Elsevier

Pedosphere

Volume 26, Issue 4, August 2016, Pages 561-566
Pedosphere

Araucaria angustifolia Aboveground Roots Presented High Arbuscular Mycorrhizal Fungal Colonization and Diversity in the Brazilian Atlantic Forest

https://doi.org/10.1016/S1002-0160(15)60065-0Get rights and content

Abstract

Almost 30 different arbuscular mycorrhizal fungi (AMF) species, distributed in different genera such as Glomus, Acaulospora, Scutellospora, Entrophospora, Ambispora, Kuklospora, Gigaspora, and Archeospora, have been identified in the root zone of Araucaria angustifolia, known as Brazil Pine. During our AMF survey in this ecosystem, our attention was called to the presence of many superficially growing Araucaria roots. Our hypothesis was that these roots were colonized with AMF because of the presence of AMF spores in organic material aboveground. Samples of these superficial roots and the organic substrate they were growing on were evaluated for their mycorrhizal status. DNA was extracted from the AMF-colonized superficial roots and submitted to polymerase chain reaction (PCR) amplification using the NS31-AM1 primer pair, followed by cloning and sequencing. We found that the root colonization percentages were between 31% and 52%, and the number of AMF spores in the substrate ranged from 27 to 164 spores per 50 g dry substrate. The phylogenetic analyses and tree construction using maximum parsimony (MP) and neighbor-joining (NJ) methods identified 13 different species of the phylum Glomeromycota belonging to the genera Glomus, Funneliformis, Rhizophagus, Gigaspora, Acaulospora, and Archaeospora, and five isolates were identified only at the genus level. To our knowledge, this is the first report on Araucaria angustifolia with roots growing aboveground, producing runner roots that develop on dead tree trunks and organic material. The higher colonization of the aboveground roots than those commonly found in belowground Araucaria roots suggests that they may present active metabolic uptake of nutrients.

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      The product was used as template DNA in a second reaction (Nested-PCR) with primers AM1 (Helgason et al., 1999) and NS31 (Simon et al., 1992). Both reactions were performed in Viriti® (Applied Biosystems), programmed to operate with an initial denaturation at 94 °C for 5 min, 35 cycles at 94 °C for 30 s, 55 °C for 30 s and 72 °C for 1 min and 30 s, followed by a final extension of 72 °C for 5 min, generating final product sizes of 1700 (NS1-NS8) (Vilela et al., 2007) and 580 (AM1-NS31) bp (Bonfim et al., 2016; Moreira et al., 2016) respectively. To confirm the amplification, PCR products were submitted to electrophoresis in agarose gel (2.0%), stained with GelRed™, visualised in a transilluminator (DNR, Bio-Imaging Systems/MiniBis Pro) under UV light and photo documented (Fig. S1, A,B).

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