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캐나다 옥수수 자식계통들에 대한 유전적 다양성 및 집단구조 분석

Analysis of Population Structure and Genetic Diversity among Canadian Maize Inbred Lines using SSR Markers

Korean Journal of Breeding Science 2017;49(2):72-79.
Published online: May 31, 2017

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*Corresponding author : (jukyonglee@kangwon.ac.kr, +82-33-250-6415, +82-33-255-5558)
• Received: April 3, 2017   • Accepted: May 9, 2017

© Korean Society of Breeding Science. All rights reserved.

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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  • We collected 32 maize inbred lines from eastern cereal and oilseed research center in Canada to develop new maize varieties. We also evaluated genetic diversity, genetic relationships, and population structure using 35 SSR markers. A total of 269 alleles were revealed in 35 loci with an average of 7.69 and a range between 3 and 15 alleles per locus. The genetic diversity values varied from 0.176 to 0.889 with an average of 0.691. The polymorphic information content varied from 0.171 to 0.879 with an average of 0.659. Population structure analysis indicated that 32 Canadian maize inbred lines comprised four major groups and one admixed group based on a membership probability threshold of 0.80. The four major groups contained 13, 2, 5 and 2 maize inbred lines, respectively. From genetic relationships analysis, the all inbred lines were divided into three main groups at 26% genetic similarity. Group I included 22 inbred lines, and Group II included 9 inbred lines. Group III consist of only one inbred line. The results in this study would be useful for the improvement and development of new cultivars, planning crosses for hybrids or development of inbred line in maize breeding program
옥수수(Zea mays L.)는 세계 3대 중요 작물 중 하나로, 1대잡종 강세를 이용하는 대표적인 타식성 작물 중 하나이다. 옥수수는 전세계적으로 식용뿐만 아니라 사료용, 공업용 등의 다양한 용도 로 이용되고 있다. 특히 우리나라의 경우 주로 식용 찰옥수수가 재배 이용되고 있지만, 최근 식생활의 서구화로 인하여 육류소비 가 증가함에 따라 사료용 옥수수의 수요 및 중요성이 점차 높아지 고 있다. 그러나 국내에서 소비되는 사료용 옥수수는 대부분 수입에 의존하고 있다. 2015년 국내 옥수수 수입량은 약 10,616 천 톤이며, 그 중에서 7,794천 톤이 사료용 옥수수로 이용되고 있어서 옥수수의 국내 곡물자급도는 약 1%에 불과하다고 한다 (농림축산식품부 2016. 농림축산식품 주요통계). 옥수수는 다른 사료작물들과 비교하여 단위 면적당 생산량이 많기 때문에 국내 에서 옥수수 수입량 감소 및 곡물자급도 향상을 위해서는 우리나 라 환경에 적합하고 수량과 품질이 우수한 신품종 개발이 필요하 다(Son et al. 2015).
옥수수 육종연구에서 수집된 유전자원들에 대한 유전적 다양 성(genetic diversity), 계통유연관계(genetic relationship) 및 집단구조(population structure)에 대한 정보는 신품종 개량을 위한 자식계통 육성 및 발굴 그리고 교배조합 예측 등에 효율적으 로 이용될 수 있고, 또한 이형 집단에서 자식계통의 선발과 품종 보호 등에도 유용한 정보를 제공할 수 있다(Hallauer et al. 1988, Pejic et al. 1998). 더욱이 다양한 자식계통들에 대한 명확한 계통분류는 품종육성에 필요한 포장실험을 줄일 수 있고, 육종가 들의 목적에 맞는 교배조합을 결정하는데 많은 도움을 줄 것이다 (Reid et al. 2011). 오늘날 개발된 다양한 분자마커(RFLP, RAPD, AFLP, SSR, SNP)들 중에서, 특히 SSRs (simple sequence repeats)은 분석이 간편하고 재현성이 매우 뛰어나며, 유전자 좌마다 풍부한 변이성으로 인하여 대립유전자의 특성을 명확하게 나타낼 수 있어 작물의 유전적 다양성, 계통유연관계 및 집단구조 분석 등에 활용되고 있다(Hu et al. 2009).
우리나라의 옥수수 육종은 주로 국내 재래종을 활용하여 수행 되면서 육종소재의 한계로 인하여 유전적 배경이 협소하고 지속 적인 우량 품종개발이 제한적이다(Park et al. 2012). 그렇기 때문에 옥수수 유전자원의 다양성 향상을 위하여 옥수수 자원의 수집 및 도입이 시급하다. 본 연구에서는 우수한 외국 옥수수 자원의 도입을 위하여 캐나다의 Eastern Cereal and Oilseed Research Center에서 개발한 옥수수 자식계통을 분양 받았으며, 대부분 종실용 옥수수 자식계통들로 구성되었다. 따라서 본 연구 는 SSR 분자마커를 이용하여, 캐나다 유전자원센터로부터 분양 받은 32계통의 옥수수 자식계통들에 대하여 유전적 다양성과 계통유연관계 분석을 통하여, 이들 자원을 활용한 신품종 개발에 유용한 정보를 얻고자 하였다.
식물 재료 및 DNA 추출
본 연구에서 분석에 이용한 32개의 캐나다 옥수수 자식계통들 의 육성내력과 기본 정보는 Table 1에 나타내었다. 이들 자식계 통들은 캐나다의 Eastern Cereal and Oilseed Research Center 에서 관리, 유지하고 있는 계통들이다. 옥수수 genomic DNA 추출은 유묘기에 어린 잎에서 Dellaporta et al. (1983)의 방법을 약간 변형하여 추출하였다.
Table 1
Derivation of 32 Canada maize inbred lines used in this study.
Table 1
Code No. Entry No. Pedigree Heterotic group

