1,522 research outputs found
Plano de Marketing da Pousada Fazenda Fogo Eterno em Urubici
No full textTCC (graduação) - Universidade Federal de Santa Catarina, Centro Sócio Econômico, Curso de Administração.O turismo rural no estado de Santa Catarina vem ganhando espaço e se desenvolvendo na atualidade. O objetivo geral desta pesquisa é elaborar estratégias de marketing para a Fazenda Fogo Eterno com base no diagnóstico de mercado realizado durante o primeiro semestre de 2006. Os objetivos específicos referentes a este trabalho são: analisar o ambiente externo e interno da organização; definir os objetivos estratégicos; sistematizar ações para inserção da Pousada no mercado de viagens e turismo; e propor ações de controle para as estratégias criadas. A metodologia utilizada foi através de um estudo de caso, com a pesquisa de dados primários (questionário aplicado) e secundários (dados já formatados). A empresa adotada para a realização deste trabalho foi a Fazenda Fogo Eterno, em Urubici, na serra catarinense. A cidade apresenta possibilidade de desenvolvimento da atividade turística, no entanto ainda possui alguns pontos a serem melhorados. A Pousada possui uma estrutura simples, suas instalações são boas, os serviços prestados são de qualidade, seus clientes são, na maioria, casais com condições financeiras favoráveis, porém, por atuar somente um ano no mercado, a situação financeira ainda não chegou na desejada. Por isso, foram estabelecidos objetivos estratégicos para melhorar sua posição no mercado, acompanhados de estratégias e ações a serem seguidas. Com a implantação das estratégias, o ideal para a Pousada é alcançar o cenário intermediário ou otimista da projeção de vendas, afim de que o plano de marketing proporcione o retorno esperado
SIMPLE AGROBACTERIUM-BASED TRANSFORMATION PROCEDURE USEFUL FOR GFP-FUSION
No full textAgrobacterium based stable transformation procedures are well established in the plant science community. In particular the floral dip method is a commonly used transformation protocol applied to the model plant, Arabidopsis thaliana. Nevertheless, often the need exists for a simpler analysis of gene-GFP fusions as control of the constructs that will be used in the creation of transgenic lines or to just obtain data from them. Analysis of GFP-fusions has become an essential step in the characterization of gene functional aspects related to tissue, cellular localization and associated protein movement. The available methodologies though are often impaired by complex protocols and technical pitfalls. Thus, a procedure based on a common and widely used system of in vitro Arabidopsis growth could be very useful. We needed a quick and simple method for analysis of gene-product-GFP fusions. Thus we decided to establish a simpler procedure for Agrobacteriummediated transformation of Arabidopsis. Results demonstrating the effectiveness of this method using a 35S-CaMV promoter-GFP construct and its validation for sub-cellular localization will be presented
Involvement of Arabidopsis thaliana Copper Amine Oxidase β in maturation of root protoxylem precursors induced by leaf wounding
No full textRoot architecture and anatomy are strictly related to crop productivity especially under environmental stress. In particular, metaxylem plasticity enhances water uptake and improves plant performance to protect yield under water stress. Xylem vessels maturation depends on coordinated events of cell wall lignification and developmental PCD, which both are triggered by a developmental- and/or stress-driven oxidative burst. Copper amine oxidases (CuAOs) oxidize polyamines to aminoaldehydes, producing H2O2 and ammonia. In this regard, the Arabidopsis AtCuAOβ is expressed in guard cells and root xylem tissues, in which it plays a role in the MeJA-induced early protoxylem differentiation. In this study, the involvement of AtCuAOβ in maturation of root protoxylem precursors induced by leaf wounding was investigated by reverse genetic approaches. An early root protoxylem differentiation was observed in leaf-cut WT plants, while atcuaoβ
insertional mutants were unresponsive in the same conditions, suggesting that the cell-wall localized AtCuAOβ plays a role in root protoxylem phenotypic plasticity triggered by systemic leaf-to-root signaling pathways after leaf wounding
