Phosphatidylinositol phospholipase C mediates carbon sensing and vegetative nuclear duplication rates in Aspergillus nidulans

In this work, we disrupted one of three putative phosphatidylinositol phospholipase C genes of Aspergillus nidulans and studied its effect on carbon source sensing linked to vegetative mitotic nuclear division. We showed that glucose does not affect nuclear division rates during early vegetative conidial germination (6-7 h) in either the wild type or the plcA-deficient mutant. Only after 8 h of cultivation on glucose did the mutant strain present some decrease in nuclear duplication. However, decreased nuclear division rates were observed in the wild type when cultivated in media amended with polypectate, whereas our plcA-deficient mutant did not show slow nuclear duplication rates when grown on this carbon source, even though it requires induction and secretion of multiple pectinolytic enzymes to be metabolized. Thus, plcA appears to be directly linked to high-molecular-weight carbon source sensing.

Identificação e diferenciação do Mycobacterium tuberculosis pela amplificação do DNA das regiões intergênicas dos operons inhA e pIC

Entre as doenças causadas por bactérias do gênero Mycobacterium, a tuberculose por M. tuberculosis é a mais conhecida. O diagnóstico da doença é feito utilizando-se um conjunto de exames que possibilitam a identificação da mesma (WATT, 2000). Contudo, sabe-se que o diagnóstico combinado de microscopia direta e com o posterior isolamento em meio de cultivo é o “padrão-ouro”. A principal desvantagem desse método é que tal bactéria possui um crescimento lento (cerca de 8 semanas). Recentemente, a detecção de doenças através da técnica de reação em cadeia da polimerase (PCR) tem proporcionado avanços significativos no diagnóstico. O uso da amplificação específica de genes, para identificar a M. tuberculosis, tais como rDNA 16S, IS6110 ou a região intergênica senX3-regX3, tem apresentado algumas restrições, ao nível de confiabilidade e sensibilidade, para a aplicação da técnica de PCR. O presente estudo mostra a construção e a aplicação de um novo alvo para a aplicação da PCR no diagnóstico da tuberculose, baseado no ensaio da diferença de organização gênica do operon plcA, B e C diferenciando a M. tuberculosis das demais micobactérias. Neste trabalho, foram examinadas 273 amostras de pacientes com suspeita de tuberculose, sendo estas submetidas ao estudo comparativo da técnica de PCR versus cultivo (padrão ouro). A PCR amplificou fragmentos de 439pb. Os resultados mostram 93,7% de acurácia para PCR/Cultivo (p<000,1), 93,1% de sensibilidade com intervalo de confiança de 88,7-96,0 e especificidade de 96,4% com intervalo de confiança de 96,4-99,4. O valor da estatística Kappa (k) foi de 0,82 com erro padrão de 0,041, demonstrando um alinhamento quase perfeito para a verificação do grau de concordância entre os testes. Desta forma, o uso desta nova região para a amplificação da PCR se mostra uma importante e confiável ferramenta no diagnóstico específico da tuberculose. Outra região que compreende parte dos genes mbaA e inhA foi utilizada para diferenciar o Complexo tuberculosis do Complexo avium. Porém, novos experimentos serão necessários para o emprego desta região como uma ferramenta de diagnóstico.

Allelopathic plants. 7. Sunflower (Helianthus annuus L.)

