通草1号虉草及其杂种后代遗传多样性的ISSR分析
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摘要
为了选育出产草量高、品质好、耐盐碱性和扩展性强,能够在内蒙古盐碱湿地大面积推广应用的优良虉草新品种,以通草1号虉草(Phalaris arundinacea L. Tongcao No.1)为母本(或父本),分别与川引3号虉草(P. arundinacea L. Chuanyin No.3)、美国虉草(P. arundinacea L.)进行杂交,得到了杂种后代(F1～F3),选择14个杂交株系和4个亲本材料,采用ISSR分子标记技术分析其遗传多样性和亲缘关系;并依据遗传相似系数,运用欧氏距离-离差平方和聚类方法对18份虉草材料进行聚类分析,为虉草新品种选育提供理论依据。结果表明,采用10条引物共扩增出176个位点,其中多态性位点148个,多态性位点百分率(PPB)为84.09%,平均有效等位基因数(Ne)为1.347 2,平均Nei's基因多样性(H)为0.228 5,平均Shannon多样性信息指数(I)为0.361 2,材料间遗传相似系数为0.602 3～0.818 2。当欧氏距离为0.57时可将18份虉草材料分为4个类群,第1类群由川引3号虉草、美国虉草和S18杂种构成;第2类群包含通草1号虉草、通辽野生虉草和S2、S17杂种构成;第3类群由6个通草1号虉草为母本(或父本)的杂交F1代构成;第4类群由4个通草1号虉草为父(母)本的杂种F2代和1个F1代构成。14份虉草杂种后代具有较丰富的遗传多样性,可作为新品种选育的基础材料。
In order to breed new variety of Phalaris arundinacea with high yield and quality, strong saline-alkaline tolerance and expansibility, and can be widely applied to saline alkali wetlands in Inner Mongolia, P. arundinacea Tongcao No.1 was selected as the female parent(or male parent) to crossbreed with P. arundinacea Chuanyin No.3 and P. arundinacea from America. Their hybrid progeny plants(F1～F3) were obtained, and 14 hybrid strains and 4 parent materials were selected, and their genetic diversity and relationship were analyzed by ISSR molecular marker technology. In addition, according to genetic similarity coefficient, clustering analysis of 18 P. arundinacea materials were carried out by using Euclidean distance-deviation square sum clustering method, which could provide theoretical basis for the breeding of new P. arundinacea varieties. The results showed that 176 locus were amplified by 10 primers, in which 148 bands were polymorphic, with the percentage of polymorphic bands(PPB)of 84.09%. The effective number of alleles(Ne) was 1.347 2, average Nei's gene diversity(H) was 0.228 5, Shannon's information index(I) was 0.361 2, and the genetic similarities coefficient among materials ranged from 0.602 3 to 0.818 2. 18 P. arundinacea materials could be divided into four groups when the Euclidean distance was 0.57,and the first group contained Chuanyin No.3, P. arundinacea from America and S18 crossbred. Group two was composed of Tongcao No.1, Tongliao wild P. arundinacea and S2 and S17 crossbred, and group three included six F1 crossbred with Tongcao No.1 as the female parent(or male parent). Moreover, group four consisted of four F2 crossbred with Tongcao No.1 as the female parent(or male parent) and 1 F1 generation. The genetic diversity of 14 P. arundinacea hybrid progenies was rich, which could be applied as the basic material for the breeding of new varieties.
In order to breed new variety of Phalaris arundinacea with high yield and quality, strong saline-alkaline tolerance and expansibility, and can be widely applied to saline alkali wetlands in Inner Mongolia, P. arundinacea Tongcao No.1 was selected as the female parent(or male parent) to crossbreed with P. arundinacea Chuanyin No.3 and P. arundinacea from America. Their hybrid progeny plants(F1～F3) were obtained, and 14 hybrid strains and 4 parent materials were selected, and their genetic diversity and relationship were analyzed by ISSR molecular marker technology. In addition, according to genetic similarity coefficient, clustering analysis of 18 P. arundinacea materials were carried out by using Euclidean distance-deviation square sum clustering method, which could provide theoretical basis for the breeding of new P. arundinacea varieties. The results showed that 176 locus were amplified by 10 primers, in which 148 bands were polymorphic, with the percentage of polymorphic bands(PPB)of 84.09%. The effective number of alleles(Ne) was 1.347 2, average Nei's gene diversity(H) was 0.228 5, Shannon's information index(I) was 0.361 2, and the genetic similarities coefficient among materials ranged from 0.602 3 to 0.818 2. 18 P. arundinacea materials could be divided into four groups when the Euclidean distance was 0.57,and the first group contained Chuanyin No.3, P. arundinacea from America and S18 crossbred. Group two was composed of Tongcao No.1, Tongliao wild P. arundinacea and S2 and S17 crossbred, and group three included six F1 crossbred with Tongcao No.1 as the female parent(or male parent). Moreover, group four consisted of four F2 crossbred with Tongcao No.1 as the female parent(or male parent) and 1 F1 generation. The genetic diversity of 14 P. arundinacea hybrid progenies was rich, which could be applied as the basic material for the breeding of new varieties.
