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Microbial diversity can be used to assess the impact of agricultural practices on the long-term sustainability of cropping systems. The aim of this study was to investigate changes in soil bacterial diversity as a result of the impact of different soil tillage and crop rotation systems in an oxisol of southern Brazil. Bacterial diversity was examined in the 0-10 cm layer in two field experiments by analyzing soil DNA using 16S rDNA-DGGE profiles. Experiment one consisted of a long-term 26-year trial with four soil tillage management systems: (1) no-tillage (NT), (2) disc plow (DP), (3) field cultivator (FC), and (4) heavy-disc harrow (DH), all under soybean (summer)/wheat (winter) crop succession. Experiment two consisted of a short-term 10-year trial with DP and NT and three crop rotations (CR) including grasses, legumes and green manures. Cluster analysis of the 16S rDNA sequences revealed that the main effect on clustering was attributed to differences in soil tillage management systems. The Shannon index confirmed greater bacterial diversity under NT, followed by the FC, DH and DP. Therefore, diversity decreased as tillage practices intensified. The evenness index demonstrated uniformity of the profiles of the bacterial communities, with dominance of a few communities, regardless of soil tillage and crop rotation. Different crop rotations had only minor effects on bacterial diversity, what could be related to a previous fallow period. The results suggest that bacterial diversity analyzed by DGGE may be useful as bioindicator of the changes caused by soil tillage.

The transgenic soybean [Glycine max (L.) Merrill] occupies about 80 % of the global area cropped with this legume, the majority comprising the glyphosate-resistant trait (Roundup Ready(®), GR or RR). However, concerns about possible impacts of transgenic crops on soil microbial communities are often raised. We investigated soil chemical, physical and microbiological properties, and grain yields in long-term field trials involving conventional and nearly isogenic RR transgenic genotypes. The trials were performed at two locations in Brazil, with different edaphoclimatic conditions. Large differences in physical, chemical and classic microbiological parameters (microbial biomass of C and N, basal respiration), as well as in grain production were observed between the sites. Some phyla (Proteobacteria, Actinobacteria, Acidobacteria), classes (Alphaproteobacteria, Actinomycetales, Solibacteres) and orders (Rhizobiales, Burkholderiales, Myxococcales, Pseudomonadales), as well as some functional subsystems (clustering-based subsystems, carbohydrates, amino acids and protein metabolism) were, in general, abundant in all treatments. However, bioindicators related to superior soil fertility and physical properties at Londrina were identified, among them a higher ratio of Proteobacteria:Acidobacteria. Regarding the transgene, the metagenomics showed differences in microbial taxonomic and functional abundances, but lower in magnitude than differences observed between the sites. Besides the site-specific differences, Proteobacteria, Firmicutes and Chlorophyta were higher in the transgenic treatment, as well as sequences related to protein metabolism, cell division and cycle. Although confirming effects of the transgenic trait on soil microbiome, no differences were recorded in grain yields, probably due to the buffering capacity associated with the high taxonomic and functional microbial diversity observed in all treatments.

Although Brazil has recently reached the position as the second largest producer of genetically modified soybean [Glycine max (L.) Merr.], there are few reports on the effects of transgenic crops and the associated use of specific herbicides on soil microbial communities, both under the edaphoclimatic conditions in Brazil, and in other producer regions in the southern hemisphere. The aim of this study was to evaluate the effects of transgenic soybean containing the ahas gene conferring resistance to herbicides of the imidazolinone group, and of the herbicides associated with transgenic soybeans on the soil microbial community. Twenty field experiments were carried out during three growing seasons (summer of 2006/2007, short-season of 2007 and summer of 2007/2008), in nine municipalities located in six Brazilian states and in the Federal District. The experiments were conducted using a completely randomized block design with four replicates and three treatments: (1) conventional (non-transgenic) soybean cultivar Conquista with conventional herbicides (bentazone + acifluorfen-sodium and other herbicides, depending on the level of infestation in each region); (2) near-isogenic transgenic Cultivance (CV127) containing the ahas gene, with conventional herbicides; (3) transgenic Cultivance with specific herbicide of the imidazolinone group (imazapyr). As the objective of the study was to verify impacts of the transgene and herbicides on the soil microbial community of the whole area and not only a punctual rhizospheric effects, samples were taken at the 0-10 cm layer prior to cropping and at R2 soybean growth stage, between plant rows. Quantitative (microbial biomass C and N, MB-C and MB-N) and qualitative (DGGE of the 16S rDNA region) parameters of soil microbial community were evaluated. No qualitative or quantitative differences were found that could be attributed to the transgene ahas. A comparison of Cultivance soybean with conventional and imidazolinone-group herbicides applications also failed to reveal differences that could be attributed to the specific use of imazapyr, even after three consecutive croppings at the same site. Finally, no differences were detected between conventional (Conquista and conventional herbicides) and transgenic soybean managements (Cultivance and imazapyr). However, marked differences were observed in MB-C and MB-N between the different sites and times of year and, for the 16S rDNA-DGGE profiles, between different sites. In conclusion, microbial community evaluations were found to be sensitive and viable for monitoring different technologies and agricultural management methods, but no differences could be attributed to the ahas transgene for three consecutive cropping seasons.

