Specific Primers For Identification and Detection of Three Major Desert Truffle Species in Iran with Special Reference to Terfezia claveryi

ABSTRACT- Desert truffles are hypogeous ascomyceteous ectomycorrhizal fungi, occurring in arid and semi-arid ecosystems. A PCR-based method was developed for the identification of 3 major desert truffles of Iran: Terfezia claveryi, Tirmania pinoyi and Tirmania nivea based on internal transcribed spacers of rDNA. Two specific PCR primers were designed for T. claveryi, 4 for T. pinoyi, and 2 for T. nivea. All primers were specific to their homologous species and neither set amplified purified DNA from other truffle species or soil-borne micromycetes. In conventional PCR, the limits of detection were 10-500 pg DNA.  ITS-Tfc1 primers detected the species in host tissues using a nested-PCR method by ITS1 and ITS4 universal primers as external set.