ABSTRACT
Microorganisms that live inside and around a plant can supply it with essential substances, such as phytohormones and essential nutrients. The present investigation aimed to isolate and characterize the phyllosphere, the endophytic, and the water tank bacteria associated with Vriesea gigantea and Tillandsia aeranthos. The bacteria were tested for siderophore and indole-3-acetic acid (IAA) production, phosphate solubilization, and presence of the nif H gene. Genetic diversity of the bacterial isolates was evaluated by rep-PCR. Sixty-eight bacterial strains were isolated from 3 different microhabitats of V. gigantea and from 2 microhabitats of T. aeranthos bromeliad plants. Gram-positive, spore-forming bacilli comprised most bacterial isolates. All isolates produced IAA in vitro in presence of very low amounts of tryptophan. More than 70% of the evaluated bacteria presented the ability of siderophore production and phosphate solubilization, and possessed the nif H gene. It was not possible to distinguish well-defined groups of isolates based on the bromeliad species and microhabitat they inhabit using genetic characterization by rep-PCR. Water tanks presented the most abundant diversity compared with phyllosphere and endophytes, probably due to the high nutrient concentration, which promotes an ideal environment for complex microbial communities.
Microrganismos que habitam o interior e a superfície podem fornecer substancias essenciais ao crescimento das plantas, como fitormônios e nutrientes essenciais. O presente trabalho teve como objetivo isolar e caracterizar as bactérias da filosfera, do ambiente endofítico e a água de tanque associadas à Vriesea gigantea e Tillandsia aeranthos. As bactérias foram submetidas a testes de verificação de produção de sideróforos e de ácido indol acético (AIA), solubilização de fosfatos, e a presença do gene nif H. A diversidade genética dos isolados bacterianos foi analisada por rep-PCR. Sessenta e oito microrganismos foram isolados de 3 microambientes distintos de V. gigantea e de 2 microambientes de T. aeranthos. A maioria das bactérias isoladas foram bacilos formadores de esporos, gram-positivos. Todos os isolados produziram AIA in vitro na presença de quantidades pequenas de triptofano. Mais de 70% das bactérias analisadas produziram sideróforos, solubilizaram fosfatos e possuíam o gene nif H. Não foi possível distinguir grupos definidos de microrganismos baseados no microhabitat e na espécie de bromélia de onde foram isolados usando rep-PCR. A água do tanque apresentou maior diversidade microbiana quando comparada com a filosfera e o ambiente endofítico, provavelmente devido à alta concentração de nutrientes, que promove um ambiente favorável para o desenvolvimento de comunidades microbianas complexas.
ABSTRACT
Bradyrhizobium elkanii is successfully used in the formulation of commercial inoculants and, together with B. japonicum, it fully supplies the plant nitrogen demands. Despite the similarity between B. japonicum and B. elkanii species, several works demonstrated genetic and physiological differences between them. In this work Representational Difference Analysis (RDA) was used for genomic comparison between B. elkanii SEMIA 587, a crop inoculant strain, and B. japonicum USDA 110, a reference strain. Two hundred sequences were obtained. From these, 46 sequences belonged exclusively to the genome of B. elkanii strain, and 154 showed similarity to sequences from B. japonicum genome. From the 46 sequences with no similarity to sequences from B. japonicum, 39 showed no similarity to sequences in public databases and seven showed similarity to sequences of genes coding for known proteins. These seven sequences were divided in three groups: similar to sequences from other Bradyrhizobium strains, similar to sequences from other nitrogen-fixing bacteria, and similar to sequences from non nitrogen-fixing bacteria. These new sequences could be used as DNA markers in order to investigate the rates of genetic material gain and loss in natural Bradyrhizobium strains.
ABSTRACT
Somatic embryos of the commercial soybean (Glycine max) cultivar IAS5 were co-transformed using particle bombardment with a synthetic form of the Bacillus thuringiensis delta-endotoxin crystal protein gene cry1Ac, the beta-glucuronidase reporter gene gusA and the hygromycin resistance gene hpt. Hygromycin-resistant tissues were proliferated individually to give rise to nine sets of clones corresponding to independent transformation events. The co-bombardment resulted in a co-transformation efficiency of 44 percent. Many histodifferentiated embryos and 30 well-developed plants were obtained. Twenty of these plants flowered and fourteen set seeds. The integration and expression of the cry1Ac, gusA and hpt transgenes into the genomes of a sample of transformed embryos and all T0, T1, T2 and T3 plants were confirmed by Gus activity, PCR, Southern and western blot, and ELISA techniques. Two T0 plants out of the seven co-transformed plants produced seeds and were analyzed for patterns of integration and inheritance until the T3 generation. Bioassays indicated that the transgenic plants were highly toxic to the velvetbean caterpillar Anticarsia gemmatalis, thus offering a potential for effective insect resistance in soybean.