雷公藤ISSR反应体系的建立与优化

【目的】建立适合雷公藤的ISSR-PCR反应体系，为雷公藤种质资源的亲缘关系鉴定和遗传多样性分析提供技术支持。【方法】采用单因素试验设计优化影响雷公藤ISSR-PCR扩增效果的TaqDNA聚合酶、dNTPs、Mg2+、引物浓度和DNA模板含量及退火温度，并以32份雷公藤样品对优化的ISSR-PCR反应体系进行验证。【结果】优化的雷公藤种质资源ISSR-PCR反应体系为：20.00μL反应液中含1.50UTaqDNA聚合酶、0.30mmol/LdNTPs、2.00mmol/LMg2+、0.40μmol/L引物、20ngDNA模板和2.00μL10×Buffer。ISSR-PCR扩增程序中最佳退火...

Full description

Saved in:
Bibliographic Details
Published in南方农业学报 Vol. 48; no. 10; pp. 1755 - 1760
Main Author 瞿印权;徐雯;李欣欣;胡迪科;何天友;荣俊冬;陈礼光;郑郁善
Format Journal Article
LanguageChinese
Published 福建农林大学 林学院,福州,350002%福建农林大学 艺术学院园林学院,福州,350002 2017
Subjects
雷公藤;ISSR;反应体系;优化;种质资源鉴定
Tripterygium wilfordii Hook. f
optimization
优化
germplasm resources identifica-tion
reaction system
反应体系
ISSR
雷公藤
种质资源鉴定
Online AccessGet full text
ISSN2095-1191
DOI10.3969/j.issn.2095-1191.2017.10.05

Cover

More Information
Summary:【目的】建立适合雷公藤的ISSR-PCR反应体系，为雷公藤种质资源的亲缘关系鉴定和遗传多样性分析提供技术支持。【方法】采用单因素试验设计优化影响雷公藤ISSR-PCR扩增效果的TaqDNA聚合酶、dNTPs、Mg2+、引物浓度和DNA模板含量及退火温度，并以32份雷公藤样品对优化的ISSR-PCR反应体系进行验证。【结果】优化的雷公藤种质资源ISSR-PCR反应体系为：20.00μL反应液中含1.50UTaqDNA聚合酶、0.30mmol/LdNTPs、2.00mmol/LMg2+、0.40μmol/L引物、20ngDNA模板和2.00μL10×Buffer。ISSR-PCR扩增程序中最佳退火温度为54.7℃。【结论】建立优化的雷公藤ISSR-PCR反应体系具有较高的稳定性、重现性和适用性，可应用于雷公藤不同地理种源间的遗传多样性鉴定。
Bibliography:45-1381/S
Tripterygium wilfordii Hook. f.;ISSR;reaction system;optimization;germplasm resources identification
Objective】This research was conducted to establish a ISSR-PCR reaction system for the paternity indentification and genetic diversity analysis of Tripterygium wilfordii Hook.f.【Method】Single-factor analysis method was applied to optimize and select the main components and annealing temperature of five PCR reaction system having effects on PCR amplification of the32T.wilfordii samples,including Mg2+,dNTPs,Taq DNA polymerase,primer concentration and template DNA content.And the32samples were used to testify the optimized ISSR-PCR reaction system.【Result】An ISSR-PCR reaction system applicable for indentifying germplasm resources of T.wilfordii was established:20.00μL reaction mixture contained2.00mmol/L Mg2+,0.30mmol/L dNTPs,1.50U Taq DNA polymerase,0.40mol/L primer,20.00ng template DNA and2.00μL10×Buffer.The optimal annealing temperature was54.7℃.【Conclusion】The T.wilfordii ISSR-PCR reaction system estab
ISSN:2095-1191
DOI:10.3969/j.issn.2095-1191.2017.10.05