A new chemolithoautotrophic arsenite-oxidizing bacterium isolated from a gold mine: Phylogenetic, physiological, and preliminary biochemical studies

A previously unknown chemolithoautotrophic arsenite-oxidizing bacterium has been isolated from a gold mine in the Northern Territory of Australia. The organism, designated NT-26, was found to be a gram-negative motile rod with two subterminal flagella. In a minimal medium containing only arsenite as the electron donor (5 mM), oxygen as the electron acceptor, and carbon dioxide-bicarbonate as the carbon source, the doubling time for chemolithoautotrophic growth was 7.6 h. Arsenite oxidation was found to be catalyzed by a periplasmic arsenite oxidase (optimum pH, 5.5). Based upon 16S rDNA phylogenetic sequence analysis, NT-26 belongs to the Agrobacterium/Rhizbium branch of the alpha-Proteobacteria and may represent a new species. This recently discovered organism is the most rapidly growing chemolithoautotrophic arsenite oxidizer known.

Application of biotechnology in breeding lentil for resistance to biotic and abiotic stress

Lentil is a self-pollinating diploid (2n = 14 chromosomes) annual cool season legume crop that is produced throughout the world and is highly valued as a high protein food. Several abiotic stresses are important to lentil yields world wide and include drought, heat, salt susceptibility and iron deficiency. The biotic stresses are numerous and include: susceptibility to Ascochyta blight, caused by Ascochyta lentis; Anthracnose, caused by Colletotrichum truncatum; Fusarium wilt, caused by Fusarium oxysporum; Sclerotinia white mold, caused by Sclerotinia sclerotiorum; rust, caused by Uromyces fabae; and numerous aphid transmitted viruses. Lentil is also highly susceptible to several species of Orabanche prevalent in the Mediterranean region, for which there does not appear to be much resistance in the germplasm. Plant breeders and geneticists have addressed these stresses by identifying resistant/tolerant germplasm, determining the genetics involved and the genetic map positions of the resistant genes. To this end progress has been made in mapping the lentil genome and several genetic maps are available that eventually will lead to the development of a consensus map for lentil. Marker density has been limited in the published genetic maps and there is a distinct lack of co-dominant markers that would facilitate comparisons of the available genetic maps and efficient identification of markers closely linked to genes of interest. Molecular breeding of lentil for disease resistance genes using marker assisted selection, particularly for resistance to Ascochyta blight and Anthracnose, is underway in Australia and Canada and promising results have been obtained. Comparative genomics and synteny analyses with closely related legumes promises to further advance the knowledge of the lentil genome and provide lentil breeders with additional genes and selectable markers for use in marker assisted selection. Genomic tools such as macro and micro arrays, reverse genetics and genetic transformation are emerging technologies that may eventually be available for use in lentil crop improvement.

Plant transformation: Problems and strategies for practical application

Plant transformation is now a core research tool in plant biology and a practical tool for cultivar improvement. There are verified methods for stable introduction of novel genes into the nuclear genomes of over 120 diverse plant species. This review examines the criteria to verify plant transformation; the biological and practical requirements for transformation systems; the integration of tissue culture, gene transfer, selection, and transgene expression strategies to achieve transformation in recalcitrant species; and other constraints to plant transformation including regulatory environment, public perceptions, intellectual property, and economics. Because the costs of screening populations showing diverse genetic changes can far exceed the costs of transformation, it is important to distinguish absolute and useful transformation efficiencies. The major technical challenge facing plant transformation biology is the development of methods and constructs to produce a high proportion of plants showing predictable transgene expression without collateral genetic damage. This will require answers to a series of biological and technical questions, some of which are defined.

Expression of Mycobacterium leprae HSP65 in tobacco and its effectiveness as an oral treatment in adjuvant-induced arthritis

Transgenic plants are able to express molecules with antigenic properties. In recent years, this has led the pharmaceutical industry to use plants as alternative systems for the production of recombinant proteins. Plant-produced recominant proteins can have important applications in therapeutics, such as in the treatment of rheumatoid arthritis (RA). In this study, the mycobacterial HSP65 protein expressed in tobacco plants was found to be effective as a treatment for adjuvant-induced arthritis (AIA). We cloned the hsp65 gene from Mycobacterium leprae into plasmid pCAMBIA 2301 under the control of the double 35S promoter from cauliflower mosaic virus. Agrobacterium tumefaciens bearing the pChsp65 plasmid was used to transform tobacco plants. Incorporation of the hsp65 gene was confirmed by PCR, reverse transcription-PCR, histochemistry, and western blot analyses in several transgenic lines of tobacco plants. Oral treatment of AIA rats with the HSP65 protein allowed them to recover body weight and joint inflammation was reduced. Our results suggest a synergistic effect between the HSP65 expressed protein and metabolites presents in tobacco plants.

