| 摘要: |
| 【目的】 探索枸杞遗传变异与亲缘关系分析的新方法,为枸杞的种质遗传多样性分析和杂交育种提供了初步理论依据。【方法】 以枸杞为试材,对ISSR-PCR反应中各主要影响因子进行优化筛选，确立枸杞ISSR-PCR的最佳反应体系；并利用该优化体系，对枸杞的8个品种(系)和5个花药培养再生植株进行种质遗传多样性分析。【结果】 枸杞ISSR-PCR的最佳反应体系为:每25 μL反应体系中含10×Buffer 2.5 μL，40 ng模板DNA，2.0 mmol/L Mg2+，0.1 mmol/L dNTPs，0.6 μmol/L引物，1 UTaqDNA聚合酶；15个引物在13份枸杞种质中共扩增出80个条带，其中多态性条带有61条，多态位点比率为76.3%；13份供试材料可分为2 类：一类群含有宁杞1号及其花培植株、宁杞2号及其花培植株、小麻叶、宁杞3号及其花培植株、06-16及其花培植株；另一类群含有04-03-32及其花培植株、04-03-29、北方。从聚类结果来看，宁杞2号、宁杞3号、04-03-32经花药培养所得到的再生植株与母株相比,在DNA水平上发生了一定程度变异。【结论】 建立的ISSR-PCR优化反应体系可用于枸杞遗传多样性和基因组研究。 |
| 关键词: 枸杞 ISSR 体系优化 遗传多样性 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金项目（30760127） |
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| Optimization of ISSR reaction system and its preliminary applicationin wolfberry（Lycium barbarumL.） |
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SUN Jing zu1a XUE Quan hong1a TANG Ming1b CAO Shu miao1a XING Sheng li2
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| Abstract: |
| 【Objective】 The study was conducted to obtain analysis means of wolfberry in genetic variation and relationship.【Method】 The optimal ISSR-PCR reaction system in wolfberry was established by studying main influential factors.It was used to assess the germplasm genetic diversity among 13 individuals of wolfberry,including eight cultivars and five regeneration plantlets by anther culture.【Result】 The optimal PCR system for ISSR analysis was as follows：10×Buffer 2.5 μL,40 ng template DNA,2.0 mmol/L Mg2+,0.1 mmo1/L dNTPs,0.6 μmol/L Primer and 1 U Taq DNA Polymerase in 25 μL reaction volume.80 total bands were amplified from 13 wolfberry with 15 ISSR primers,among which 61 were polymorphic bands.The polymorphic percentage was 76.3%.In dendrograms,they could be divided into two groups：one group containing 9 individuals such as Ningqi No.1,Huapei No.1,Ningqi No.2,Huapei No.2,Xiaomaye,Ningqi No.3,Huapei No.3,Gouqi 06-16,Huapei 06-16；the other consisting of 4 ones such as Gouqi 04-03-32,Huapei 04-03-32,Gouqi 04-03-29,Beifang.It indicated that Huapei No.2,Huapei No.3 and Huapei 04-03-32 were detected the DNA level variations.【Conclusion】 The results obtained can be used for wolfberry genetics and genome research and give the theoretical basis for the breeding of wolfberry. |
| Key words: Lycium barbarum L. ISSR reaction system genetic diversity |
