Chen, Wei

Liu, Xiaoxia

Mu, Qun

Tang, Chengbin

Wang, Xiaoli

Wu, Jiahai

Zhong, Li

English

University of Kentucky

University of Kentucky UKnowledge International Grassland Congress Proceedings XXI International Grassland Congress / VIII International Rangeland Congress ISSR Analysis on Genetic Diversity of Wild Agrostis stolonifera L. Germplasm Resources Xiaoli Wang Guizhou Institute of Prataculture, China Chengbin Tang Guizhou Institute of Prataculture, China Li Zhong Guizhou Institute of Prataculture, China Wei Chen Guizhou Institute of Prataculture, China Jiahai Wu Guizhou Institute of Prataculture, China See next page for additional authors Follow this and additional works at: https://uknowledge.uky.edu/igc  Part of the Plant Sciences Commons, and the Soil Science Commons This document is available at https://uknowledge.uky.edu/igc/21/12-2/15 The XXI International Grassland Congress / VIII International Rangeland Congress took place in Hohhot, China from June 29 through July 5, 2008. Proceedings edited by Organizing Committee of 2008 IGC/IRC Conference Published by Guangdong People's Publishing House This Event is brought to you for free and open access by the Plant and Soil Sciences at UKnowledge. It has been accepted for inclusion in International Grassland Congress Proceedings by an authorized administrator of UKnowledge. For more information, please contact UKnowledge@lsv.uky.edu. Presenter Information Xiaoli Wang, Chengbin Tang, Li Zhong, Wei Chen, Jiahai Wu, Qun Mu, and Xiaoxia Liu This event is available at UKnowledge: https://uknowledge.uky.edu/igc/21/12-2/15 瞯 ]396　 瞯 　 Multifunctional Grasslands in a Changing World 　 Volume Ⅱ 　Grasslands/Rangelands Production Systems——— Developing Improved PlantsISSR analysis on genetic diversity of wild Agrostis stoloni f era L . Germplasm resourcesW ang X iao‐L i ,Tang Cheng‐Bin ,Zhong L i ,Chen Wei ,W u Jia‐Hai ,Mu Qun ,L iu X iao‐X iaGuiz hou Institute o f p rataculture ,Dushan ,558200 ,China ;TEL : 0854‐3231250 ; E‐mail :w angx iaoliz henyuan＠ 126 .comKey words :A grostis stoloni f era L ,genetic diversity ,genetic structure ,ISSR ,germplasm resourcesIntroduction A grostis stoloni f era L . is a major cool season turf grass in China .ISSR( Zietkiewicz E ,１９９４ ) molecular markerswere used to detect the genetic diversity among three populations of wild and one of cultivated A grostis stoloni f era L . collectedfrom Guizhou .Materials and methodsPlant material The plants used in this study were sampled from three natural populations and one cultivated population ofA grostis stoloni f era L . located in Guizhou . Each populations had several subpopulations with the exception of the cultivatedpopulation KROMI . The locations of sampled populations shown in Table １ .Table 1 Natural and cultiv ated populations o f Agrostis stolonifera L . used in this study .Name of populations origins No . of subpopulationsNatural populations BSE Southeastern of Biji ６  BNW Northwestern of Biji ７  LPS Liu Panshui ５  Cultivated population KROMI Dushan １  DNA extraction and ISSR amplification Fifteen individuals were selected from each subpopulation for ex traction of mixed DNA .PCR amplifications were then performed in a final volume of ２５μL containing ５０ng DNA templates , １μL ( ２mmol/ L ) dNTPs ,２μL１０ × PCR buffer ,２μL ( ２５mmol/ L ) Mg２ ＋ ,０ .４U ( ２ .５U /L ) Taq polymerase ,１μL Primer ( １０pmo１ ) .Reactions were carriedout under the following regime : ９４ ℃ for ７ min ; Followed by ３５ cycles of for ３０s ,５O ℃ for ４５s ,７２ ℃ for１min ;and ended with７２ ℃ for７min .Results and discussionGenetic diversity Nine primer pairs produced ６６ polymorphic bands , the average percentage of polymorphic bands was ８１ .４８ .At population level , the percentage of polymorphic bands ranged from ４３ .２１ to ５９ .２６ , w ith an average of ５０ .２１ .The Nei摧sgene diversity index was ０ .２４１４ and Shannon diversity index was ０ .３７１９ . These results suggested a rich genetic diversityamong the natural populations of Agrostis stolonifera L .Amova The genetic differentiation coefficient was ０ .４９２ and the gene flow was ０ .５１６４ among three wild populations , Itrevealed a significant genetic differentiation .Genetic relatives The Nei摧s genetic similarity coefficient of the sub‐populations ranged from ０ .４０７４ to ０ .９１２３ , w ith three clustergroups . Moreover , the findings implied a correlation among the populations . A significantly high correlation betweengeographical and genetic distance was observed( r ＝ ０ .４９４ , ０ .０５ ＞ P ＞ ０ .０１) .Further analysis on genetic relationship suggestedrelatively high genetic similarity among populations ,ranging from ０ .７００３ to ０ .９４０９ .Conclusion There was a rich genetic diversity and significant genetic differentiation among the natural populations of Agrostisstolonifera L . and the cultivated populations showed a significant genetic difference from natural populations .ReferenceZietkiewicz E ,Rafalaki A ,Labuda D , １９９４ .Genome fingerprinting by simple sequence repeat ( SSR) － anchored polymerasechain reaction amplification . Genomics Jourmal , ２０ ,１７６ ～ １８３ .

