菌草种质资源AFLP多样性分析
|
摘要
利用AFLP技术对48份菌草种质资源进行了遗传多样性分析。首先,在海南象草等4份菌草材料上检测了20个AFLP引物组合的扩增情况。然后,从中筛选出6个扩增效果较好的组合用来进一步分析48份菌草种质材料的DNA,共产生了208个多态性条带。用NTSYS-pc 2.1软件结合UPGMA聚类分析方法对数据进行分析,结果显示,48份种质材料间遗传相似系数(SM)在0.48~0.98,在遗传相似系数为0.57时可将48份材料分成4个类群。以上研究结果表明,AFLP分子标记技术的分析结果能直观地表现菌草种质材料之间的亲缘关系,为菌草种质资源的收集和评价提供了一项新的技术手段。
In this study, AFLP technique was used to study the genetic diversity of 48 JUCAO germplasms. A total of 20 combinations of AFLP primer pairs were firstly tested on a subset of four samples. Then six combinations of AFLP primer pairs were selected and analyzed the DNA of 48 JUNCAO germplasms and a total of 208 polymorphic DNA fragments were obtained. The genetic similarity coefficient(SM) was estimated among all accessions by NTSYS-pc 2.1 software and ranged from 0.48 to 0.98. The 48 JUNCAO materials were separated into four clusters when the SM was 0.57.The results showed that the AFLP technique could more directly reveal the genetic difference among different accessions, and be served as a new technique for the collection and evaluation of JUNCAO germplasms.
In this study, AFLP technique was used to study the genetic diversity of 48 JUCAO germplasms. A total of 20 combinations of AFLP primer pairs were firstly tested on a subset of four samples. Then six combinations of AFLP primer pairs were selected and analyzed the DNA of 48 JUNCAO germplasms and a total of 208 polymorphic DNA fragments were obtained. The genetic similarity coefficient(SM) was estimated among all accessions by NTSYS-pc 2.1 software and ranged from 0.48 to 0.98. The 48 JUNCAO materials were separated into four clusters when the SM was 0.57.The results showed that the AFLP technique could more directly reveal the genetic difference among different accessions, and be served as a new technique for the collection and evaluation of JUNCAO germplasms.
引文
[1]林占熺.菌草学(第三版)[M].北京:国家行政学院出版社,2013.
[2]林兴生,林占熺,林冬梅,等.低温胁迫5种菌草的抗寒性评价[J].草业学报,2013,22(2):227-234.
[3]阳宴清,王咏,朱美兰,等.芦竹愈伤组织诱导及再生体系的建立[J].草业科学,2016,33(7):1332-1341.
[4]WANG Yong,ZHU Meilan,YANG Yanqing,et al.Exogenous Application of Abscisic Acid(ABA)Enhances Chilling Tolerance in Seedlings of Napier Grass(Pennisetum purpureum Schum.)[J].Agricultural Science&Technology,2017,18(3):417-423.
[5]赵凡.发展菌草业造福全人类——记菌草技术发明人林占熺[J].科技成果管理与研究,2015,12:49-50.
[6]梅嘉洺,黄小霞,王咏,等.46份菌草种质资源PEPC基因的PCR-FLP多样性分析[J].热带农业科学,2015,35(11):45-50,60.
[7]Li C,Zhao Y H,Mei J M,et al.Genetic Diversity of JUNCAO Germplasms Detected by i PBS Markers[J].A gricultural Science&Technology,2016,17(9):2000-2 004,2 058.
[8]赵永红,李婵,梅嘉洺,等.菌草种质资源SCo T遗传多样性分析[J].热带农业工程,2016,40(2):29-34.
[9]Huang Xiaoxia,Liu Cuicui,Ge Hongliu,et al.Analysis of Genetic Diversity of JUNCAO Germplasms by SRAP Markers[J].Agricultural Science&Technology,2017,18(9):1 595-1 600.
[10]Vos P,Hogers R,Bleeker M,et a1.AFLP:a new technique for DNA fingerprinting[J].Nucleic Acids Research,1995,23(21):4 407-4 414.
[11]罗海燕,应东山,黄建峰等.78份杧果种质资源AFLP分析[J].热带作物学报,2015,36(12):2130-2137.
[12]纵丹,员涛,周安佩等.滇杨优树遗传多样性的AFLP分析[J].西北林学院学报,2014,29(4):104-108.
[13]劳方业,刘睿,何慧怡,等.崖城系列甘蔗亲本遗传多样性的AFLP标记分析[J].分子植物育种,2008,6(3):517-522
[14]杜艳,王娟,陈冲,等.耧斗菜野生资源的AFLP遗传多样性分析[J].北方园艺,2017,5:85-88.
[15]孙凤梅,沈晓霞,沈宇峰,等.薏苡AFLP体系的建立与优化[J].浙江农业学报,2014,26(1):14-19.
[16]昝逢刚,应雄美,吴才文,等.98份甘蔗种质资源遗传多样性的AFLP分析[J].中国农业科学,2015,48(5):1 002-1 010.
