基于SRAP分子标记的冰草遗传连锁图谱构建
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摘要
构建高密度遗传连锁图谱是冰草抗性、品质、产量等重要性状QTL精细定位及标记辅助育种研究的基础。该试验以四倍体杂交冰草F2群体的202个分离单株及其亲本为材料,利用SRAP分子标记技术和Join Map 4.0作图软件对冰草的遗传连锁图谱进行了构建。结果表明:(1)共筛选出22对多态性好、标记位点清晰稳定的SRAP适宜引物,对冰草杂种F2分离单株的基因组DNA进行PCR扩增,共获得510个SRAP多态性标记位点,其比率占88.2%。(2)偏分离分析表明,偏分离标记比率仅为14.12%,符合遗传作图的要求。(3)成功构建了冰草的SRAP分子标记遗传连锁图谱,该图谱有14个连锁群、510个标记,连锁群间长度范围86.4～179.0cM,覆盖基因组总长度1 912.9cM,标记间平均间距3.75cM,为高密度遗传图谱。
To construct a high-density genetic linkage map of crested wheatgrass is the basis of fine mapping for important QTLs such as resistance,quality and yield,as well as the marker-assisted breeding research.In this research,with 202 individuals from F2 segregation population of tetraploid hybrid crested wheatgrass and their parents as materials,we constructed a genetic linkage map of crested wheatgrass based on SRAP molecular marker technique and Join map 4.0 mapping software.The results revealed that:(1)a total of 22 SRAP suitable primers with good polymorphism,clear and stable bands were screened to make a wide range of PCR amplification on the genomic DNA of F2 individuals.A total of 510 SRAP polymorphic markers were obtained,accounting for 88.2%.(2)The analysis of segregation distortion showed that the ratio of distorted markers was only 14.12%,which met the requirements of genetic mapping.(3)The genetic linkage map of SRAP molecular markers of crested wheatgrass was successfully constructed.The map consisted of 14 linkage groups,including 510 SRAP markers,and covered the whole genome of 1 912.9 cM.The map length of linkage groups ranged from 86.4 cM to 179.0 cM,and the average distance between markers is 3.75 cM.It is a high-density genetic map.
To construct a high-density genetic linkage map of crested wheatgrass is the basis of fine mapping for important QTLs such as resistance,quality and yield,as well as the marker-assisted breeding research.In this research,with 202 individuals from F2 segregation population of tetraploid hybrid crested wheatgrass and their parents as materials,we constructed a genetic linkage map of crested wheatgrass based on SRAP molecular marker technique and Join map 4.0 mapping software.The results revealed that:(1)a total of 22 SRAP suitable primers with good polymorphism,clear and stable bands were screened to make a wide range of PCR amplification on the genomic DNA of F2 individuals.A total of 510 SRAP polymorphic markers were obtained,accounting for 88.2%.(2)The analysis of segregation distortion showed that the ratio of distorted markers was only 14.12%,which met the requirements of genetic mapping.(3)The genetic linkage map of SRAP molecular markers of crested wheatgrass was successfully constructed.The map consisted of 14 linkage groups,including 510 SRAP markers,and covered the whole genome of 1 912.9 cM.The map length of linkage groups ranged from 86.4 cM to 179.0 cM,and the average distance between markers is 3.75 cM.It is a high-density genetic map.
引文
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[6]杜丽媛,刘伟华,杨欣明,等.小麦-冰草新种质普冰2011姊妹系的育种效应分析[J].植物遗传资源学报,2016,17(3):395-403.DU L Y,LIU W H,YANG X M,et al.Breeding achievement analysis of novel wheat-Agropyron cristatum germplasm sister lines Pubing 2011[J].Journal of Plant Genetic Resources,2016,17(3):395-403.
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[8]于卓,张志成,马艳红,等.四倍体杂交冰草新品系植株形态及生长特性分析[J].草业科学,2013,30(11):1 762-1 766.YU Z,ZHANG Z C,MA Y H,et al.Analysis on morphology and growth of new lines of tetraploid hybrid wheatgrass[J].Pratacultural Science,2013,30(11):1 762-1 766.
[9]解新明,云锦凤,尹俊,等.蒙古冰草遗传多样性的RAPD分析[J].西北植物学报,2002,22(1):56-62.XIE X M,YUN J F,YIN J,et al.RAPD analysis of genetic diversity of Agropyron mongolicum[J].Acta Botanica Boreali-Occidentalia Sinica,2002,22(1):56-62.
