A set of conserved PCR primers for the analysis of simple sequence repeat polymorphisms in chloroplast genomes of dicotyledonous angiosperms.
A set of conserved PCR primers for the analysis of simple sequence repeat polymorphisms in chloroplast genomes of dicotyledonous angiosperms.
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DOI：
10.1139/g98-104
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发表时间：
1999-02
期刊：
影响因子：
3.1
通讯作者：
K. Weising;R. Gardner
中科院分区：
文献类型：
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作者：
K. Weising;R. Gardner
在高等植物的叶绿体基因组中存在短串的单核苷酸重复序列。在大豆、水稻和松树中，用侧翼引物进行的PCR（聚合酶链反应）表明，在这些重复序列中A或T残基的数量在密切相关的类群中是可变的。在此，我们描述了一组引物，用于研究被子植物叶绿体DNA中的单核苷酸重复序列变异，其中数据库信息有限。在烟草叶绿体基因组中总共识别出39个（A）n和（T）n重复序列（n >或= 10），并将涵盖这39个区域的DNA序列与数据库中的直系同源DNA序列进行比对。构建了共有引物对并用于扩增被子植物等级集的总基因组DNA。所有10个引物对产生的PCR产物的茄科的成员，和10个中的8个也在大多数其他被子植物物种的功能。烟草属，番茄属（茄科），猕猴桃属（猕猴桃科）内的种间多态性水平被证明是高的，而种内变异烟草，番茄，猕猴桃是有限的。从三个引物对的PCR产物的序列分析显示，在猕猴桃的多态性的主要原因是在单核苷酸序列的A，G和T残基的可变数目。我们的研究结果表明，通用引物针对单核苷酸重复序列可以作为一般的工具，研究被子植物叶绿体变异。
Short runs of mononucleotide repeats are present in chloroplast genomes of higher plants. In soybean, rice, and pine, PCR (polymerase chain reaction) with flanking primers has shown that the numbers of A or T residues in such repeats are variable among closely related taxa. Here we describe a set of primers for studying mononucleotide repeat variation in chloroplast DNA of angiosperms where database information is limited. A total of 39 (A)n and (T)n repeats (n > or = 10) were identified in the tobacco chloroplast genome, and DNA sequences encompassing these 39 regions were aligned with orthologous DNA sequences in the databases. Consensus primer pairs were constructed and used to amplify total genomic DNA from a hierarchical set of angiosperms. All 10 primer pairs generated PCR products from members of the Solanaceae, and 8 of the 10 were also functional in most other angiosperm species. Levels of interspecific polymorphism within the genera Nicotiana, Lycopersicon (both Solanaceae), and Actinidia (Actinidiaceae) proved to be high, while intraspecific variation in Nicotiana tabacum, Lycopersicon esculentum, and Actinidia chinensis was limited. Sequence analysis of PCR products from three primer pairs revealed variable numbers of A, G, and T residues in mononucleotide arrays as the major cause of polymorphism in Actinidia. Our results suggest that universal primers targeted to mononucleotide repeats may serve as general tools to study chloroplast variation in angiosperms.