1 CO416 (A632 x CO125) CO125 (2) E.Flint
2 CO417 CB3 x CM383 P3994
3 CO418 Ottawa Cold Tolerant Syn CO E.Flint
4 CO419 24-44-1 Minn13
5 CO420 CM423(DOR x A) Iodent
6 CO421 DEA Iodent
7 CO423 Unknow Commercial Hybrid E.Butler
8 CO425 (B87 x CB8) CB8 P3994
9 CO428 OH43 x H99 Lanc
10 CO429 Pioneer 3707 Lanc
11 CO430 Fusarium Resistant Synthetic P3990
12 CO431 Fusarium Resistant Synthetic Iodent
13 CO432 Fusarium Resistant Synthetic Minn13
14 CO433 Pride K127 Minn13
15 CO434 CM105 x A632 BSSS
16 CO435 A632 x A634 BSSS
17 CO436 CO275 x CO300 P3994
18 CO437 European Synthetic E.Flint
19 CO438 CB3 x CL29 P3994
20 CO439 Nebraska BSSS BSSS
21 CO440 Pride 5 x CO258 Minn13
22 CO441 Jacques 7700 x CO298 Lanc
23 CO442 Iodent/NSS Iodent
24 CO443 B104 x CO272 BSSS/E.Butler
25 CO444 S1381 x CO382 E.Flint
26 CO445 CO386 x W64AHT Lanc
27 CO446 CO341 x CO328 BSSS
28 CO447 CO352 x CO328 BSSS/Minn
29 CO448 CO273 x CO431 P3990/Iodent
30 CO449 CO432 x CO433 Minn13
31 CO450 Eyespot Resistant Synthetic (99ESR) BSSS/Mix
32 CO451 CO309 x CO328 BSSS/Minn
SSR 분석 및 PCR 증폭
32개의 옥수수 자식계통들에 대한 유전적 다양성 및 집단구조 분석 을 위해서 SSR primer들은 MaizeGDB (http://www.maizegdb.org/) 정보를 기초하여, 총 35개의 SSR primer를 이용하였다(Table 2).
Table 2
Characteristics of 35 SSR loci in 32 Canada maize inbred lines.
Table 2
SSR Loci Chr Allele size range(bp) No. of Allele MAFz GDy PICx