Involvement of Arabidopsis Copper Amine Oxidase β in MeJA/wounding-induced stomatal closure
No full textThe developmentally-regulated and stress-induced copper amine oxidases (CuAOs) oxidize polyamines to aminoaldehydes producing the plant signal molecule hydrogen peroxide (H2O2) and ammonia. The Arabidopsis thaliana CuAOβ encodes an apoplastic CuAO expressed in root protoxylem and in guard cells. In this study, its role in MeJA/wounding-induced stomatal closure was explored by pharmacological and genetic approaches. MeJA treatments induced stomatal closure in WT plants after 15’. Leaf/root wounding induced both local and systemic signaling leading to stomatal closure after 5’ (local response) as well as 5’ and 60’ (systemic responses to root or distal leaf wounding), while atcuaoβ insertional mutants were unresponsive.No differences in stomatal closure between WT and mutants were observed in physiological conditions. Treatment with the H2O2-scavenger N,N1-dimethylthiourea reversed the MeJA/wounding-induced stomatal closure in WT plants, suggesting a key role of the AtCuAOβ-delivered H2O2 in these event. Results suggest the AtCuAOβ involvement in MeJA/wounding-induced stomatal closure, with a dynamic implying extremely rapid long-distance leaf-to-leaf or root-to-leaf communication
Stress-triggered long-distance communication leads to phenotypic plasticity: The case of the early root protoxylem maturation induced by leaf wounding in arabidopsis
No full textRoot architecture and xylem phenotypic plasticity influence crop productivity by affecting water and nutrient uptake, especially under those environmental stress, which limit water supply or imply excessive water losses. Xylem maturation depends on coordinated events of cell wall lignification and developmental programmed cell death (PCD), which could both be triggered by developmental-and/or stress-driven hydrogen peroxide (H2O2) production. Here, the effect of wounding of the cotyledonary leaf on root protoxylem maturation was explored in Arabidopsis thaliana by analysis under Laser Scanning Confocal Microscope (LSCM). Leaf wounding induced early root protoxylem maturation within 3 days from the injury, as after this time protoxylem position was found closer to the tip. The effect of leaf wounding on protoxylem maturation was independent from root growth or meristem size, that did not change after wounding. A strong H2O2 accumulation was detected in root protoxylem 6 h after leaf wounding. Furthermore, the H2O2 trap N,N1-dimethylthiourea (DMTU) reversed wound-induced early protoxylem maturation, confirming the need for H2O2 production in this signaling pathway
Enzyme activity inhibition of the ABA-inducible copper amine oxidase AtCuAOδ reverses most of the ABA-mediated stomatal closure in Arabidopsis.
No full textPlant copper amine oxidases (CuAOs) catalyze the oxidation of polyamines to aminoaldehydes producing ammonia and hydrogen peroxide (H2O2). The involvement of H2O2 derived by CuAO-driven polyamine oxidation in stomatal closure induced by stress hormones has been observed in different plants. Previous study showed that two loss-of-function mutants of the Arabidopsis thaliana CuAOδ (At4g12290) were unresponsive to ABA-induced stomatal closure. Here we investigated the role of the vacuolar AtCuAOδ in the ABA-mediated stomatal closure by pharmacological approaches. Present data based on RT-qPCR studies show that AtCuAOδ expression is up-regulated by ABA treatments with a 2-to 4-fold increase of expression levels depending on ABA concentration (from 1 to 100 μM) as soon as 3 h from the onset of treatment. This induction peaked at 6 h with a 4-fold increase at 100 μM ABA. The well-known inhibitors of CuAO enzyme activity, 2-bromoethylamine and aminoguanidine, as well as the H2O2 scavenger N,N1-dimethylthiourea reversed most of the ABA-induced stomatal closure in WT plants. Our results, consistently with previous data, suggest that the vacuolar AtCuAOδ is a necessary component in the network orchestrating the ABA-mediated regulation of stomatal closure via H2O2 production, cooperating with the other ABA-induced ROS enzymatic sources involved in this event
LEAF SAPORIN ISOFORMS: CELLULAR LOCALIZATION BY GFP-FUSION ANALYSIS.
