ABSTRACT Soybean looper (Chrysodeixis includens) and Spodoptera group are important defoliators insects responsible for significant yield losses in soybean, cotton and maize in Brazil. Bioinsecticides and transgenic plants expressing insecticidal proteins from Bacillus thuringiensis are among the most used sustainable control methods for pest control in agriculture, especially for several lepidopteran species. To provide new components for insect control, this study aimed to select and characterize B. thuringiensis strains toxic to C. includens, S. cosmioides, S. eridania, and S. frugiperda. We performed initial bioassays with fifty B. thuringiensis strains for C. includens and selected four strains – 1608A, 726, 773, and 775E – that caused high larval mortality (100%) to be tested against Spodoptera species. These strains harbored cry insecticidal genes, megaplasmids, absence of β-exotoxin and showed two major proteins about 65 and 130 kDa in SDS-page attributed to Cry protein classes toxic to lepidopteran species. The 1608A and 775E strains also showed high toxicity to Spodoptera species that demonstrate great potential for B. thuringiensis-based bioproduct development and as sources of new insecticidal genes for transgenic crops for multiple pest control relevant in the Brazilian agricultural system.

Abstract Bacterial endophytes are considered to have a beneficial effect on host plants, improving their growth by different mechanisms. The objective of this study was to investigate the capacity of four endophytic Bacillus strains to solubilize iron phosphate (Fe-P), produce siderophores and indole-acetic acid (IAA) in vitro, and to evaluate their plant growth promotion ability in greenhouse conditions by inoculation into pearl millet cultivated in a P-deficient soils without P fertilization, with Araxá rock phosphate or soluble triple superphosphate. All strains solubilized Fe-P and three of them produced carboxylate-type siderophores and high levels of IAA in the presence of tryptophan. Positive effect of inoculation of some of these strains on shoot and root dry weight and the N P K content of plants cultivated in soil with no P fertilization might result from the synergistic combination of multiple plant growth promoting (PGP) traits. Specifically, while B1923 enhanced shoot and root dry weight and root N P content of plants cultivated with no P added, B2084 and B2088 strains showed positive performance on biomass production and accumulation of N P K in the shoot, indicating that they have higher potential to be microbial biofertilizer candidates for commercial applications in the absence of fertilization.

Marcadores microssatélite têm sido utilizados na determinação da pureza genética em programas de melhoramento e de produção de sementes, visando garantir a qualidade genética das sementes. Objetivou-se neste trabalho demonstrar, com auxílio de marcadores moleculares microssatélites, que variações na coloração do hilo em sementes de soja, muitas vezes não constituem variação genética. Foram avaliadas sementes de soja da cultivar CD 222 que apresenta sementes com hilo preto, e de duas linhagens (CD 02RV-8444 e CD 01RV-7618), as quais apresentam sementes com hilo marrom. As sementes com variações na coloração e tonalidade do hilo encontradas em cada genótipo foram separadas visualmente em grupos. O DNA de cada semente foi extraído e analisado com marcadores microssatélites em "bulks" contendo DNA de cinco sementes. As variações encontradas na coloração do hilo da cultivar CD 222 e da linhagem CD 02RV-8444 não foram devidas à variação genética, considerando 16 locos microssatélites. Para a linhagem CD 01RV-7618, os bulks de sementes com hilo marrom pigmentado de preto (B1), com hilo marrom mais intenso (B2) e intermediário (B3), foram considerados iguais. Para esta mesma linhagem foi detectada diferença no bulk de sementes com hilo marrom menos intenso (B4), com o primer Satt 070. Duas sementes deste bulk apresentaram o mesmo padrão molecular observado para os outros bulks, no mesmo loco. As demais sementes foram consideradas misturas. A utilização de marcadores microssatélites demonstra que variações na cor ou tonalidade do hilo da soja nem sempre correspondem à variação genética.

Microsatellite markers have been used in determining genetic purity in breeding programs and seed production to ensure seed genetic quality. The objective of this study was to demonstrate, with the aid of microsatellite molecular markers, that changes in the hilum color of soybean seeds are not necessarily related to genetic variation. Seeds of the CD 222 cultivar, showing a black hilum, and of two breeding lines (CD 02RV-8444 and CD 01RV-7618) showing a brown hilum, were evaluated. Seeds with color variation in the hilum of each genotype were visually separated into groups. The DNA of each seed was extracted and analyzed with microsatellite markers in DNA "bulks" of five seeds. The color variation found in the hilum of CD 222 and CD 02RV-8444 seeds did not show any genetic variation based on 16 microsatellite loci. For the CD 01RV-7618 line, the bulk of seeds with a brown-black hilum (B1), with an intense brown hilum (B2) and with an intermediary brown hilum (B3) showed the same molecular pattern. The seeds from the same line with a faint brown hilum (B4) showed a difference with the locus Satt070. Of the five seeds of this bulk, two showed the same molecular pattern of the other bulks at the same locus. The remaining seeds were considered mixtures. The use of microsatellite markers demonstrated that variations in color or tone of the soybean hilum do not always correspond to genetic variation.