bnlg1564 1 95~120 8 0.375 0.785 0.760
dupssr12 1 115~138 7 0.438 0.707 0.664
phi056 1 250~270 6 0.375 0.715 0.663
phi094 1 160~200 6 0.688 0.502 0.478
umc2012 1 70~120 8 0.406 0.705 0.659
phi10964 2 135~145 3 0.594 0.561 0.496
umc1551 2 150~165 8 0.406 0.760 0.731
umc1042 2 85~120 9 0.250 0.826 0.805
dupssr21 2 95~130 7 0.281 0.791 0.761
mmc0111 2 145~220 8 0.438 0.736 0.706
bnlg1182 3 70~190 11 0.344 0.783 0.757
mmc0022 3 125~160 8 0.344 0.781 0.752
mmc0251 3 120~190 15 0.219 0.889 0.879
umc1639 3 90~105 3 0.813 0.320 0.294
umc1394 3 95~185 4 0.906 0.176 0.171
umc1086 4 85~105 8 0.406 0.748 0.716
umc1720 4 120~175 5 0.438 0.693 0.642
nc005 4 120~190 12 0.281 0.834 0.815
phi021 4 85~135 6 0.438 0.693 0.642
bnlg1621 4 165~215 15 0.219 0.885 0.875
bnlg1695 5 105~170 13 0.250 0.848 0.832
bnlg565 5 55~130 11 0.531 0.689 0.673
phi008 5 100~115 4 0.750 0.400 0.395
umc1225 5 90~135 10 0.219 0.861 0.846
phi024 5 160~175 5 0.531 0.594 0.522
bnlg249 6 105~155 11 0.375 0.789 0.766
phi123 6 145~155 4 0.344 0.701 0.642
umc1178 6 145~165 4 0.844 0.277 0.262
bnlg1371 6 80~135 6 0.313 0.783 0.751
nc013 6 100~125 7 0.281 0.814 0.789
bnlg657 7 75~115 8 0.500 0.707 0.684
phi116 7 160~180 3 0.469 0.639 0.567
umc1066 7 135~160 5 0.531 0.645 0.600
umc1359 7 220~315 9 0.500 0.703 0.678
umc1863 8 120~205 12 0.219 0.871 0.858
Total 269
Average 7.69 0.438 0.691 0.659

