Mycorrhizal fungi arbuscular in organic and conventional sugarcane systems.

Organic production systems are increasingly gaining market share; however, there are still few studies on their influence on the activity of soil microorganisms in sugarcane. Arbuscular mycorrhizal fungi are extremely sensitive to environmental changes, and their activity can be used as a parameter of comparison and quality between organic and conventional systems. The objective of this work was to evaluate mycorrhizal activity in different varieties of sugarcane under two production systems. This work was carried out in a commercial plantation of the Jalles Machado plant in the municipality of Goianésia in Goiás, Brazil. The values of spore density in the soil, mycorrhizal colonization rate in the roots and easily extractable glomalin were evaluated, and the associated fungal species were identified. There was no effect of sugarcane variety on the number of spores or the glomalin content in the soil. The conventional system presented significantly lower mycorrhizal colonization rates than did the organic system. The varieties cultivated under the conventional planting system showed a greater diversity of arbuscular mycorrhizal fungi, where 12 of the 13 different species of mycorrhizal fungi found in both cultivation systems occurred.

Association of mycoheterotrophic Gentianaceae with specific Glomus lineages.

Some plant species took an alternative evolutionary pathway in which they lost their photosynthetic capacity to depend exclusively on carbon supplied by arbuscular mycorrhizal fungi (AMF) in an association called mycoheterotrophy. Among them is Voyriella parviflora, a species of the family Gentianaceae, which is found in tropical regions such as the Amazon basin. Here, we assessed the identity of AMF symbionts associated with this species. DNA was isolated from eight Gentianaceae specimens and from litter and surrounding roots of photosynthetic plants. The atp1 gene was amplified by Sanger sequencing to determine the taxonomic affiliation of the mycoheterotrophic plants. A 280 bp region of the 18S rRNA gene of AMF was amplified with primers NS31/AML2 by high-throughput sequencing. The mycoheterotrophic specimens were assigned to V. parviflora with a bootstrap support of 72%. Glomus was the most abundant AMF genus, both in the mycoheterotrophic plants and in the litter and roots of photosynthetic plants. In addition, a few Glomus genotypes were abundantly enriched in the mycoheterotrophic plants, with only a few specimens colonized by Gigaspora, Acaulospora, and Scutellospora in a low proportion. These genotypes formed a cluster within a larger clade, suggesting that V. parviflora shows a preferential association with a narrow Glomus lineage which is not phylogenetically close to a previously identified V. parviflora's associated lineage. Furthermore, detecting fungi from other families suggests that V. parviflora is colonized by other genera, although with low frequency. These findings provide new insights into the association between AMF and mycoheterotrophic species and highlight the importance of considering trap culture-independent approaches in understanding this symbiosis.

Pioneer Tree Bellucia imperialis (Melastomataceae) from Central Amazon with Seedlings Highly Dependent on Arbuscular Mycorrhizal Fungi.

Bellucia imperialis is one of the most abundant pioneer tree species in anthropized areas of the Central Amazon, and has ecological importance for the environmental resilience of phosphorus (P)-depleted areas. Thus, we investigated whether B. imperialis depends on symbiosis with arbuscular mycorrhizal fungi (AMF) to grow and establish under the edaphic stresses of low nutrient content and low surface moisture retention capacity of the substrate. We tried three AMF inoculation treatments: (1) CON-no mycorrhizae; (2) MIX-with AMF from pure collection cultures, and (3) NAT-with native AMF, combined with five doses of P via a nutrient solution. All CON treatment seedlings died without AMF, showing the high mycorrhizal dependence of B. imperialis. Increasing P doses significantly decreased the leaf area and shoot and root biomass growth for both the NAT and MIX treatments. Increasing P doses did not affect spore number or mycorrhizal colonization, but decreased the diversity of AMF communities. Some species of the AMF community showed plasticity, enabling them to withstand shortages of and excess P. B. imperialis was shown to be sensitive to excess P, promiscuous, dependent on AMF, and tolerant of scarce nutritional resources, highlighting the need to inoculate seedlings to reforest impacted areas.

Arbuscular mycorrhizal fungi in the soil using cover crops with and without nitrogen addition.

This study aimed to evaluate the occurrence of mycorrhizal fungi and glomalin content in soil under different cover crops with and without the application of nitrogen in the cover. The following cover plants were used: Crotalaria juncea (Crotalaria juncea L.), wild beans from Ceará (Canavalia brasiliensis Mart. ex Benth.), Guandú 'BRS mandarin' [Cajanus cajan (L.) Millsp.], millet 'BR05' [Pennisetum glaucum (L.) R.Br.] and sorghum 'BR 304' [Sorghum bicolor (L.) Moench]. The absolute control of the experiment was the treatment without the use of cover crops, that is, the vegetation of spontaneous occurrence in the area. The experimental design was randomized blocks in subplots with three replications. Spore density, mycorrhizal colonization rate, easily extractable glomalin, and species present in the rhizosphere of the cover crops were determined. No differences were found in the diversity of mycorrhizal fungi associated with the different cover crops studied or in the values of spore density, root colonization, or glomalin content. Nitrogen application did not influence the mycorrhizal activity in the investigated cover crops. The most frequent species associated with cover crops were Scutellospora pellucida and Scutellospora persica in C. juncea; Gigaspora sp. on Sorghum; Glomus macrocarpum in Guandu; G. macrocarpum and Glomus clavisporum in millet; and Glomus microaggregatum and Glomus tortuosum in Spontaneous Vegetation.