Sunflower is an annual dicotyledonous plant, herbaceous, erect and native of North America. It is thermo- and photo-insensitive, hence, can be grown round the year in sub-tropical and tropical countries. Only two spp. H. annuus and H. tuberosum are cultivated for food, remaining spp. are ornamentals, weeds and wild plants. However, H. annuus is allelopathic and inhibit the growth and development of other plants thus reducing their productivity. Much information is available about the allelopathic effects of sunflower crop on following crops in crop rotations. Although it is harmful to all crops, but, is less harmful to crops of Graminae family than other families. It seems that the harmful effects of sunflower in crop rotations are due to release and accumulation of root exudates during crop growth in soil. Soil incorporation of its fresh (green manure) or dry biomass in soil is inhibitory to both crops and weed spp. Several allelochemicals have been characterized from the H. annuus, which inhibit the seed germination and seedling growth of A. albus, A. viridis, Agropyron repens (Elymus repens), Ambrosia artemsiifolia, Avena fatua, Celosia crustata, Chenopodium album, Chloris barbara, Cynodon dactylon, D. sanguinalis, Dactyloctenium ageyptium, Digitaria ciliaris, Echinochloa crus-galli, Flaveria australasica, Parthenium hysterophorus, Portulaca oleracea, Sida spinosa, Trianthema portulacastrum, Veronica perisca the inhibitory effects of this crop may be used for weed management with less herbicides for sustainable agriculture.

Photoacoustics as a tool for the diagnosis of radicular stress: Measurements in eucalyptus seedlings

In reforesting companies (cellulose industry), eucalyptus is usually cultivated in small plastic containers (50 mL). As seedlings remain for about 120 days in these containers-until transplantation-their roots become space restricted, with consequent limitations in water and nutrient absorption. These restrictions may lead to plant stress, decreasing productivity. In this work, we used the photoacoustic technique to evaluate the photosynthetic activity of Eucalyptus grandis, E. urophylla and E. urograndis seedlings subjected to this limited space availability, seeking a correlation with morphological parameters and fluorescence measurements in these seedlings. Photoacoustic, fluorescence, and morphological analysis were conducted every 15 days, from 45 to 120 days after sowing. Fluorescence and photosynthetic rate were evaluated in vivo and in situ, the latter one using the open photoacoustic technique. Data show that root dry matter diminished markedly at 90 and 120 days after sowing; this behavior showed a high correlation with the gas exchange component of the photoacoustic signal, as well as with the fluorescence ratio Fv/Fm. These results indicate that the soil volume of the container becomes insufficient for the roots after 90 days, probably leading to a nutritional deficiency in plants, which explains the decrease observed in the photosynthetic rate of seedlings. (C) 2003 American Institute of Physics.

Weed interference on maize (Zea mays) under no tillage system

A field trial was carried out in Brazil in March 2002 with the aim to evaluate the effects of different timing and extension of weedy period on maize productivity. The hybrid Pioneer 30K75 was sowed under 7 t ha(-1) mulching promoted by glyphosate spraying. The treatments were divided in two groups: In the first group, weeds were maintained since the maize sowing until different periods in the crop cycle: 0, 14, 28, 42, 56, 70, and 150 days (harvesting time). In the second group, the maize crop was kept weed free for the same periods of the first group. Weed control was done through hand hoeing. A complete randomized blocks experimental design with five replications was used for plots distribution in the field. Nonlinear regression model was used to study the effects of weedy or weedfree periods on maize productivity. Weed community included 13 families and 31 species. Asteraceae, Poaceae, and Euphorbiaceae were the most abundant families. Results showed that under no tillage condition with 7 t ha-1 mulching at sowing time, the maize crop could cohabit with weed community for 54 days without any yield lost. on the other hand, if the crop was kept weed free for 27 days, the weed interference was not enable to reduce maize production. According to these results one weed control measure between 27 and 54 days after crop emergence could be enough to avoid any reduction in maize productivity.

Photosynthesis in eucalyptus studied by the open photoacoustic technique: Effects of irradiance and temperature

This work demonstrates the usefulness of the Open Photoacoustic Cell Technique to study the effects of irradiance and temperature on photosynthesis. bl vivo and ill situ photosynthetic induction measurements were performed in three different species of eucalyptus plants (E. grandis, E. urophylla, and E, urograndis) previously dark-adapted at different temperatures. Photosynthetic activity curves were built as a function of light intensity, indicating the occurrence of photosynthesis saturation. E. urograndis presented higher photosynthetic activity than the other species, especially at low temperature, indicating its tolerance to stress conditions. The incidence of background saturation light of various intensities allowed the irt situ study of photoinhibition in eucalyptus plants through open photoacoustics. (C) 2001 MAIK Nauka/Interperiodica.