zMAF: Major Allele Frequency

yGD: Gene Diversity

xPIC: Polymorphic Information Content

PCR 증폭에서 용액 조성은 총 30μL로 20ng의 genomic DNA, 1x PCR buffer, 0.3mM의 forward와 reverse primers, 0.2mM dNTPs, 1 unit의 Taq Polymerase (Biotools)로 구성하 였다. PCR 반응은 처음 94°C에서 5분간 최초 denaturation을 한 후, 다음 94°C에서 1분간 denaturation, 65°C에서 1분간 annealing, 72°C에서 2분간 extension을 하였으며, 2번째 cycle 부터 annealing 온도를 1°C씩 낮추어 최종 55°C까지 20cycle을 반복한 후, final extension을 72°C에서 10분간 실시하였다.
전기영동 및 silver-staining
PCR 증폭이 끝난 product 5μl를 10 μl의 loading-buffer (98% formamide, 0.02% BPH, 0.02% Xylene C, 5 mM of NaOH)와 혼합하였다. 그 후 denaturation 처리를 한 후 얼음에 냉각하여, 6% denaturing(7.5M urea) acrylamide-bisacrylamide gel (19:1)에 1x TBE buffer를 처리 후 2μl의 sample을 로딩하고, 1800V, 60W에서 2시간 동안 전기영동을 실시하였다. 전기영동 이 끝난 gel은 silver-staining kit (Promega, USA)를 이용하여 DNA band를 확인하였다.
Data 통계분석
통계분석은 SSR primer들에서 증폭된 DNA 단편을 대립단편 (allele)의 유무에 따라 1(유)과 0(무)으로 기록하였다. 유전적 유사성(genetic similarities, GS)값은 Dice similarity index (Dice 1945)를 이용하여 계산하였고, Dendrogram (SAHN-Clustering)은 NTSYS-pc.V.2.1 프로그램을 이용하여 UPGMA (un-weighted pair group methods using arithmetic averages algorithm)방법에 따라 작성하였다(Rohlf 1998). 그리 고 대립단편 수, major allele frequency (MAF), gene diversity (GD), polymorphic information content (PIC)는 PowerMarker V3.25 프로그램을 이용하여 분석하였다(Liu & Muse 2005). 유전적 다양성 값(gene diversity, GD)은 다음과 같이
Gene diversity (GD) = (1u=1kplu2)/(1+1+fn)
의 공식으로 계산하였으며, 여기서 plu는 u번째 allele의 빈도, f는 inbreeding coefficient, n은 sample size를 나타낸다. 그리고 PIC값은 Bostein et al. (1980)의 계산 방식에 따라 아래와 같이
PIC = 1u=1kplu2u=1k1v=u+1k12Plu2Plv2
의 공식을 이용하였으며, 여기서 plu는 u번째의 allele 빈도, plv는 v번째의 allele 빈도를 나타낸다.
한편 32개의 캐나다 옥수수 자식계통들에 대한 집단구조 분석 은 model-based program인 STRUCTURE 2.2를 이용하여 Pritchard et al. (2003)가 제안한 방법에 의해 계산하였다. STRUCTURE program을 이용한 분석에서 admixture model 은 100,000의 burn-in, 100,000의 run length로 1~10의 범위에 서 각 K값 당 5반복을 실시하여 평균 likelihood value인 LnP(D) 값을 계산하였으며, STRUCTURE program에서 측정된 데이터 의 log probability [LnP(D)]는 정확한 subgroup 수를 평가할 수 없어서, Evanno et al. (2005)가 제안한 ad hoc criterion (ΔK)를 이용하여 K값을 결정하였다.
캐나다 옥수수 자식계통들의 SSR 변이성
본 연구는 캐나다 Eastern Cereal and Oilseed Research Center에서 분양 받은 32개의 옥수수 자식계통들에 대해서 총 35개의 SSR primer들을 이용하여 유전적 변이성을 분석하였다. 그 결과 35개의 SSR primer들은 대립단편의 크기가 55bp에서 315bp의 범위에서 총 269개의 대립단편을 증폭시켰다. 35개의 SSR primer들에서 증폭된 대립단편의 수는 최소 3개(phi10964, umc1639, phi116)에서 최대 15개(mmc0251, bnlg1621)까지 로 다양하게 나타나, SSR primer 당 평균 7.69개가 확인되었다 (Table 2). 그리고 35개의 SSR primer들에서 측정된 MAF (major allele frequency)는 0.219(umc1225, umc1863, mmc0251, bnlg1621)에서 0.906(umc1394)의 범위로 나타나, SSR primer 당 평균 0.438로 나타났고, 유전적 다양성(GD)값은 0.176에서 0.889의 범위로 나타나, SSR primer 당 평균 0.691의 값을 나타내었다. PIC값은 0.171에서 0.879의 범위로 나타나, SSR primer 당 평균 0.659 의 값을 나타내었다(Table 2).