Arbuscular mycorrhizal fungi community in coffee agroforestry, consortium and monoculture systems.

Understanding the effects of different production systems on arbuscular mycorrhizal fungi (AMF) can help to interpret interactions between their components and to define management strategies. As a result, our study was conducted on soils under three coffee production systems (one homogeneous and two heterogeneous) and in a native forest located in the Bahia state, Brazil. This study aimed to answer the following questions: 1) Does the organization and management of the coffee production system affect the occurrence and diversity of AMF?; and 2) Is the seasonality effect similar between systems? To do so, soil samples (0-10 cm depth) were collected at two times of the year (rainy and dry). Number of spores (NS) and average richness did not show differences between the systems, only between seasons. There was a reduction in NS in the dry season (1.4 and 2.7 spores g-1 soil) in relation to the rainy season (3.8 to 12.5 spores g-1 soil). The influence of coffee production systems was observed in the presence and absence of some AMF species. The AMF community was shown to be related to the plant species composition of the system, which was reflected in the dissimilarity of heterogeneous systems in relation to the coffee monoculture system.

Development of a taxon-discriminating molecular marker to trace and quantify a mycorrhizal inoculum in roots and soils of agroecosystems.

Crop inoculation with Glomus cubense isolate (INCAM-4, DAOM-241198) promotes yield in banana, cassava, forages, and others. Yield improvements range from 20 to 80% depending on crops, nutrient supply, and edaphoclimatic conditions. However, it is difficult to connect yield effects with G. cubense abundance in roots due to the lack of an adequate methodology to trace this taxon in the field. It is necessary to establish an accurate evaluation framework of its contribution to root colonization separated from native arbuscular mycorrhizal fungi (AMF). A taxon-discriminating primer set was designed based on the ITS nrDNA marker and two molecular approaches were optimized and validated (endpoint PCR and quantitative real-time PCR) to trace and quantify the G. cubense isolate in root and soil samples under greenhouse and environmental conditions. The detection limit and specificity assays were performed by both approaches. Different 18 AMF taxa were used for endpoint PCR specificity assay, showing that primers specifically amplified the INCAM-4 isolate yielding a 370 bp-PCR product. In the greenhouse, Urochloa brizantha plants inoculated with three isolates (Rhizophagus irregularis, R. clarus, and G. cubense) and environmental root and soil samples were successfully traced and quantified by qPCR. The AMF root colonization reached 41-70% and the spore number 4-128 per g of soil. This study demonstrates for the first time the feasibility to trace and quantify the G. cubense isolate using a taxon-discriminating ITS marker in roots and soils. The validated approaches reveal their potential to be used for the quality control of other mycorrhizal inoculants and their relative quantification in agroecosystems.

Mycorrhizal networks facilitate the colonization of legume roots by a symbiotic nitrogen-fixing bacterium.

Arbuscular mycorrhizal fungi (AMF) absorb and translocate nutrients from soil to their host plants by means of a wide network of extraradical mycelium (ERM). Here, we assessed whether nitrogen-fixing rhizobia can be transferred to the host legume Glycine max by ERM produced by Glomus formosanum isolate CNPAB020 colonizing the grass Urochloa decumbens. An H-bridge experimental system was developed to evaluate the migration of ERM and of the GFP-tagged Bradyrhizobium diazoefficiens USDA 110 strain across an air gap compartment. Mycorrhizal colonization, nodule formation in legumes, and occurrence of the GFP-tagged strain in root nodules were assessed by optical and confocal laser scanning microscopy. In the presence of non-mycorrhizal U. decumbens, legume roots were neither AMF-colonized nor nodulated. In contrast, G. formosanum ERM crossing the discontinuous compartment connected mycorrhizal U. decumbens and G. max roots, which showed 30-42% mycorrhizal colonization and 7-11 nodules per plant. Fluorescent B. diazoefficiens cells were detected in 94% of G. max root nodules. Our findings reveal that, besides its main activity in nutrient transfer, ERM produced by AMF may facilitate bacterial translocation and the simultaneous associations of plants with beneficial fungi and bacteria, representing an important structure, functional to the establishment of symbiotic relationships.

Vegetative compatibility and anastomosis formation within and among individual germlings of tropical isolates of arbuscular mycorrhizal fungi (Glomeromycota).

