Isolation, selection and characterization of root-associated growth promoting bacteria in Brazil Pine (Araucaria angustifolia)

Araucaria angustifolia, a unique species of this genus that occurs naturally in Brazil, has a high socio-economic and environmental value and is critically endangered of extinction, since it has been submitted to intense predatory exploitation during the last century. Root-associated bacteria from A. angustifolia were isolated, selected and characterized for their biotechnological potential of growth promotion and biocontrol of plant pathogenic fungi. Ninety-seven strains were isolated and subjected to chemical tests. All isolates presented at least one positive feature, characterizing them as potential PGPR. Eighteen isolates produced indole-3-acetic acid (IAA), 27 were able to solubilize inorganic phosphate, 21 isolates were presumable diazotrophs, with pellicle formation in nitrogen-free culture medium, 83 were phosphatases producers, 37 were positive for siderophores and 45 endospore-forming isolates were antagonistic to Fusarium oxysporum, a pathogen of conifers. We also observed the presence of bacterial strains with multiple beneficial mechanisms of action. Analyzing the fatty acid methyl ester (FAME) and partial sequencing of the 16S rRNA gene of these isolates, it was possible to characterize the most effective isolates as belonging to Bacillaceae (9 isolates), Enterobacteriaceae (11) and Pseudomonadaceae (1). As far as we know, this is the first study to include the species Ewingella americana as a PGPR. (C) 2011 Elsevier GmbH. All rights reserved.

Efeito de diferentes sistemas de manejo de plantas invasoras sobre o controle biológico e incidência de Cinara atlantica (Hemiptera: Aphididae) em Pinus taeda e biologia de Coccinelídeos (Coleoptera)

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

PHOSPHORUS DOSES DETERMINE THE PREVALENCE OF NATIVE ARBUSCULAR MYCORRHIZAL FUNGI IN Araucaria angustifolia

A greenhouse experiment was installed with bait cultures to extract the AMF species present in a rhizosphere soil sample of a native Araucaria angustifolia forest in Campos do Jordao, Brazil. The experimental design was completely randomized, with four increasing phosphorus doses (0, 20, 50, and 150 mg kg(-1), as triple superphosphate), with five replicates, the bait plant was araucaria, and all pots were inoculated with 100 g of rhizospheric soil collected in an araucaria forest. After twelve months the spores were extracted, counted and identified, and the percent root colonization was also determined. When taking all four P doses into account, eleven AMF species could be identified: Acaulospora bireticulata, Acaulospora morrowiae, Acaulospora sp., Entrophospora colombiana, Gigaspora margarita, Glomus diaphanum, Glomus etunicatum, Glomus macrocarpum, Scutellospora calospora, Scutellospora gilmorei, and Scutellospora pellucida. There was no effect of the P dose on the total amount of spores neither on the percent root colonization. However, the correspondence analysis showed that the different AMF species were selectively associated mostly to either one or another P dose.

Ambrosia and bark beetles (Scolytidae : Coleoptera) in pine and eucalypt stands in southern Brazil

More than 95% of the reforested area in Brazil is covered by exotic Eucalyptus and Pinus plantations. Native Scolytidae, mostly ambrosia beetles, appear to be rapidly adapting to these exotic trees, and reports of economic damage are becoming frequent. The objectives of our research were to survey, characterize and compare the Scolytidae fauna present in a P. taeda and an E. grandis stand in Telemaco Borba, Parana state, Brazil. Beetles were caught in ethanol baited ESALQ-84 vane traps in weekly collections from July 1995 until July 1997. In all, 87 species were trapped, 62 in the pine and 75 in the eucalypt stand. The most abundant beetle species in the pines were Hypothenemus eruditus, Xyleborinus gracilis, Cryptocarenus sp. and Xylosandrus retusus, while the most frequent were H. eruditus, Cryptocarenus sp., H. obscurus, Ambrosiodmus obliquus, and X. gracilis. In the eucalypt stand, H. eruditus, X. retusus, H. obscurus, X. ferrugineus and Microcorthylus minimus were the most abundant species, and H. eruditus, H. obscurus and M. minimus were the most frequently trapped. The majority of the species, regardless of the forest community, were most active between August (end of winter) and October (mid-spring). Significantly more H. eruditus, X. gracilis, Cryptocarenus sp., Corthylus obliquus, Hypothenemus bolivianus, A. obliquus, Sampsonius dampfi and Xyleborus affinis were trapped in the pine stand, while X. retusus, H. obscurus, X. ferrugineus, Xyleborinus linearicollis, Corthylus sp, and Corthylus convexicauda were caught in higher numbers in the eucalypt stand. Approximately 50% of the species trapped were found in both communities. Morisita's similarity index indicates the composition of the two communities is very similar, suggesting that most of the beetles are polyphagous. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Attraction of ambrosia beetles (Coleoptera : Scolytidae) to different tropical pine species in Brazil