본 연구에서는 35개의 SSR primer를 이용하여 캐나다 옥수수 자식계통들에서 확인된 전체 269개의 대립단편들에 대하여 대 립단편들의 빈도(private/rare/intermediate/abundant alleles) 를 확인하였다(Fig. 1). 그 결과 32개의 캐나다 옥수수 자식계통 에서 증폭된 269개의 대립단편들 중에서 112개(41.6%)의 대립 단편들은 private allele로 확인되었으며, 179개(66.5%)의 대립 단편들은 rare allele(빈도 <0.1)를, 81개(30.1%)의 대립단편들 은 intermediate allele(빈도 0.1∼0.5)를, 그리고 9개(3.3%)의 대립단편들은 abundant allele(빈도 >0.5)를 각각 나타내었다.
Fig. 1
Histograms of allele frequencies for the 269 alleles in 32 accessions of Canada maize inbred lines.
KJBS-49-72_F1.gif
이상의 결과에 의하면, 분석에 이용된 35개의 SSR 마커들 중에서 특히 nc005, bnlg1695, umc1225, umc1863, bnlg1621, mmc0251 마커는 캐나다 옥수수 자식계통들에서 비교적 많은 수의 대립단편과 높은 GD값을 나타냈으며, 반면에 umc1394, umc1178, umc1639, phi008은 비교적 적은 수의 대립단편과 낮은 GD값을 나타내었다. 더욱이 Reid et al. (2011)는 본 연구에 서 분석된 캐나다 계통들이 일부 포함한 총 129개 옥수수 자식계 통들에 대하여 105개의 SSR 마커로 유전적 다양성을 분석하였 으며, 본 연구와 비교하여 평균 대립단편의 수 및 PIC 값에서 차이를 나타내었다. 이것은 Reid et al. (2011)의 연구에서 더 많은 계통들을 이용하였지만, 분석된 계통들의 대부분이 유사한 pedigree 및 genetic background를 갖기 때문에 많은 계통 수에 도 불구하고 낮은 유전적 다양성을 나타내었을 것으로 생각된다. 더욱이 본 연구와 이전의 연구에서 동일한 SSR 마커를 이용하였 음에도 불구하고 다른 개수의 대립단편을 나타내었는데, 이것은 분석에 이용된 실험재료 또는 실험조건의 차이로 인한 결과로 생각된다.
따라서 본 연구의 32개의 캐나다 옥수수 자식계통들에서 측정 된 유전적 다양성과 대립단편들의 빈도, 비교적 높은 다양성을 나타낸 SSR 마커들에 대한 정보는 앞으로 옥수수 계통들에 대한 계통증식 및 유지 그리고 분자표지마커 개발에 유용한 정보를 제공할 것으로 생각된다.
캐나다 옥수수 자식계통들의 집단구조와 계통유연관계
본 연구에서는 32개의 캐나다 옥수수 자식계통들에 대하여 집단구조 및 계통유연관계를 분석하였다. STRUCTURE 프로 그램을 이용한 집단구조 분석 결과 ΔK 값이 K=4에서 가장 높은 값이 확인되었다(Fig. 2). 더욱이 Wang et al. (2008)이 제시한 membership probability threshold 0.8에 근거하여 분석 한 결과, 32개의 캐나다 옥수수 자식계통들은 groups I, II, III, IV, admixed group으로 구분되었다(Fig. 3). Group I은 13계통 (CO417, CO419, CO420, CO423, CO428, CO429, CO435, CO438, CO439, CO443, CO444, CO445, CO450)이 포함되었 고, Group II는 2계통(CO446, CO451), Group III에는 5계통 (CO430, CO431, CO433, CO441, CO449)이 포함되었다. 그리 고 Group IV에는 2계통(CO436, CO437)이 포함되었으며, 마지 막으로 admixed group에는 총 10개의 자식계통(CO416, CO418, CO421, CO425, CO432, CO434, CO440, CO442, CO447, CO448)으로 구성되어 있다.
Fig. 2
Rate of change in the log probability of data between true K values(ΔK) described by Evanno et al. (2005).
KJBS-49-72_F2.gif
Fig. 3
Assignment of 32 Canada maize inbred lines to K=4 by STRUCTURE program.
KJBS-49-72_F3.gif
더욱이 UPGMA법을 이용하여 dendrogram을 작성한 결과, 32개의 캐나다 옥수수 자식계통들은 유전적 유사성 약 26%의 수준에서 크게 3개 그룹으로 나누어졌다. Group I은 22개의 계통(CO416, CO433, CO441, CO449, CO430, CO431, CO418, CO425, CO432, CO421, CO442, CO448, CO434, CO435, CO443, CO429, CO439, CO417, CO438, CO445, CO444, CO450)을 포함하였고, Group II는 9개의 계통(CO419, CO436, CO437, CO420, CO423, CO440, CO446, CO451, CO447)을 포함하였다. 마지막으로 Group III는 CO428 계통만 을 포함하였다. 본 연구에서 확인된 각 계통들간 유전적 유사성 분석 결과, 대부분의 자식계통들은 계통들 사이에서 비교적 낮은 유전적 유사성을 나타내었다. 그러나 CO433과 CO441, CO436 과 CO437, CO430과 CO431 계통들에서 높은 수준의 유전적 유사성(약 80%)를 나타내었다.
