Fluorescent in situ RT-PCR to visualise the expression of a phosphate transporter gene from an ectomycorrhizal fungus.
Expression of a mycorrhizal fungal-specific phosphate (P) transporter gene (HcPT1) was studied in mycelium of the ectomycorrhizal fungus Hebeloma cylindrosporum, by in situ reverse transcriptase polymerase chain reaction using amplification of complementary DNA sequences. The expression of HcPT1 was visualised under two different P treatments. Mycelium was transferred to liquid medium with or without P and incubated for 5 days. Under P starvation, mycelium growth and vitality was reduced and the expression of HcPT1 up regulated. Enzyme-labelled fluorescent substrate was used to detect gene expression in situ with epi-fluorescence microscopy and to visualise it at the level of the individual hyphae both in starved and non-starved hyphae. Up-regulation of HcPT1 was observed as a more intense fluorescent signal and from the larger proportion of hyphae that showed expression.
van Aarle
Ingrid M
IM
Biogéochimie du Sol et de la Rhizosphère, UMR 1222 INRA/SupAgro, 2 Place Viala, 34060, Montpellier Cedex 1, France.
Viennois
Gaëlle
G
Biochimie et Physiologie Moléculaire des Plantes, UMR 5004 INRA/CNRS/SupAgro/UM II, 2 Place Viala, 34060, Montpellier Cedex 1, France.
Amenc
Laurie K
LK
Biogéochimie du Sol et de la Rhizosphère, UMR 1222 INRA/SupAgro, 2 Place Viala, 34060, Montpellier Cedex 1, France.
Tatry
Marie-Violaine
MV
Biogéochimie du Sol et de la Rhizosphère, UMR 1222 INRA/SupAgro, 2 Place Viala, 34060, Montpellier Cedex 1, France.|Biochimie et Physiologie Moléculaire des Plantes, UMR 5004 INRA/CNRS/SupAgro/UM II, 2 Place Viala, 34060, Montpellier Cedex 1, France.
Luu
Doan T
DT
Biochimie et Physiologie Moléculaire des Plantes, UMR 5004 INRA/CNRS/SupAgro/UM II, 2 Place Viala, 34060, Montpellier Cedex 1, France.
Plassard
Claude
C
Biogéochimie du Sol et de la Rhizosphère, UMR 1222 INRA/SupAgro, 2 Place Viala, 34060, Montpellier Cedex 1, France. plassard@supagro.inra.fr.
D000363:Agaricales / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D025301:Hyphae / Q000254:growth & development / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D025282:Mycelium / Q000254:growth & development / Q000378:metabolism*; D038821:Mycorrhizae; D028061:Phosphate Transport Proteins / Q000235:genetics / Q000378:metabolism*; D010710:Phosphates / Q000378:metabolism / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D028061:Phosphate Transport Proteins; D010710:Phosphates