Genomic-proteomic analysis of a novel Bacillus thuringiensis strain: toxicity against two lepidopteran pests, abundance of Cry1Ac5 toxin, and presence of InhA1 virulence factor.

Bacillus thuringiensis (Bt) is a biological alternative to the indiscriminate use of chemical insecticides in agriculture. Due to resistance development on insect pests to Bt crops, isolating novel Bt strains is a strategy for screening new pesticidal proteins or strains containing toxin profile variety that can delay resistance. Besides, the combined genomic and proteomic approaches allow identifying pesticidal proteins and virulence factors accurately. Here, the genome of a novel Bt strain (Bt TOL651) was sequenced, and the proteins from the spore-crystal mixture were identified by proteomic analysis. Toxicity bioassays with the spore-crystal mixture against larvae of Diatraea saccharalis and Anticarsia gemmatalis, key pests of sugarcane and soybean, respectively, were performed. The toxicity of Bt TOL651 varies with the insect; A. gemmatalis (LC50 = 1.45 ng cm-2) is more susceptible than D. saccharalis (LC50 = 73.77 ng cm-2). Phylogenetic analysis of the gyrB gene indicates that TOL651 is related to Bt kenyae strains. The genomic analysis revealed the presence of cry1Aa18, cry1Ac5, cry1Ia44, and cry2Aa9 pesticidal genes. Virulence factor genes such as phospholipases (plcA, piplc), metalloproteases (inhA), hemolysins (cytK, hlyIII, hblA, hblC, hblD), and enterotoxins (nheA, nheB, nheC) were also identified. The combined use of the genomic and proteomic data indicated the expression of Cry1Aa18, Cry1Ac5, and Cry2Aa9 proteins, with Cry1Ac5 being the most abundant. InhA1 also was expressed and may contribute to Bt TOL651 pathogenicity. These results provide Bt TOL651 as a new tool for the biocontrol of lepidopteran pests.

Pseudomonas aeruginosa and Bacillus cereus Isolated from Brazilian Cerrado Soil Act as Phosphate-Solubilizing Bacteria.

The phosphate-solubilizing microorganism is essential for soil quality and plant development and can serve as an alternative to reduce such Brazilian needs for importing phosphate overseas. Here, we isolated and selected bacteria from Brazilian Cerrado soils capable of solubilize phosphate. We obtained 53 bacteria isolates, of which 23 could solubilize phosphate at a pH of 7.0, 17 could solubilize phosphate at a pH of 6.0, and 8 could solubilize at a pH of 5.5. Using 16S rRNA gene sequences, we identified nine bacteria species clustered in four groups: Bacillus sp., Pseudomonas sp., Priestia sp., and Klebsiella sp. Our results revealed that the UFT01 (P. aeruginosa) and UFT42 (B. cereus) isolates exhibited the best phosphate solubilization performance at all tested pH values. We further recorded higher levels of solubilization and phosphate availability six days after the soil inoculation with P. aeruginosa, and enzymatic analysis of the soil samples revealed that the P. aeruginosa-inoculated samples resulted in four-fold higher enzymatic activities when compared to non-inoculated soils. The B. cereus soil inoculation increased ß-glucosidase activities and resulted in reduced the activities of arylsulfatase. Altogether, our findings demonstrated that P. aeruginosa and B. cereus isolated from Cerrado soils showed high phosphate solubilization potential.

Antagonism of Trichoderma isolates against Leucoagaricus gongylophorus (Singer) Möller.

Filamentous fungi from the genus Trichoderma are commonly found in soil. They are considered facultative mycoparasites, and are antagonists of other fungi such as the cultivar of leaf-cutting ants (Leucoagaricus gongylophorus). The aim of the present study was to bioprospect Trichoderma spp. from different soils collected from Gurupi, Tocantins, Brazil, for antagonistic effects against the mutualistic fungus of leaf-cutting ants. To isolate filamentous fungi, samples were collected from six locations. Preliminarily, isolates were identified by morphological analysis as belonging to Trichoderma. Trichoderma spp. had their internal transcribed spacer region (ITS) of ribosomal RNA genes (rRNA) sequenced to confirm species-level taxonomy. L. gongylophorus was isolated from a laboratory ant colony. Antagonistic properties of seven isolates of Trichoderma against L. gongylophorus were measured using paired disks in Petri dishes with potato dextrose agar medium (PDA). All Trichoderma isolates inhibited the growth of L. gongylophorus in Petri dishes. Isolate 2 of Trichoderma spirale group exhibited slow mycelial growth in the Petri dish, and a high rate of inhibition against L. gongylophorus. This isolate is a promising fungus for field tests of biological control methods for leaf-cutting ants.

Contributions of γ-Aminobutyric Acid (GABA) Receptors for the Activities of Pectis brevipedunculata Essential Oil against Drosophila suzukii and Pollinator Bees.