引文
Bao M.L.,Yu Z.,Ma Y.H.,Su H.,and Li X.Y.,2010,ISSR analysis on chromosome doubling plants F2of reciprocal hybrids between Agropyron mongolicum and A.cristatum cv.Fairway,Zhongguo Caodi Xuebao(Chinese Journal of Grassland),32(5):31-35(包美莲,于卓,马艳红,苏和,李晓宇,2010,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,中国草地学报,32(5):31-35)
Emine A.L.,and Ahmet T.K.,2011,The application of ISSR-PCRto determine the genetic relationship and genetic diversity between narrow leaved bluegrass(Poa angustifolia)and rough bluegrass(Poa trivialis)accessions,Turkish J.Biol.,35:415-423
Guo J.W.,Li D.M.,Tian X.H.,and Du W.H.,2017,Purity identification and genetic diversity studies on the F1hybrid generation of triticale,Mailei Zuowu Xuebao(Journal of Triticeae Crops),37(8):1031-1037(郭建文,李冬梅,田新会,杜文华,2017,小黑麦杂交F1代真假杂种的ISSR标记鉴定及遗传多样性分析,麦类作物学报,37(8):1031-1037)
Hu F.R.,He G.R.,Wang F.,and Ren C.,2015,The identification of the hyacinth hybrid progeny by the method of ISSR molecular marker,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),13(6):1336-1342(胡凤荣,何国仁,王斐,任翠,2015,利用ISSR分子标记方法鉴定风信子杂种后代,分子植物育种,13(6):1336-1342)
Huang F.,Wang M.J.,He L.J.,Chen L.L.,and Wei C.Q.,2012,ISSR analysis of Trifolium ambiguum Bieb.,T.repens L.and their hybrid F1,Zhongguo Caodi Xuebao(Chinese Journal of Grassland),34(6):107-112(黄帆,王明玖,何丽君,陈丽丽,魏春秋,2012,高加索三叶草、白三叶及其杂种F1代的ISSR分析,中国草地学报,34(6):107-112)
Li X.L.,Bao H.C.,and Yu Z.,2016,ISSR analysis of Elymus sibiricus,Elymus purpuraristatus and their hybrid F1and BC1,Zhongzi(Seed),35(12):17-20(李小雷,鲍红春,于卓,2016,老芒麦和紫芒披碱草及其杂种F1代与BC1代的ISSR分析,种子,35(12):17-20)
Li Y.Y.,and Dong K.H.,2014,ISSR analysis of genetic diversity of germplasm resources of Bothriochloa ischaemum in Shanxi,Caoye Xuebao(Acta Prataculture Sinica),23(1):217-222(李钰莹,董宽虎,2014,山西白羊草种质资源遗传多样性的ISSR分析,草业学报,23(1):217-222)
Liu G.,Zhang C.B.,Li D.X.,Liu B.,Zhang Y.,You M.H.,and Bai S.Q.,2008,Primary study on feed value of Phalaris arundinacea L.Chuancao No.3,Caoye Yu Xumu(Prataculture and Animal Husbandry),157(12):11-12,26(刘刚,张昌兵,李达旭,刘斌,张玉,游明鸿,白史且,2008,‘川草3号’虉草饲用价值初步研究,草业与畜牧,157(12)11-12,26)
Liu H.,Mu P.,and Zhao G.Q.,2012,A study on genetic diversity of Avena germplasm resources detected by ISSR,Caoye Xuebao(Acta Prataculture Sinica),21(4):116-224(刘欢慕平,赵桂琴,2012,燕麦种质资源遗传多样性ISSR研究,草业学报,21(4):116-224)
McGregor C.E.,Lambert C.A.,Greyling M.M.,Louw J.H.,and Warnich L.,2000,A comparative assessment of DNA ingerprinting techniques(RAPD,ISSR,AFLP and SSR)in tetraploidpotato(Solanum tuberosum L.)germplasm,Euphytica,113(2):135-144
Miao M.M.,Zhao M.M.,Liu X.M.,Hou Y.,and Fan J.P.,2016Analysis on genetic relationships in lily(Lilium spp.)hybrids based on ISSR molecular markers,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(2):437-441(苗美美,赵明明,刘绪明,侯悦,樊金萍,2016,百合(Lilium spp.)杂交后代ISSR分析亲缘关系,分子植物育种,14(2):437-441)
Sahramaa M.,2003,Evaluation of reed canary grass for differen enduses and in breeding,Agr.Food Sci.Finland,12(3-4)227-241