Ambrosia beetles are the predominant Scolytidae in Brazil. Little is known about the attractiveness of exotic conifer tree volatiles to native scolytids. Objectives were to compare the attractiveness of logs with and without bark of Pinus oocarpa Schiede, P. caribaea variety bahamensis Barrett & Golfari, P, car. variety caribaea Barrett & Golfari and P. car. variety hondurensis Barrett & Golfari over time to native scolytids in different pine stands, to compare the relative attractiveness of logs relative to ethanol traps, to determine how long it takes for logs to become attractive to ambrosia beetles and when attraction peaks occur, and to determine if volatiles released by live standing trees would mask volatiles released by logs of the same species. In young stands, Hypothenemus was the predominant insect genus, whereas in older stands Xyleborus predominated. Debarked logs trapped more beetles than logs with bark. Pine log species attractiveness was not influenced by volatiles present in the stand. Beetles were divided into the following 3 groups, based on response to log volatiles and ethanol: (1) species attracted to ethanol and not responding to pine terpenes Ambrosiodmus hagedorni (Iglesia), A. retusus (Eichhoff), X. spinulosus Blandford, Corthylus schaufussi Schiede, Cryptocarenus heveae (Hagedorn), H. obscurus (F.), (2) species attracted to ethanol but responding to pine terpenes Xyleborinus gracilis (Eichhoff), X. affinis Eichhoff, H. eruditus Westwood, Premnobius cavipennis Eichhoff, and (3) species more attracted to pine terpenes and less responsive to ethanol, A. obliquus (Le Conte), X. ferrugineus F,, X. catulus Blandford. Pinus car. ;variety bahamensis was the least attractive pine, P. oocarpa the most attractive. The attraction peak varied according to the season; logs were not attractive to beetles 10 wk after cutting.

Can native vegetation recover after slash pine cultivation in the Brazilian Savanna?

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Charcoal production using tropical pine thinnings

In Brazil, the wood from pine-plantation thinnings is difficult to sell. However, these wood thinnings could be used to produce charcoal. The physical and chemical properties of charcoal produced from logs or first and fourth thinnings of Pinus caribaea var. hondurensis and Pinus oocarpa were determined. Similar properties were also determined for sawmill slabs cut from first and fourth thinnings. Properties were compared to charcoal made from pine materials and 7-yr-old Eucalyptus grandis. Results show that Pinus spp had better chemical properties than did E. grandis. In contrast, charcoal obtained from E. grandis had higher values of apparent density and lower values of bulk density than did Pinus spp. © 1992.

Rhizospheric streptomycetes as potential biocontrol agents of Fusarium and Armillaria pine rot and as PGPR for Pinus taeda

Pinus taeda is one of the main timber trees in Brazil, occupying 1.8 million ha with an annual productivity of 25-30 m(3) ha(-1). Another important species is Araucaria angustifolia, belonging to the fragile Rainforest biome, which for decades has been a major source of timber in Brazil. Some diseases that affect the roots and/or the stem of these trees and cause "damping-off" of the seedlings, with economic and environmental losses for the forest sector, are caused by the plant pathogenic fungi Fusarium sp. or Armillaria sp. This research project intended to isolate actinobacteria from the Araucaria rhizosphere, which present an antagonistic effect against these fungi. After the selection of the best pathogen inhibitors, morphologic characteristics, enzyme production, and their effect on the growth of Pinus taeda were studied. The actinobacteria were tested for their antagonistic capacity against Fusarium sp. in Petri plates with PDA as substrate. The inhibition zone was measured after 3, 5, 7, and 10 days. Of all the isolates tested, only two of them maintained inhibition zones up to 4 mm for 10 days. The inhibition of Armillaria sp. was tested in liquid medium and also in Petri dishes through the evaluation of the number of the fungal rhizomorphs in dual culture with the actinobacteria. It was found that all five isolates were able to inhibit the rhizomorph production, with the best performance of the isolate A43, which was capable of inhibiting both fungi, Fusarium and Armillaria. In a greenhouse experiment, the effect of five isolates on the growth of Pinus taeda seedlings was tested. Plant height, stem diameter, root and shoot dry matter were determined. The Streptomyces isolate A43 doubled plant growth. These results may lead to the development of new technologies in the identification of still unknown bacterial metabolites and new management techniques to control forest plant diseases.

Detection of hormones in surface and drinking water in Brazil by LC-ESI-MS/MS and ecotoxicological assessment with Daphnia magna

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Cloning and expression of embryogenesis-regulating genes in Araucaria angustifolia (Bert.) O. Kuntze (Brazilian Pine)

Angiosperm and gymnosperm plants evolved from a common ancestor about 300 million years ago. Apart from morphological and structural differences in embryogenesis and seed origin, a set of embryogenesis-regulating genes and the molecular mechanisms involved in embryo development seem to have been conserved alike in both taxa. Few studies have covered molecular aspects of embryogenesis in the Brazilian pine, the only economically important native conifer in Brazil. Thus eight embryogenesis-regulating genes, viz.,ARGONAUTE 1, CUP-SHAPED COTYLEDON 1, WUSCHEL-related WOX, S-LOCUS LECTIN PROTEIN KINASE, SCARECROW-like, VICILIN 7S, LEAFY COTYLEDON 1, and REVERSIBLE GLYCOSYLATED POLYPEPTIDE 1, were analyzed through semi-quantitative RT-PCR during embryo development and germination. All the eight were found to be differentially expressed in the various developmental stages of zygotic embryos, seeds and seedling tissues. To our knowledge, this is the first report on embryogenesis-regulating gene expression in members of the Araucariaceae family, as well as in plants with recalcitrant seeds.

