Low-temperature-induced changes in trehalose, mannitol and arabitol associated with enhanced tolerance to freezing in ectomycorrhizal basidiomycetes ( Hebeloma spp.)

Ectomycorrhizal fungi have been shown to survive sub-zero temperatures in axenic culture and in the field. However, the physiological basis for resistance to freezing is poorly understood. In order to survive freezing, mycelia must synthesise compounds that pro-tect the cells from frost damage, and certain fungal-spe-cific soluble carbohydrates have been implicated in this role. Tissue concentrations of arabitol, mannitol and trehalose were measured in axenic cultures of eight Hebeloma strains of arctic and temperate origin grown at 22, 12, 6 and 2°C. In a separate experiment, mycelia were frozen to –5°C after pre-conditioning at either 2°C or 22°C. For some, especially temperate strains, there was a clear increase in specific soluble carbohydrates at lower growth temperatures. Trehalose and mannitol were present in all strains and the highest concentrations (close to 2.5% and 0.5% dry wt.) were recorded only after a cold period. Arabitol was found in four strains only when grown at low temperature. Cold pre-condi-tioning enhanced recovery of mycelia following freez-ing. In four out of eight strains, this was paralleled by increases in mannitol and trehalose concentration at low temperature that presumably contribute towards cryopro-tection. The results are discussed in an ecological con-text with regard to mycelial overwintering in soil.

Temperature regulation of extracellular proteases in ectomycorrhizal fungi (Hebeloma spp.) grown in axenic culture

Strains of Hebeloma representative of different climatic zones were grown in axenic culture at either 2 °C and 22° or 6° and 22°. Culture filtrates were assayed for proteolytic activity using FITC labelled BSA as a substrate. Assays were run between 0–37°. Growth at low temperature induced greater proteolytic activity (g−1 D.W. mycelium). Many of the strains produced protease(s) which retained significant activity at temperatures as low as 0°, and a thermal optimum between 0–6° with a second optimum at higher temperature. The results are discussed in relation the nutrient acquisition potential of ectomycorrhizal fungi at low temperature and the contribution such cold active proteases might make to the soil enzyme pool.

The effect of temperature and inorganic phosphorus supply on growth and acid phosphatase production in arctic and temperate strains of ectomycorrhizal Hebeloma spp. in axenic culture

Acid phosphatase production by 12 Hebeloma strains was usually derepressed when inorganic phosphorus in the growth medium was limited, but appeared to be constitutive in some strains. At low temperatures (≤ 12°) arctic strains produced more extracellular and wall-bound acid phosphatase, yet grew more slowly than the temperate strains. We suggest that low growth rates in arctic strains may be a physiological response to cold whereby resources are diverted into carbohydrate accumulation for cryoprotection. At near freezing temperatures, increased extracellular phosphatase production may compensate for a loss of enzyme activity at low temperature and serve to hydrolyse organic phosphorus in frozen soil over winter.

Utilization of organic nitrogen by ectomycorrhizal fungi (Hebeloma spp.) of arctic and temperate origin

Arctic and temperate strains of Hebeloma spp. were grown in axenic culture on glutamic acid, alanine, lysine and NH4+ as sole sources of nitrogen (N), with excess carbon (C) or deficient C (supplied as glucose). Their ability to utilize seed protein as a natural N source was also assessed. All strains tested had the capacity to assimilate amino acids and generally utilized alanine and glutamic acid more readily than NH4+. Some strains were able to utilize amino C when starved of glucose C, and could mineralize amino-N to NH3-N. Arctic strains, in particular, appeared to be pre-adapted to the utilization of seed protein N and glutamic acid N, which is often liberated in high concentrations after soil freezing. The results are discussed in relation to their possible ecological importance.

Induction of cold active acid phosphomonoesterase activity at low temperature in psychrotrophic ectomycorrhizal Hebeloma spp.

Hebeloma strains of arctic and temperature origin, grown at 22° or 6°, were assayed for wall-bound and extracellular acid phosphomonoesterase (pNPPase) across a temperature range 2-37°. Only when grown at 6° was a cold active extracellular pNPPase induced in all the arctic strains and most of the temperature strains tested. Such enzymes are suggested to be a adaptation to low soil temperatures, and are discussed in the context of ectomycorrhizal access to soil PO4− monoesters at low temperature.

Long-term storage of ectomycorrhizal basidiomycetes (Hebeloma spp.) at low temperature

A method for maintaining viable cultures of ectomycorrhizal Hebeloma strains in cold liquid culture medium is described. Isolates of Hebeloma spp., collected over a wide geographic range, were stored at 2 °C for a period of three years. All cultures survived this storage period, a greater time period and success rate than has previously been reported for the long term storage of ectomycorrhizal basidiomycetes. The method may prove useful for long-term storage of other basidiomycete genera.