[17]李伟,林富荣,郑勇奇,等.10个南方皂荚群体遗传多样性的AFLP分析[J].林业科学研究,2017,30(1):46-52.
[18]田松杰,石云素,宋燕春,等.利用AFLP技术研究玉米及其野生近缘种的遗传关系[J].作物学报,2004,30(4):354-359.
[19]李鸿雁,李志勇,辛霞,等.49份野生扁蓿豆种质资源的AFLP遗传多样性分析[J].中国草地学报,2016,38(4):20-26.
[20]杨春生,卢永彬,林燕芳,等.广西毛竹种质资源AFLP分析[J].广西植物,2014,34(6):742-746,787.
[21]何桂芳,刘玉皎,袁宏.青海蚕豆种质资源的AFLP多样性分析[J].安徽农业科学,2011,39(7):3824-3 826.
[22]王春梅,任健,兰平秀,等.山蚂蝗属植物遗传多样性的AFLP分析[J].热带作物学报,2017,3:521-528.
[23]Costa R,Pereira G,Garrido I,et al.Comparison of RAPD,ISSR,and AFLP Molecular Markers to Reveal and Classify Orchardgrass(Dactylis glomerata L.)Germplasm Variations[J].PLo S One,2016,11(4):e0152972.
[24]El-Esawi M A,Germaine K,Bourke P,et al.AFLP analysis of genetic diversity and phylogenetic relationships of Brassica oleracea in Ireland[J].C R Biol,2016,339(5-6):163-170.
[25]Ma H Y,Li S J,Feng N N,et al.First genetic linkage map for the mud crab(Scylla paramamosain)constructed using microsatellite and AFLP markers[J].Genet Mol Res.2016,15(2):gmr.15026929.
[26]Arias M,Hernandez M,Remondegui N,et al.First genetic linkage map of Taraxacum koksaghyz Rodin based on AFLP,SSR,COS and EST-SSR markers[J].Sci Rep,2016,6:31 031.
[27]Aichi-Yousfi H,Bahri B A,Medini M,et al.Genetic diversity and population structure of six species of Capparis in Tunisia using AFLP markers[J].C R Biol,2016,339(11-12):442-453.
[28]Blanco E Z,Bajay M M,Siqueira M V,et al.Genetic diversity and structure of Brazilian ginger germplasm(Zingiber of ficinale)revealed by AFLP markers[J].Genetica,2016,144(6):627-638.
[29]Sun M,Zhang C,Zhang X,et al.AFLP assessment of genetic variability and relationships in an Asian wild germplasm collection of Dactylis glomer ata L[J].C R Biol,2017,340(3):145-155.
[30]Sun J,Xu H M,Zhou R C,et al.Genetic diver sity and population structure of Myoporum bontioides(Myoporaceae)in China revealed by AFLP analysis[J].Aquatic Botany,2017,138:1-7.
[31]Moscoe L J,Emshwiller E.Diversity of Oxalis tuberosa Molina:a comparison between AFLP and microsatellite markers[J].Genetic Resources and Crop Evolution,2015,62(3):335-347.
[32]Pometti C L,Bessega C F,Vilardi J C,et al.Genetic variation in natural populations of Acacia visco(Fabaceae)belonging to two sub-regions of Argentina using AFLP[J].Plant Systematics and Evolution,2016,302(7):901-910.
[33]Murray H G,Thompson W F.Rapid isolation of high molecular weight DNA[J].Nucleic Acids Research,1980,8:4 321-4 325.
[34]Chalhoub B A,Thibault S,Lauco V.Silver staining and recovery of AFLP amplification products on large denaturing polyacrylamide gels[J].Biotechniques,1997,22(2):216-218.
[2]林兴生,林占熺,林冬梅,等.低温胁迫5种菌草的抗寒性评价[J].草业学报,2013,22(2):227-234.
[3]阳宴清,王咏,朱美兰,等.芦竹愈伤组织诱导及再生体系的建立[J].草业科学,2016,33(7):1332-1341.
[4]WANG Yong,ZHU Meilan,YANG Yanqing,et al.Exogenous Application of Abscisic Acid(ABA)Enhances Chilling Tolerance in Seedlings of Napier Grass(Pennisetum purpureum Schum.)[J].Agricultural Science&Technology,2017,18(3):417-423.
[5]赵凡.发展菌草业造福全人类——记菌草技术发明人林占熺[J].科技成果管理与研究,2015,12:49-50.
[6]梅嘉洺,黄小霞,王咏,等.46份菌草种质资源PEPC基因的PCR-FLP多样性分析[J].热带农业科学,2015,35(11):45-50,60.
[7]Li C,Zhao Y H,Mei J M,et al.Genetic Diversity of JUNCAO Germplasms Detected by i PBS Markers[J].A gricultural Science&Technology,2016,17(9):2000-2 004,2 058.
[8]赵永红,李婵,梅嘉洺,等.菌草种质资源SCo T遗传多样性分析[J].热带农业工程,2016,40(2):29-34.