Hyphal anastomoses which play a key role in the formation of interconnected mycorrhizal networks and in genetic exchange among compatible individuals have been studied in a limited number of species and isolates of arbuscular mycorrhizal fungi (AMF), mainly in symbiotic mycelium. In this work, the occurrence and frequency of anastomosis between hyphae of the same and different germlings were assessed in tropical isolates belonging to Acaulospora, Claroideoglomus, Gigaspora, Glomus, Rhizophagus and Scutellospora. Germlings belonging to Acaulospora, Claroideoglomus, Glomus and Rhizophagus formed perfect hyphal fusions, with frequencies ranging from 9.29 ± 3.01 to 79.84 ± 4.39 % within the same germling and from 14.02 ± 7.36 to 91.41 ± 3.92 % between different germlings. Rare fusions, occurring within the same hypha, were detected in Gigaspora species, and no anastomoses were observed in Scutellospora species. The consistent detection of nuclei in perfect fusions suggests that nuclear migration is active both within and between germlings. Present data on anastomosis formation, nuclear migration and germling viability in tropical isolates of AMF widen our knowledge on the extensive and consistent occurrence of successful hyphal fusions in this group of beneficial symbionts. The ability to anastomose and establish protoplasm flow, fundamental for the maintenance of physiological and genetic continuity, may produce important fitness consequences for the obligately biotrophic AMF.

Absorção de fósforo em doze genótipos de milho inoculados com fungo micorrízico arbuscular em solo de cerrado / Phosphorus absorption in twelve corn genotypes inoculated with arbuscular mycorrhizal fungus in cerrado soil

Este trabalho foi realizado com o objetivo de avaliar a eficiência de absorção de fósforo em doze genótipos de milho na presença e na ausência de inoculação com fungo micorrízico arbuscular. Para tanto, foi instalado um experimento em casa de vegetação no Centro de Ciências Agrárias e Biológicas da Universidade Federal de Goiás, em Jataí, Goiás, com delineamento em blocos ao acaso com três repetições em fatorial 12x2x2. Os genótipos foram: AG9010, P30K75, AG8060, P30P70, AG405, P30F33, AG7000, BANDEIRANTE, EMGOPA501, BALU178, DINA657 e 2C599, cultivados sob duas doses de fósforo (10 e 100mg kg-1 de P na solução do solo) na presença ou na ausência de micorrização com Glomus etunicatum. Foi verificada diferença entre os genótipos na produção de matéria seca na raiz, na razão entre matéria seca na raiz e matéria seca na parte aérea, na colonização com fungo micorrízico e na razão fósforo acumulado na parte aérea por matéria seca na raiz. A micorrização alterou a classificação dos genótipos quanto à produção de matéria seca de parte aérea sob baixo P e à resposta ao aumento do suprimento de P.

The aim of this research was to evaluate the efficiency of phosphorus absorvation in twelve corn genotypes. The study was performed in presence and absence of arbuscular mycorrhizal fungus inoculation. The experiment took place at the Agrarian and Biological Center greenhouse, in the Federal University of Goiás, Jataí, Brazil. The genotypes were randomly distributed in blocks with three replicates in a 12x2x2 factorial. The genotypes were AG9010, P30K75, AG8060, P30P70, AG405, P30F33, AG7000, BANDEIRANTES, EMGOPA501, BALU178, DINA657, 2C599, cultivated under two doses of phosphorus (10 and 100mg kg-1 of P in soil solution) in presence or absence of Glomus etunicatum mycorrization. The study verified the difference among genotypes in dry root matter production. The ratio among root dry and shoot dry matter colonized with mycorrizal fungus was observed. Ratio accumulated phosphorus among shoot and dry root matter was also seen. The mycorrization changed classification of genotypes regarding to dry matter shoot production under low P and the response for higher P supply.

Arbuscular mycorrhizal fungi in a semiarid copper mining area in Brazil.

The occurrence of arbuscular mycorrhizal fungi (AMF) in a copper mining area was investigated. Soil samples were collected from six sites at the Mineração Caraiba, Bahia State, northeastern Brazil, comprising: (1) a site that receives the waste product; (2) a site that receives low grade deposits; (3) the interface between the caatinga and site 1; (4) the surroundings of the industrial area; (5) the site for extracting topsoil for land filling; (6) the preserved caatinga. Thirty-two plant species were identified around the collection locations. Trap cultures were maintained in the greenhouse for 3 months, using bahia grass ( Paspalum notatum Flügge) as the host plant. Spores were extracted from soil and 21 AMF species (15 Glomus and one of each of Acaulospora, Archaeospora, Entrophospora, Gigaspora , Paraglomus and Scutellospora) were identified. In site 1, plants or AMF were not found during the dry season. Site 6, with native vegetation, had the highest number of plants and AMF species. The disturbed sites showed less plant diversification, with the community of AMF being quantitative and qualitatively affected by disturbance.