Notes on Acaulospora bireticulata Rothwell & Trappe and first record of Acaulospora koskei Blask. from Brazil)

(Notes on Acaulospora bireticulata Rothwell & Trappe and first record of Acaulospora koskei Blask. from Brazil). Our knowledge of species distribution in arbuscular mycorrhizal fungi (AMF) is still limited and morphological variations found in isolates of one species may provide additional insight for understanding the characters used in the taxonomy of this group. The aims of this work were to expand biogeographical data regarding Acaulospora koskei Blaszk. and Acaulospora bireticulata Rothwell & Trappe, both found in an Araucaria Forest in Sao Paulo state, Brazil, as well as to compare the descriptions of these species with those of other similar AMF.

Genome of an arbuscular mycorrhizal fungus provides insight into the oldest plant symbiosis.

The mutualistic symbiosis involving Glomeromycota, a distinctive phylum of early diverging Fungi, is widely hypothesized to have promoted the evolution of land plants during the middle Paleozoic. These arbuscular mycorrhizal fungi (AMF) perform vital functions in the phosphorus cycle that are fundamental to sustainable crop plant productivity. The unusual biological features of AMF have long fascinated evolutionary biologists. The coenocytic hyphae host a community of hundreds of nuclei and reproduce clonally through large multinucleated spores. It has been suggested that the AMF maintain a stable assemblage of several different genomes during the life cycle, but this genomic organization has been questioned. Here we introduce the 153-Mb haploid genome of Rhizophagus irregularis and its repertoire of 28,232 genes. The observed low level of genome polymorphism (0.43 SNP per kb) is not consistent with the occurrence of multiple, highly diverged genomes. The expansion of mating-related genes suggests the existence of cryptic sex-related processes. A comparison of gene categories confirms that R. irregularis is close to the Mucoromycotina. The AMF obligate biotrophy is not explained by genome erosion or any related loss of metabolic complexity in central metabolism, but is marked by a lack of genes encoding plant cell wall-degrading enzymes and of genes involved in toxin and thiamine synthesis. A battery of mycorrhiza-induced secreted proteins is expressed in symbiotic tissues. The present comprehensive repertoire of R. irregularis genes provides a basis for future research on symbiosis-related mechanisms in Glomeromycota.

Ecology and evolution of multigenomic arbuscular mycorrhizal fungi.

Arbuscular mycorrhizal fungi (AMF) form extremely important mutualistic symbioses with most plants. Their role in nutrient acquisition, plant community structure, plant diversity, and ecosystem productivity and function has been demonstrated in recent years. New findings on the genetics and biology of AMF also give us a new picture of how these fungi exist in ecosystems. In this article, I bring together some recent findings that indicate that AMF have evolved to contain multiple genomes, that they connect plants together by a hyphal network, and that these different genomes may potentially move around in this network. These findings show the need for more intensive studies on AMF population biology and genetics in order to understand how they have evolved with plants, to better understand their ecological role, and for applying AMF in environmental management programs and in agriculture. A number of key features of AMF population biology have been identified for future studies and most of these concern the need to understand drift, selection, and genetic exchange in multigenomic organisms, a task that has not previously presented itself to evolutionary biologists.

Micorrizas arbusculares no crescimento pós-transplantio de mudas de árvores em solo com excesso de metais pesados

