Isolation and study of morphology and phylogeny of arbuscular mycorrhizal fungi coexisting with the roots of some medicinal plants in Kerman province

Abstract

Objective Plants that have a symbiotic relationship with arbuscular mycorrhizal fungi (AMF) compared with non-mycorrhizal plants have a higher ability to resist against various biotic and abiotic environmental stresses such as drought, salinity, pests, and plant diseases. This study aim is studying different species of AMFs in Kerman province. The identification of these microorganisms is to provide the basis for further studies on the use of these microorganisms in germination, growth, and proliferation of medicinal plants. Methods To molecular and morphological identification of arbuscular mycorrhizal fungi, sampling was done from the rhizosphere of medicinal plants in some areas of Kerman province. Arbuscular mycorrhizal fungi spores were isolated from the sampled soils by the wet sieve method. The grouping was done based on spore microscopic slides and morphological characteristics such as spore color, shape, surface ornamentation, size, wall structure and spore characteristics in water. DNA was extracted from single spores. A Part of ribosomal DNA was amplified by nested PCR using SSUmaf and LSUmAr primers in the first step, SSUmCf, and LSUmBr in the second step. After horizontal agarose gel electrophoresis and see the related band the PCR product was sent for sequencing. Then sequencing results were analyzed using BLAST, Gene Runner and Clustal Omega Softwares, and the phylogenetic tree was plotted using MEGA 7 software. Results According to morphological and molecular studies, Viscospora viscosa, Septoglomus deserticola, Funneliformis caledonium, Funneliformis mosseae, and Diversispora spurca were identified. Finally, the existence and manner of species identity were investigated in the phylogeny tree. Conclusion Since the morphological features used in the identification and classification of mycorrhizal fungi are limited, molecular methods can be used for safer classification, and in this way, Recognize differences of Similar species morphologically and similarity of different species in appearance.

Keywords

• : classification
• nested pcr
• primer
• ribosomal dna
• symbiosis