| 摘要: |
| 【目的】利用ISSR标记和ITS序列分析方法，对11份柴胡种质进行鉴定和亲缘关系分析，为柴胡种质资源利用和进化分析提供理论依据。【方法】以取自不同省份的11份柴胡干品为材料，提取其基因组DNA，进行ISSR分析，利用DPS（V7.5）软件计算遗传距离，利用UPGMA方法进行聚类分析，构建11份柴胡样品的系统进化树。同时，利用ITS引物对柴胡样品进行PCR扩增和测序，利用DNAMAN软件分析11份柴胡种质的ITS序列，对其遗传相似性进行鉴定。【结果】从100条ISSR引物中筛选出21条扩增谱带清晰、多态性高的引物，利用这21条引物从11份柴胡样品中共获得185条扩增条带，其中156条条带呈现多态性，多态性条带比例为84.3%。11份柴胡样品间遗传距离为0.458 8～0.782 2，表明这11份柴胡样品间遗传基础较狭窄；聚类结果将11份柴胡样品分成2大类，大部分来源相同或相近的柴胡样品聚在一起。ITS序列分析结果表明， 11份柴胡样品的ITS序列均长321 bp,也可以分为2类，部分柴胡样品存在同质异名现象。【结论】利用ITS技术或将其与ISSR标记相结合对柴胡种质资源进行鉴定和分析，可以提高柴胡种质资源鉴定的准确性和效率。 |
| 关键词: 柴胡 ISSR ITS 种质资源 遗传多样性 聚类分析 |
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| 基金项目:国家科技部子课题项目（2011BAI07B05-4）；河北省科技厅项目（10965511D）；唐山市科技重大项目（12130203A）；唐山师范学院博士基金项目（11A01） |
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| ISSR-PCR and ITS sequence analysis of Bupleurum germplasm resources |
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MA Yan-zhi,KE Shao-ying
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| Abstract: |
| 【Objective】This paper aimed to identify and analyze germplasm resources of 11 Bupleurum varieties using ISSR PCR and ITS sequence analysis methods,which would provide a theoretical basis for resource utilization and evolution analysis of Bupleurum germplasm.【Method】DNA samples were extracted from 11 dry Bupleurum cultivars.With ISSR PCR,genetic similarity coefficients (GS) were calculated by DPS software and cluster analysis was conducted by UPGMA.The ITS sequences of 11 Bupleurum cultivars were obtained after PCR and sequencing and the DNAMAN software was employed to align the ITS sequences.The genetic similarity was also identified.【Result】21 out of 100 primers with clear amplification spectrum and high polymorphism were selected,and a total of 185 fragments were amplified,among which 156 fragments were polymorphic with a ratio of 84.3%.The GS varied from 0.458 8 to 0.782 2,indicating that the genetic diversity of these cultivars was relatively low.Both UPGMA cluster and ITS sequence analysis showed that the tested 11 Bupleurum cultivars could be divided into two categories.The average sequence length of the samples was 321 bp.Some cultivars were homogeneous.【Conclusion】Using ITS and ISSR methods increased the accuracy and efficiency of Bupleurum germplasm resources analysis. |
| Key words: Bupleurum L. ISSR ITS germplasm resources genetic diversity cluster analysis |
