河南省黄颡鱼和鲇自然居群遗传多样性分析-水生生物学专业论文.docxVIP

ABSTRACTGenetic ABSTRACT Genetic diversity wag onc of the COre in the conservation biology,which refers to the level of genetic variation be{tlwecli various groups in one population and the degree of variation of different individuals in the same group．ISSR molecular markets technology could be called preferable method to explore the genetic diversity,on account of the advantage of speediness，efficiency and sensitiveness．In this research，ISSR molecular markers technology was devoted to make comprehensive analysis of genetic diversity and genetic structure in Pelteobagrusfuividraco and S／／urus asotus，which belongs to$iluriformes． The work for Peiteobagrus如M讥co and S／／urus asotus in Hamn has significant development in prot酬ion ofspies re$O嘲，sustainable development and cultivation ofpopulation． Muscular tissue of Pelteobagrus fulvidraco and Silurus asotus we他conserved in 95％ethyl alcohol， they could be印q加∞酬with four methods and then compared the consequence of genomic DNA extraction．The results showed that the pretreatment sample of STE buffet was the desired pro-treatment to ex臼llCt muscular ti剐Kle in elhyl alcohol，bl础lluse after pretreatment of STE buffet,the banof DNA Were clear,orderliness and out ofdegradation。in the meantime，the values ofA260／A2so w讹berw∞ll 1．8-2．0． Fifteen ISSR primer we他respectively singled out by literature and consult relatives to discuss the level of gene in Pelteobagrusfuividraco and Silurus． The orthognml test design method and re-optimization in Pelteobagrus fulvidraco and Silurus asotus，which were applied to establish optimal of ISSR-PCR reaction condition．The 25此system in Peiteobagrus fulvidraco and Silurus asotus included 45ng DNA template，1．5mmol／L M92+，0．4mmoFL dNTPa,0．4pmoFL primer,0．5U砌DNA polym6釉ae．Not only that，the optimal procedure of ISSR-PCR wag also d晚曩如嘁including initial denaturation at 94C for 5min，unwinding at 9412 for 36，thermal denaturation at 45-石0C for 40s，prolonged at 72(2 for 90s，re-prolonged at 72 for 5min，conserved