Gene expression during early somatic embryogenesis in Brazilian pine (Araucaria angustifolia (Bert) O. Ktze)

Brazilian pine (Araucaria angustifolia (Bert) O. Ktze) is the only native conifer species with economic importance in Brazil. Recently, due to intensive exploitation Brazilian pine was included in the official list of endangered Brazilian plants, under the "vulnerable" category. Biotechnology tools like somatic embryogenesis (SE) are potentially useful for mass clonal propagation and ex situ conservation strategies of commercial and endangered plant species. In spite of that, numerous obstacles still hamper the full application of SE technology for a wider range of species, including Brazilian pine. To enhance somatic embryogenesis in Brazilian pine and to gain a better understanding of the molecular events associated with somatic embryo development, we analyzed the steady-state transcript levels of genes known to regulate somatic embryogenesis using semiquantitative reverse transcription polymerase chain reaction (sqRT-PCR). These genes included Argonaute (AaAGO), Cup-shaped cotyledon1 (AaCUC), wushel-related WOX (AaWOX), a S-locus lectin protein kinase (AaLecK), Scarecrow- like (AaSCR), Vicilin 7S (AaVIC), Leafy Cotyledon 1 (AaLEC), and a Reversible glycosylated polypeptide (AaRGP). Expression patterns of these selected genes were investigated in embryogenic cultures undergoing different stages of embryogenesis, and all the way to maturation. Up-regulation of AaAGO, AaCUC, AaWOX, AaLecK, and AaVIC was observed during transition of somatic embryos from stage I to stage II. During the maintenance phase of somatic embryogenesis, expression of AaAGO and AaSCR, but not AaRPG and AaLEC genes was influenced by presence/ absence of plant growth regulators, both auxins and cytokinins. The results presented here provide new insights on the molecular mechanisms responsible for somatic embryo formation, and how selected genes may be used as molecular markers for Brazilian pine embryogenesis.

Finding hy-Brazil : eugenics and modernism in the Pacific

In the 1930s a number of Australian women writers took issue with material and symbolic aspects of public health discourse and in particular challenged institutional endorsement of eugenicist movements. This paper discusses the response of inter-war women writers to the national discussion of science and medicine and, in particular, eugenics.

Conifer defence against insects: microarray gene expression profiling of Sitka spruce (Picea sitchensis) induced by mechanical wounding or feeding by spruce budworms (Choristoneura occidentalis) or white pine weevils (Pissodes strobi) reveals large

Conifers are resistant to attack from a large number of potential herbivores or pathogens. Previous molecular and biochemical characterization of selected conifer defence systems support a model of multigenic, constitutive and induced defences that act on invading insects via physical, chemical, biochemical or ecological (multitrophic) mechanisms. However, the genomic foundation of the complex defence and resistance mechanisms of conifers is largely unknown. As part of a genomics strategy to characterize inducible defences and possible resistance mechanisms of conifers against insect herbivory, we developed a cDNA microarray building upon a new spruce (Picea spp.) expressed sequence tag resource. This first-generation spruce cDNA microarray contains 9720 cDNA elements representing c. 5500 unique genes. We used this array to monitor gene expression in Sitka spruce (Picea sitchensis) bark in response to herbivory by white pine weevils (Pissodes strobi, Curculionidae) or wounding, and in young shoot tips in response to western spruce budworm (Choristoneura occidentalis, Lepidopterae) feeding. Weevils are stem-boring insects that feed on phloem, while budworms are foliage feeding larvae that consume needles and young shoot tips. Both insect species and wounding treatment caused substantial changes of the host plant transcriptome detected in each case by differential gene expression of several thousand array elements at 1 or 2 d after the onset of treatment. Overall, there was considerable overlap among differentially expressed gene sets from these three stress treatments. Functional classification of the induced transcripts revealed genes with roles in general plant defence, octadecanoid and ethylene signalling, transport, secondary metabolism, and transcriptional regulation. Several genes involved in primary metabolic processes such as photosynthesis were down-regulated upon insect feeding or wounding, fitting with the concept of dynamic resource allocation in plant defence. Refined expression analysis using gene-specific primers and real-time PCR for selected transcripts was in agreement with microarray results for most genes tested. This study provides the first large-scale survey of insect-induced defence transcripts in a gymnosperm and provides a platform for functional investigation of plant-insect interactions in spruce. Induction of spruce genes of octadecanoid and ethylene signalling, terpenoid biosynthesis, and phenolic secondary metabolism are discussed in more detail.