The γ-aminobutyric acid (GABA) receptors play pivotal roles in the transmission of neuronal information in the nervous system of insects, which has led these proteins to be targeted by synthetic and natural products. Here, we assessed the insecticidal potential of the essential oil of Pectis brevipedunculata (Gardner) Sch. Bip., a neotropical Asteraceae plant used in traditional medicine, for controlling Drosophila suzukii (Matsumura) adults by feeding exposure. By using in silico approaches, we disentangle the contribution of GABA receptors and other potential neuronal targets (e.g., acetylcholinesterase, glutathione-S-transferases) in insects that may explain the essential oil differential activities against D. suzukii and two essential pollinator bees (Apis mellifera Linnaeus and Partamona helleri Friese). Neral (26.7%) and geranial (33.9%) were the main essential oil components which killed D. suzukii with an estimated median lethal concentration (LC50) of 2.25 µL/mL. Both pollinator forager bee species, which would likely contact this compound in the field, were more tolerant to the essential oil and did not have their diet consumptions affected by the essential oil. Based on the molecular predictions for the three potential targets and the essential oil main components, a higher affinity of interaction with the GABA receptors of D. suzukii (geranial -6.2 kcal/mol; neral -5.8 kcal/mol) in relation to A. mellifera (geranial -5.2 kcal/mol; neral -4.9 kcal/mol) would contribute to explaining the difference in toxicities observed in the bioassays. Collectively, our findings indicated the involvement of GABA receptors in the potential of P. brevipedunculata essential oil as an alternative tool for controlling D. suzukii.

Suitability of the predatory mites Iphiseiodes zuluagai and Euseius concordis in controlling Polyphagotarsonemus latus and Tetranychus bastosi on Jatropha curcas plants in Brazil.

One of the most promising plant species for biofuel production in Brazil is the physic nut Jatropha curcas. Major phytosanitary problems include the attack of two pest mite species, the broad mite Polyphagotarsonemus latus and the spider mite Tetranychus bastosi. Owing to pesticide-related problems, there is an increasing demand for sustainable environmental-friendly control methods such as biological control. In this study we evaluated the suitability of the predatory mite species Iphiseiodes zuluagai and Euseius concordis in controlling P. latus and T. bastosi on J. curcas. The number of T. bastosi killed by I. zuluagai was lower than the number of P. latus consumed. Euseius concordis preyed upon both T. bastosi and P. latus but the number of prey killed was always lower in comparison with I. zuluagai. However, P. latus and T. bastosi are suitable for the development of I. zuluagai and E. concordis as oviposition of both predators did not differ in relation to prey species. The preference of I. zuluagai for leaves of plants infested by either P. latus or T. bastosi, combined with the higher values for predation obtained by this predatory mite when fed on P. latus, compared to those values obtained by E. concordis, suggests that I. zuluagai can be more efficient than E. concordis in reducing populations of P. latus and T. bastosi under field conditions. Furthermore, we report here on the first record of predatory mites associated with P. latus and T. bastosi on native J. curcas plants in Brazil. In conclusion, we emphasize the crucial importance of predatory mites as agents of natural biological control of mite pests on J. curcas in small farms.

The complete chloroplast genome of Artocarpus altilis (Moraceae) and phylogenetic relationships.

The chloroplast (cp) is an essential organelle in higher plants. The genes of the plastome are well suited to infer phylogenetic relationships among taxa. In this study, we report the assembly of the cp genome of Artocarpus altilis and its phylogeny among species from Moraceae family. The cp genome of A. altilis was 160,822 base pair (bp) in length, comprising one large single-copy region of 88,692 bp, one small single-copy region of 19,290 bp, and a pair of inverted repeat regions (IRs) of 26,420 bp. A total of 113 different genes were predicted, including 79 protein-coding genes, 30 tRNA genes, and four rRNA genes. The phylogenetic analysis of 19 species belonging to the Moraceae family confirmed the phylogenetic proximity of the genus Artocarpus and Morus and the genetic similarity of A. camansi and A. altilis.

Geostatistical Assessment of Frankliniella schultzei (Thysanoptera: Thripidae) Spatial Distribution in Commercial Watermelon Crops.

Spatial distribution studies of insect pests make it possible to determine their colonization and dispersal patterns. Watermelon (Citrullus lanatus (Thunb.) Matsum. et Nakai) is among the most frequently consumed fruits in the world, and the common blossom thrips, Frankliniella schultzei (Trybom) (Thysanoptera: Thripidae), is one of the most important insect pests of this plant. The objective of this study was to determine the spatial distribution of F. schultzei in commercial watermelon crops using geostatistics. The studied F. schultzei populations presented an aggregated distribution. The colonization of thrips began at the borders of the crops, especially in the areas located in the opposite direction to that of the prevailing winds. The highest densities of thrips occurred in crops that had cucurbits in the surrounding areas. When monitoring for F. schultzei populations, greater attention should be given to sampling that part of the crop located in the opposite direction to that of the prevailing winds because this is where colonization begins. Even at low densities, the aggregation points of thrips in the crop should be located and controlled so that they do not cause damage. In sampling programs for F. schultzei, samples should be taken at distances greater than 9 m apart because this is the distance up to which densities of this species show spatial dependence. Planting watermelon crops close to other cucurbits should be avoided, as these alternate hosts may act as a source of infestation by this pest.

