熊蜂ISSR-PCR体系的建立与优化及其在亲缘关系分析中的应用  

Establishment and optimization of ISSR-PCR reaction system and its application in the genetic coefficient analysis of Bombus hypocrita Pérez(Hymenoptera:Apidae)

作　　者：耿金虎[1] 黄强[1,2] 徐希莲[1] 曾志将[2] 

机构地区：[1]北京市农林科学院农业科技信息研究所,北京100097 [2]江西农业大学蜜蜂研究所,南昌330045

出　　处：《昆虫学报》2009年第2期223-227,共5页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(30600453);北京市自然科学基金项目(6072008)

摘　　要：以小峰熊蜂Bombus hypocrita Pérez为实验材料,建立并优化简单序列重复区间(inter-simple sequence repeat, ISSR)扩增多态反应体系,并运用ISSR标记和NTSYS聚类软件分析群内亲缘关系和亚家系组成。结果表明:在10μL的PCR反应体系中各组分的适宜终浓度为1×PCR缓冲液、0.3 mmoL/L反应底物、1.25μmol/L引物、3 mmol/L镁离子、2.5 U Taq酶和8.18～23.76μg/mL DNA模板。用此优化后的反应体系对小峰熊蜂基因组DNA进行ISSR-PCR扩增,能有效分析群内亲缘关系和亚家系组成。In this study, the ISSR-PCR reaction system was established, optimized and then used to analyze the genetic coefficient and the subfamily composition of Bombus hypocrita Perez. The optimized reaction system was established as follows： 1×PCR buffer, 0.3 mmol/L dNTP, 1.25μmol/L primer, 3 mmol/L Mg^2＋ , 2.5U Taq polymerase and 8. 18 - 23.76 μg/mL DNA. Our results indicated that ISSR markers obtained with this optimized system could be effectively used to analyze the genetic coefficient and subfamily composition.

关 键 词：熊蜂 小峰熊蜂 ISSR—PCR 聚类分析 亲缘关系 交配 

分 类 号：Q961[生物学—昆虫学]