槲寄生药材亲缘关系的RAPD分析
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摘要
本研究以半寄生性药材-槲寄生为研究对象,利用RAPD分子标记技术,以生长环境和寄主植物为要素,旨在探讨这两个因素对槲寄生药材亲缘关系的影响。研究表明:10个RAPD引物在15个槲寄生样本中共扩增出190条带,其中多态性带143条,多态性比率高达75.26%。采用NTSYSpc软件的UPGMA法构建聚类系统树型图,当相似性系数为0.657时,以产地为基础,可以将15个槲寄生样本分为3类。槲寄生样本间的遗传多样性有显著差异:来自河北和黑龙江的槲寄生亲缘关系较近,辽宁地区的槲寄生样本与河北及黑龙江的槲寄生相似性小,亲缘关系相对较远;而来源于不同寄主种属的槲寄生亲缘关系错综复杂,对聚类系谱的划分影响较小。分析结果表明,不同槲寄生药材通过RAPD方法聚类谱系,未能与其寄主种属形成对应关系,但很可能与其生长的地域之间存在一定联系。这为进一步研究槲寄生药材与寄主和产地间的复杂关系提供了更有利的现代研究手段和理论参考。
In this study, the se mi-parasitic medicinal material-mistletoe was used as the material, and RAPD molecular marker technology was applied to investigate the effect of growth environment and the host plant on the genetic relationship of mistletoe. The results showed that 10 RAPD primers could amplify 190 bands in 15 mistletoe samples, in which 143 were polymorphic bands, with the polymorphic ratio of 75.26%. The UPGMA method of NTSYSpc software was used to construct the tree map of clustering system. When the similarity coefficient was0.657, 15 mistletoe samples could be divided into 3 categories based on the origin. The genetic diversity of these mistletoe samples had significant differences: the genetic relationship of the mistletoe from Hebei and Heilongjiang was closer, and the mistletoe samples from Liaoning had little similarity to the mistletoe of Hebei and Heilongjiang, and the genetic relationship was relatively distant. While the genetic relationship of different host species was complex, which had little influence on the classification of the pedigree. The analysis results indicated that the different mistletoe herbs clustered by RAPD method, and they did not form the one-to-one relations with their host species, but there was probably a certain relationship with their regions. This research could provide more favorable modern research methods and a theoretical basis for further study on the complex relationship between mistletoe herbs, host and origin.
In this study, the se mi-parasitic medicinal material-mistletoe was used as the material, and RAPD molecular marker technology was applied to investigate the effect of growth environment and the host plant on the genetic relationship of mistletoe. The results showed that 10 RAPD primers could amplify 190 bands in 15 mistletoe samples, in which 143 were polymorphic bands, with the polymorphic ratio of 75.26%. The UPGMA method of NTSYSpc software was used to construct the tree map of clustering system. When the similarity coefficient was0.657, 15 mistletoe samples could be divided into 3 categories based on the origin. The genetic diversity of these mistletoe samples had significant differences: the genetic relationship of the mistletoe from Hebei and Heilongjiang was closer, and the mistletoe samples from Liaoning had little similarity to the mistletoe of Hebei and Heilongjiang, and the genetic relationship was relatively distant. While the genetic relationship of different host species was complex, which had little influence on the classification of the pedigree. The analysis results indicated that the different mistletoe herbs clustered by RAPD method, and they did not form the one-to-one relations with their host species, but there was probably a certain relationship with their regions. This research could provide more favorable modern research methods and a theoretical basis for further study on the complex relationship between mistletoe herbs, host and origin.
引文
Hu Y.,Huang Y.Z.,Li Y.,and Liu P.W.,2016,Genetic diversity evaluation of sugarcane varieties using SSR and RAPDmarkers,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(9):2494-2503(胡杨,黄有总,李赟,刘平武,2016,利用SSR与RAPD分子标记评估甘蔗品种的遗传多样性,基因组学与应用生物学,35(9):2494-2503)
Iacumin L.,Comi G.,Cantoni C.,and Cocolin L.,2006,Molecular and technological characterization of Staphylococcus xylosus isolated from naturally fermented Italian sausages by RAPD,Rep-PCR and Sau-PCR analysis,Meat Science,74(2):281-288
Mullis K.,Faloona F.,Scharf S.,Saiki R.K.,Horn G.T.,and Erlich H.,1986,Specific enzymatic amplification of DNA in vitro:the polymerase chain reaction,Cold Spring Harb Symp.Quant Biol.,51:263-273
Nei M.,and Li W.H.,1979,Mathematical model for studying genetic variation in terms of restriction endonucleases,Proc Natl.Acad.Sci.USA,76(10):5269-5273