Tahir M.H.N.,Casler M.D.,Moore K.J.,and Brummer E.C.2011,Biomass yield and quality of reed canarygrass under five harvest management systems for bioenergy production Bioenerg.Res.,4(2):111-119
Zeng L.,Yuan Q.H.,Wang F.,and Wang Y.,2013,Genetic diversity analysis of Agropyron germlasm resources by ISSRCaoye Xuebao(Acta Prataculture Sinica),22(1):260-267(曾亮,袁庆华,王方,王瑜,2013,冰草属植物种质资源遗传多样性的ISSR分析,草业学报,22(1):260-267)
Zhang H.S.,Xia Z.R.,Li M.F.,Wang C.M.,and Yang S.Z.,2014,Genetic diversity of Pennisetun longissimum var.intermedium germplasm resources using ISSR markers,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),34(2):256-264(张怀山,夏曾润,栗孟飞,王春梅,杨世柱,2014,中型狼尾草种质资源遗传多样性的ISSR分析,西北植物学报,34(2):256-264)
Zhang X.,Liu Q.S.,Shi W.G.,Li Z.Y.,and Liu L.,2014,Analysis of genetic diversity of 25 Bromus inermis accessions based on ISSR,Zhongguo Caodi Xuebao(Chinese Journal of Grassland),36(3):104-107(张雪,刘青松,师文贵,李志勇,刘磊,2014,25份无芒雀麦种质资源遗传多样性的ISSR分析,中国草地学报,36(3):104-107)
Zhang Y.,Lan Z.Q.,and Wu T.,2018,ISSR analysis of Camellia nitidissima germplasm resources from Dawei mountain in Yunnan,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),16(2):649-655(张玥,蓝增全,吴田,2018,云南大围山金花茶种质资源的ISSR分析,分子植物育种,16(2):649-655)
Zhang Y.L.,and Zhang H.,2013,Effect of row spacing and nitrogen fertilizer application on tiliers and yield of Phalaris arundinacaea,Zhongguo Caodi Xuebao(Chinese Journal of G-rassland),32(2):43-47(张永亮,张浩,2013,行距与氮肥对虉草分蘖和产量的影响,中国草地学报,32(2):43-47)
Zhang Y.L.,Luo X.M.,Liu P.,and Liu Y.,2017,Genetic diversity analysis of 22 reed canarygrass germplasm resources based on ISSR markers,Caodi Xuebao(Acta Agrestia Sinica),25(2):366-372(张永亮,骆秀梅,刘鹏,刘杨,2017,22份虉草种质资源遗传多样性的ISSR分析,草地学报,25(2):366-372)
Emine A.L.,and Ahmet T.K.,2011,The application of ISSR-PCRto determine the genetic relationship and genetic diversity between narrow leaved bluegrass(Poa angustifolia)and rough bluegrass(Poa trivialis)accessions,Turkish J.Biol.,35:415-423
Guo J.W.,Li D.M.,Tian X.H.,and Du W.H.,2017,Purity identification and genetic diversity studies on the F1hybrid generation of triticale,Mailei Zuowu Xuebao(Journal of Triticeae Crops),37(8):1031-1037(郭建文,李冬梅,田新会,杜文华,2017,小黑麦杂交F1代真假杂种的ISSR标记鉴定及遗传多样性分析,麦类作物学报,37(8):1031-1037)
Hu F.R.,He G.R.,Wang F.,and Ren C.,2015,The identification of the hyacinth hybrid progeny by the method of ISSR molecular marker,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),13(6):1336-1342(胡凤荣,何国仁,王斐,任翠,2015,利用ISSR分子标记方法鉴定风信子杂种后代,分子植物育种,13(6):1336-1342)
Huang F.,Wang M.J.,He L.J.,Chen L.L.,and Wei C.Q.,2012,ISSR analysis of Trifolium ambiguum Bieb.,T.repens L.and their hybrid F1,Zhongguo Caodi Xuebao(Chinese Journal of Grassland),34(6):107-112(黄帆,王明玖,何丽君,陈丽丽,魏春秋,2012,高加索三叶草、白三叶及其杂种F1代的ISSR分析,中国草地学报,34(6):107-112)
Li X.L.,Bao H.C.,and Yu Z.,2016,ISSR analysis of Elymus sibiricus,Elymus purpuraristatus and their hybrid F1and BC1,Zhongzi(Seed),35(12):17-20(李小雷,鲍红春,于卓,2016,老芒麦和紫芒披碱草及其杂种F1代与BC1代的ISSR分析,种子,35(12):17-20)