Os efeitos adversos dos metais pesados para as diversas formas de vida dificultam a recuperação de solos contaminados por estes elementos. Neste trabalho, avaliaram-se os efeitos da inoculação com fungos micorrízicos arbusculares no crescimento e absorção de metais de mudas de cinco espécies arbóreas, transplantadas para misturas que continham diferentes proporções de um solo contaminado (PSC). Mudas de Senna multijuga (L.C. Rich.) Irwin et Barneby (cássia verrugosa), Luehea grandiflora Mart. et Zucc. (açoita-cavalo), Enterolobium contortisiliquum (Vell.) Morong (tamboril), Albizia lebbeck (L.) Benth. (albizia) e Senna macranthera (Collard.) Irwin et Barneby (fedegoso), inoculadas e sem inoculação, foram transplantadas para as misturas de solos e desenvolvidas por 180 dias, no período de abril a novembro de 1996, em vasos, em casa de vegetação do Departamento de Ciência do Solo da UFLA, Lavras (MG). Verificou-se que a elevação na PSC na mistura reduziu o desenvolvimento das mudas e a colonização micorrízica (CM), sendo isto causado pela elevada absorção de metais pelas plantas, especiamente, de Cd e Zn. A inoculação favoreceu o crescimento das mudas após transplantio, sendo esse efeito mais evidente nas misturas de solo com baixa PSC. A CM foi reduzida de 70 a 90% no solo não contaminado para valores próximos de zero na mistura com alta PSC. Os níveis críticos de toxidez (redução de 10% na matéria seca das plantas inoculadas) dos metais no solo foram, em mg dm-3, de 83, 57, 153, 256 e 16, para o Zn, e de 1,3; 0,9; 0,8; 4,0 e 1,6, para Cd, para açoita-cavalo, cássia verrugosa, fedegoso, tamboril e albizia, respectivamente. Observando esses níveis críticos, as plantas não inoculadas apresentaram produção de matéria seca relativa, média para todas as espécies, de apenas 39%, evidenciando os benefícios da inoculação para o crescimento pós-transplantio das mudas. Esses benefícios relacionaram-se com menores teores de metais na parte aérea. Mesmo desconhecendo os mecanismos envolvidos nestas respostas, os resultados deste trabalho evidenciaram a importância das micorrizas arbusculares para o crescimento de mudas de árvores e para a recuperação de áreas tropicais contaminadas com metais pesados.

Manganês e germinação de esporos de fungos micorrízicos arbusculares in vitro

A alta disponibilidade de íons metálicos no solo, dentre eles o Mn2+, pode inibir os fungos micorrízicos arbusculares (FMA), retardando a germinação dos esporos e, conseqüentemente, a formação de micorriza, o que reduz a eficiência simbiótica. O objetivo deste trabalho foi avaliar o efeito do íon Mn2+ sobre a germinação de esporos de seis espécies de FMA dos gêneros Glomus, Acaulospora, Gigaspora e Scutellospora em experimento in vitro Em substrato constituído por areia lavada, adicionaram-se 15; 30 e 75 mg kg-1 de Mn2+, na forma de MnCl2.4H2O, mantendo-se um controle sem adição de Mn. Acondicionaram-se os esporos em envelopes membranosos, introduzindo-os entre duas camadas de areia com diferentes níveis de Mn, em placas de Petri. Após 30 dias de incubação, avaliou-se a germinação dos esporos. Houve decréscimo médio de 32, 49 e 75 % na germinação dos esporos, à medida que se aumentaram as doses de Mn, em comparação ao controle. A germinação de esporos do gênero Glomus foi totalmente inibida na maior dose. O gênero Acaulospora sofreu decréscimos de até 50 % já na dose de 15 mg kg-1 de Mn2+, enquanto os gêneros Scutellospora e Gigaspora apresentaram os maiores índices de germinação de esporos, tendo havido tolerância no caso de Gigaspora, mesmo na maior dose de Mn2+, e estímulo à germinação no caso de Scutellospora até à dose 30 mg kg-1 de Mn2+.

The ecological significance of arbuscular mycorrhizal fungal effects on clonal reproduction in plants

The population ecology of clonal plants depends on the number and distribution of ramets formed during growth. Variation in clonal reproduction has previously been explained by variation in effects of abiotic resource heterogeneity and by plant genotypic variation. Different co-occurring species of the mutualistic arbuscular mycorrhizal fungi (AMF) have been shown to differentially alter growth traits of Prunella vulgaris which we hypothesize would lead to changes in clonal reproduction. Two experiments were carried out to test whether different co-occurring mycorrhizal fungi significantly influence clonal reproduction of P. vulgaris whether this effect also occurs when P. vulgaris is growing in an artificial plant community and how the effects compare with plant genotype effects on clonal growth of P. vulgaris. In the first experiment the number of ramets of P. vulgaris growing in a plant community of simulated calcareous grassland was significantly affected by inoculation with different mycorrhizal fungi. The number of ramets produced by P. vulgaris differed by a factor of up to 1.8 with different mycorrhizal fungi. The fungal effects on the number of new ramets were independent of their effects on the biomass of P. vulgaris. In a second experiment 17 different genotypes of P. vulgaris were inoculated with different mycorrhizal fungi. There were significant main effects of genotypes and mycorrhizal fungi on clonal reproduction of P. vulgaris. The effect of different mycorrhizal fungi contributed more than the effect of plant genotype to variation in size and ramet production. However mean stolon length and spacer length which determine the spatial arrangement of ramets were only significantly affected by plant genotype. There were no mycorrhizal fungal X plant genotype interactions on clonal growth of P. vulgaris indicating that there is no obvious evidence that selection pressures would favor further coevolution between P. vulgaris and mycorrhizal fungal species. In natural communities plants can be colonized by several different AMF at the same time. The effect of the mixed AMF treatment on the growth and clonal reproduction of P. vulgaris could not be predicted from the responses of the plants to the single AMF To what extent however the patterns of colonization by different AMF differ among plants in a natural community is unknown. Since the effects of AMF on growth and clonal reproduction occur on a population of P. vulgaris in a microcosm plant community and because the effects are also as great as those caused by plant genotypic variation we conclude that the effects are strong enough to potentially affect population size and variation of clonal plants in communities.