ISSR Analysis on Genetic Diversity of Wild \u3ci\u3eAgrostis stolonifera\u3c/i\u3e L. Germplasm Resources

University of Kentucky UKnowledge International Grassland Congress Proceedings XXI International Grassland Congress / VIII International Rangeland Congress ISSR Analysis on Genetic Diversity of Wild Agrostis stolonifera L. Germplasm Resources Xiaoli Wang Guizhou Institute of Prataculture, China Chengbin Tang Guizhou Institute of Prataculture, China Li Zhong Guizhou Institute of Prataculture, China Wei Chen Guizhou Institute of Prataculture, China Jiahai Wu Guizhou Institute of Prataculture, China See next page for additional authors Follow this and additional works at: https://uknowledge.uky.edu/igc  Part of the Plant Sciences Commons, and the Soil Science Commons This document is available at https://uknowledge.uky.edu/igc/21/20-1/17 The XXI International Grassland Congress / VIII International Rangeland Congress took place in Hohhot, China from June 29 through July 5, 2008. Proceedings edited by Organizing Committee of 2008 IGC/IRC Conference Published by Guangdong People's Publishing House This Event is brought to you for free and open access by the Plant and Soil Sciences at UKnowledge. It has been accepted for inclusion in International Grassland Congress Proceedings by an authorized administrator of UKnowledge. For more information, please contact UKnowledge@lsv.uky.edu. Presenter Information Xiaoli Wang, Chengbin Tang, Li Zhong, Wei Chen, Jiahai Wu, Qun Mu, and Xiaoxia Liu This event is available at UKnowledge: https://uknowledge.uky.edu/igc/21/20-1/17 　 Multifunctional Grasslands in a Changing World 　 Volume Ⅱ 　 瞯 ]1013　 瞯Grasslands/Rangelands People and Policies ——— Institutional Innovations for the Conservation of Grasslands/Rangelands BiodiversityISSR analysis on genetic diversity of wild Agrostis stoloni f era L . germplasm resourcesW ang X iao‐L i ,Tang Cheng‐Bin ,Zhong L i ,Chen Wei ,W u Jia‐Hai ,Mu Qun ,L iu X iao‐X iaGuiz hou Institute o f p rataculture ,Dushan ,558200 ,China ; TEL : 0854‐3231250 ; E‐mail :w angx iaoliz henyuan＠ 126 .comKey words : A grostis stoloni f era L . ,Genetic Diversity ,Genetic structure ,ISSR ,Germplasm resourcesIntroduction A grostis stoloni f era L . is a major cool season turf grass in China .ISSR(Zietkiewicz E ,１９９４) . Molecular markerswere used to detect the genetic diversity among three populations of wild and one of cultivated Agrostis stolonifera L . collectedfrom GuizhouMaterials and methodsPlant materialThe plants used in this study were sampled from three natural populations and one cultivated population of A grostis stoloni f eraL . located in Guizhou . And every populations had several subpopulations ( except KROMI ) . The locations of sampledpopulations were reported in Table１ .Table 1 Natural and cultiv ated populations o f Agrostis stoloniferaL . used in this studyName of populations origins No . of subpopulationsNatural BSE Southeastern of Biji ６ 北populations BNW Northwestern of Biji ７ 北LPS Liu Panshui ５ 北Cultivated population KROMI Dushan １ 北DNA extraction and ISSR amplification Within each subpopulation we selected １５ individuals to extract and mixed DNA , thenPCR amplifications were performed in a final volume of ２５μL containing ５０ng DNA templates , １μL ( ２mmol/ L ) dNTPs ,２μL１０× PCR buffer ,２μL (２５mmol/ L ) Mg２ ＋ ,０ .４U ( ２ .５U /L ) Taq polymerase ,１μL Primer ( １０pmo１ ) .Reactions were conducted byfollowing program :９４ ℃ for７ min ; Followed by ３５cycles of for３０s ,５O ℃ for４５s ,７２ ℃ for１min ;and ended with ７２ ℃ for７min .Results and discussionsGenetic diversity Nine primer pairs produced ６６ polymorphic bands , the average percentage of polymorphic bands was ８１ .４８ .At the population level , the percentage of polymorphic bands ranged from ４３ .２１ to ５９ .２６ , with the average of ５０ .２１ .TheNei摧s gene diversity index was ０ .２４１４ and Shannon diversity index was ０ .３７１９ . These results suggested that there was a richgenetic diversity among the natural populations of Agrostis stolonifera L .Amova The genetic differentiation coefficient was ０ .４９２ and the gene flow was ０ .５１６４ among three wild populations , Itrevealed a significant genetic differentiation .Genetic relatives The Nei摧s genetic similarity coefficient of the sub‐populations ranged from ０ .４０７４ to ０ .９１２３ , can be clusteredinto three groups . Moreover , the findings implied that a correlation among the populations . The significantly high correlationbetween geographical and genetic distance were observed( r ＝ ０ .４９４ , ０ .０５ ＞ P ＞ ０ .０１) . Further analysis on genetic relationshipsuggested relatively high genetic similarity among populations ,ranged from ０ .７００３ to ０ .９４０９ .Conclusions There was a rich genetic diversity and significant genetic differentiation among the natural populations of Agrostisstolonifera L . and the cultivated populations showed a significant genetic difference from natural populations .ReferenceZietkiewicz E ,Rafalaki A ,Labuda D , １９９４ .Genome fingerprinting by simple sequence repeat( SSR)‐anchored polymerase chainreaction amplification . Genomics Jourmal , ２０ ,１７６ ～ １８３ .

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ISSR Analysis on Genetic Diversity of Wild \u3ci\u3eAgrostis stolonifera\u3c/i\u3e L. Germplasm Resources

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