[9]Huang Xiaoxia,Liu Cuicui,Ge Hongliu,et al.Analysis of Genetic Diversity of JUNCAO Germplasms by SRAP Markers[J].Agricultural Science&Technology,2017,18(9):1 595-1 600.
[10]Vos P,Hogers R,Bleeker M,et a1.AFLP:a new technique for DNA fingerprinting[J].Nucleic Acids Research,1995,23(21):4 407-4 414.
[11]罗海燕,应东山,黄建峰等.78份杧果种质资源AFLP分析[J].热带作物学报,2015,36(12):2130-2137.
[12]纵丹,员涛,周安佩等.滇杨优树遗传多样性的AFLP分析[J].西北林学院学报,2014,29(4):104-108.
[13]劳方业,刘睿,何慧怡,等.崖城系列甘蔗亲本遗传多样性的AFLP标记分析[J].分子植物育种,2008,6(3):517-522
[14]杜艳,王娟,陈冲,等.耧斗菜野生资源的AFLP遗传多样性分析[J].北方园艺,2017,5:85-88.
[15]孙凤梅,沈晓霞,沈宇峰,等.薏苡AFLP体系的建立与优化[J].浙江农业学报,2014,26(1):14-19.
[16]昝逢刚,应雄美,吴才文,等.98份甘蔗种质资源遗传多样性的AFLP分析[J].中国农业科学,2015,48(5):1 002-1 010.
[17]李伟,林富荣,郑勇奇,等.10个南方皂荚群体遗传多样性的AFLP分析[J].林业科学研究,2017,30(1):46-52.
[18]田松杰,石云素,宋燕春,等.利用AFLP技术研究玉米及其野生近缘种的遗传关系[J].作物学报,2004,30(4):354-359.
[19]李鸿雁,李志勇,辛霞,等.49份野生扁蓿豆种质资源的AFLP遗传多样性分析[J].中国草地学报,2016,38(4):20-26.
[20]杨春生,卢永彬,林燕芳,等.广西毛竹种质资源AFLP分析[J].广西植物,2014,34(6):742-746,787.
[21]何桂芳,刘玉皎,袁宏.青海蚕豆种质资源的AFLP多样性分析[J].安徽农业科学,2011,39(7):3824-3 826.
[22]王春梅,任健,兰平秀,等.山蚂蝗属植物遗传多样性的AFLP分析[J].热带作物学报,2017,3:521-528.
[23]Costa R,Pereira G,Garrido I,et al.Comparison of RAPD,ISSR,and AFLP Molecular Markers to Reveal and Classify Orchardgrass(Dactylis glomerata L.)Germplasm Variations[J].PLo S One,2016,11(4):e0152972.
[24]El-Esawi M A,Germaine K,Bourke P,et al.AFLP analysis of genetic diversity and phylogenetic relationships of Brassica oleracea in Ireland[J].C R Biol,2016,339(5-6):163-170.
[25]Ma H Y,Li S J,Feng N N,et al.First genetic linkage map for the mud crab(Scylla paramamosain)constructed using microsatellite and AFLP markers[J].Genet Mol Res.2016,15(2):gmr.15026929.
[26]Arias M,Hernandez M,Remondegui N,et al.First genetic linkage map of Taraxacum koksaghyz Rodin based on AFLP,SSR,COS and EST-SSR markers[J].Sci Rep,2016,6:31 031.
[27]Aichi-Yousfi H,Bahri B A,Medini M,et al.Genetic diversity and population structure of six species of Capparis in Tunisia using AFLP markers[J].C R Biol,2016,339(11-12):442-453.
[28]Blanco E Z,Bajay M M,Siqueira M V,et al.Genetic diversity and structure of Brazilian ginger germplasm(Zingiber of ficinale)revealed by AFLP markers[J].Genetica,2016,144(6):627-638.
[29]Sun M,Zhang C,Zhang X,et al.AFLP assessment of genetic variability and relationships in an Asian wild germplasm collection of Dactylis glomer ata L[J].C R Biol,2017,340(3):145-155.
[30]Sun J,Xu H M,Zhou R C,et al.Genetic diver sity and population structure of Myoporum bontioides(Myoporaceae)in China revealed by AFLP analysis[J].Aquatic Botany,2017,138:1-7.
[31]Moscoe L J,Emshwiller E.Diversity of Oxalis tuberosa Molina:a comparison between AFLP and microsatellite markers[J].Genetic Resources and Crop Evolution,2015,62(3):335-347.
[32]Pometti C L,Bessega C F,Vilardi J C,et al.Genetic variation in natural populations of Acacia visco(Fabaceae)belonging to two sub-regions of Argentina using AFLP[J].Plant Systematics and Evolution,2016,302(7):901-910.
[33]Murray H G,Thompson W F.Rapid isolation of high molecular weight DNA[J].Nucleic Acids Research,1980,8:4 321-4 325.
[34]Chalhoub B A,Thibault S,Lauco V.Silver staining and recovery of AFLP amplification products on large denaturing polyacrylamide gels[J].Biotechniques,1997,22(2):216-218.