A molecular marker diagnostic of a specific isolate of an arbuscular mycorrhizal fungus,<i>Gigaspora margarita</i>

Yokoyama, Keiichi; Tateishi, Takahiro; Marumoto, Takuya; Saitô, Masanori

doi:10.1111/j.1574-6968.2002.tb11262.x

Cited by 6 publications

(7 citation statements)

References 16 publications

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“…Some molecular tools to differentiate among AMF species and strains have been developed. However these new technologies remain to be tested for a variety of AM fungal strains and species (26,29,42,53). Sexual reproduction has not yet been observed for these symbiotic fungi; therefore they are considered asexual.…”

Section: Propagation Cycle Of Arbuscular Mycorrhizal Fungimentioning

confidence: 99%

Arbuscular Mycorrhiza Inoculum to Support Sustainable Cropping Systems

Dalpé

Monreal

2004

Crop Management

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Arbuscular mycorrhizae (AM) are symbiotic associations, formed between plants and soil fungi that play an essential role in plant growth, plant protection, and soil quality. The AM fungi expand their filaments in soil and plant roots. This filamentous network promote bi‐directional nutrient movement where soil nutrients and water move to the plant and plant photosynthates flow to the fungal network. AM fungi are ubiquitous in the soil and can form symbiosis with most terrestrial plants including major crops, cereals, vegetables, and horticultural plants. In agriculture, several factors, such as host crop dependency to mycorrhizal colonization, tillage system, fertilizer application, and mycorrhizal fungi inoculum's potential can affect plant response and plant benefits from mycorrhizae. Due to their obligate symbiotic status, AM fungi need to associate with plant for growth and proliferation. Consequently, the cultivation of AM fungal strains and the maintenance of reference collections require methodologies and infrastructures quite different from those used with other microbial collections and inoculum production. Interest in AM fungi propagation for agriculture is increasing due to their role in the promotion of plant health, in soil nutrition improvement, and soil aggregate stability. The comprehensive life cycle of AM fungi and methods currently used for the propagation of inoculum and the maintenance of in vivo and in vitro source collections are described. Methods and regulations of large‐scale production of commercial inoculum that provide users with products of high quality and efficiency are discussed.

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Section: Propagation Cycle Of Arbuscular Mycorrhizal Fungimentioning

confidence: 99%

Arbuscular Mycorrhiza Inoculum to Support Sustainable Cropping Systems

Dalpé

Monreal

2004

Crop Management

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“…Their analysis also revealed intraspore heterogeneity, which caused ambiguities in the signatures they found. Lanfranco et al (32) continued the molecular characterization of selected Gigaspora species and proposed a number of species-specific primer sets, and Yokoyama et al (58) developed primers based on satellite fragments to identify specific isolates of G. margarita. Thus, while important strides have been made recently, rapid and reliable methods to assess Gigaspora diversity are still lacking.…”

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confidence: 99%

PCR-Denaturing Gradient Gel Electrophoresis Profiling of Inter- and Intraspecies 18S rRNA Gene Sequence Heterogeneity Is an Accurate and Sensitive Method To Assess Species Diversity of Arbuscular Mycorrhizal Fungi of the GenusGigaspora

Souza

Kowalchuk

Leeflang

et al. 2004

Appl Environ Microbiol

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Despite the importance of arbuscular mycorrhizal fungi in the majority of terrestrial ecosystems, their ecology, genetics, and evolution are poorly understood, partly due to difficulties associated with detecting and identifying species. We explored the inter-and intraspecies variations of the 18S rRNA genes of the genus Gigaspora to assess the use of this marker for the discrimination of Gigaspora isolates and of Gigasporaceae populations from environmental samples. Screening of 48 Gigaspora isolates by PCR-denaturing gradient gel electrophoresis (DGGE) revealed that the V3-V4 region of the 18S rRNA gene contained insufficient variation to discriminate between different Gigaspora species. In contrast, the patterns of 18S ribosomal DNA (rDNA) heterogeneity within the V9 region of this marker could be used for reliable identification of all recognized species within this genus. PCR-DGGE patterns provided insight into some putative misidentifications and could be used to differentiate geographic isolates of G. albida, G. gigantea, and G. margarita but not G. rosea. Two major clusters were apparent based upon PCR-DGGE ribotype patterns, one containing G. albida, G. candida, G. ramisporophora, and G. rosea and the other containing G. decipiens and G. margarita. Dissection of the DGGE patterns by cloning, DGGE screening, and sequencing confirmed these groupings and revealed that some ribotypes were shared across species boundaries. Of the 48 isolates examined, only two displayed any spore-to-spore variation, and these exceptions may be indicative of coisolation of more than one species or subspecies within these cultures. Two Brazilian agricultural soils were also analyzed with a Gigasporaceaespecific nested PCR approach, revealing a dominance of G. margarita within this family.

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“…Roots DNA preparation. Total DNA was extracted from five or ten spores with a similar size to those of G. margarita (350 to 450 pm in diameter) independently, as described in our previous study (Yokoyama et al 2002). After thawing, plant roots were soaked in sterilized water to avoid desiccation.…”

Section: Methodsmentioning

confidence: 99%

“…The determination of the sequence as a marker was carried out through PCR analysis and probing. This was a simple method for the identification of the fungus at both sporal and infection stages in pot cultures (Yokoyama et al 2002). This technique may enable to trace the isolate in field ecosystems.…”

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confidence: 99%

Application of a Molecular Method for the Identification of aGigaspora margaritaIsolate Released in a Field

Yokoyama

Tateishi

Saitô

et al. 2005

Soil Science and Plant Nutrition

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A molecular technique for the identification of the Gigaspora margarita isolate CK based on the detection of a DNA sequence of 236 bp as its diagnostic marker was evaluated to investigate the survival and establishment of introduced arbuscular mycorrizal Pungi (AMF) in a field ecosystem. In March 2001, roots and rhizosphere soil of Eragrostis curuula and Mbcanthus sinensis were collected from the Mizunashi River at Mt. Fugendake (Nagasaki Prefecture, Japan), where plant seeds and AMF including G. margarita CK had been introduced for reforestation after the occurrence of repeated pyroclastic flows. We detected the marker sequence from DNA preparations of E. curuula roots and Gigaspora spores in the rhizosphere. This clearly showed that the isolate occurred at both hyphal and sporal stages. It was shown that the isolate survived and developed a life cycle in the revegetation area for 4 years. It was confhned that the method was effective for tracing the isolate in samples collected from field ecosystems.

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A molecular marker diagnostic of a specific isolate of an arbuscular mycorrhizal fungus,Gigaspora margarita

Cited by 6 publications

References 16 publications

Arbuscular Mycorrhiza Inoculum to Support Sustainable Cropping Systems

Arbuscular Mycorrhiza Inoculum to Support Sustainable Cropping Systems

PCR-Denaturing Gradient Gel Electrophoresis Profiling of Inter- and Intraspecies 18S rRNA Gene Sequence Heterogeneity Is an Accurate and Sensitive Method To Assess Species Diversity of Arbuscular Mycorrhizal Fungi of the GenusGigaspora

Application of a Molecular Method for the Identification of aGigaspora margaritaIsolate Released in a Field

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