Spatial monitoring of gene activity in extraradical and intraradical developmental stages of arbuscular mycorrhizal fungi by direct fluorescent in situ RT-PCR

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RESEARCH PRODUCT

Spatial monitoring of gene activity in extraradical and intraradical developmental stages of arbuscular mycorrhizal fungi by direct fluorescent in situ RT-PCR

Christine Arnould Vivienne Gianinazzi-pearson Marie Tollot Cecilia M. Arias Pascale M. A. Seddas

subject

0106 biological sciences MYCORHIZES A ARBUSCULES GENE EXPRESSION Hypha GLOMUS INTRARADICES DIRECT FLUORESCENT IN SITU RT-PCR 01 natural sciences Microbiology Plant Roots ARBUSCULAR MYCORRHIZAL FUNGI Fungal Proteins SUPEROXIDE DISMUTASE 03 medical and health sciences Fungal Structures Gene Expression Regulation Fungal Mycorrhizae Botany Gene expression Genetics Medicago CONFOCAL MICROSCOPY Gene 030304 developmental biology DNA Primers Fluorescent Dyes Peptidylprolyl isomerase 0303 health sciences [SDV.GEN]Life Sciences [q-bio]/Genetics Microscopy Confocal biology PEPTIDYLPROPYL ISOMERASE Reverse Transcriptase Polymerase Chain Reaction Gene Expression Profiling fungi SYMBIOSIS Gene Expression Regulation Developmental Peptidylprolyl Isomerase biology.organism_classification Medicago truncatula Cell biology Arbuscular mycorrhiza Gene expression profiling STEAROYL-CoA-DESATURASE Xanthenes MEDICAGO TRUNCATULA Stearoyl-CoA Desaturase 010606 plant biology & botany

description

International audience; Gene expression profiling based on tissue extracts gives only limited information about genes associated with complex developmental processes such as those implicated in fungal interactions with plant roots during arbuscular mycorrhiza development and function. To overcome this drawback, a direct fluorescent in situ RT-PCR methodology was developed for spatial mapping of gene expression in different presymbiotic and symbiotic structures of an arbuscular mycorrhizal fungus. Transcript detection was optimized by targeting the LSU rRNA gene of Glomus intraradices and monitoring expression of a stearoyl-CoA-desaturase gene that is consistently expressed at high levels in spores, hyphae, arbuscules and vesicles. This method was further validated by localizing expression of fungal peptidylprolyl isomerase and superoxide dismutase genes, which are expressed to different extents in fungal structures. Direct fluorescent in situ RT-PCR offers new perspectives for the sensitive analysis of fungal developmental processes that occur during functional differentiation in symbiotic arbuscular mycorrhiza interactions.

year	journal	country	edition	language
2008-01-01

10.1016/j.fgb.2008.04.013 https://hal.inrae.fr/hal-02662900