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摘要: 基于山核桃Carya cathayensis脂肪代谢相关cDNA文库的部分测序结果,发现43条cDNA序列具有简单重复序列(SSR)碱基重复特点,用Primer Premier 5设计了34对表达序列标签-简单重复序列(EST-SSR)引物;建立了适用于山核桃的SSR分析体系,并用设计的SSR引物对对天然群体山核桃40个样品进行了初步的应用性试验。结果表明:34对引物中有4对无扩增产物,3对引物的扩增产物超过了设计范围,27对引物在供试的40个山核桃材料中有较清晰且稳定的目标扩增产物,其中有18对引物的扩增产生了22个多态性位点,占总扩增位点数的47.8%。将SSR标记分析结果与山核桃已有的显性标记分析结果进行比较,发现SSR标记多态性位点的比例较高,且能够区分显性纯合位点及显性杂合位点。结论:设计开发山核桃的EST-SSR标记是可行有效的。图2表3参36Abstract: Codominant markers are better than dominant ones in genetic studies for their ability to distinguish homozygotes and heterozygotes,but their development is usually costly. There has been no report on application of codominant markers to Carya cathayensis so far,but the cDNA library associated with fat metabolism in Carya cathayensis revealed 43 cDNA sequences characterized by base repeats of simple sequence repeat (SSR) markers,from which 34 pairs of expressed sequence tag (EST)-SSR primers were designed with Primer Premier 5. With an SSR protocol established,these SSR primers were used to analyze DNA of 40 C. cathayensis trees from a natural population in Linan of Zhejiang Province,which are at least 100 m apart from each other. A native PAGE (polyacrylamide gel electrophoresis) design was used to analyze the applicability of the SSR primers in C. cathayensis. Results showed four pairs of primers had no amplification product,three pairs had an amplification product beyond the pre-designed product length,and 27 pairs had clear and stable amplification products. From the 27 pairs,18 pairs amplified 22 polymorphic loci accounting for 47.8% of the total loci amplified. Compared to other dominant markers (RAPD,ISSR,SRAP and AFLP) developed for C. cathayensis,the SSR markers were only high in the percentage of polymorphic loci. Thus,it would be feasible to develop EST-SSR markers in C. cathayensis.[Ch,2 fig. 3 tab. 36 ref.]
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https://zlxb.zafu.edu.cn/article/doi/10.11833/j.issn.2095-0756.2012.04.023
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