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Arbuscular mycorrhizal fungi (AMF) are an important element of the plant microbiome as they establish an endosymbiotic relationship with the roots of most plant species. This association enhances access to nutrients and water for plants, and provides the fungus with plant-derived organic carbon. In this chapter, I describe a range of methods to work with AMF including: soil sampling; isolation of AMF propagules (spores, sporocarps, roots, and mycelium) by a wet sieving and centrifugation in a sucrose solution; trap (from field soil with AMF spores) and one-species pot cultures (from AMF spores divided into morphotypes); staining of mycorrhizae in plant roots; and production of diagnostic slides. These methods are widely used in taxonomic and ecological studies to characterize the morphology of AMF.

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Basil (Ocimum L.) species are used as medicinal plants due to their essential oils exhibiting specific biological activity. The present work demonstrated that both the variety and season/conditions of cultivation had a significant effect on (i) the produced amount (extraction yield), (ii) qualitative, as well as (iii) quantitative profile of basil essential oil. Among studied basil varieties, a new variety, 'Mánes', was characterized for the first time. Based on our quantitative evaluation of GC-MS profiles, the following chemotypes and average concentrations of a main component were detected in the studied basil varieties: 'Ohre', 'Lettuce Leaf', 'Purple Opaal', 'Dark Green' (linalool, 5.99, 2.49, 2.34, 2.01 mg/mL, respectively), and 'Mammolo Genovese', 'Mánes', 'Red Rubin' (eucalyptol, 1.34, 0.96, 0.76 mg/mL, respectively). At the same time, when considering other compounds identified in GC-MS profiles, all the studied varieties, except from 'Lettuce Leaf', were methyl eugenol-rich with a strong dependence of the eugenol:methyl eugenol ratio on the seasonal changes (mainly solar irradiation, but also temperature and relative humidity). More complex and/or variable (depending on the season and cultivation) chemotypes were observed with 'Lettuce Leaf' (plus estragole, 2.27 mg/mL), 'Dark Green' (plus eucalyptol, 1.36 mg/mL), 'Mammolo Genovese' (plus eugenol, 1.19 mg/mL), 'Red Rubin' (plus linalool and eugenol, 0.46 and 0.56 mg/mL, respectively), and 'Mánes' (plus linalool and eugenol, 0.58 and 0.40 mg/mL, respectively). When considering superior extraction yield (ca. 17 mL·kg-1, i.e., two to five times higher than other examined varieties) and consistent amounts (yields) of essential oil when comparing inter-seasonal or inter-year data (RSD and inter-year difference in mean yield values ˂2.5%), this new basil variety is very promising for use in the pharmaceutical, food, and cosmetic industries.

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The aim of this work was to evaluate the occurrence of arbuscular mycorrhizal fungi (AMF) species diversity in soil samples from the Amazon region under distinct land use systems (Forest, Old Secondary Forest, Young Secondary Forest, Agroforestry systems, Crops and Pasture) using two distinct trap cultures. Traps established using Sorghum sudanense and Vigna unguiculata (at Universidade Regional de Blumenau - FURB) and Brachiaria decumbens and Neonotonia wightii (at Universidade Federal de Lavras - UFLA) were grown for 150 days in greenhouse conditions, when spore density and species identification were evaluated. A great variation on species richness was detected in several samples, regardless of the land use systems from where samples were obtained. A total number of 24 AMF species were recovered using both methods of trap cultures, with FURB';s traps yielding higher number of species. Acaulospora delicata, A. foveata, Entrophospora colombiana and two undescribed Glomus species were the most abundant and frequent species recovered from the traps. Number of species decreased in each genus according to this order: Acaulospora, Glomus, Entrophospora, Gigaspora, Archaeospora, Scutellospora and Paraglomus. Spore numbers were higher in Young Secondary Forest and Pastures. Our study demonstrated that AMF have a widespread occurrence in all land use systems in Amazon and they sporulate more abundantly in trap cultures from land uses under interference than in the pristine Forest ecosystem.

O objetivo deste trabalho foi avaliar a ocorrência de espécies de fungos micorrízicos arbusculares (FMAs) em amostras de solos da região Amazônica sob diferentes sistemas de uso da terra (Floresta, Capoeira Velha, Capoeira Nova, Sistema Agroflorestal, Culturas e Pastagens) usando dois métodos de culturas armadilhas. Culturas armadilhas estabelecidas com Sorghum sudanense e Vigna unguiculata (na Universidade Regional de Blumenau - FURB) e Brachiaria decumbens e Neonotonia wightii (na Universidade Federal de Lavras - UFLA) foram cultivadas por 150 dias em condições de casa-de-vegetação e avaliadas para o número de esporos e identificação das espécies. Uma grande variação na riqueza de espécies foi detectada em várias amostras, independente do sistema de uso da terra de onde as amostras foram obtidas. Um total de 24 espécies de FMAs foram recuperadas usando ambas metodologias de culturas armadilhas e as culturas estabelecidas na FURB produziram um número maior de espécies. Acaulospora delicata, A. foveata, Entrophospora colombiana e duas espécies não descritas de Glomus foram as espécies mais abundantes e freqüentes recuperadas das culturas armadilhas. O número de espécies diminui em cada gênero na seguinte ordem: Acaulospora, Glomus, Entrophospora, Gigaspora, Archaeospora, Scutellospora e Paraglomus. Número de esporos foi maior na Capoeira jovem e Pastagens. Nosso estudo demonstrou que os FMAs possuem uma ampla dispersão em todos os sistemas de uso na Amazônia e que eles esporulam mais abundantemente em culturas armadilhas estabelecidas de solos sob interferência antrópica do que em solos de Floresta nativa.

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The aim of this work was to evaluate the occurrence of arbuscular mycorrhizal fungi (AMF) species diversity in soil samples from the Amazon region under distinct land use systems (Forest, Old Secondary Forest, Young Secondary Forest, Agroforestry systems, Crops and Pasture) using two distinct trap cultures. Traps established using Sorghum sudanense and Vigna unguiculata (at Universidade Regional de Blumenau -FURB) and Brachiaria decumbens and Neonotonia wightii (at Universidade Federal de Lavras - UFLA) were grown for 150 days in greenhouse conditions, when spore density and species identification were evaluated. A great variation on species richness was detected in several samples, regardless of the land use systems from where samples were obtained. A total number of 24 AMF species were recovered using both methods of trap cultures, with FURB's traps yielding higher number of species. Acaulospora delicata, A. foveata, Entrophospora colombiana and two undescribed Glomus species were the most abundant and frequent species recovered from the traps. Number of species decreased in each genus according to this order: Acaulospora, Glomus, Entrophospora, Gigaspora, Archaeospora, Scutellospora and Paraglomus. Spore numbers were higher in Young Secondary Forest and Pastures. Our study demonstrated that AMF have a widespread occurrence in all land use systems in Amazon and they sporulate more abundantly in trap cultures from land uses under interference than in the pristine Forest ecosystem.