Open AccessExpressed genes in the extraradical hyphae of an arbuscular mycorrhizal fungus, Glomus intraradices , in the symbiotic phase
Author(s) -
Sawaki Hiromichi,
Saito Masanori
Publication year - 2001
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2001.tb10506.x
Subject(s) - hypha , biology , fungus , glomus , complementary dna , mycorrhiza , rapid amplification of cdna ends , botany , microbiology and biotechnology , symbiosis , gene , biochemistry , bacteria , spore , molecular cloning , genetics
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.