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Genotypic and phenotypic characterization of Bacillus spp. from polluted freshwater has been poorly addressed. The objective of this research was to determine the diversity and enzymatic potentialities of Bacillus spp. strains isolated from the Almendares River. Bacilli strains from a polluted river were characterized by considering the production of extracellular enzymes using API ZYM. 14 strains were selected and identified using 16S rRNA, gyrB and aroE genes. Genotypic diversity of the Bacillus spp. strains was evaluated using pulsed field gel electrophoresis. Furthermore, the presence of genetic determinants of potential virulence toxins of the Bacillus cereus group and proteinaceous crystal inclusions of Bacillus thuringiensis was determined. 10 strains were identified as B. thuringiensis, two as Bacillus megaterium, one as Bacillus pumilus and one as Bacillus subtilis. Most strains produced proteases, amylases, phosphatases, esterases, aminopeptidases and glucanases, which reflect the abundance of biopolymeric matter in Almendares River. Comparison of the typing results revealed a spatio-temporal distribution among B. thuringiensis strains along the river. The results of the present study highlight the genotypic and phenotypic diversity of Bacillus spp. strains from a polluted river, which contributes to the knowledge of genetic diversity of Bacilli from tropical polluted freshwater ecosystems.

The effect of chitosan was evaluated in three isolates of Rhizopus stolonifer from infected tomatoes that were harvested in different regions of the state of Morelos, Mexico. Changes in the membrane integrity of spores, modifications in pH media and the proteins released were analysed in each isolate against three different molecular weights of chitosan at three fixed concentrations. There was an observed decline in the integrity of spores in the presence of chitosan at all the concentrations evaluated. The protein released was different depending on the isolate, kind of chitosan and used concentration. There was a significant difference in the pH changes on the growth media for all the tested isolates. Chitosan of high molecular weight showed the best results to inhibit the infection caused by R. stolonifer on the tomato fruits. The severity of symptoms of soft rot was not related with the molecular weight of chitosan.

Introduction Theobroma cacao L. is economically speaking the most important species of the genus Theobroma . Cacao is cultured in tropical regions and its yield is affected by several diseases, such as black pod. Black pod rot in cacao . Cacao black pod, particularly, is an economically serious problem in all areas of the world where cacao is grown, causing significant pod losses of up to 30% and killing up to 10% of the trees annually. The disease is caused by different species of the stramenopile genus Phytophthora and, once it has infected a cacao field, its control is fairly difficult. Black pod rot control strategies . Several approaches are used to manage black pod: chemical control, phytosanitary and cultural methods, genetic resistance, and biological control. Losses in yield due to black pod could be reduced through integrated management practices, although the results may vary for each cacao-growing region. Main challenges and new approaches Black pod control could be achieved if an integrated management strategy is established, with the combination of biological and chemical methods, genetic control, and adequate cultural methods in an integrated program.

The saprophytic bacterium Burkholderiacepacia has been shown to play an active role as plant growth promoting bacteria (PGPB). In this study, the ability of cell-free culture medium (CFCM) of B. cepacia to improve early developmental stages of plants has been assessed on two agronomically important crops, maize (Zeamays) and rice (Oryzasativa). Treating maize and rice seeds for 45 min before germination significantly improved seed germination and consequent seedling growth. The effect of CFCM was confirmed by the increased biomass of the shoot and, mainly, the root systems of treated seedlings. Chromatographic characterization of the CFCM revealed that the spent culture medium of B. cepacia is a complex mix of different classes of metabolites including, among others, salicylic acid, indole-3-acetic acid (IAA) and several unidentified phenolic compounds. Fractionation of the CFCM components revealed that the impressive development of the root system of CFCM-treated seedlings is due to the synergistic action of several groups of components rather than IAA alone. The data presented here suggest that a CFCM of B. cepacia can be used to improve crop germination.

Plants growth promoting bacteria (PGPB) are capable of increasing directly or indirectly crop yield. Burkholderia and Pseudomonas have been found in large populations, which inhabit in the rhizosphere of several agriculture crops. The aim of this study was to determine the antagonistic activity in vitro and in vivo of Cuban native plant growth promoting rhizobacteria against Fusarium verticillioides M1, and to assess their ability to produce IAA and to promote plant growth. The bacterial strains were tested to assess their fungal growth inhibition and plant growth stimulation. The plant growth assays were done in a modified microcosm experiment and in greenhouse conditions. The results demonstrated that these rhizobacteria produce IAA and exhibit antagonistic effect on F. verticillioides M1 inoculated in the substrate. Burkholderia sp. MBf21, MBp1, MBf15 and Pseudomonas fluorescens MPp4 stood out for their plant growth stimulation in maize and for the biological control exerted on F. verticillioides M1. The strains Burkholderia sp. MBf21 and MBf15 showed the best results in disease suppression, which was achieved up to 80%. These results have practical relevance, since potentially effective bacterial strains were selected and could be used in agriculture, for the enhancement of plant growth and disease control in maize crop.

