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 ۳ 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, ۴ for T. pinoyi, and ۲ 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 ۱۰-۵۰۰ pg DNA.  ITS-Tfc۱ primers detected the species in host tissues using a nested-PCR method by ITS۱ and ITS۴ universal primers as external set.