Inhibition of Snake Venoms and Phospholipases A(2) by Extracts from Native and Genetically Modified Eclipta alba: Isolation of Active Coumestans

We genetically modified Eclipta alba using Agrobacterium rhizogenes LBA 9402, with the aim of producing secondary metabolites with pharmacological properties against phospholipase A(2) and the myotoxic activities of snake venom. Extracts from in natura aerial parts and roots, both native and genetically modified (in vitro), were prepared and analysed by high-performance liquid chromatography. In natura materials showed the coumestan wedelolactone at higher concentration in the aerial parts, while demethylwedelolactone appeared at higher concentration in roots. Among the modified roots, clone 19 showed higher concentrations of these coumestans. Our results show that the in natura extracts of plants collected from Botucatu and Ribeirao Preto were efficient in inhibiting snake venom phospholipase A(2) activity. Regarding in vitro material, the best effect against Crotalus durissus terrificus venom was that of clone 19. Clone 19 and isolated coumestans (wedelolactone and demethylwedelolactone) inhibited the myotoxic activity induced by basic phospholipases A(2) isolated from the venoms of Crotalus durissus terrificus (CB) and Bothrops jararacussu (BthTX-I and II). The search for antivenom is justified by the need of finding active principles that are more efficient in neutralizing snake venoms and also as an attempt to complement serum therapy.

Quebra de dormência de sementes da videira cv. niágara rosada sem estratificação

O objetivo deste trabalho foi verificar o efeito de três fatores na germinação de sementes de uva cv. niágara rosada: tratamentos físicos, tratamentos químicos e ambientes de germinação. Os tratamentos físicos consistiram de sementes com corte transversal na região mediana e de sementes com corte na micrópila. Os tratamentos químicos foram aplicados mediante imersão das sementes inteiras em solução aquosa de ácido giberélico (GA3) nas concentrações de 0 e 4000 mg L-1. Os ambientes avaliados foram in vitro e ex vitro. O experimento foi um fatorial do tipo 2 x 2 x 2 no delineamento em blocos ao acaso, com quatro repetições, sendo a parcela experimental constituída de um Gerbox®, contendo 25 sementes ou uma grade com 25 tubos de ensaio. Os parâmetros avaliados foram porcentagem e velocidade de germinação das sementes. Os resultados indicaram que o corte das sementes é imprescindível à germinação de sementes não estratificadas. A combinação entre cortes na micrópila, 4000 mg L-1 de GA3 e ambiente in vitro, proporcionou a germinação de 77% das sementes, sendo o ambiente in vitro significativamente superior ao ex vitro.

Teores de antocianinas, fenóis totais, taninos e ácido ascórbico em uva 'bordô' sobre diferentes porta-enxertos

Este trabalho teve como objetivo avaliar o conteúdo de antocianinas, fenóis totais, taninos e ácido ascórbico, em variedade de uva 'Bordô', cultivada sobre os porta-enxertos '420A', 'IAC-766' e 'Ripária-do-Traviú', em três períodos de colheita distintos. O experimento foi realizado em vinhedo de uva 'Bordô', no município de Missal, no Oeste do Paraná, na safra 2007/2008. O efeito dos porta-enxertos foi avaliado em três períodos de colheita, a partir da época de floração: 90 dias, com bagas em amadurecimento, tendendo ao roxo definitivo; 100 dias, com bagas totalmente roxas; e 110 dias, com bagas em sobrematuração. Foi utilizado o delineamento em blocos casualizados, em parcelas subdivididas no tempo. As variáveis analisadas foram sólidos solúveis, acidez titulável, índice de fenóis totais, antocianinas, taninos e teor de ácido ascórbico. Os resultados mostraram que, nos porta-enxertos '420A' e 'IAC-766', a uva 'Bordô' apresentou respostas similares em acúmulos de antocianinas, taninos, acidez titulável e sólidos solúveis, mas os acúmulos de fenóis totais, tanto na polpa, quanto na epiderme das bagas, foram maiores no '420A'. Ocorreram menores acúmulos de antocianinas, fenóis totais e taninos da epiderme da uva 'Bordô', quando enxertada em 'Ripária-do-Traviú'. Acúmulos superiores de ácido ascórbico foram verificados nas uvas enxertadas em 'IAC-766'. Há evidências que apontam o porta-enxerto 'Ripária-do-Traviú' como inferior aos demais, nos aspectos nutricional e enológico.

