Expressed genes in the extraradical hyphae of an arbuscular mycorrhizal fungus, Glomus intraradices, in the symbiotic phase.
To collect extraradical hyphae of arbuscular mycorrhizal (AM) fungi for RNA isolation, a PVDF membrane was laid on the hyphal compartment of a two-compartment culture system of transformed carrot hairy roots and Glomus intraradices. Extraradical hyphae free from host tissue were easily collected, and their RNA was rapidly extracted with a modified acid guanidinium thiocyanate-phenol-chloroform method. A 3'-RACE (rapid amplification of cDNA ends) of a known gene indicated that this protocol enabled the isolation of mRNA molecules as small as 2.3 kb. The cDNA libraries of an AM fungus from the aseptic extraradical hyphae in a symbiotic state were constructed for the first time. Three-fourth of 150 ESTs (expressed sequence tags) indicated low or no similarities to known sequences from other organisms.
Sawaki
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Bio-oriented Technology Research Advancement Institution, Department of Ecology, National Grassland Research Institute, Tochigi, Japan.
Saito
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D018076:DNA, Complementary / Q000235:genetics; D018552:Daucus carota / Q000382:microbiology; D020224:Expressed Sequence Tags; D005658:Fungi / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D015870:Gene Expression; D008969:Molecular Sequence Data; D018517:Plant Roots / Q000382:microbiology; D012331:RNA, Fungal / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D013559:Symbiosis
D018076:DNA, Complementary; D012331:RNA, Fungal