이상의 집단구조와 계통유연관계 분석결과에 의하면 일부 계통을 제외하고 대부분의 옥수수 계통들은 그들의 pedigree에 따라 명확하게 구분 되지 않았다(Fig. 3, 4). 예를 들면, 자식계통 인 CO446, CO447, CO451은 동일한 교배부본(CO328)을 이용 하여 육성되었으며, CO446과 CO451은 비교적 높은 유전적 유사성(66.7%)을 나타내고 집단구조 분석 결과에서 Group II에 포함되어 있으나 나머지 계통인 CO447은 다른 Group에 포함되 었다. 더욱이 CO431은 CO448의 부본으로 이용되었고, 계통유 연관계 분석에서 같은 Group I에 포함되었지만, 비교적 낮은 유전적 유사성(41.2%)을 나타내었다. 이에 반하여 CO449는 CO432와 CO433의 교잡으로 육성된 자식계통이며 계통유연관 계 분석에서 동일한 Group에 포함되었고, CO449와 CO432 및 CO433 간 유전적 유사성은 각각 50.7%와 55.4%로 비교적 높은 유전적 유사성을 나타내었다. 또한 집단구조 분석에서도 CO433과 CO449는 높은 membership probability로 Group III에 포함되었다. 비록 CO432가 admixed group이었지만 membership probability가 Group III에서 78.7%로 근소한 차이 로 Group III에 포함되지 못하였다는 것을 감안하면 이들의 pedigree 정보와 일치하는 결과로 생각된다.
Fig. 4
UPGMA dendrogram base on the 35 SSR markers.
KJBS-49-72_F4.gif
본 연구는 옥수수 신품종 육성을 위하여 캐나다에서 수집한 32개의 옥수수 자식계통들에 대하여 35개의 SSR primer들을 이용하여 유전적 다양성, 집단구조 및 계통유연관계 분석을 실시 하여, 32개 옥수수 자식계통들에서 비교적 높은 대립단편의 개수 및 유전적 다양성값을 갖는 SSR 마커들을 확인하였고, 분석에 이용된 자식계통들에 대한 계통 분류를 완료하였다. 결론적으로 본 연구에서 확인된 캐나다 자식계통들에 대한 유전적 다양성 및 집단구조, 계통유연관계의 결과들은 이후 이들 자식계통을 이용하여 우리나라에 적합한 종실용 옥수수 및 사료용 옥수수 육종을 위한 계통 육성 및 교배조합 구성, 예측에 유용한 정보를 제공할 것으로 기대된다.
본 연구는 총 35개의 SSR 마커를 이용하여, 캐나다 옥수수 자식계통들의 유전적 다양성, 집단구조 및 계통유연관계를 분석 하였다. 그 결과 55bp에서 315bp 크기의 범위로 총 269개의 대립단편들을 증폭시켰다. 35개의 SSR primer들에서 증폭된 대립단편의 수는 최소 3개에서부터 최대 15개까지의 범위로 나타났고, 평균 7.69개가 증폭되었다. 그리고 SSR primer들에서 측정된 MAF는 0.219에서 0.906의 범위로 평균 0.438로 나타났 고, GD값은 0.176에서 0.889의 범위로 평균 0.691의 값을 나타 내었다. PIC값은 0.171에서 0.879의 범위로 나타났으며, 평균 0.659 의 값을 나타내었다. 더욱이 32개의 캐나다 옥수수 자식계 통들의 집단구조를 분석한 결과, 13개의 계통은 group I에 포함 되었고, Group II는 2개의 자식계통들이 포함되었다. Group III에는 5개 자식계통이, Group IV에는 2개 자식계통이 포함되 었다. 나머지 10개의 자식계통들은 admixed group에 포함되었 다. 또한 UPGMA법에 의한 계통유연관계 분석 결과, 32개 캐나 다 옥수수 자식계통들은 유전적 유사성 약 26% 수준에서 크게 3개의 그룹으로 나누어졌다. 22개의 자식계통들이 Group I에 포함되었고, Group II는 9개의 자식계통을, 그리고 Group III은 단지 1개의 자식계통을 포함하고 있었다. 따라서 본 연구의 결과 는 이들 자식계통을 이용한 종실용 및 사료용 옥수수 육종을 위한 계통 육성 및 교배조합 구성, 예측에 유용한 정보를 제공할 것으로 기대된다.
본 논문은 골든씨드프로젝트(식량종자사업단, 213009-05-1- WT821, PJ012650012017)의 지원과 2016년도 강원대학교 학 술연구조성비로 연구하였음(과제번호-520160352).
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Analysis of Population Structure and Genetic Diversity among Canadian Maize Inbred Lines using SSR Markers
Korean. J. Breed. Sci.. 2017;49(2):72-79.   Published online June 1, 2017
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Analysis of Population Structure and Genetic Diversity among Canadian Maize Inbred Lines using SSR Markers
Korean. J. Breed. Sci.. 2017;49(2):72-79.   Published online June 1, 2017
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Analysis of Population Structure and Genetic Diversity among Canadian Maize Inbred Lines using SSR Markers
Image Image Image Image
Fig. 1 Histograms of allele frequencies for the 269 alleles in 32 accessions of Canada maize inbred lines.
Fig. 2 Rate of change in the log probability of data between true K values(ΔK) described by Evanno et al. (2005).
Fig. 3 Assignment of 32 Canada maize inbred lines to K=4 by STRUCTURE program.
Fig. 4 UPGMA dendrogram base on the 35 SSR markers.
Analysis of Population Structure and Genetic Diversity among Canadian Maize Inbred Lines using SSR Markers

