Characterization of bacterial endophytes from the roots of native and cultivated Brazil nut trees (Bertholletia excelsa)

Brazil nut is a very important nontimber forest product in the Amazon region. Propagation of this tree still represents a challenge due to slow and uneven seed germination. In this context, plant growth-promoting bacteria can facilitate the process of propagation. The aims of this study were to isolate and characterize endophytic bacteria from the roots of Brazil nut trees in native terra firme forest and cultivation areas in northern Brazil, and to identify mechanisms by which bacteria act in plant growth promotion. Overall, 90 bacterial isolates were obtained from the roots of Brazil nut trees in monoculture, agroforestry and native forest areas by using different semisolid media. The isolates were characterized by sequencing the 16S rRNA gene. Plant growth-promoting characteristics were evaluated by the presence of the nifH gene, aluminum phosphate solubilization and the production of indole compounds. The isolates were affiliated with 18 genera belonging to 5 different classes (α-Proteobacteria, ß-Proteobacteria, γ-Proteobacteria, Bacilli and Actinobacteria). The genus Bacillus was predominant in the forest and monoculture areas. Fourteen isolates presented the nifH gene. Most of the bacteria were able to solubilize aluminum phosphate and synthetize indole compounds. The results indicated high diversity of endophytic bacteria present among the roots of Brazil nut trees, mainly in the agroforestry area, which could be related to soil attributes. Among the 90 isolates, the 22 that presented the best results regarding plant growth promotion traits were good candidates for testing in seedling production of Brazil nut trees. (AU)

Contribution of dark septate fungi to the nutrient uptake and growth of rice plants

ABSTRACT The use of dark septate fungi (DSE) to promote plant growth can be beneficial to agriculture, and these organisms are important allies in the search for sustainable agriculture practices. This study investigates the contribution of dark septate fungi to the absorption of nutrients by rice plants and their ensuing growth. Four dark septate fungi isolates that were identified by Internal transcribed spacer phylogeny were inoculated in rice seeds (Cv. Piauí). The resulting root colonization was estimated and the kinetic parameters Vmax and Km were calculated from the nitrate contents of the nutrient solution. The macronutrient levels in the shoots, and the NO3--N, NH4+-N, free amino-N and soluble sugars in the roots, sheathes and leaves were measured. The rice roots were significantly colonized by all of the fungi, but in particular, isolate A103 increased the fresh and dry biomass of the shoots and the number of tillers per plant, amino-N, and soluble sugars as well as the N, P, K, Mg and S contents in comparison with the control treatment. When inoculated with isolates A103 and A101, the plants presented lower Km values, indicating affinity increases for NO3--N absorption. Therefore, the A103 Pleosporales fungus presented the highest potential for the promotion of rice plant growth, increasing the tillering and nutrients uptake, especially N (due to an enhanced affinity for N uptake) and P.

Diversity and capacity to promote maize growth of bacteria isolated from the Amazon region / Diversidade e capacidade de promoção de crescimento de milho de bactérias isoladas da região Amazônica

Maize plants can establish beneficial associations with plant growth-promoting bacteria. However, few studies have been conducted on the characterization and inoculation of these bacteria in the Amazon region. This study aimed to characterize endophytic bacteria isolated from maize in the Amazon region and to assess their capacity to promote plant growth. Fifty-five bacterial isolates were obtained from maize grown in two types of ecosystems, i.e., a cerrado (savanna) and a forest area. The isolates were characterized by the presence of the nifH gene, their ability to synthesize indole-3-acetic acid (IAA) and solubilize calcium phosphate (CaHPO4), and 16S rRNA partial gene sequencing. Twenty-four bacteria contained the nifH gene, of which seven were isolated from maize plants cultivated in a cerrado area and seventeen from a forest area. Fourteen samples showed the capacity to synthesize IAA and only four solubilized calcium phosphate. The following genera were found among these isolates: Pseudomonas; Acinetobacter; Enterobacter; Pantoea; Burkholderia and Bacillus. In addition, eight isolates with plant growth-promoting capacity were selected for a glasshouse experiment involving the inoculation of two maize genotypes (a hybrid and a variety) grown in pots containing soil. Inoculation promoted the development of the maize plants but no significant interaction between maize cultivar and bacterial inoculation was found. A high diversity of endophytic bacteria is present in the Amazon region and these bacteria have potential to promote the development of maize plants.

