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...
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| Published in | 南方农业学报 Vol. 48; no. 10; pp. 1755 - 1760 |
|---|---|
| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| 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
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| Subjects | |
| Online Access | Get full text |
| ISSN | 2095-1191 |
| DOI | 10.3969/j.issn.2095-1191.2017.10.05 |
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| Abstract | 【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。 |
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| AbstractList | 【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。 S567.1; [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Taq DNA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1.50 U Taq DNA聚合酶、0.30 mmol/L dNTPs、2.00 mmol/L Mg2+、0.40μmol/L引物、20 ng DNA模板和2.00μL 10×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. |
| Abstract_FL | [Objective]This research was conducted to establish a ISSR-PCR reaction system for the paternity indenti-fication 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 the 32 T. wilfordii samples,including Mg2+,dNTPs,Taq DNA polymerase,primer concentration and template DNA content. And the 32 samples 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 reac-tion mixture contained 2.00 mmol/L Mg2+,0.30 mmol/L dNTPs,1.50 U Taq DNA polymerase,0.40 mol/L primer,20.00 ng template DNA and 2.00μL 10 × Buffer. The optimal annealing temperature was 54.7℃.[Conclusion]The T. wilfordii ISSR-PCR reaction system established is of high stability,repeatability and applicability,indicating this reaction system is suitable for the genetic diversity analysis of T. wilfordii from different geographical provenances. |
| Author | 瞿印权;徐雯;李欣欣;胡迪科;何天友;荣俊冬;陈礼光;郑郁善 |
| AuthorAffiliation | 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 |
| AuthorAffiliation_xml | – name: 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 |
| Author_FL | HE Tian-you QU Yin-quan XU Wen LI Xin-xin CHEN Li-guang RONG Jun-dong ZHENG Yu-shan HU Di-ke |
| Author_FL_xml | – sequence: 1 fullname: QU Yin-quan – sequence: 2 fullname: XU Wen – sequence: 3 fullname: LI Xin-xin – sequence: 4 fullname: HU Di-ke – sequence: 5 fullname: HE Tian-you – sequence: 6 fullname: RONG Jun-dong – sequence: 7 fullname: CHEN Li-guang – sequence: 8 fullname: ZHENG Yu-shan |
| Author_xml | – sequence: 1 fullname: 瞿印权;徐雯;李欣欣;胡迪科;何天友;荣俊冬;陈礼光;郑郁善 |
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| ClassificationCodes | S567.1 |
| ContentType | Journal Article |
| Copyright | Copyright © Wanfang Data Co. Ltd. All Rights Reserved. |
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| DOI | 10.3969/j.issn.2095-1191.2017.10.05 |
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| DocumentTitleAlternate | Establishment and optimization of ISSR reaction system for Tripterygium wilfordii Hook.f |
| DocumentTitle_FL | Establishment and optimization of ISSR reaction system for Tripterygium wilfordii Hook. f |
| EndPage | 1760 |
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| Keywords | 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 |
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| Notes | 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 |
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| PublicationTitle | 南方农业学报 |
| PublicationTitleAlternate | Journal of Southern Agriculture |
| PublicationTitle_FL | Journal of Southern Agriculture |
| PublicationYear | 2017 |
| Publisher | 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 |
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| Snippet | ... S567.1; [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Taq... |
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| SubjectTerms | 雷公藤;ISSR;反应体系;优化;种质资源鉴定 |
| Title | 雷公藤ISSR反应体系的建立与优化 |
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