Potential vectors and hosts of rickettsia spp: epidemiological studies in the Vale do Paraiba, state of Rio de Janeiro/Brazil

FAPESP

Trypanosoma cruzi in Brazilian Amazonia: Lineages TCI and TCIIa in wild primates, Rhodnius spp. and in humans with Chagas disease associated with oral transmission

In this study, we provide phylogenetic and biogeographic evidence that the Trypanosomo cruzi lineages T. cruzi I (TCI) and T. cruzi IIa (TCIIa) circulate amongst non-human primates in Brazilian Amazonia, and are transmitted by Rhodnius species in overlapping arboreal transmission cycles, sporadically infecting humans. TO presented higher prevalence rates, and no lineages other than TCI and TCIIa were found in this study in wild monkeys and Rhodnius from the Amazonian region. We characterised TO and TCIIa from wild primates (16 TO and five TCIIa), Rhodnius spp, (13 TCI and nine TCIIa), and humans with Chagas disease associated with oral transmission (14 TO and five TCIIa) in Brazilian Amazonia. To our knowledge, TCIIa had not been associated with wild monkeys until now. Polymorphisms of ssrDNA, cytochrome b gene sequences and randomly amplified polymorphic DNA (RAPD) patterns clearly separated TCIIa from TCIIb-e and TCI lineages, and disclosed small intra-lineage polymorphisms amongst isolates from Amazonia. These data are important in understanding the complexity of the transmission cycles, genetic structure, and evolutionary history of T cruzi populations circulating in Amazonia, and they contribute to both the unravelling of human infection routes and the pathological peculiarities of Chagas disease in this region. (C) 2008 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

Selectivity of the herbicide flazasulfuron applied after postemergence in sugarcane (Saccharum spp. L.) crop

The selectivity of the herbicide flazasulfaron was evaluated when applied at two rates (50 and 100 g/ha) with and without surfactants (Aterbane or Agral at 0.2% v/v). The treatment was applied at early (three leaves) and late (five to six leaves) stages of the postemergence of sugarcane plants (var. RB845257) grown in two soils (sandy and clay) with good moisture status before and after application. Despite the toxicity symptoms, especially at the late stage with the higher rate of application in the sandy soil, the herbicide did not affect the growth nor the stalk yield. The presence of the surfactants had no effect on the toxicity symptoms.

Efeitos do período e volume de aplicação na segurança dos tratoristas aplicando herbicidas na cultura de cana-de-açúcar (Saccharum spp.)

O objetivo deste trabalho foi avaliar os efeitos do período e volume de aplicação na segurança da atividade de tratoristas aplicando herbicidas na cultura de cana-de-açúcar com o pulverizador de barra montado em trator e a eficiência de equipamentos de proteção individual (EPIs) e de uma cabina acoplada ao trator. As exposições dérmicas de 13 condições de trabalho foram avaliadas e analisadas estatisticamente por meio do delineamento inteiramente ao acaso e do esquema fatorial 3 x 2 x 2 + 1. O fator A foi a condição de exposição: 1) exposição dérmica potencial (EDP) - sem nenhuma medida de segurança; 2) exposição com cabina no trator (Cabina); e 3) exposição com as vestimentas (EPI). O fator B foi o volume de aplicação: 1) 200 L ha-1 e 2) 100 L ha-1; e o fator C foi o período de aplicação: 1) diurno e 2) noturno. Como testemunha foi avaliada a EDP do tratorista aplicando na atividade usual de 300 L de calda ha-1, no período diurno. As exposições dérmicas (EDs) aos herbicidas considerados nessas condições de trabalho foram estimadas por meio de dados substitutos das EDs avaliadas ao cátion Cu+2 adicionado como traçador nas caldas aplicadas. O pulverizador utilizado foi do modelo PJ 600, com barra de 12 m de comprimento e 24 bicos de jato plano TT 110 04 ou TT 110 02. As 13 condições de trabalho avaliadas foram classificadas como seguras (MS>1) para o tratorista aplicando os herbicidas glyphosate (48% i.a.), MSMA (48%), diuron (46,8%) + hexazinone (13,2%), clomazone (50%), sulfentrazone (50%), ametryne (50%), diuron (50%), isoxaflutole (75%), metribuzin (48%), 2,4-D (80,6%), ametryne (30%) + clomazone (20%), ametryne (73,25%) + trifloxysulfuron (1,85%) e tebuthiuron (80 %) e inseguras para o herbicida atrazine (50%), nos dois períodos e nos três volumes de aplicação, e ametryne (50%), na aplicação diurna e 100 L de calda ha-1. As aplicações noturnas e os volumes de aplicação reduzidos tornaram as condições de trabalho mais seguras, exceto para o atrazine. A eficiência dos EPIs para a aplicação de 300 L ha-1 variou de 69,5 a 89,3%, e a da cabina do trator variou entre 76,4 e 83,3%, em relação aos volumes reduzidos de 100 e 200 L ha-1.

Seletividade de imazapic às soqueiras de cana-de-açúcar (Saccharum spp.)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Relações filogenéticas e diversidade de isolados de Guignardia spp oriundos de diferentes hospedeiros nas regiões ITS1-5,8S-ITS2

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Transferability of microsatellite loci from exotic Cervidae to Brazilian brocket deer (Mazama spp, Mammalia: Cervidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Evaluation of resistance to Tomato severe rugose virus (ToSRV) in Capsicum spp. genotypes

Tomato severe rugose virus (ToSRV) is the predominant species of begomovirus in São Paulo State, Brazil, and infects primarily tomato and pepper plants. There is no information about genetic resistance of pepper to this virus, so in this work the reaction of 29 genotypes of Capsicum spp. was evaluated by inoculation of two ToSRV isolates: ToSRV-Sk (isolated from a tomato plant) and ToSRV-PJU (isolated from a pepper plant). For both isolates, two C. annuun genotypes (Catarino Cascabel - México and Silver) showed no symptoms 30 days after inoculation (d.a.i). In a second experiment, these two genotypes were evaluated for 150 d.a.i and, again, no symptoms could be observed. However, the virus was detected by RCA-PCR, indicating that both genotypes are susceptible, but less affected by ToSRV infection. Catarino Cascabel - México and Silver can be indicated for use in breeding programs for resistance of pepper to ToSRV.