菠菜AFLP多态性引物组合的筛选  

The Adaptive AFLP Primer Combinations Screening of Spinacia oleracea L

作　　者：吴娅妮[1,2] 刘玲[2,3,4] 王文科[1] 孟淑春[2,3,4] 

机构地区：[1]山西师范大学生命科学学院,山西临汾041000 [2]北京市农林科学院蔬菜研究中心,北京100097 [3]农业部华北地区园艺作物生物学与种质创制重点实验室,北京100097 [4]农业部都市农业(北方)重点实验室,北京100097

出　　处：《中国食品学报》2014年第7期207-213,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：北京市科技新星计划项目(2008B37);北京市科委重大课题(D131100000413001);"十二五"国家科技支撑计划项目(2012BAK26B03)

摘　　要：利用改良的CTAB法提取高质量的DNA,对菠菜AFLP反应体系中酶切与连接、预扩增、选择性扩增等关键试验条件进行优化,建立了适合菠菜AFLP分析的反应体系:酶切连接采用一步法,即模板DNA150 ng,EcoRⅠ/MseⅠ各0.25 U,37℃酶切5 h,16℃连接6 h;预扩增体系浓度dNTPs 0.2 mmol/L,Mg2+1.5 mmol/L,预扩增产物稀释200倍作为选择性扩增体系模板。利用优化的反应体系,从256对E+ANN/M+ANN引物组合中,筛选出20对条带清晰、多态性丰富的引物组合,为菠菜群体的遗传多样性、种质鉴定等研究提供了理论基础。In this research, we have optimized AFLP reaction system for Spinacia oleracea L. High quality genomic DNA have been isolated by modified method of CTAB, and the critical process of AFLP have been optimized. The AFLP reaction system established in Spinacia oleracea L was as follows： template DNA 150 ng, digestion 37 ℃ for 5 h, ligation16 ℃ for 6 h and digestion-ligation finished by single step; 0.2 mmol/L dNTPs, 1.5mmol/L Mg2＋in pre-amplification system; pre-amplification products has been diluted 200 times used for selection amplification system. Using the optimized system, 20 pairs of E＋ANN/M＋ANN primers combination were screened out from 256 pairs of AFLP primers. The present results provided a strong foundation for the further study of population genetic diversity and germplasm identify of Spinacia oleracea L by AFLP method.

关 键 词：菠菜 AFLP 反应体系 引物筛选 

分 类 号：TS255.1[轻工技术与工程—农产品加工及贮藏工程]