通过随机扩增多态性DNA区分马铃薯孢囊线虫罗氏 Globodera rostochiensis 和苍白球孢囊线虫 G. pallida 的单个幼虫。

Single juveniles of the potato cyst nematodes Globodera rostochiensis and G. pallida differentiated by randomly amplified polymorphic DNA.

作者信息

Roosien J, Van Zandvoort P M, Folkertsma R T, Van der Voort J N, Goverse A, Gommers F J, Bakker J

机构信息

Department of Nematology, Wageningen Agricultural University, The Netherlands.

出版信息

Parasitology. 1993 Dec;107 ( Pt 5):567-72. doi: 10.1017/s0031182000068153.

DOI:10.1017/s0031182000068153

PMID:8295795

Abstract

Random amplified polymorphic DNA (RAPD) offers a potential basis for the development of a diagnostic assay to differentiate the potato cyst nematode species Globodera rostochiensis and G. pallida. Nine decamer primers have been tested for their ability to amplify species-specific DNA sequences. Primer OPG-05 produced 2 discrete DNA fragments, which were consistently present in 5 G. rostochiensis populations and absent in 5 G. pallida populations. These fragments were detectable in single females as well as in single 2nd-stage juveniles. Their amplification is extremely efficient, and reproducible over a wide range of template concentrations. One-fifth of a single juvenile is sufficient to generate reproducible RAPD markers. The amplification from single juveniles requires no DNA isolation. The use of a crude homogenate does not impair the polymerase chain reaction.

摘要

随机扩增多态性DNA（RAPD）为开发一种诊断检测方法以区分马铃薯胞囊线虫物种罗斯托克球孢囊线虫和苍白球孢囊线虫提供了潜在基础。已测试了9种十聚体引物扩增物种特异性DNA序列的能力。引物OPG - 05产生了2个离散的DNA片段，这些片段始终存在于5个罗斯托克球孢囊线虫种群中，而在5个苍白球孢囊线虫种群中不存在。这些片段在单个雌虫以及单个第二阶段幼虫中均可检测到。它们的扩增极其高效，并且在广泛的模板浓度范围内具有可重复性。单个幼虫的五分之一就足以产生可重复的RAPD标记。从单个幼虫进行扩增无需进行DNA分离。使用粗匀浆不会损害聚合酶链反应。