속(Zoysia) 들잔디와 갯잔디의 구별

홍민지(Min-Ji Hong),양대화(Dae-Hwa Yang),정옥철(Ok-Cheol Jeong),김양지(Yang-Ji Kim),박미영(Mi-Young Park),강홍규(Hong-Gyu Kang),선현진(Hyeon-Jin Sun),권용익(Yong-Ik Kwon),박신영(Shin-Young Park),양바오로(Paul Yang),송필순(Pill-Soon So 한국원예학회 2017 원예과학기술지 Vol.35 No.3

Zoysia 속 잔디는 학교운동장 및 공원, 골프장, 스포츠경기장과 같이 다양한 장소에 식재되고 있는 중요한 잔디이다. 해안가에서 자생하는 Zoysia 속 들잔디와 갯잔디는 외부 형태적 특성이 유사하여 외부 형태적 분류 뿐 만 아니라 분자생물학적 분류도 필요하다. 본 연구에서는 nrDNA ̵ ITS(Internal Transcribed Spacer)의 DNA 바코드 분석을 통해서 자생하는 들잔디와 갯잔디의 분자생물학적 신속한 분류체계를 확립하고자 하였다. 이를 위해 난지형 잔디인 Zoysia 속 들잔디(Z. japonica) 및 갯잔디(Z. sinica)와 한지형 대표 잔디인 크리핑 벤트그라스(A. stolonifera) 및 켄터키 블루그라스(P. pratensis)의 nrDNA - ITS 염기서열을 확보하였다. 확보된 들잔디및 갯잔디, 크리핑 벤트그라스, 켄터키 블루그라스의 ITS 염기서열 전체 구간은 각 686bp와 687bp, 683bp, 681bp으로 확인되었으며, nrDNA - ITS 내부 염기서열구간 분석 결과, ITS1의 크기는 248 ̵ 249bp, ITS2는 270 - 274bp, 5.8S rDNA는 163 - 164bp의 차이로, 각 4종의 잔디가 ITS 염기서열을 이용하여 식별되었다. 특히, 들잔디와 갯잔디 nrDNA-ITS 염기서열은 19 염기(2.8%) 차이를 나타냈으며, ITS1과 ITS2의 G + C 함량은 55.4 ̵ 63.3% 임을 확인하였다. 이러한 들잔디와 갯잔디의 ITS 염기서열 차이를 바탕으로 CAPS 마커로 전환하여 대조구 및 수집된 자생 Zoysia 속 잔디 영양체 62개체를 분석한 결과, 외부형태학적 분류법으로 들잔디 개체, 갯잔디 개체로 동정되었지만, ITS CAPS 마커를 이용한 분자생물학적 분류법으로 들잔디 36개체와 갯잔디 22개체 뿐만 아니라 들잔디와 갯잔디간의 자연교배종 4개체도 식별하였다. 이상의 결과에서 들잔디와 갯잔디는 ITS 염기서열 및 ITS 기반 CAPS를 통하여 식별할 수 있을 것으로 판단된다. Zoysiagrasses are important turf plants used for school playgrounds, parks, golf courses, and sports fields. The two most popular zoysiagrass species are Zoysia japonica and Zoysia sinica. These are widely distributed across different growing zones and are morphologically distinguishable from each other; however, it is phenotypically difficult to differentiate those that grow along the coastal line from those in beach area habitats. A combination of morphological and molecular approaches is desirable to efficiently identify these two plant cultivars. In this study, we used a rapid identification system based on DNA barcoding of the nrDNA-internal transcribed spacer (ITS) regions. The nrDNA-ITS regions of ITS1, 5.8S nrDNA, and ITS2 from Z. japonica , Z. sinica , Agrostis stolonifera , and Poa pratensis were DNA barcoded to classify these grasses according to their molecular identities. The nrDNA-ITS sequences of these species were found at 686 bp, 687 bp, 683 bp, and 681 bp, respectively. The size of ITS1 ranged from 248 to 249 bp, while ITS2 ranged from 270 to 274 bp. The 5.8S coding region ranged from 163 - 164bp. Between Z. japonica and Z. sinica , nineteen (2.8%) nucleotide sites were variable, and the G+C content of the ITS region ranged from 55.4 to 63.3%. Substitutions and insert/deletion (indel) sites in the nrDNA-ITS sequence of Z. japonica and Z. sinica were converted to cleaved amplified polymorphic sequence (CAPS) markers, and applied to the Zoysia grasses sampled to verify the presence of these markers. Among the 62 control and collected grass samples, we classified three groups: 36 Z. japonica , 22 Z. sinica , and 4 Z. japonica /Z. sinica hybrids. Morphological classification revealed only two groups; Z. japonica and Z. sinica . Our results suggest that used of the nrDNA-ITS barcode region and CAPS markers can be used to distinguish between Z. japonica and Z. sinica at the species level.

Microsatellite를 이용한 자포니카 벼의 다양성 분석

나소(Luo Xiao),상세티(Sangshetty Balkunde),양바오로(Paul Yang),이현숙(Hyun-Sook Lee),안상낙(Sang-Nag Ahn) 충남대학교 농업과학연구소 2012 농업과학연구 Vol.39 No.1

The study was conducted to evaluate the genetic similarity among commercial japonica rice varieties in Korea and China and to develop markers to differentiate between japonica cultivars developed in Korea and China. The genetic similarity and cluster of 38 accessions were analyzed using 47 SSR(simple sequence repeat) markers. The number of alleles by 47 SSR markers ranged from 2 to 9 with an average of 3.6. A total of 169 alleles were detected among these tested rice varieties. The PIC value varied from 0.05 to 0.79 with an average of 0.44. The Chinese japonica cultivars could be differentiated from the japonica cultivars in Korea by combining 2 SSR markers, RM223 and RM266. Cluster analysis showed that 38 tested varieties could be distinguished into japonica and indica based on the genetic distance.

벼 일품벼/모로베레칸 조합의 이입계통을 이용한 내건성 유전자 탐지

강주원(Ju-Won Kang),구홍광(Hong-Guang Ju),양바오로(Paul Yang),안상낙(Sang-Nag Ahn) 충남대학교 농업과학연구소 2011 농업과학연구 Vol.38 No.2

This study was conducted to map quantitative trait loci (QTLs) related to drought stress tolerance. An introgression line population derived from a cross, “Ilpum” / “Moroberekan” was used in this study. F₁ plants were backcrossed three times to Ilpum to produce BC₃F₁ plants. These plants were advanced by selfing for four generation and a total of 117 BC₃F? introgression lines were developed. These lines were evaluated for percent seed set and spikelets per panicle under the control (field) and drought condition. To identify QTLs related to drought tolerance, 134 SSR markers showing polymorph isms between the parents were genotyped for the 117 BC₃F? lines. A total of 6 QTLs associated with drought stress were detected on chromosomes 1, 3, 4, 7 and 10. These include two QTLs for phenotypic acceptability, two QTLs for percent seed set (R² = 19.0 - 20.9%), and two QTLs for spikelets per panicle (R² = 22.3 - 23.10%). The Moroberekan alleles at three loci contributed the positive effect for drought tolerance. The SSR markers linked to drought stress tolerance can not only facilitate the selection of valuable genes from Moroberekan, but also allow identification of lines with drought tolerance.