Phenotypic and molecular characteristics of rhizobia isolated from nodules of peanut (Arachis hypogaea L.) grown in Brazilian Spodosols

Peanut (Arachis hypogaea L.) is an important crop that can fix nitrogen through symbiosis with rhizobia. Rhizobia populations with diverse characteristics than those traditionally described as peanut microsymbionts have been found in tropical soils. With the objective of studying the diversity and phylogeny of these rhizobia, 22 bacterial strains were isolated from nodules of seven peanut varieties grown in Spodosols of Pernambuco State, Brazil. The isolates were examined in culture medium by means of some of their phenotypic characteristics and tested for intrinsic antibiotic resistance (IAR). DNA profiles were determined with the BOX-PCR and compared with 19 reference strains. All isolates showed rapid growth, and most of them acidified the culture medium. In general, high antibiotic resistance was observed to penicillin G, chloramphenicol and tetracycline and susceptibility was observed to neomycin, erythromycin and rifampicin. The analysis of their phenotypic characteristics, that is, colony morphology and IAR, provided little information about the phylogeny of the isolates. However, using compilation of phenotypic and molecular characteristics, we were able to observe a great diversity of these rhizobia and to reveal the presence of new species.Keywords: Antibiotic resistance, Arachis hypogaea L., BOX-PCR, colony morphology, diversityAfrican Journal of Biotechnology Vol. 12(17), pp. 2147-215

Avaliação do estado microbiológico do solo em área preservada e impactada por mineração no semiárido de Pernambuco

A mineração remove a camada orgânica do solo, causando grave impacto ambiental nas populações microbianas (bactérias e fungos) e nos fungos micorrízicos arbusculares (FMA). Análises microbiológicas constituem ferramentas importantes no monitoramento da poluição do solo. Da mesma forma que, a utilização de técnicas moleculares pode ser útil para esclarecer a identidade e variabilidade de FMA em ambientes nativos e degradados. O objetivo deste trabalho foi avaliar e comparar o estado biológico de solos e determinar a ocorrência de FMA em áreas nativa e impactada por mineração de gesso. Coletas de solo foram realizadas em 4 áreas: 1- nativa (AN); 2 – arredores da mina (AM); 3 – rejeito (AR); 4 – interface entre a área de rejeito e a área degradada (AI) em dois períodos de coleta: chuvoso e seco. No estudo molecular foram identificadas algumas espécies de FMA, que mostram capacidade para tolerar as condições de estresse da área minerada. Em geral, os valores do número mais provável de propágulos (NMP) de fungos, bactérias e actinomicetos foram maiores em área nativa quando comparada com as áreas impactadas. A atividade mineradora produziu impacto negativo na microbiota do solo, reduzindo o NMP de micro-organismos

Characterization of Metarhizium anisopliae using amplifed ribossomal DNA restriction analysis (ARDRA) and internal transcribed spacer (ITS) sequence analysis

Molecular methods have revolutionized systematic entomology in the genus Metarhizium. In Brazil, the importance of these entomopathogenic fungi in agriculture is critical to the control of the sugar cane spittlebug (Mahanarva posticata) (Homoptera: Cercopidae). Other important factors for the selection of isolates of Metarhizium anisopliae for use in the biological control of particular pests are virulence, reproductive characteristics and the media of artificial cultures. In addition, the genetic variability of these entomopathogenic fungi indicates a great potential for pest control and a lack of a link between isolates from specific host sites and fungal virulence. We studied five Metarhizium hosts including the sugar cane spittlebug (IPA213, IPA215, IPA219 and IPA216) and the grasshopper (IPA217). It is noteworthy that these five isolates are used as biological control for growing sugar cane on an industrial scale for the Brazilian Northeast. Hence, the importance to genetically characterize these isolates of Metarhizium cannot be over emphasized. Amplifed ribossomal DNA restriction analysis (ARDRA) did not differentiate any of the isolates that were studied following digestion with three endonuclease namely; HindIII, HinfI and HpaI. The data that were generated using ITS1 and ITS4 markers revealed that although these isolates infect the same host, they are genetically distinct perhaps due to the unknown origin of the hosts. However, the isolate IPA217, which was isolated from the grasshopper, did not behave differently from the others. These results show that the M. anisopliae complex is monophyletic and it was concluded that the use of markers and morphological studies yielded results that may corroborate the idea that investigation of the phenotypic and molecular characteristics of this genus may indentify a new species of Metarhizium.Key words: Cordyceps brittebankisoides, genetic identity, ITS1, ITS4, phylogeny, sugar cane spittlebug

Isolation and Characterization of Plant Growth-Promotion Diazotrophic Endophytic Bacteria Associated to Sugarcane (Saccharum officinarum L.) Grown in Paraíba, Brazil

