Generation of RAPD‐PCR primers for the identification of isolates of Glomus mosseae, an arbuscular mycorrhizal fungus

Abstract

Mycorrhizal fungi are usually identified on the basis of the morphological characters shown by fruit bodies, spores, vegetative mycelia or symbiotic structures. The development of molecular techniques provides a valuable and alternative approach to identify mycorrhizal fungi, especially when it is difficult to gather a sufficient number of data on morphological features. Short arbitrary oligonucleotides were used as primers for the amplification of genomic DNA extracted from spores of arbuscular fungi. The RAPD fingerprints showed banding patterns which allowed us to distinguish between species and even isolates within Glomales. In order to identify mycorrhizal fungi during their symbiotic phase, a nonpolymorphic RAPD band identified as marker for some isolates of Glomus mosseae was purified from agarose gels and cloned in a bluescript vector. The fragment was sequenced and specific primers (PO-M3) were designed for the mycorrhizal fungus. They specifically and successfully amplified the DNA not only from G. mosseae spores, but also from roots of pea, clover, leek and onion plants when they were colonized by G. mosseae isolates.

Tansley Review No. 82. Strategies of arbuscular mycorrhizal fungi when infecting host plants

“…27). The first contributions of molecular biology to the field of arbuscular mycorrhizas have covered topics more related to taxonomy, molecular ecology and phylogeny (Simon et al, 1993;Wyss & Bonfante, 1993;Lanfranco et al, 1994). The use of molecular techniques to search for functional genes controlling fungal morphogenesis, the infection process and the metabolism of mycorrhizal roots is still in its infancy.…”

Quantificação de micélio extrarradicular de fungos micorrízicos arbusculares em plantas cítricas: II. Comparação entre diferentes espécies cítricas e endófitos

“…Isto vem comprovar que a micorrizosfera difere profundamente da rizosfera de plantas não micorrizadas, como já discutido por Silveira (1992), e que a única maneira realmente confiável para identificar espécies de FMAs seria por meio de métodos como imunofluorescência (Kough et al, 1983;Wilson et al, 1983) ou moleculares (Lanfranco et al, 1993;Di Bonito et al, 1995;Lanfranco et al, 1995). Para todos os FMAs de laranja caipira, observou-se redução acentuada de comprimento de MEA com o aumento das doses de P, cuja causa pode estar associada aos nutrientes acrescentados quando da adubação com KH 2 PO 4 , principalmente o K + .…”

Molecular identification of arbuscular mycorrhizal fungi in roots: Perspectives and problems

“…In only a few studies, other regions were used, e.g, a RAPD fragment of unknown function (LANFRANCO et al 1995). The first three 18S small subunit (SSU) sequences from ribosomal DNA of these fungi were determined by SIMON et al (1992).…”