Derivation of 32 Canada maize inbred lines used in this study.

Code No. Entry No. Pedigree Heterotic group

1 CO416 (A632 x CO125) CO125 (2) E.Flint
2 CO417 CB3 x CM383 P3994
3 CO418 Ottawa Cold Tolerant Syn CO E.Flint
4 CO419 24-44-1 Minn13
5 CO420 CM423(DOR x A) Iodent
6 CO421 DEA Iodent
7 CO423 Unknow Commercial Hybrid E.Butler
8 CO425 (B87 x CB8) CB8 P3994
9 CO428 OH43 x H99 Lanc
10 CO429 Pioneer 3707 Lanc
11 CO430 Fusarium Resistant Synthetic P3990
12 CO431 Fusarium Resistant Synthetic Iodent
13 CO432 Fusarium Resistant Synthetic Minn13
14 CO433 Pride K127 Minn13
15 CO434 CM105 x A632 BSSS
16 CO435 A632 x A634 BSSS
17 CO436 CO275 x CO300 P3994
18 CO437 European Synthetic E.Flint
19 CO438 CB3 x CL29 P3994
20 CO439 Nebraska BSSS BSSS
21 CO440 Pride 5 x CO258 Minn13
22 CO441 Jacques 7700 x CO298 Lanc
23 CO442 Iodent/NSS Iodent
24 CO443 B104 x CO272 BSSS/E.Butler
25 CO444 S1381 x CO382 E.Flint
26 CO445 CO386 x W64AHT Lanc
27 CO446 CO341 x CO328 BSSS
28 CO447 CO352 x CO328 BSSS/Minn
29 CO448 CO273 x CO431 P3990/Iodent
30 CO449 CO432 x CO433 Minn13
31 CO450 Eyespot Resistant Synthetic (99ESR) BSSS/Mix
32 CO451 CO309 x CO328 BSSS/Minn

Characteristics of 35 SSR loci in 32 Canada maize inbred lines.

SSR Loci Chr Allele size range(bp) No. of Allele MAFz GDy PICx

bnlg1564 1 95~120 8 0.375 0.785 0.760
dupssr12 1 115~138 7 0.438 0.707 0.664
phi056 1 250~270 6 0.375 0.715 0.663
phi094 1 160~200 6 0.688 0.502 0.478
umc2012 1 70~120 8 0.406 0.705 0.659
phi10964 2 135~145 3 0.594 0.561 0.496
umc1551 2 150~165 8 0.406 0.760 0.731
umc1042 2 85~120 9 0.250 0.826 0.805
dupssr21 2 95~130 7 0.281 0.791 0.761
mmc0111 2 145~220 8 0.438 0.736 0.706
bnlg1182 3 70~190 11 0.344 0.783 0.757
mmc0022 3 125~160 8 0.344 0.781 0.752
mmc0251 3 120~190 15 0.219 0.889 0.879
umc1639 3 90~105 3 0.813 0.320 0.294
umc1394 3 95~185 4 0.906 0.176 0.171
umc1086 4 85~105 8 0.406 0.748 0.716
umc1720 4 120~175 5 0.438 0.693 0.642
nc005 4 120~190 12 0.281 0.834 0.815
phi021 4 85~135 6 0.438 0.693 0.642
bnlg1621 4 165~215 15 0.219 0.885 0.875
bnlg1695 5 105~170 13 0.250 0.848 0.832
bnlg565 5 55~130 11 0.531 0.689 0.673
phi008 5 100~115 4 0.750 0.400 0.395
umc1225 5 90~135 10 0.219 0.861 0.846
phi024 5 160~175 5 0.531 0.594 0.522
bnlg249 6 105~155 11 0.375 0.789 0.766
phi123 6 145~155 4 0.344 0.701 0.642
umc1178 6 145~165 4 0.844 0.277 0.262
bnlg1371 6 80~135 6 0.313 0.783 0.751
nc013 6 100~125 7 0.281 0.814 0.789
bnlg657 7 75~115 8 0.500 0.707 0.684
phi116 7 160~180 3 0.469 0.639 0.567
umc1066 7 135~160 5 0.531 0.645 0.600
umc1359 7 220~315 9 0.500 0.703 0.678
umc1863 8 120~205 12 0.219 0.871 0.858
Total 269
Average 7.69 0.438 0.691 0.659

zMAF: Major Allele Frequency

yGD: Gene Diversity

xPIC: Polymorphic Information Content

Table 1 Derivation of 32 Canada maize inbred lines used in this study.
Table 2 Characteristics of 35 SSR loci in 32 Canada maize inbred lines.

MAF: Major Allele Frequency

GD: Gene Diversity

PIC: Polymorphic Information Content