Ni L.,Yao Y.S.,Gao L.,Wu Z.M.,Tan N.Z.,and Zhu J.S.,2014Density-weighted algorithms for similarity computation and cluster tree construction in the RAPD analysis of natural Cordyceps sinensis,Am.J.Biomed.Sci.,6(2):82-104
Orhan D.D.,Aslan M.,Sendogdu N.,Ergun F.,and Yesilada E.2005,Evaluation of the hypoglycemic effect and antioxidant activity of three Viscum album subspecies(European mistletoe)in streptozotocin-diabetic rats,J.Ethnopharmacol.,9 8(1-2):95-102
Rahman M.O.,2010,Use of Random PCR(RAPD)technology to analyze systematic relationships in terrestrial bladderworts(Utricularia L.),Bangladesh J.Bot.,39(1):97-102
Ramesh M.,Vijayakumar K.P.,Karthikeyan A.,and Pandian S.K.,2011,RAPD based genetic stability analysis among micropropagated,synthetic seed derived and hardened plants of Bacopa monnieri(L.):a threatened Indian medicinal herb,Acta Physiol.Plant,33(1):163-171
Sesli M.,and Yegenoglu E.D.,2010,Comparison of similarity coefficients used for cluster analysis based on RAPD markers in wild olives,Genet.Mol.Res.,9(4):2248-2253
Venkatachalam P.,Jinu U.,Sangeetha P.,Geetha N.,and Sahi S.V.,2018,High frequency plant regeneration from cotyledonary node explants of Cucumis sativus L.cultivar'Green Long'via adventitious shoot organogenesis and assessment of genetic fidelity by RAPD-PCR technology,3 Biotech,8(1):60
Welsh J.,and McClelland M.,1990,Fingerprinting genomes using PCR with arbitrary primers,Nucleic Acids Res.,18(24):7213-7218
Williams J.G.K.,Kubelik A.R.,Livak K.J.,Rafalski J.A.,and Tingey S.V.,1990,DNA polymorphisms amplified by arbitrary primers are useful as genetic markers,Nucleic Acids Res.,18(22):6531-6535
Yang C.L.,Wu X.P.,Chen B.,Deng S.S.,Chen Z.E.,Huang Y.Y.,and Jin S.S.,2017,Comparative analysis of genetic polymorphisms among Monascus strains by ISSR and RAPDmarkers,J.Sci.Food Agric.,97(2):636-640
Iacumin L.,Comi G.,Cantoni C.,and Cocolin L.,2006,Molecular and technological characterization of Staphylococcus xylosus isolated from naturally fermented Italian sausages by RAPD,Rep-PCR and Sau-PCR analysis,Meat Science,74(2):281-288
Mullis K.,Faloona F.,Scharf S.,Saiki R.K.,Horn G.T.,and Erlich H.,1986,Specific enzymatic amplification of DNA in vitro:the polymerase chain reaction,Cold Spring Harb Symp.Quant Biol.,51:263-273
Nei M.,and Li W.H.,1979,Mathematical model for studying genetic variation in terms of restriction endonucleases,Proc Natl.Acad.Sci.USA,76(10):5269-5273
Ni L.,Yao Y.S.,Gao L.,Wu Z.M.,Tan N.Z.,and Zhu J.S.,2014Density-weighted algorithms for similarity computation and cluster tree construction in the RAPD analysis of natural Cordyceps sinensis,Am.J.Biomed.Sci.,6(2):82-104
Orhan D.D.,Aslan M.,Sendogdu N.,Ergun F.,and Yesilada E.2005,Evaluation of the hypoglycemic effect and antioxidant activity of three Viscum album subspecies(European mistletoe)in streptozotocin-diabetic rats,J.Ethnopharmacol.,9 8(1-2):95-102
Rahman M.O.,2010,Use of Random PCR(RAPD)technology to analyze systematic relationships in terrestrial bladderworts(Utricularia L.),Bangladesh J.Bot.,39(1):97-102
Ramesh M.,Vijayakumar K.P.,Karthikeyan A.,and Pandian S.K.,2011,RAPD based genetic stability analysis among micropropagated,synthetic seed derived and hardened plants of Bacopa monnieri(L.):a threatened Indian medicinal herb,Acta Physiol.Plant,33(1):163-171
Sesli M.,and Yegenoglu E.D.,2010,Comparison of similarity coefficients used for cluster analysis based on RAPD markers in wild olives,Genet.Mol.Res.,9(4):2248-2253
Venkatachalam P.,Jinu U.,Sangeetha P.,Geetha N.,and Sahi S.V.,2018,High frequency plant regeneration from cotyledonary node explants of Cucumis sativus L.cultivar'Green Long'via adventitious shoot organogenesis and assessment of genetic fidelity by RAPD-PCR technology,3 Biotech,8(1):60
Welsh J.,and McClelland M.,1990,Fingerprinting genomes using PCR with arbitrary primers,Nucleic Acids Res.,18(24):7213-7218
Williams J.G.K.,Kubelik A.R.,Livak K.J.,Rafalski J.A.,and Tingey S.V.,1990,DNA polymorphisms amplified by arbitrary primers are useful as genetic markers,Nucleic Acids Res.,18(22):6531-6535
Yang C.L.,Wu X.P.,Chen B.,Deng S.S.,Chen Z.E.,Huang Y.Y.,and Jin S.S.,2017,Comparative analysis of genetic polymorphisms among Monascus strains by ISSR and RAPDmarkers,J.Sci.Food Agric.,97(2):636-640