Similar mycorrhizal fungal communities associated with epiphytic and lithophytic orchids of Coelogyne corymbosa

Mycorrhizal fungi are essential for the growth and development of both epiphytic(growing on trees)and lithophytic(growing on rocks)orchids.Previous studies indicate that in lowland tropical areas,orchid mycorrhizal fungal compositions are correlated with the life form(i.e.,epiphytic,lithophytic,or terrestrial)of their host plants.We therefore tested if a similar correlation exists in an orchid distributed at higher elevations.Coelogyne corymbosa is an endangered ornamental orchid species that can be found as a lithophyte and epiphyte in subtropical to subalpine areas.Based on high-throughput sequencing of the fungal internal transcribed spacer 2(ITS2)-rDNA region of mycorrhizae of C.corymbosa,we detected 73 putative mycorrhizal fungal Operational Taxonomic Units(OTUs).The OTUs of two dominant lineages(Cantharellales and Sebacinales)detected from C.corymbosa are phylogenetically different from those of other species within the genus Coelogyne,indicating that different orchid species prefer specific mycorrhizal fungi.We also found that the Non-metric multidimensional scaling(NMDS)plots of orchid mycorrhizal fungi were not clustered with life form,the variations among orchid mycorrhizal fungal communities of different life forms were not significant,and most of the OTUs detected from epiphytic individuals were shared by the lithophytic plants,suggesting that orchid mycorrhizal associations of C.corymbosa were not affected by life form.These findings provide novel insights into mycorrhizal associations with endangered ornamental orchids.

Screening for differentially expressed genes in Anoectochilus roxburghii (Orchidaceae) during symbiosis with the mycorrhizal fungus Epulorhiza sp.

Mycorrhizal fungi promote the growth and development of plants, including medicinal plants. The mechanisms by which this growth promotion occurs are of theoretical interest and practical importance to agriculture. Here, an endophytic fungus （AR-18） was isolated from roots of the orchid Anoectochilus roxburghii growing in the wild, and identified as Epulorhiza sp. Tissue-cultured seedlings of A. roxburghii were inoculated with AR-l 8 and co-cultured for 60 d. Endotrophic mycorrhiza formed and the growth of A. roxburghii was markedly promoted by the fungus. To identify genes in A. roxburghii that were differentially expressed during the symbiosis with AR-18, we used the differential display reverse transcription polymerase chain reac- tion （DDRT-PCR） method to compare the transcriptomes between seedlings inoculated with the fungus and control seedlings. We amplified 52 DDRT-PCR bands using 15 primer combinations of three anchor primers and five arbitrary primers, and nine bands were re-amplified by double primers. Reverse Northern blot analyses were used to further screen the bands. Five clones were up-regulated in the symbiotic interaction, including genes encoding a uracil phosphoribosyltransferase （UPRTs; EC 2.4.2.9） and a hypothetical protein. One gene encoding an amino acid transmembrane transporter was down-regulated, and one gene encoding a tRNA-Lys （trnK） and a maturase K （matK） pseudogene were expressed only in the inoculated seedlings. The possible roles of the above genes, especially the UPRTs and marK genes, are discussed in relation to the fungal interaction. This study is the first of its type in A. roxburghii.