Sugarcane is an important Brazilian commodity, being usually cultivated in soils with low natural fertility. This study aimed to isolate diazotrophic endophytes from sugarcane tissues and evaluate the morphological and physiological characteristics of their colonies as well as their plant growth-promoting (PGP) traits in select diazotrophic endophytic bacteria. Fifty-six bacterial isolates were identified in the sugarcane tissues, and these isolates presented distinct morphological and physiological traits. A total of thirty-five bacterial isolates were biochemically evaluated. Overall, Bacillus was the dominant genus. Isolates of Methylobacterium spp. and Brevibacillus agri were present only in leaves, while Herbaspirillum seropedicae occurred only in stems. Except to IPA-CF45A, all isolates were nitrogenase positive. All endophytes exhibit production of indol 3-acetic acid. Over 50% of endophytes solubilize phosphate, release N-acyl homoserine lactones, and present the activity of 1-aminocyclopropane-1-carboxylic acid deaminase, catalase, lipase and protease. The network analysis showed that isolates belonged to Burkholderia, Herbaspirillum, and Methylobacterium interact with Bacillus. Bacterial endophytes exhibited distinct morphological, physiological, and PGP traits that are useful for sustainable agriculture, highlighting the isolates IPA-CC33, IPA-CF65, IPA-CC9 and IPA-CF27. Further studies on the effects of these diazotrophic endophytes and their potential for providing microbial inoculants for improving sugarcane fields will provide valuable information to maintain the sustainability and environment quality.National Council for Scientific and Technological Development 426655/2018-

The complete genome sequence of Chromobacterium violaceum reveals remarkable and exploitable bacterial adaptability

Chromobacterium violaceum is one of millions of species of free-living microorganisms that populate the soil and water in the extant areas of tropical biodiversity around the world. Its complete genome sequence reveals (i) extensive alternative pathways for energy generation, (ii) ≈500 ORFs for transport-related proteins, (iii) complex and extensive systems for stress adaptation and motility, and (iv) wide-spread utilization of quorum sensing for control of inducible systems, all of which underpin the versatility and adaptability of the organism. The genome also contains extensive but incomplete arrays of ORFs coding for proteins associated with mammalian pathogenicity, possibly involved in the occasional but often fatal cases of human C. violaceum infection. There is, in addition, a series of previously unknown but important enzymes and secondary metabolites including paraquat-inducible proteins, drug and heavy-metal-resistance proteins, multiple chitinases, and proteins for the detoxification of xenobiotics that may have biotechnological applications

SYMBIOTIC EFFECTIVENESS AND COMPETITIVENESS OF CALOPO RHIZOBIAL ISOLATES IN AN ARGISSOLO VERMELHO-AMARELO UNDER THREE VEGETATION COVERS IN THE DRY FOREST ZONE OF PERNAMBUCO

Biological N fixation in forage legumes is an important alternative to reduce pasture degradation, and is strongly influenced by the inoculant symbiotic capability. This paper evaluates the effectiveness of Calopo (Calopogonium mucunoides) rhizobial isolated from soil under three vegetation covers of an Argissolo Vermelho-Amarelo of the Dry Forest Zone of Pernambuco. An experiment was conducted evaluating 25 isolates, aside from 5 uninoculated controls with 0; 309; 60; 90 and 120 kg ha-1 N, and a treatment inoculated with the SEMIA 6152 strain. The first cut was performed 45 days after inoculation and a second and third cut after 45-day-intervals. Shoot N content was quantified at all cuts. Shoot dry mass was affected by N rates at all cuts. Shoot dry mass increased from the first to the second cut in inoculated plants. There was no difference between rhizobial isolates from the different plant covers for any of the variables. Most variables were significantly and positively correlated

Efetividade da inoculação com rizóbio e fungos micorrízicos arbusculares em mudas de sabiá submetidas a diferentes níveis de fósforo

Visando otimizar a produção de mudas de sabiá (Mimosa caesalpiniifolia Benth), foi conduzido um experimento para avaliar a efetividade da dupla inoculação com fungos micorrízicos arbusculares (FMA) e rizóbio. Os tratamentos, arrumados em esquema fatorial consistiram de presença e ausência de Rhizobium sp. e de FMA (Glomus etunicatum, Acaulospora morrowae e A. longula), e de três níveis de P (0, 20 e 40 kg/ha de P2O5, na forma de superfosfato triplo). A aplicação de P na ausência e na presença dos fungos não favoreceu o desenvolvimento das plantas. As mudas com a dupla inoculação apresentaram valores significativos no crescimento, área foliar, altura das plantas, atividade da enzima nitrogenase, porcentagem de colonização radicular e outros parâmetros analisados, independentemente do nível de P usado. A nodulação do sabiá foi favorecida pela micorrização, uma vez que as mudas inoculadas apenas com Rhizobium apresentaram nodulação significativamente menor. Houve aumento da colonização micorrízica e diminuição da esporulação na presença de Rhizobium

Efetividade da inoculação com rizóbio e fungos micorrízicos arbusculares em mudas de sabiá submetidas a diferentes níveis de fósforo

Visando otimizar a produção de mudas de sabiá (Mimosa caesalpiniifolia Benth), foi conduzido um experimento para avaliar a efetividade da dupla inoculação com fungos micorrízicos arbusculares (FMA) e rizóbio. Os tratamentos, arrumados em esquema fatorial consistiram de presença e ausência de Rhizobium sp. e de FMA (Glomus etunicatum, Acaulospora morrowae e A. longula), e de três níveis de P (0, 20 e 40 kg/ha de P2O5, na forma de superfosfato triplo). A aplicação de P na ausência e na presença dos fungos não favoreceu o desenvolvimento das plantas. As mudas com a dupla inoculação apresentaram valores significativos no crescimento, área foliar, altura das plantas, atividade da enzima nitrogenase, porcentagem de colonização radicular e outros parâmetros analisados, independentemente do nível de P usado. A nodulação do sabiá foi favorecida pela micorrização, uma vez que as mudas inoculadas apenas com Rhizobium apresentaram nodulação significativamente menor. Houve aumento da colonização micorrízica e diminuição da esporulação na presença de Rhizobium