O objetivo deste trabalho foi avaliar a diversidade funcional e genética de bactérias associadas à rizosfera de genótipos de milho contrastantes quanto à eficiência de uso de fósforo, por meio do teste de fontes de carbono no sistema EcoPlate e da eletroforese em gel de gradiente desnaturante (DGGE) dos fragmentos amplificados dos genes 16S ribossomais (rDNA) das bactérias. Foram coletadas amostras de solo da rizosfera de linhagens e híbridos contrastantes quanto à eficiência de uso de fósforo, cultivados em Latossolo Vermelho-Escuro fase cerrado, com baixo e alto teor de P. Bactérias da rizosfera de híbridos e linhagens eficientes, sob estresse de P, analisadas pelo sistema EcoPlate, tenderam a se agrupar conforme a análise de componentes principais, o que indica que utilizaram fontes de carbono semelhantes. Não houve diferença na diversidade bacteriana, analisada pela DGGE, entre bactérias associadas a genótipos eficientes e ineficientes no uso de P. Com base no sequenciamento do 16S rDNA, foi verificado que a rizosfera de genótipos de milho sob estresse de P parece selecionar grupos específicos de bactérias. A estrutura populacional genética e metabólica de bactérias da rizosfera foi mais influenciada pelo teor de fósforo no solo do que pela eficiência das plantas em usar o fósforo.

The objective of this work was to evaluate the functional and genetic diversity of bacteria associated to the rhizosphere of maize genotypes contrasting for phosphorus use efficiency by means of the EcoPlate carbon source test and denaturing gradient gel electrophoresis (DGGE) of amplified 16S ribosomal DNA (rDNA) fragments of bacteria. Rhizosphere soil samples of maize genotypes (hybrids and lineages) contrasting for phosphorus use efficiency cultivated in an Oxisol with high and low P content were collected. Bacteria from the rhizosphere of P-efficient maize genotypes under P stress conditions analyzed by the EcoPlate system tended to group together according to main components analysis, which indicates that these bacteria used similar carbon sources. Microbial diversity analyzed by DGGE did not differ between P-efficient and inefficient maize genotypes. Based on the 16S rDNA fragment sequencing, the rhizosphere of maize genotypes growing on low-P soils seemed to select specific bacteria species. The genetic and metabolic structure of the rhizosphere bacterial community was more strongly influenced by the level of P in the soil than by the genotypes with contrasting P-use efficiency.

Bactérias endofíticas desempenham papel importante na agricultura, melhorando a performance e adaptação de plantas contra estresses bióticos e abióticos. No presente estudo, métodos moleculares foram empregados para identificar bactérias endofíticas do gênero Bacillus isoladas de cultivares de milho doce brasileiro. SDS-PAGE de extratos protéicos totais de quarenta e dois isolados revelaram elevado número de bandas escrutináveis. Vinte e quatro isolados formaram nove grupos diferentes de réplicas bactérianas e dezoito foram considerados como únicos. Entre os isolados, alguns polipeptídios, de tamanhos variados, foram altamente acumulados. Seqüenciamento parcial do gene ribosomal 16S revelou que todos os isolados pertencem ao gênero Bacillus e que, entre treze isolados com padrão protéico similar, dois eram linhagens diferentes. Entre os quarenta e dois isolados identificados por seqüenciamento de rDNA, Bacillus subtilis e B. pumilus foram mais frequentes (15 e 12 isolados, respectivamente), seguido por, B. licheniformes (7 isolados), B. cereus (5 isolados) e B. amiloliquefascens (3 isolados). Baseado nos resultados, conclui-se que a técnica de SDS-PAGE poderá ser usada como primeiro procedimento, rápido e barato, para identificar variação inter-específica em coleções de bactérias endofíticas isoladas do milho, enquanto o método de seqüenciamento de rDNA poderá ser aplicado para analisar variações intra-específica entre isolados com padões similares de proteínas e estudos de taxonomia.

Endophytic bacteria play an important role in agriculture by improving plant performance and adaptation against biotic and abiotic stresses. In the present study molecular methods were used for identifying Bacillus endophytic bacteria isolated from Brazilian sweet corn. SDS-PAGE of wholecell protein extract of fortytwo isolates revealed a high number of scrutinable bands. Twenty-four isolates were identified in nine different groups of duplicated bacteria and eighteen were identified as unique. Some high-accumulated polipeptides with variable length were observed in almost isolates. Partial sequencing of 16S ribosomal gene revealed that all isolates are Bacillus sp. and among thirteen isolates with similar protein profiles, two were different strains. Among the forty-two isolates identified by rDNA sequencing, Bacillus subitilis and B. pumilus were the most frequenty species (15 and 12 isolates, respectively) followed by B. licheniformes (7 isolates), B. cereus (5 isolates) and B. amiloliquefascens (3 isolates). According to present results, SDS-PAGE technique could be used as a fast and cheap first tool for identifying interspecific variation in maize endophytic bacterial collections while rDNA sequencing could be applied for analyzing intraspecific variation among isolates with similar protein profile as well as for taxonomic studies.