Department of Molecular Biology, Agricultural University, Dreijenlaan 3, NL-6703, HA Wageningen, The Netherlands.
Theor Appl Genet. 1991 Nov;83(1):108-14. doi: 10.1007/BF00229232.
A new DNA polymorphism assay was developed in 1990 that is based on the amplification by the polymerase chain reaction (PCR) of random DNA segments, using single primers of arbitrary nucleotide sequence. The amplified DNA fragments, referred to as RAPD markers, were shown to be highly useful in the construction of genetic maps ("RAPD mapping"). We have now adapted the RAPD assay to tomato. Using a set of 11 oligonucleotide decamer primers, each primer directed the amplification of a genome-specific "fingerprint" of DNA fragments. The potential of the original RAPD assay to generate polymorphic DNA markers with a given set of primers was further increased by combining two primers in a single PCR. By comparing "fingerprints" of L. esculentum, L. pennellii, and the L. esculentum chromosome 6 substitution line LA1641, which carries chromosome 6 from L. pennellii, three chromosome 6-specific RAPD markers could be directly identified among the set of amplified DNA fragments. Their chromosomal position on the classical genetic map of tomato was subsequently established by restriction fragment length polymorphism (RFLP) linkage analysis. One of the RAPD markers was found to be tightly linked to the nematode resistance gene Mi.
1990 年开发了一种新的 DNA 多态性分析方法，该方法基于聚合酶链反应（PCR）扩增随机 DNA 片段，使用任意核苷酸序列的单引物。扩增的 DNA 片段被称为 RAPD 标记，在构建遗传图谱（“RAPD 图谱”）中非常有用。我们现在已经将 RAPD 分析方法应用于番茄。使用一组 11 个寡核苷酸十聚体引物，每个引物指导基因组特异性 DNA 片段“指纹”的扩增。通过在单个 PCR 中组合两个引物，可以进一步提高原始 RAPD 分析方法用给定引物组生成多态性 DNA 标记的潜力。通过比较 L. esculentum、L. pennellii 和携带 L. pennellii 染色体 6 的 L. esculentum 染色体 6 替代系 LA1641 的“指纹”，可以在扩增的 DNA 片段中直接鉴定出三个染色体 6 特异性 RAPD 标记。随后通过限制性片段长度多态性（RFLP）连锁分析确定它们在番茄经典遗传图谱上的染色体位置。其中一个 RAPD 标记被发现与线虫抗性基因 Mi 紧密连锁。
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