Development of primer sets for PCR amplification of the PgiC gene in ferns
Development of primer sets for PCR amplification of the PgiC gene in ferns
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DOI：
10.1007/s102650200010
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发表时间：
2002-02-01
影响因子：
2.8
通讯作者：
Kurita, S
中科院分区：
文献类型：
--
作者：
Ishikawa, H;Watano, Y;Kurita, S
建立了基于聚合酶链式反应(PCR)的蕨类植物核DNA标记。我们首先测定了鳞毛虫胞质磷酸葡萄糖异构酶编码基因pGIC的部分核苷酸序列，然后设计了用于外显子引发内含子交叉(EPIC)扩增的聚合酶链式反应引物。EPIC引物普遍适用于鳞毛蕨科、鳞毛蕨科、木犀科等衍生程度最高的工业化蕨类植物。含有两个内含子的引物14F/16R的扩增产物大小适中(534个碱基-约534个碱基)。1000个碱基)，可能在特定和属水平上的系统发育重建中有价值。利用14F/15R和15F/16R两对引物，通过单链构象多态性(SSCP)和EPIC扩增相结合的方法，开发出了可用于交配系统评价和其他群体遗传学研究的共显性核DNA标记。为了提供一个利用这些标记的案例研究，利用15F/16R对蜘蛛种群的等位基因变异进行了检测。
Polymerase chain reaction (PCR)-based nuclear DNA markers were developed for fern species. We first determined the partial nucleotide sequence of cDNA of the pgiC gene encoding cytosolic phosphoglucose isomerase from Dryopteris caudipinna, and then PCR primers for exon-primed, intron-crossing (EPIC) amplifications were designed. The EPIC primers are universally applicable to the most derived indusiate fern families such as Dryopteridaceae, Thelypteridaceae, and Woodsiaceae. The PCR products of primers 14F/16R containing two introns are moderate in size (534 bp-ca. 1000 bp) and are possibly of value in phylogenetic reconstruction at specific and generic levels. Codominant nuclear DNA markers applicable to the estimation of mating systems and other population genetic studies were also developed by a combination of single-strand conformation polymorphism (SSCP) and EPIC amplification using primers 14F/15R and 15F/16R. In order to provide a case study using these markers, allelic variation of PCR products using 15F/16R was examined in populations of Arachniodes standishii (Dryopteridaceae).