[10]包美莲,于卓,马艳红,等.蒙古冰草与航道冰草正反交杂种染色体加倍植株F2的ISSR分析[J].中国草地学报,2010,32(5):31-35.BAO M L,YU Z,MA Y H,et al.ISSR analysis on chromosome doubling plants F2of reciprocal hybrids between Agropyron mongolicumand A.cristatumcv.Fairway[J].Chinese Journal of Grassland,2010,32(5):31-35.
[11]于卓,马艳红,李小雷,等.蒙古冰草与航道冰草正、反交F1及其染色体加倍植株AFLP分析[J].中国草地学报,2009,31(6):14-19.YU Z,MA Y H,LI X L,et al.AFLP analysis on reciprocal hybrids F1between Agropyron mongolicumand A.cristatum cv.Fairway and its chromosome doubling plants[J].Chinese JournalofGrassland,2009,31(6):14-19.
[12]LI G,QUIROS C F.Sequence-related amplified polymorphism(SRAP),a new marker system based on a simple PCRreaction:its application to mapping and gene tagging in Brassica[J].Theoretical and Applied Genetics,2001,103(2/3):455-461.
[13]GRODZICKER T,WILLIAMS J,SHARP P,et al.Physical mapping of temperature-sensitive mutations of adenoviruses[J].Cold Spring Harbor Symposia on Quantitative Biology,1975,97(3):369-390.
[14]TAUTZ D.Hypervariabflity of simple sequences as a general source for polymorphic DNA markers[J].Nucleic Acids Research,1989,17(16):6 463-6 471.
[15]唐茜,何凤发,王季春,等.甘薯SRAP遗传图谱构建及淀粉含量QTL初步定位[J].西南大学学报(自然科学版),2010,32(6):40-45.TANG Q,HE F F,WANG J C,et al.Construction of a linkage map using SRAP markers and QTL mapping for starch content in sweetpotato[J].Journal of Southwest University(Natural Science Edition),2010,32(6):40-45.
[16]王刚,潘俊松,李效尊,等.黄瓜SRAP遗传连锁图的构建及侧枝基因定位[J].中国科学C辑:生命科学,2004,34(6):510-516.WANG G,PAN J S,LI X Z,et al.Construction of SRAP genetic linkage map of cucumber and location of lateral branches[J].Science in China Ser.C Life Sciences,2004,34(6):510-516.
[17]林忠旭,张献龙,聂以春,等.棉花SRAP遗传连锁图构建[J].科学通报,2003,48(15):1 676-1 679.LIN Z X,ZHANG X L,NIE Y C,et al.Construction of cotton SRAP genetic linkage map[J].Chinese Science Bulletin,2003,48(15):1 676-1 679.
[18]姜志艳,于肖夏,于卓,等.四倍体杂交冰草AFLP遗传连锁图谱的构建[J].麦类作物学报,2015,35(4):457-463.JIANG Z Y,YU X X,YU Z,et al.Construction of an AFLP-based genetic linkage map of tetraploid hybrid wheatgrass[J].Journal of Triticeae Crops,2015,35(4):457-463.
[19]PEZZOTTI G,YAMADA K.Genetic diversity of oilseed Brassica napus inbred lines based on sequence-related amplified polymorphism and its relation to hybrid performance[J].Plant Breeding,2010,120(5):411.
[20]LIN Z,HE D,ZHANG X,et al.Linkage map construction and mapping QTL for cotton fibre quality using SRAP,SSRand RAPD[J].Plant Breeding,2010,124(2):180-187.
[21]NEI M,TAJIMA F.DNA polymorphism detectable by restriction endonucleases[J].Genetics,1981,97(1):145.
[22]宋宪亮,孙学振,张天真.偏分离及对植物遗传作图的影响[J].农业生物技术学报,2006,14(2):140-146.SONG X L,SUN X Z,ZHANG T Z.Segregation distortion and its effect on genetic mapping in plants[J].Journal of Agricultueal Biotechnology,2006,14(2):140-146.
[23]郭利建,王竹林,汪世娟,等.基于SRAP和SSR标记的小麦品质相关性状的QTL定位[J].麦类作物学报,2016,36(10):1 275-1 282.GUO L J,WANG Z L,WANG S J,et al.QTL mapping of wheat grain quality traits based on SRAP and SSR marker[J].Journal of Triticeae Crops,2016,36(10):1 275-1 282.