Disponibilidade de nitrogênio de fontes minerais e orgânicas aplicadas em um Argissolo cultivado com videira

As videiras em produção tendem a aproveitar pequena quantidade do N, derivado do fertilizante mineral, mesmo cultivadas em solos com textura arenosa e baixo teor de matéria orgânica, o que confere a esses solos baixa capacidade de fornecimento de nitrogênio (N). O objetivo deste trabalho foi avaliar, por meio de incubação, a disponibilidade de nitrogênio (N) em solo com histórico de cultivo com videiras e submetido à aplicação de fontes minerais e orgânicas de nutrientes. Foram coletadas amostras de um Argissolo Vermelho, em um vinhedo e em uma área sob campo natural, no município de Rosário do Sul (RS). Após o preparo do solo, foram montados os seguintes tratamentos: solo do campo nativo; solo do vinhedo; solo do vinhedo + composto orgânico; solo do vinhedo + ureia revestida com polímeros e solo do vinhedo + ureia. Os solos foram incubados por 141 dias, e aos 0, 38, 70, 102 e 141 dias de incubação, foram determinados os teores de N-NH4+ e N-NO3-, e calculados os teores de N mineral, mineralização líquida, N mineralizado em relação ao total e o N mineral acumulado. No solo de vinhedo submetido à aplicação de ureia revestida com polímeros e de ureia, a maior mineralização de N aconteceu no período inicial. A liberação de N do composto orgânico tem um maior sincronismo com a necessidade da videira pelo nutriente. As reservas de N total potencialmente mineralizável do solo podem suprir a demanda da videira pelo N.

Contributions à la paleontology du Miocène moyen continental du basin du tage

The lignite-clays of Póvoa de Santarém dated as Upper «Vindobonian» (mammalian zone MN6), fielded abundant remains of animals and plants (spores, pollens, seeds, etc.). The forms identified are indicative of several environments. Plants, either aquatic or belonging to swampy areas are predominant (Nuphar sp., Sparganium sp., Stratiotes kaltennordheimensis, cf. Ranunculus sp.). There are also remains of plants characteristiques of a humid rather than a swampy soil such as Polypodiaceae, Myrica ceriferiformis, Toddalia maii, Spirematospermum wetzeleri. The genera Vitis and Ephedra, although rare, point fowards the existence of drier regions in the neighbourhood. The presence of polens such as Picea indicate the presence at some distance less warm upland forest areas.

Contributions à la paleontology du Miocène moyen continental du basin du Tage; (V) les végètaux de Póvoa de Santarém (note préliminaire)

The lignite-clays of Póvoa de Santarém dated as Upper «Vindobonian» (mammalian zone MN6), fielded abundant remains of animals and plants (spores, pollens, seeds, etc.). The forms identified are indicative of several environments. Plants, either aquatic or belonging to swampy areas are predominant (Nuphar sp., Sparganium sp., Stratiotes kaltennordheimensis, cf. Ranunculus sp.). There are also remains of plants characteristiques of a humid rather than a swampy soil such as Polypodiaceae, Myrica ceriferiformis, Toddalia maii, Spirematospermum wetzeleri. The genera Vitis and Ephedra, although rare, point fowards the existence of drier regions in the neighbourhood. The presence of polens such as Picea indicate the presence at some distance less warm upland forest areas.

Biochemistry of grape berries: post-genomics approaches to uncover the effects of water deficits on ripening

Dissertation presented to obtain the Ph.D degree in Biochemistry, Plant Physiology

The grapevine defences

Grapevine is one of the most cultivated fruit crop worldwide with Vitis vinifera being the species with the highest economical importance due to the high quality standards of its berries. Nonetheless, it is also the most susceptible Vitis species to fungal pathogens. Among others, relevant fungal diseases currently threatening grapevine cultures are powdery mildew, caused by Erysiphe necator, and esca, a disease complex comprised of several fungi in which Phaeomoniella chlamydospora and Phaeoacremonium aleophilum participate.(...)

The influence of kaolin application on key metabolic pathways associated with grape berry quality: a molecular and biochemical analysis

Dissertação de mestrado em Biologia Molecular, Biotecnologia e Bioempreendedorismo em Plantas

Biotechnological approaches for yield improvement of anticancer alkaloids in the plant Catharanthus roseus

Dissertação de mestrado em Biologia Molecular, Biotecnologia e Bioempreendedorismo em Plantas