Evidence of recombination in putative ancient asexuals.

Ancient asexuals have been considered to be a contradiction of the basic tenets of evolutionary theory. Barred from rearranging genetic variation by recombination, their reduced number of gene arrangements is thought to hamper their response to changing environments. For the same reason, it should be difficult for them to avoid the build-up of deleterious mutations. Several groups of taxonomically diverse organisms are thought to be ancient asexuals, although clear evidence for or against the existence of recombination events is scarce. Several methods have recently been developed for predicting recombination events by analyzing aligned sequences of a given region of DNA that all originate from one species. The methods are based on phylogenetic, substitution, and compatibility analyses. Here we present the results of analyses of sequence data from different loci studied in several groups of evolutionarily distant species that are considered to be ancient asexuals, using seven different types of analysis. The groups of organisms were the arbuscular mycorrhizal fungi (Glomales), Darwinula stevensoni (Darwinuloidea crustacean ostracods) and the bdelloid rotifers (Bdelloidea), which are thought to have been asexual for the last 400, 25-100, and 35-40 Myr, respectively. The seven different analytical methods evaluated the evolutionary relationships among haplotypes, and these methods had previously been shown to be reliable for predicting the occurrence of recombination events. Despite the different degree of genetic variation among the different groups of organisms, at least some evidence for recombination was found in all species groups. In particular, predictions of recombination events in the arbuscular mycorrhizal fungi were frequent. Predictions of recombination were also found for sequence data that have previously been used to infer the absence of recombination in bdelloid rotifers. Although our results have to be taken with some caution because they could signal very ancient recombination events or possibly other genetic variation of nonrecombinant origin, they suggest that some cryptic recombination events may exist in these organisms.

Potencial de infectividade de fungos micorrízicos arbusculares oriundos de área de caatinga nativa e degradada por mineração, no Estado da Bahia, Brasil

Os objetivos deste trabalho foram quantificar o número de esporos e o número mais provável de propágulos infectivos de FMA em solos da mineradora Caraíba, verificando influências sazonais na dinâmica desses propágulos e determinando os efeitos da mineração sobre o potencial de infectividade micorrízica. Foram realizadas coletas de solo na estação seca (agosto/98) e na chuvosa (fevereiro/99), em seis sub áreas da mineradora de cobre: 1 - local onde é depositado o rejeito; 2 - arredores da área industrial; 3 - local onde são depositados restos de rocha com pouco minério; 4 - caatinga nativa, não impactada; 5 - interface entre a caatinga e o rejeito; 6 - local onde foi retirada a camada superficial do solo. Foram identificadas 32 espécies de plantas num raio de dois metros, a partir dos pontos de coleta de solo. Maior diversidade (21 espécies) foi encontrada na sub área 4 e menor (2 espécies) na sub área 3. As sub áreas 1, 3 e 5 apresentaram o menor número de esporos (< 1 por g de solo), possivelmente devido aos elevados valores de Cu e Fe e ao pH mais alcalino. Em geral a densidade dos esporos e o número de propágulos infectivos foram baixos (< 2 por g de solo). Não houve diferença significativa entre o número de esporos nas estações seca e chuvosa, a não ser para a sub área 6. Entretanto, houve variação entre as sub áreas, com diferenças significativas nas duas estações do ano.