[24]王茂芊,李博,王华忠.甜菜遗传连锁图谱初步构建[J].作物学报,2014,40(2):222-230.WANG M Q,LI B,WANG H Z.Construction of molecular genetic linkage map of sugarbeet[J].Acta Agronomica Sinica,2014,40(2):222-230.
[25]石悦,于肖夏,南志标,等.高丹草SRAP和SSR分子遗传连锁图谱的构建[J].中国草地报,2018,40(5):11-17.SHI Y,YU X X,NAN Z B,et al.Construction of genetic linkage map of sorghum-sudangrass based on SRAP and SSRmolecular markers[J].Chinese Journal of Grassland,2018,40(5):11-17.
[26]刘恩英,王源秀,徐立安,等.基于SSR和SRAP标记的簸箕柳×绵毛柳遗传框架图[J].林业科学,2011,47(5):23-30.LIU E Y,WANG Y X,XU L A,et al.A genetic frame map of Salix suchowensis×S.eriocladabased on SSR and SRAPmarkers[J].Scientla Silvae Sinicae,2011,47(5):23-30.
[27]金梦阳,刘列钊,付福友,等.甘蓝型油菜SRAP、SSR、AFLP和TRAP标记遗传图谱构建[J].分子植物育种,2006,4(4):520-526.JIN M Y,LIU L Z,FU F Y,et al.Construction of a genetic linkage map in Brassica napus based on SRAP,SSR,AFLPand TRAP[J].Molecular Plant Breeding,2006,4(4):520-526.
[28]武耀龙,李德芳,李建军,等.利用SRAP和SSR标记构建红麻遗传连锁图谱[J].中国麻业科学,2016,38(6):249.WU Y L,LI D F,LI J J,et al.Constructing agenetic linkage map by SRAP and SSR marker for kenaf(Hibiscus cannabinus L.)[J].Plant Fiber Sciences in China,2016,38(6):249.
[29]张丽,姜树坤,张喜娟,等.粳稻SRAP分子标记遗传群的构建与分析[J].植物生理学通讯,2007,43(3):443-447.ZHANG L,JIANG S K,ZHANG X J,et al.Construction and analysis of a SRAP molecular genetic map of japonica rice(Oryza sativa L.sp.japonica)[J].Plant Physiology Communications,2007,43(3):443-447.
[30]陈晖,陈美霞,陶爱芬,等.长果种黄麻SRAP标记遗传连锁图谱的构建及3个质量性状基因定位[J].中国农业科学,2011,44(12):2 422-2 430.CHEN H,CHEN M X,TAO A F,et al.Construction of a molecular linkage map using SRAP and mapping of three qualitative traits in Corchorus olitorius L.[J].Scientia Agricultura Sinica,2011,44(12):2 422-2 430.
[31]李爱贤,刘庆昌,王庆美,等.利用SRAP标记构建甘薯分子连锁图谱[J].作物学报,2010,36(8):1 286-1 295.LI A X,LIU Q C,WANG Q M,et al.Establishment of molecular linkage maps using SRAP markers in sweet potato[J].Acta Agronomica Sinica,2010,36(8):1 286-1 295.
[2]于肖夏,姜志艳,于卓,等.四倍体杂交冰草新品种-蒙杂冰草1号的选育[J].草业科学,2015,32(5):738-744.YU X X,JIANG Z Y,YU Z,et al.The new variety breeding of tetraploid hybrid wheatgrass-Mengza No.1[J].Pratacultural Science,2015,32(5):738-744.
[3]JENSEN K B.Cytogenetics of Elymus magellanicus and its intra-and inter-generic hybrids with Pseudoroegneria spicata,Hordeum violaceum,E.trachycaulus,E.lanceolatus,and E.glaucus(Poaceae:Triticeae)[J].Genome,1993,36(1):72-76.
[4]李立会,杨欣明,李秀全,等.通过属间杂交向小麦转移冰草优异基因的研究[J].中国农业科学,1998,31(6):1-5.LI L H,YANG X M,LI X Q,et al.Introduction of desirable genes fromAgropyron cristatuminto common wheat by intergeneric hybridization[J].Scientia Agricultura Sinica,1998,31(6):1-5.
[5]FATIH A M B.Analysis of the breeding potential of wheatAgropyron and wheat-Elymus derivatives[J].Hereditas,2010,98(2):287-295.