-The no-tillage (NT) system of soil management is recognized as more sustainable than conventional tillage (CT), with an important role played by soil microorganisms. The objective of this study was to estimate differences in soil microbial diversity under NT and CT at different soil depths. For that, bacterial (16S rDNA) and fungal (18S rDNA) communities were evaluated by denaturing gradient gel electrophoresis (DGGE) in a 20-year field experiment established on an Oxisol in southern Brazil on which soybean has been grown in the summer and wheat in the winter. Soil samples were collected at the depths of 0-5, 5-10, 10-20 and 20-30 cm, and submitted to DGGE analyses. The results revealed lower similarity (28%) between bacterial communities in the NT and the CT systems at the 0-5 cm layer. The Shannon index (H) confirmed higher bacterial diversity with NT at all depths, when compared to CT. In relation to fungal communities, higher diversity was detected with CT, especially at the 0-5-cm depth. The results indicate that fungal communities can be more tolerant to environmental stresses related to soil disturbance than bacteria. More emphasis should be given for understanding processes affecting the diversity of microorganisms in agricultural soils, with particular emphasis on tillage systems.Key words: Bacterial diversity. Fungal diversity. PCR-DGGE. Soil tillage.RESUMO -O sistema de plantio direto (PD) é reconhecido como mais sustentável que o plantio convencional (PC), desempenhando um importante papel na atividade dos microrganismos do solo. O objetivo deste estudo foi estimar diferenças na diversidade microbiana do solo sob PD e PC, em diferentes profundidades. Para isso, foram avaliadas as comunidades bacteriana (16S rDNA) e fúngica (18S rDNA) através da eletroforese em gel com gradiente desnaturante (DGGE), em um experimento de campo estabelecido há 20 anos em um Latossolo Vermelho no sul do Brasil, com a soja cultivada no verão e trigo no inverno. As amostras de solo foram coletadas nas profundidades de 0-5; 5-10; 10-20 e 20-30 cm, e foram submetidas a análises de DGGE. Os resultados revelaram baixa similaridade (28%) na profundidade de 0-5 cm para as comunidades bacterianas entre o sistema de PD e PC. O índice de Shannon (H) confirmou diversidade bacteriana superior no PD em todas as profundidades, quando comparado ao PC. Em relação às comunidades fúngicas, a maior diversidade foi detectada no sistema de PC, especialmente na profundidade de 0-5 cm. Os resultados indicam que as comunidades fúngicas podem ser mais tolerantes a estresses ambientais relacionados com a perturbação do solo que as bactérias. Maior ênfase deve ser dada para compreender os processos que afetam a diversidade dos microrganismos em solos agrícolas, com particular destaque para os sistemas de preparo do solo. Palavras-chave: Diversidade bacteriana. Diversidade fúngica. PCR-DGGE. Manejo do Solo

ABSTRACT. Glyphosate-tolerant (Roundup Ready ® , RR) soybean is the most widely cultivated genetically modified crop in the world. The aim of this study was to evaluate, in a long-term field experiment, the chemical composition of grains in a pair of cultivars composed of the conventional parent (BRS 133) and the nearly isogenic transgenic (BRS 245RR) soybean. Plants were cropped under two different edaphoclimatic conditions in Brazil. The experiments began during the 2003/2004 growing season; samples were collected during the 2011/2012 harvest. Were quantified the contents of isoflavones (high performance liquid chromatography), fatty acids (gas chromatography), lipid and proteins (near infrared spectroscopy in the NIR -Near Infrared Reflectance) and minerals (atomic absorption flame spectrophotometry). All samples were analyzed in three replications. Major differences were observed between the two cultivation locations. In Ponta Grossa, which has lower temperatures and higher rainfall during the grain filling period, the contents of lipids and isoflavones were higher. In Londrina, which hassoil with higher fertility, the contents of minerals, proteins and carbohydrates in soybean grains were higher. Significant differences were observed between the cultivars. The conventional parent had higher protein contents and lower lipid contents. The transgenic soybean line had higher isoflavone contents.Keywords: Glycine max, transgenic soybean crop, isoflavones, fatty acids, macro and micro minerals.Composição química de grãos de soja resistente ao glifosato e sua respectiva parental convencional em diferentes condições edafoclimáticas do Brasil RESUMO. A soja resistente ao glifosato (Roundup Ready ® , RR) representa a principal cultura geneticamente modificada (GM) cultivada no mundo. O objetivo deste estudo foi avaliar, em um experimento de campo de longa duração, a composição química dos grãos das cultivares de soja parental convencional (BRS 133) e sua respectiva transgênica quase isogênica (BRS 245RR). Os experimentos foram realizados em duas condições edafoclimáticas distintas e tiveram início em 2003/2004 e as amostras foram coletadas na safra 2011/2012. Foram quantificados os teores de isoflavonas (cromatografia líquida de alta eficiência), ácidos graxos (cromatografia em fase gasosa), lipídeos e proteínas (espectroscopia no infravermelho próximo na NIR -refletância de infravermelho próximo) e minerais (espectrofotometria de absorção atômica de chama). Todas as amostras foram analisadas em triplicata. Foram observadas diferenças significativas entre os locais de cultivo. Em Ponta Grossa, que possui temperaturas menores e maior precipitação durante o enchimento de grãos, o conteúdo de lipídeos e isoflavonas foram maiores. Em Londrina, com solo de maior fertilidade, o conteúdo de minerais, proteínas e carboidratos foram maiores. Diferenças significativas foram associadas ao transgene RR. A cultivar BRS 133 apresentou maiores teores de proteínas e a BRS 245 RR maiores teores de lipídeos e isoflavonas.Palavras-chav...