Glomus clarum e G. etunicatum: cultivo em solo e aeroponia

Foi avaliada em casa de vegetação, a produção de inóculo dos fungos micorrízicos arbusculares (FMA) Glomus clarum e G. etunicatum em cultura aeropônica e em solo. O experimento foi conduzido em delineamento inteiramente casualizado com arranjo fatorial de 2 × 2 × 5, sendo: 2 tratamentos relativos à inoculação (Glomus clarum e G. etunicatum) × 2 sistemas de cultivo (solo e aeroponia) × 5 períodos de avaliação (0, 30, 60, 90 e 120 dias) e cinco repetições. Ramas de batata-doce desinfestadas com hipoclorito de sódio foram plantadas em 120 potes com solo + vermiculita, sendo a metade inoculada com 50 esporos de G. clarum e o restante com 50 esporos de G. etunicatum. Após 64 dias, metade das plantas foi transferida para câmaras aeropônicas (uma para cada fungo) contendo água destilada + solução nutritiva a pH 6,0. A solução nutritiva foi aplicada por microaspersão por um minuto com intermitência de três minutos. A colonização das raízes e a produção de esporos foram avaliadas aos 0, 30, 60, 90 e 120 dias de crescimento. G. clarum promoveu extensiva colonização das raízes quando comparado a G. etunicatum,com densidade de esporos semelhantes. Maiores níveis de esporulação e colonização foram obtidos no cultivo em solo, com melhor tempo de cultivo aos 60 dias, independentemente do FMA. No entanto, a maior produção de inóculo foi obtida no solo, apesar do sistema aeropônico também ter sido bastante promissor aos FMA, principalmente aos 90 dias de crescimento.

Glomus clarum e G. etunicatum: cultivo em solo e aeroponia

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Congruence of fatty acid methyl ester profiles and morphological characters of arbuscular mycorrhizal fungi in Gigasporaceae.

Arbuscular mycorrhizal (AM) fungi (Order Glomales, Class Zygomycetes) are a diverse group of soil fungi that form mutualistic associations with the roots of most species of higher plants. Despite intensive study over the past 25 years, the phylogenetic relationships among AM fungi, and thus many details of evolution of the symbiosis, remain unclear. Cladistic analysis was performed on fatty acid methyl ester (FAME) profiles of 15 species in Gigaspora and Scutellospora (family Gigasporaceae) by using a restricted maximum likelihood approach of continuous character data. Results were compared to a parsimony analysis of spore morphological characters of the same species. Only one tree was generated from each character set. Morphological and developmental data suggest that species with the simplest spore types are ancestral whereas those with complicated inner wall structures are derived. Spores of those species having a complex wall structure pass through stages of development identical to the mature stages of simpler spores, suggesting a pattern of classical Haeckelian recapitulation in evolution of spore characters. Analysis of FAME profiles supported this hypothesis when Glomus leptotichum was used as the outgroup. However, when Glomus etunicatum was chosen as the outgroup, the polarity of the entire tree was reversed. Our results suggest that FAME profiles contain useful information and provide independent criteria for generating phylogenetic hypotheses in AM fungi. The maximum likelihood approach to analyzing FAME profiles also may prove useful for many other groups of organisms in which profiles are empirically shown to be stable and heritable.

Lack of mycorrhizal autoregulation and phytohormonal changes in the supernodulating soybean mutant nts1007

Autoregulatory mechanisms have been reported in the rhizobial and the mycorrhizal symbiosis. Autoregulation means that already existing nodules or an existing root colonization by an arbuscular mycorrhizal fungus systemically suppress subsequent nodule formation/root colonization in other parts of the root system. Mutants of some legumes lost their ability to autoregulate the nodule number and thus display a supernodulating phenotype. On studying the effect of pre-inoculation of one side of a split-root system with an arbuscular mycorrhizal fungus on subsequent mycorrhization in the second side of the split-root system of a wild-type soybean (Glycine max L.) cv. Bragg and its supernodulating mutant nts1007, we observed a clear suppressional effect in the wild-type, whereas further root colonization in the split-root system of the mutant nts1007 was not suppressed. These data strongly indicate that the mechanisms involved in supernodulation also affect mycorrhization and support the hypothesis that the autoregulation in the rhizobial and the mycorrhizal symbiosis is controlled in a similar manner. The accumulation patterns of the plant hormones IAA, ABA and Jasmonic acid (JA) in non-inoculated control plants and split-root systems of inoculated plants with one mycorrhizal side of the split-root system and one non-mycorrhizal side, indicate an involvement of IAA in the autoregulation of mycorrhization. Mycorrhizal colonization of soybeans also resulted in a strong induction of ABA and JA levels, but on the basis of our data the role of these two phytohormones in mycorrhizal autoregulation is questionable.