Microbes and Environments
Online ISSN : 1347-4405
Print ISSN : 1342-6311
ISSN-L : 1342-6311
Regular Papers
PCR-denaturing Gradient Gel Electrophoresis as a Simple Identification Tool of Arbuscular Mycorrhizal Fungal Isolates
Ryo Ohtomo Norikuni OkaSho Morimoto
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2019 年 34 巻 4 号 p. 356-362

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Due to their obligate symbiotic nature and lack of long-term storage methods, the strain collection of arbuscular mycorrhizal (AM) fungi requires periodic proliferation using a pot culture with host plants. Therefore, a method to evaluate the purity of proliferated AM fungal cultures is critical for the quality control of their collection. In a simple evaluation of the purity and identity of a proliferated AM fungal culture, DNA extracted from the culture was amplified using AM fungi-specific PCR followed by an analysis with denaturing gradient gel electrophoresis (PCR-DGGE). The present results showed that the DGGE band patterns of AM fungal strains differed according to their phylogenetic positions, allowing for the rapid and easy identification of the proliferated AM fungal strains. When a culture was contaminated with another AM fungal strain, the DGGE pattern became a mixture of those strains. A contaminant strain was detectable even when its ratio was 1/9 of the main strain. It was also possible to confirm the purity of the culture by comparing whether the DGGE band pattern of the proliferated culture was identical to that obtained from single spores isolated from the culture. Therefore, PCR-DGGE is useful as a quality control tool for maintaining culture collections of AM fungi.

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© 2019 by Japanese Society of Microbial Ecology / Japanese Society of Soil Microbiology / Taiwan Society of Microbial Ecology / Japanese Society of Plant Microbe Interactions / Japanese Society for Extremophiles.
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