Plantas de milho podem estabelecer associações benéficas com bactérias promotoras do crescimento vegetal. No entanto, poucos estudos de caracterização e inoculação com essas bactérias foram realizados na região Amazônica. O objetivo deste estudo foi caracterizar bactérias endofíticas isoladas de milho na região Amazônica e avaliar sua capacidade de promover o crescimento. Cinquenta e cinco isolados bacterianos foram obtidos de plantas de milho cultivadas em diferentes tipos de ecossistemas, cerrado (savana) e área de floresta. Estes foram caracterizados pela presença do gene nifH, capacidade de sintetizar ácido indol-3-acético (AIA), solubilizar fosfato de cálcio (CaHPO4) e sequenciamento parcial do gene 16S rRNA. Vinte e quatro bactérias apresentaram o gene nifH. Destas, sete são oriundas de plantas cultivadas em uma área de cerrado e 17 em área de floresta. Quatorze bactérias demonstraram a capacidade de solubilizar AIA e quatro de solubilizar fosfato de cálcio. Entre estas bactérias, foram encontrados os seguintes gêneros: Pseudomonas; Acinetobacter; Enterobacter; Pantoea; Burkholderia e Bacillus. Oito isolados com capacidade de promoção do crescimento vegetal foram selecionados para experimento em casa de vegetação com a inoculação em dois diferentes genótipos de milho (um híbrido e uma variedade) em vasos com solo. Quanto à promoção de crescimento vegetal, a inoculação promoveu o desenvolvimento de plantas de milho, mas não foi encontrada interação significativa entre a cultivar de milho e a inoculação bacteriana. Na região Amazônica há uma diversidade de bactérias endofíticas e estas possuem potencial para promover o desenvolvimento de plantas de milho.

Dinâmica de rizóbios em solo do cerrado de Roraima durante o período de estiagem / Dynamic of rhizobia in the soil during the dry season in cerrado of Roraima

The biological nitrogen fixation in legumes is performed by a group of bacteria known as rhizobia. The survival of these bacteria in soils is affected by several factors, such as temperature, drought and soil fertility. This study was performed to evaluate the dynamics of rhizobia in the soil after soybean cultivation and during a dry season in the cerrado of Roraima. Three areas were sampled: i) native cerrado as reference; ii) an area previously cultivated with soybean for one season; and iii) another one cultivated for two seasons also with soybean. The soil was sampled at a depth of 0-10 cm in five times (0, 45, 90, 135 and 180 days) during the dry season (September 2006 to March 2007). The rhizobial density in the soil was evaluated by the most probable number method with infection of soybean and cowpea plants. It was observed very low number of soybean nodulating bacteria in the reference area, but a high density, of up to several hundred rhizobia capable to nodulate cowpea was measured in this same area. Cropping of soybean with inoculated seeds increased rhizobial density evaluated by both trapping hosts. In cropped areas, an intense reduction of rhizobium density was observed just after soybean harvest, and this reduction continued until the end of the period of evaluation. It was concluded that soybean cultivation increases the density of rhizobial in the cerrado soil; however, this density is drastically reduced, during the dry season, by 99% at the end of the dry period.

A fixação biológica de nitrogênio que ocorre em leguminosas é realizada por um grupo de bactérias conhecidas como rizóbios. A sobrevivência destas bactérias no solo é influenciada por diversos fatores como a temperatura, umidade e fertilidade do solo. O objetivo deste trabalho foi avaliar a dinâmica da população de rizóbios em solo após o cultivo de soja, durante o período de estiagem no cerrado de Roraima. Foram amostradas três áreas: i) cerrado nativo como referência; ii) área cultivada uma vez com soja inoculada com rizóbio; iii) e área cultivada duas vezes com soja inoculada com rizóbio em anos consecutivos. O solo foi coletado na profundidade de 0-10 cm em cinco períodos a partir do inicio da estiagem no mês de setembro de 2006 coincidindo com a época de colheita da soja e prolongando-se até março de 2007 (0, 45, 90, 135 e 180 dias). A população de rizóbios no solo foi avaliada pela técnica do número mais provável (NMP) utilizando plantas de soja e de feijão-caupi como espécies isca. Foi observado que na área nativa praticamente não existiam bactérias nodulantes de soja, mas havia uma população capaz de nodular o feijão-caupi de até algumas centenas de rizóbios por gramas de solo. O cultivo da soja utilizando sementes inoculadas elevou a população de rizóbios no solo que foi constatada por ambas às espécies de plantas isca. Nas áreas cultivadas, constatou-se uma intensa redução da população de rizóbios no solo, em especial logo após a colheita da soja, continuando o decréscimo até o último período de avaliação. Conclui-se que o cultivo da soja inoculada com rizóbio eleva a densidade de rizóbios em solo do cerrado, mas durante a estiagem ocorre uma drástica redução dessa população, que pode chegar a mais de 99% considerando o início e final do período.