[6]杜丽媛,刘伟华,杨欣明,等.小麦-冰草新种质普冰2011姊妹系的育种效应分析[J].植物遗传资源学报,2016,17(3):395-403.DU L Y,LIU W H,YANG X M,et al.Breeding achievement analysis of novel wheat-Agropyron cristatum germplasm sister lines Pubing 2011[J].Journal of Plant Genetic Resources,2016,17(3):395-403.
[7]于卓,李造哲,云锦凤.几种小麦族禾草及其杂交后代农艺特性的研究[J].草业学报,2003,12(3):83-89.YU Z,LI Z Z,YUN J F.Study on the agronomic characters of several perennial species in Triticeae and their hybrids[J].Acta Prataculturae Sinica,2003,12(3):83-89.
[8]于卓,张志成,马艳红,等.四倍体杂交冰草新品系植株形态及生长特性分析[J].草业科学,2013,30(11):1 762-1 766.YU Z,ZHANG Z C,MA Y H,et al.Analysis on morphology and growth of new lines of tetraploid hybrid wheatgrass[J].Pratacultural Science,2013,30(11):1 762-1 766.
[9]解新明,云锦凤,尹俊,等.蒙古冰草遗传多样性的RAPD分析[J].西北植物学报,2002,22(1):56-62.XIE X M,YUN J F,YIN J,et al.RAPD analysis of genetic diversity of Agropyron mongolicum[J].Acta Botanica Boreali-Occidentalia Sinica,2002,22(1):56-62.
[10]包美莲,于卓,马艳红,等.蒙古冰草与航道冰草正反交杂种染色体加倍植株F2的ISSR分析[J].中国草地学报,2010,32(5):31-35.BAO M L,YU Z,MA Y H,et al.ISSR analysis on chromosome doubling plants F2of reciprocal hybrids between Agropyron mongolicumand A.cristatumcv.Fairway[J].Chinese Journal of Grassland,2010,32(5):31-35.
[11]于卓,马艳红,李小雷,等.蒙古冰草与航道冰草正、反交F1及其染色体加倍植株AFLP分析[J].中国草地学报,2009,31(6):14-19.YU Z,MA Y H,LI X L,et al.AFLP analysis on reciprocal hybrids F1between Agropyron mongolicumand A.cristatum cv.Fairway and its chromosome doubling plants[J].Chinese JournalofGrassland,2009,31(6):14-19.
[12]LI G,QUIROS C F.Sequence-related amplified polymorphism(SRAP),a new marker system based on a simple PCRreaction:its application to mapping and gene tagging in Brassica[J].Theoretical and Applied Genetics,2001,103(2/3):455-461.
[13]GRODZICKER T,WILLIAMS J,SHARP P,et al.Physical mapping of temperature-sensitive mutations of adenoviruses[J].Cold Spring Harbor Symposia on Quantitative Biology,1975,97(3):369-390.
[14]TAUTZ D.Hypervariabflity of simple sequences as a general source for polymorphic DNA markers[J].Nucleic Acids Research,1989,17(16):6 463-6 471.
[15]唐茜,何凤发,王季春,等.甘薯SRAP遗传图谱构建及淀粉含量QTL初步定位[J].西南大学学报(自然科学版),2010,32(6):40-45.TANG Q,HE F F,WANG J C,et al.Construction of a linkage map using SRAP markers and QTL mapping for starch content in sweetpotato[J].Journal of Southwest University(Natural Science Edition),2010,32(6):40-45.
[16]王刚,潘俊松,李效尊,等.黄瓜SRAP遗传连锁图的构建及侧枝基因定位[J].中国科学C辑:生命科学,2004,34(6):510-516.WANG G,PAN J S,LI X Z,et al.Construction of SRAP genetic linkage map of cucumber and location of lateral branches[J].Science in China Ser.C Life Sciences,2004,34(6):510-516.
[17]林忠旭,张献龙,聂以春,等.棉花SRAP遗传连锁图构建[J].科学通报,2003,48(15):1 676-1 679.LIN Z X,ZHANG X L,NIE Y C,et al.Construction of cotton SRAP genetic linkage map[J].Chinese Science Bulletin,2003,48(15):1 676-1 679.
[18]姜志艳,于肖夏,于卓,等.四倍体杂交冰草AFLP遗传连锁图谱的构建[J].麦类作物学报,2015,35(4):457-463.JIANG Z Y,YU X X,YU Z,et al.Construction of an AFLP-based genetic linkage map of tetraploid hybrid wheatgrass[J].Journal of Triticeae Crops,2015,35(4):457-463.
