Polymerase chain reaction and hybridization on DNA extracted from soil as a tool for Frankia spp. population distribution studies in soil

Polymerase chain reaction and hybridization on DNA extracted from soil as a tool for Frankia spp. population distribution studies in soil
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DOI：
10.1139/b99-065
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发表时间：
1999-12
期刊：
Botany
影响因子：
1.1
通讯作者：
R. Nalin;P. Normand;P. Simonet;A. Domenach
R. Nalin;P. Normand;P. Simonet;A. Domenach
中科院分区：
生物学4区
文献类型：
--
作者：
R. Nalin;P. Normand;P. Simonet;A. Domenach

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土壤中Frankia种群的分布通常通过植物诱捕结合遗传特征来研究。我们比较了这种方法与聚合酶链反应（PCR）和杂交的人口分布的评价。Frankia spp.菌株，其特征在于由HaeIII限制的nifD-nifK扩增片段（nif-HaeIII谱），被发现在60 cm深的土柱中具有不同的分布。对nifD-nifK基因间间隔区和nifK基因的5 '-部分进行测序，以设计特异性寡核苷酸探针。从10-20 cm、30-40 cm、50-60 cm三个土层中直接提取DNA，经PCR扩增和特异性探针杂交，对Frankia菌群分布的研究结果与植物诱捕法观察到的分布一致。
The distribution of Frankia populations in soil is usually studied by plant trapping in combination with genetic characterization. We compared this approach with the evaluation of population distribution by polymerase chain reaction (PCR) and hybridization. Seven populations of Frankia spp. strains, characterized by restriction by HaeIII of the nifD-nifK amplified fragments (nif-HaeIII profiles), were found to have different distribution in a soil column of 60 cm in depth. The nifD-nifK intergenic spacer and the 5'-part of the nifK gene were sequenced for one representative of the seven nif-HaeIII profiles to design specific oligonucleotide probes. The results on the distribution of Frankia populations by PCR amplification on DNA extracted directly from samples from three soil depths (10-20 cm; 30-40 cm; 50-60 cm) and hybridization with specific probes are congruent with the distribution observed by plant trapping.Key words: nif genes, soil, detection, Frankia, PCR, hybridization, plant trapping.