Identification of Arbuscular Mycorrhizal Fungi Associated with Different Plants in Rafsanjan Based on Morphological Characteristics and ß-tubulin Gene Sequence

Abstract

Introduction: Arbuscular Mycorrhizal Fungi (AMF), as the most abundant mutual symbiotic association between fungi and plants in the soil, are of great ecological and economic importance. The present study aimed to identify AMF species associated with different plant roots based on morphological and molecular characteristics of ß-tubulin gene sequences of collected samples in Raviz and Daveran regions of Rafsanjan city. Materials and methods: Samples were collected from the rhizosphere of different plants and AMF spores were isolated from the soil samples using the wet sieve method. The identification was performed based on morphological characteristics of the spores such as the color, shape, surface ornamentation, size, wall structure as well as characterization of the spores in water. The DNA of the fungi was extracted from single spores and partial sequences of ß-tubulin gene were amplified by three nestedPCR using appropriate primers. The PCR products of the selected samples were sent for Sanger sequencing. The phylogenetic relationship of the selected sequences with other available isolates in GenBank was analyzed using the Maximum likelihood method. Results: Based on morphological characteristics and phylogenetic analysis of the partial sequence of ß-tubulin gene, the following species were detected: Funneliformis mosseae, Funneliformis coronatum, Gigaspora gigantean, Scutellospora calospora, andSeptoglomus constrictum. F. mosseae followed by F. coronatum were recognized as the dominant species in the present study. Discussion and conclusion: The partial sequence of ß-tubulin gene was well separated from the studied orders, families, and genera of arbuscular mycorrhiza fungi. All the identified species in this study were placed in the expected taxonomic level. The results of the molecular analysis confirmed the classification of species based on morphological characterization.

Keywords

• symbiosis
• arbuscular mycorrhizal fungi
• phylogeny
• alignment
• ß-tubulin