Li Y.Y.,and Dong K.H.,2014,ISSR analysis of genetic diversity of germplasm resources of Bothriochloa ischaemum in Shanxi,Caoye Xuebao(Acta Prataculture Sinica),23(1):217-222(李钰莹,董宽虎,2014,山西白羊草种质资源遗传多样性的ISSR分析,草业学报,23(1):217-222)
Liu G.,Zhang C.B.,Li D.X.,Liu B.,Zhang Y.,You M.H.,and Bai S.Q.,2008,Primary study on feed value of Phalaris arundinacea L.Chuancao No.3,Caoye Yu Xumu(Prataculture and Animal Husbandry),157(12):11-12,26(刘刚,张昌兵,李达旭,刘斌,张玉,游明鸿,白史且,2008,‘川草3号’虉草饲用价值初步研究,草业与畜牧,157(12)11-12,26)
Liu H.,Mu P.,and Zhao G.Q.,2012,A study on genetic diversity of Avena germplasm resources detected by ISSR,Caoye Xuebao(Acta Prataculture Sinica),21(4):116-224(刘欢慕平,赵桂琴,2012,燕麦种质资源遗传多样性ISSR研究,草业学报,21(4):116-224)
McGregor C.E.,Lambert C.A.,Greyling M.M.,Louw J.H.,and Warnich L.,2000,A comparative assessment of DNA ingerprinting techniques(RAPD,ISSR,AFLP and SSR)in tetraploidpotato(Solanum tuberosum L.)germplasm,Euphytica,113(2):135-144
Miao M.M.,Zhao M.M.,Liu X.M.,Hou Y.,and Fan J.P.,2016Analysis on genetic relationships in lily(Lilium spp.)hybrids based on ISSR molecular markers,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(2):437-441(苗美美,赵明明,刘绪明,侯悦,樊金萍,2016,百合(Lilium spp.)杂交后代ISSR分析亲缘关系,分子植物育种,14(2):437-441)
Sahramaa M.,2003,Evaluation of reed canary grass for differen enduses and in breeding,Agr.Food Sci.Finland,12(3-4)227-241
Tahir M.H.N.,Casler M.D.,Moore K.J.,and Brummer E.C.2011,Biomass yield and quality of reed canarygrass under five harvest management systems for bioenergy production Bioenerg.Res.,4(2):111-119
Zeng L.,Yuan Q.H.,Wang F.,and Wang Y.,2013,Genetic diversity analysis of Agropyron germlasm resources by ISSRCaoye Xuebao(Acta Prataculture Sinica),22(1):260-267(曾亮,袁庆华,王方,王瑜,2013,冰草属植物种质资源遗传多样性的ISSR分析,草业学报,22(1):260-267)
Zhang H.S.,Xia Z.R.,Li M.F.,Wang C.M.,and Yang S.Z.,2014,Genetic diversity of Pennisetun longissimum var.intermedium germplasm resources using ISSR markers,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),34(2):256-264(张怀山,夏曾润,栗孟飞,王春梅,杨世柱,2014,中型狼尾草种质资源遗传多样性的ISSR分析,西北植物学报,34(2):256-264)
Zhang X.,Liu Q.S.,Shi W.G.,Li Z.Y.,and Liu L.,2014,Analysis of genetic diversity of 25 Bromus inermis accessions based on ISSR,Zhongguo Caodi Xuebao(Chinese Journal of Grassland),36(3):104-107(张雪,刘青松,师文贵,李志勇,刘磊,2014,25份无芒雀麦种质资源遗传多样性的ISSR分析,中国草地学报,36(3):104-107)
Zhang Y.,Lan Z.Q.,and Wu T.,2018,ISSR analysis of Camellia nitidissima germplasm resources from Dawei mountain in Yunnan,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),16(2):649-655(张玥,蓝增全,吴田,2018,云南大围山金花茶种质资源的ISSR分析,分子植物育种,16(2):649-655)
Zhang Y.L.,and Zhang H.,2013,Effect of row spacing and nitrogen fertilizer application on tiliers and yield of Phalaris arundinacaea,Zhongguo Caodi Xuebao(Chinese Journal of G-rassland),32(2):43-47(张永亮,张浩,2013,行距与氮肥对虉草分蘖和产量的影响,中国草地学报,32(2):43-47)
Zhang Y.L.,Luo X.M.,Liu P.,and Liu Y.,2017,Genetic diversity analysis of 22 reed canarygrass germplasm resources based on ISSR markers,Caodi Xuebao(Acta Agrestia Sinica),25(2):366-372(张永亮,骆秀梅,刘鹏,刘杨,2017,22份虉草种质资源遗传多样性的ISSR分析,草地学报,25(2):366-372)