title: Identification of mycorrhiza-regulated genes with arbuscule development-related expression profile.
abstract: Suppressive subtractive hybridisation was applied to the analysis of late stage arbuscular mycorrhizal development in pea. 96 cDNA clones were amplified and 81, which carried fragments more than 200 nt in size, were sequence analysed. Among 67 unique fragments, 10 showed no homology and 10 were similar to sequences with unknown function. RNA accumulation of the corresponding 67 genes was analysed by hybridisation of macro-arrays. The cDNAs used as probes were derived from roots of wild type and late mutant pea genotypes, inoculated or not with the AM fungus Glomus mosseae. After calibration, a more than 2.5-fold mycorrhiza-induced RNA accumulation was detected in two independent experiments in the wild type for 25 genes, 22 of which seemed to be induced specifically during late stage AM development. Differential expression for 7 genes was confirmed by RT-PCR using RNA from mycorrhiza and from controls of a different pea cultivar. In order to confirm arbuscule-related expression, the Medicago truncatula EST data base was screened for homologous sequences with putative mycorrhiza-related expression and among a number of sequences with significant similarities, a family of trypsin inhibitor genes could be identified. Mycorrhiza-induced RNA accumulation was verified for five members by real-time PCR and arbuscule-related activation of the promoter could be shown in transgenic roots for one of the genes, MtTi 1.
authors.affiliation: Max-Planck Institut für terrestrische Mikrobiologie, and Laboratorium für Mikrobiologie, Philipps-University, Karl-von-Frisch-Strasse, Marburg, Germany.
authors.forename: Ulf
authors.initials: U
authors.lastname: Grunwald
authors.forename: Oyunbileg
authors.initials: O
authors.lastname: Nyamsuren
authors.forename: M'Barek
authors.initials: M
authors.lastname: Tamasloukht
authors.forename: Laurence
authors.initials: L
authors.lastname: Lapopin
authors.forename: Anke
authors.initials: A
authors.lastname: Becker
authors.forename: Petra
authors.initials: P
authors.lastname: Mann
authors.forename: Vivienne
authors.initials: V
authors.lastname: Gianinazzi-Pearson
authors.forename: Franziska
authors.initials: F
authors.lastname: Krajinski
authors.forename: Philipp
authors.initials: P
authors.lastname: Franken
mesh_terms: D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D046913:Medicago truncatula / Q000235:genetics / Q000382:microbiology; D038821:Mycorrhizae / Q000254:growth & development*; D018532:Pisum sativum / Q000235:genetics* / Q000382:microbiology; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D018749:RNA, Plant