Aim: To isolate and characterize rhizobacteria from Theobroma cacao with antagonistic activity against Phytophthora palmivora, the causal agent of the black pod rot, which is one of the most important diseases of T. cacao. Methods and Results: Among 127 rhizobacteria isolated from cacao rhizosphere, three isolates (CP07, CP24 and CP30) identified as Pseudomonas chlororaphis, showed in vitro antagonistic activity against P. palmivora. Direct antagonism tested in cacao detached leaves revealed that the isolated rhizobacteria were able to reduce symptom severity upon infection with P. palmivora Mab1, with Ps. chlororaphis CP07 standing out as a potential biocontrol agent. Besides, reduced symptom severity on leaves was also observed in planta where cacao root system was pretreated with the isolated rhizobacteria followed by leaf infection with P. palmivora Mab1. The production of lytic enzymes, siderophores, biosurfactants and HCN, as well as the detection of genes encoding antibiotics, the formation of biofilm, and bacterial motility were also assessed for all three rhizobacterial strains. By using a mutant impaired in viscosin production, derived from CP07, it was found that this particular biosurfactant turned out to be crucial for both motility and biofilm formation, but not for the in vitro antagonism against Phytophthora, although it may contribute to the bioprotection of T. cacao. Conclusions: In the rhizosphere of T. cacao, there are rhizobacteria, such as Ps. chlororaphis, able to protect plants against P. palmivora. Significance and Impact of the Study: This study provides a theoretical basis for the potential use of Ps. chlororaphis CP07 as a biocontrol agent for the protection of cacao plants from P. palmivora infection. © 2015 The Society for Applied Microbiology.

The genetic diversity of 29 endophytic bacterial strains isolated from apoplastic sap of the medullary parenchym of the stem of healthy sugarcane plants grown in Cuba was analysed by Two Primers-Ramdom Amplified Polymorphic DNA fingerprinting (TP-RAPD) and 16S rRNA gene sequencing. The strains were distributed into 17 groups on the basis of their TP-RAPD patterns, and a representative strain from each group was subjected to 16S rRNA gene sequencing. Analysis of these sequences showed that the isolates belong to a wide variety of phylogenetic groups being closely related to species of genera Bacillus and Staphylococcus from Firmicutes, Microbacterium, Micrococcus and Kokuria from Actinobacteria, Rhizobium and Gluconacetobacter from α -Proteobacteria, Comamonas and Xanthomonas from β -Proteobacteria, and Acinetobacter and Pantoea from γ -Proteobacteria. These results show the complexity of the bacterial populations present in inner tissues of sugarcane, and indicate the interest and relevance of the studies on microbial diversity to improve our knowledge on the plant endophytic bacterial communities. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Aislamiento de Bacillus solubilizadores de fosfatos asociados al cultivo del arroz. El objetivo de este trabajo fue determinar la capacidad de bacterias del género Bacillus pro-venientes del cultivo del arroz (Oryza sativa L.) en solubilizar fosfatos. Los aislamientos se obtuvieron utilizando el modelo Microcosmos de los cultivares Iacuba 30 y Perla, en el año 2008 en Bauta, La Habana, Cuba. Hubo 58 aislados, tanto rizosféricos como endófitos de la raíz y de la parte aérea, que presentaban características distintivas del género Bacillus, como respuesta positiva a la tinción de Gram, forma bacilar y formación de endospora. Se realizó la detección cualitativa en cuanto a la capacidad de solubilización de fosfatos. Solo 19 aislamientos mostraron solubilización en medio sólido NBRIP y en algunos casos cambiaron la coloración del medio de azul a amarillo, lo que indica la producción de ácidos. Se seleccionaron los positivos para la cuantificación en medio líquido. Las cepas mostraron un comportamiento diferente cuando se midió el fosfato solubilizado a distintos tiempos, variando el máximo de solubilización. Phosphate solubilization by rice associate Bacillus. The objective pursued with this study was to isolate Bacillus bacteria from rice (Oryza sativa L.) and determine its potential for phosphate solubilization. Isolates were obtained using the Microcosm model from Iacuba 30 and Perla cultivar, established by Bauta (2008) in La Habana, Cuba. Fifty eight isolates were obtained from the rice rhizosphere and inside of the roots and leaves with characteristics typical of Bacillus, such as Gram positive, rod cells and endospore formation. The qualitative capacity of these isolates for phosphate solubilization was assayed in solid NBRIP medium; the results revealed that only 19 strains solubilized inorganic phosphate and some of them changed the medium color from blue to yellow, presumably due to acid excretion. Positives were selected for assay in liquid medium, and they showed different responses in terms of time and maximum phosphate solubilization.