Photosynthesis induction in Eucalyptus urograndis seedlings and cuttings measured by Open Photoacoustics

In this work we study the photosynthetic induction in Eucalyptus urograndis leaves using the Open Photoacoustic Cell Technique. In vivo and in situ measurements were performed in leaves of four months-old E. urograndis seedlings and C041 cuttings previously dark-adapted for at least 10 h. Experimental results for the gas exchange component of the photoacoustic (PA) signal are interpreted considering that a gas uptake component would have a phase angle nearly opposite to that of the oxygen evolution component. Analysis of the photosynthetic induction data shows that seedlings present a net oxygen evolution before cuttings, but cuttings reach a higher steady-state photosynthetic activity.

Photosynthetic induction in Eucalyptus urograndis seedlings and cuttings measured by an open photoacoustic cell

Photosynthetic induction in leaves of four-month-old Eucalyptus urograndis seedlings and of cuttings obtained from adult trees that were previously dark-adapted was studied by the in vivo and in situ Open Photoacoustic Cell Technique, Results for the gas exchange component of the photoacoustic (PA) signal were interpreted considering that the gas uptake component would have a phase angle nearly opposite to that of the oxygen evolution component. By subtracting the thermal component from the total PA signal, we studied the competition between gas uptake and oxygen evolution during the photosynthetic induction. Seedlings presented a net oxygen evolution prior to cuttings, but cuttings reached a higher steady-state photosynthetic activity. The chlorophyll (Chl) a/b ratio and the Chl fluorescence induction characteristic F-v/F-m were significantly higher for cuttings, while there was no difference between samples in stomata density and leaf thickness. Thus the differences in PA signals of seedlings and cuttings are associated to differences between the photosystem 2 antenna systems of these samples.

Identification and characterization of different accessions of itchgrass (Rottboellia cochinchinensis)

Studies were conducted to identify and characterize different accessions of itchgrass. Seeds were collected in the counties of Aramina, Campinas, Dumont, Igarapava, Jaboticabal, and Ribeirao Preto, all in the state of São Paulo, Brazil. Accessions were characterized based on dimensions of their stomata, stomatal index (SI), and length and width of their seed (caryopses and husk). Chromosome number and length also were determined, and accessions were further differentiated using molecular markers (polymerase chain reaction [PCR]). Itchgrass from Ribeirao Preto had much longer and narrower seeds than those from the other locations, and their husks were longer as well. Accessions had similar SIs, both on the abaxial and adaxial leaf surfaces. Stomata from Campinas and Igarapava accessions were longer and wider, whereas those from Dumont and Ribeirao Preto were similar and smaller than all others. The accession from Ribeirao Preto is diploid (2n = 20); the rest are polyploid, with the total length of chromosomes smaller than all others. These differences were confirmed by molecular differentiation (PCR).

Listeria monocytogenes infection of P388D1 macrophages results in a biphasic NF-κB (RelA/p50) activation induced by lipoteichoic acid and bacterial phospholipases and mediated by IκBα and IκBβ degradation

As previously reported, Listeria monocytogenes infection of P388D1 macrophages results in a rapid induction of NF-κB DNA-binding activity. Here we show that this induction of NF-κB activity occurs in a biphasic mode: first, a transient, IκBα degradation-dependent phase of activity, also induced by the nonvirulent species Listeria innocua, which is mediated by binding of the bacteria to the macrophage, or by adding Listeria-derived lipoteichoic acid to the macrophage; the second persistent phase of activation is only markedly induced when the bacteria enter the cytoplasm of the host cell and express the virulence genes plcA and plcB, encoding two phospholipases. We suggest that products of the enzymatic activity of phospholipases directly interfere with host cell signal transduction pathways, thus leading to persistent NF-κB activation via persistent IκBβ degradation.