[19]PEZZOTTI G,YAMADA K.Genetic diversity of oilseed Brassica napus inbred lines based on sequence-related amplified polymorphism and its relation to hybrid performance[J].Plant Breeding,2010,120(5):411.
[20]LIN Z,HE D,ZHANG X,et al.Linkage map construction and mapping QTL for cotton fibre quality using SRAP,SSRand RAPD[J].Plant Breeding,2010,124(2):180-187.
[21]NEI M,TAJIMA F.DNA polymorphism detectable by restriction endonucleases[J].Genetics,1981,97(1):145.
[22]宋宪亮,孙学振,张天真.偏分离及对植物遗传作图的影响[J].农业生物技术学报,2006,14(2):140-146.SONG X L,SUN X Z,ZHANG T Z.Segregation distortion and its effect on genetic mapping in plants[J].Journal of Agricultueal Biotechnology,2006,14(2):140-146.
[23]郭利建,王竹林,汪世娟,等.基于SRAP和SSR标记的小麦品质相关性状的QTL定位[J].麦类作物学报,2016,36(10):1 275-1 282.GUO L J,WANG Z L,WANG S J,et al.QTL mapping of wheat grain quality traits based on SRAP and SSR marker[J].Journal of Triticeae Crops,2016,36(10):1 275-1 282.
[24]王茂芊,李博,王华忠.甜菜遗传连锁图谱初步构建[J].作物学报,2014,40(2):222-230.WANG M Q,LI B,WANG H Z.Construction of molecular genetic linkage map of sugarbeet[J].Acta Agronomica Sinica,2014,40(2):222-230.
[25]石悦,于肖夏,南志标,等.高丹草SRAP和SSR分子遗传连锁图谱的构建[J].中国草地报,2018,40(5):11-17.SHI Y,YU X X,NAN Z B,et al.Construction of genetic linkage map of sorghum-sudangrass based on SRAP and SSRmolecular markers[J].Chinese Journal of Grassland,2018,40(5):11-17.
[26]刘恩英,王源秀,徐立安,等.基于SSR和SRAP标记的簸箕柳×绵毛柳遗传框架图[J].林业科学,2011,47(5):23-30.LIU E Y,WANG Y X,XU L A,et al.A genetic frame map of Salix suchowensis×S.eriocladabased on SSR and SRAPmarkers[J].Scientla Silvae Sinicae,2011,47(5):23-30.
[27]金梦阳,刘列钊,付福友,等.甘蓝型油菜SRAP、SSR、AFLP和TRAP标记遗传图谱构建[J].分子植物育种,2006,4(4):520-526.JIN M Y,LIU L Z,FU F Y,et al.Construction of a genetic linkage map in Brassica napus based on SRAP,SSR,AFLPand TRAP[J].Molecular Plant Breeding,2006,4(4):520-526.
[28]武耀龙,李德芳,李建军,等.利用SRAP和SSR标记构建红麻遗传连锁图谱[J].中国麻业科学,2016,38(6):249.WU Y L,LI D F,LI J J,et al.Constructing agenetic linkage map by SRAP and SSR marker for kenaf(Hibiscus cannabinus L.)[J].Plant Fiber Sciences in China,2016,38(6):249.
[29]张丽,姜树坤,张喜娟,等.粳稻SRAP分子标记遗传群的构建与分析[J].植物生理学通讯,2007,43(3):443-447.ZHANG L,JIANG S K,ZHANG X J,et al.Construction and analysis of a SRAP molecular genetic map of japonica rice(Oryza sativa L.sp.japonica)[J].Plant Physiology Communications,2007,43(3):443-447.
[30]陈晖,陈美霞,陶爱芬,等.长果种黄麻SRAP标记遗传连锁图谱的构建及3个质量性状基因定位[J].中国农业科学,2011,44(12):2 422-2 430.CHEN H,CHEN M X,TAO A F,et al.Construction of a molecular linkage map using SRAP and mapping of three qualitative traits in Corchorus olitorius L.[J].Scientia Agricultura Sinica,2011,44(12):2 422-2 430.
[31]李爱贤,刘庆昌,王庆美,等.利用SRAP标记构建甘薯分子连锁图谱[J].作物学报,2010,36(8):1 286-1 295.LI A X,LIU Q C,WANG Q M,et al.Establishment of molecular linkage maps using SRAP markers in sweet potato[J].Acta Agronomica Sinica,2010,36(8):1 286-1 295.