A new Geopora species (Pyronemataceae), Geopora ramila was described and illustrated from the soil, under or in the vicinity of Helianthemum ledifolium var. ledifolium annual plant in Fars province, Iran. Morphologically, G. ramila is similar to G. pinyonensis and G. arenicola but distinguished from both by a combination of morphological characters including, color and size of ascocarps, size and shape of ascospores, habit and associated host. The ribosomal DNA internally transcribed spacer (rDNA ITS) sequence of the new species (Acc. No. MT108930 to MT108934) showed 87.82% identity with G. pinyonensis in the BLAST search in GenBank. ITS-based phylogenetic analysis clearly supports G. ramila is a new and distinctive species lacking close relatives among described species of Geopora.
Truffle farming does not exist in Iran, and no formal studies have been conducted on its culture. For commercial production of Tirmania pinoyi, it is necessary to accurate detection of it in the roots of the host plants. In this study, for the first time, mycorrhizal association of T. pinoyi with several perennial Cistaceae including Helianthemum lippii, H. almeriense, Cistus albidus, and Cistua ladanifer were investigated. In all plant species inoculated with Tirmania pinoyi, an ectendomycorrhizal association with varying degrees of sheath development was observed. Our results shows that H. lippii is suitable plant host for Tirmania pinoyi, and mycorrhization rate and the relative mycorrhizal dependency (RMD) in this native plant was about 90% and 57.85%, respectively, and higher than the other three plant species. The effect of the mycorrhizal fungus on host growth (root weight, root height, shoot weight, shoot height, plant height, and plant weight) was statistically significant compared to non-inoculated plants. To detect the roots colonized by T. pinoyi, a PCR based diagnostic method was developed with the species-specific primer pairs FTiPi and RTiPi designed from the sequence of the internal transcribed space regions (ITS1 and ITS2). The specificity of the primers was verified by PCR analysis of DNA from T. pinoyi specimens and other desert truffles. The method detected T. pinoyi in artificialy infected root plants, and no cross-reactions were observed with any other tested desert truffles. This species-specific PCR method is suitable for quick, simple, and reliable detection of T. pinoyi mycorrhizas.
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