Detection of Colletotrichum gloeosporioides in native cashew species in Brazil.

The savanna cashew (Anacardium humile A. St. Hil.) nut is a native shrub species distributed in the savanna biome of Central Western Brazil. This species is similar to the cashew (Anacardium occidentale L.) nut, and its edible pseudo-fruits have been used for various purposes. However, A. humile is at risk of extinction. One reason for this is the phytosanitary issues. Therefore, phytosanitary studies of the serious plant diseases caused by fungal pathogens are necessary. The aim of this study was to survey the savanna cashew nut (Anacardium humile) in Central Western Brazil from August 2013 to October 2014. Serious damage caused by anthracnose to the plant's leaves and fruits was consistently detected. The pathogen fungal species was Colletotrichum gloeosporioides. It was identified by morphological characteristics and molecular analyses. Additional experiments showed evidence that C. gloeosporioides can be transmitted by cashew seeds. We observed anthracnose symptoms on the surfaces of the floral peduncle, leaves, and peduncle of the A. humile cashew nuts. The genomic DNA sequences of the selected strains for molecular characterization have had 99% identity with the analogous sequences of C. gloeosporioides. Naturally occurring C. gloeosporioides infection of the leaves of A. humile was low (20.3%) compared with that of the nuts (79.7%). Our results can be used to design strategies to prevent the introduction and establishment of C. gloeosporioides in new areas. They can also be helpful in monitoring programs in areas with a current occurrence of C. gloeosporioides. Finally, these results can be used in future research plans of C. gloeosporioides infection management.

Characterization of the Catalytic Structure of Plant Phytase, Protein Tyrosine Phosphatase-Like Phytase, and Histidine Acid Phytases and Their Biotechnological Applications.

Phytase plays a prominent role in monogastric animal nutrition due to its ability to improve phytic acid digestion in the gastrointestinal tract, releasing phosphorus and other micronutrients that are important for animal development. Moreover, phytase decreases the amounts of phytic acid and phosphate excreted in feces. Bioinformatics approaches can contribute to the understanding of the catalytic structure of phytase. Analysis of the catalytic structure can reveal enzymatic stability and the polarization and hydrophobicity of amino acids. One important aspect of this type of analysis is the estimation of the number of ß-sheets and α-helices in the enzymatic structure. Fermentative processes or genetic engineering methods are employed for phytase production in transgenic plants or microorganisms. To this end, phytase genes are inserted in transgenic crops to improve the bioavailability of phosphorus. This promising technology aims to improve agricultural efficiency and productivity. Thus, the aim of this review is to present the characterization of the catalytic structure of plant and microbial phytases, phytase genes used in transgenic plants and microorganisms, and their biotechnological applications in animal nutrition, which do not impact negatively on environmental degradation.

Efeito da adubação orgânica na produção de biomassa e bioatividade do óleo essencial do capim citronela / Effect of organic fertilization on biomass production and bioactivity of citronella grass essential oil

RESUMO Este trabalho teve como objetivo analisar o efeito da adubação orgânica no crescimento e na produção de biomassa do capim citronela (Cymbopogon nardus), assim como avaliar o efeito do óleo essencial do capim citronela e do composto citronelal na inibição do crescimento micelial do fungo Didymella bryoniae. Na avaliação do efeito da adubação orgânica no crescimento do capim citronela, foi utilizado o delineamento em blocos casualizados em esquema de parcela subdividida. As parcelas foram constituídas por quatro doses de adubação orgânica de esterco bovino curtido (0, 3, 6 e 9 Kg cova-1) e as subparcelas por cinco épocas de amostragem (80, 108, 136, 164, 192 dias após o transplante). Para avaliar a fungitoxicidade do óleo essencial do capim citronela na inibição do crescimento micelial do fungo D. bryoniae, foi instalado no delineamento inteiramente casualizado em esquema fatorial. Os tratamentos foram compostos por cinco alíquotas (5, 10, 15, 20 e 25 µL) do óleo essencial do capim citronela e do composto citronelal, em cinco épocas de amostragem. Verificou-se no tratamento de adubação orgânica de 9 Kg cova-1 os maiores valores em todas as variáveis analisadas na última época de amostragem. Constatou-se maior efeito de inibição do crescimento micelial utilizando o citronelal em comparação com o óleo essencial. Na alíquota de 25 µL do citronelal ocorreu inibição total do crescimento micelial do fungo D. bryoniae.