No full textRibosome-inactivating proteins (RIPs) are a large group of plant enzymes present in a great variety of species, which inhibit protein synthesis through a site-specific deadenylation of the large ribosomal RNA at level of the conserved alpha-sarcin/ricin loop. Saporins are highly basic single chain RIPs present in different organs of the plant Saponaria officinalis (Caryiophyllacae). Previously in our laboratory, two leaf isoforms, saporin L3 and saporin S6-like, characterized by different toxicity profiles were isolated. These isoforms were isolated from intra and extracellular fractions, respectively. We constructed GFP fusions to obtain more detailed information on the localization of these two saporins. Transformation of Arabidopsis thaliana seedling roots using Agrobacterium and of onion epidermal tissues using biolistics were performed. In roots, our preliminary data on the fusions with the NH-terminus portion of two saporins (first 100 aa of the immature protein with signal peptide) suggests this region is sufficient for intra- and extracellular targeting. A potential tissue dependent targeting of saporin S6-like derived from onion epidermal data is presented
CHARACTERIZATION OF ARABIDOPSIS INSERTIONAL MUTANTS FOR COPPER-CONTAINING AMINE OXIDASES.
No full textIn plants, polyamines are catabolised by amine oxidases, which are present in different sub-cellular compartments. The chemical identity of AO-catalyzed reaction products depends on mode of polyamine oxidation, hydrogen peroxide representing a shared compound in all the AO-mediated reactions. It has been suggested that AO-mediated hydrogen peroxide production in the apoplast is involved in signalling pathways leading to stress and defence responses. Herein, a study on the physiological roles of cell wall-localized copper containing- (Cu)-AOs has been carried out. In Arabidopsis, ten putative CuAO genes have been identified, four of which (AtCuAO 1-4) encoding for proteins with a deduced extracellular localization. Insertional mutant lines for these genes were genotypically characterised by PCR, RT-PCR and Southern-blot analysis in order to identify the homozygous plants for single-locus T-DNA insertion and to confirm the absence of the full-length gene transcripts. Plant phenotype analysis revealed that methyl jasmonate (MeJA) treatments differently affects root growth of mutant and wild type plants, i.e. root growth of atcuao1 plants is less inhibited than wild type plants
Developmental, hormone- and stress-modulated expression profiles of four members of the Arabidopsis copper amine oxidase gene family.
No full textEight genes encoding copper amine oxidases (AtCuAOs) have been identified in the Arabidopsis thaliana genome. Here, a comprehensive investigation of the expression pattern of four genes encoding AtCuAOs from two different phylogenetic subfamilies, namely the two peroxisomal AtCuAOα2 (At1g31690) and AtCuAOα3 (At1g31710) and the two apoplastic AtCuAOγ1 (At1g62810) and AtCuAOγ2 (At3g43670) has been carried out by both RT-qPCR and promoter::GFP-β-glucuronidase fusion. Analysis were performed at different developmental stages, under stress conditions (dehydration and wounding) as well as after hormone (abscisic acid, salicylic acid, methyl jasmonate and indoleacetic acid) and polyamine (putrescine and spermidine) treatments. AtCuAOα2/α3 and AtCuAOγ1/γ2 are expressed in hydathodes (α2, γ1, γ2) and vascular tissues (α3, γ1, γ2) as well as in differentiating tissues (α2, γ1, γ2). RT-qPCR analysis and GUS-staining revealed that dehydration and wounding as well as indoleacetic acid and putrescine treatments induced all the four analyzed AtCuAOs. The expression of AtCuAOα2/3 and AtCuAOγ1/2 in tissues and structures involved in water supply and water loss such as vascular tissues and hydathodes suggests a role in water balance homeostasis and immune system. Moreover, their expression in zones where an auxin maximum has been observed support a role in tissue and xylem maturation events
Differential expression of saporin genes upon wounding, ABA treatment and leaf development
No full textSaporins are type 1 ribosome-inactivating proteins (RIPs: EC 3.2.2.22)
produced in various organs of Saponaria officinalis L. Two distinct saporin
types, saporin-L and saporin-S isoforms, were respectively purified from
the intra- and extra-cellular fractions of soapwort leaves. The saporin-L
isoform was lowly identical, differed for toxicity, molecular mass and amino
acid composition from saporin-S proteins forming a new monophyletic
group. Genes encoding both L- and S-type isoforms were cloned from
leaf-specific cDNA library; the encoded products included the N-terminal
diversity observed by protein sequencing and showed compatible weights
with those from mass spectra. These genes were intron-less belonging to small
gene families. Reverse transcription polymerase chain reaction/quantitative
reverse transcription polymerase chain reaction experiments evidenced their
differential expression during leaf development, wounding and abscisic acid
treatment. These results suggest that the saporin-L and -S proteins may play
diversified roles during stress responses
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