All Green, But Equal? Morphological Traits And Ecological Implications On Spores Of Three Species Of Mosses In The Brazilian Atlantic Forest.

Spores of the tropical mosses Pyrrhobryum spiniforme, Neckeropsis undulata and N. disticha were characterized regarding size, number per capsule and viability. Chemical substances were analyzed for P. spiniforme and N. undulata spores. Length of sporophyte seta (spore dispersal ability) was analyzed for P. spiniforme. Four to six colonies per species in each site (lowland and highland areas of an Atlantic Forest; Serra do Mar State Park, Brazil) were visited for the collection of capsules (2008 - 2009). Neckeropsis undulata in the highland area produced the largest spores (ca. 19 µm) with the highest viability. The smallest spores were found in N. disticha in the lowland (ca. 13 µm). Pyrrhobryum spiniforme produced more spores per capsule in the highland (ca. 150,000) than in lowland (ca. 40,000); longer sporophytic setae in the lowland (ca. 64 mm) than in the highland (ca. 43 mm); and similar sized spores in both areas (ca. 16 µm). Spores of N. undulata and P. spiniforme contained lipids and proteins in the cytoplasm, and acid/neutral lipids and pectins in the wall. Lipid bodies were larger in N. undulata than in P. spiniforme. No starch was recorded for spores. Pyrrhobryum spiniforme in the highland area, different from lowland, was characterized by low reproductive effort, but presented many spores per capsule.

Indoor and outdoor atmospheric fungal spores in the São Paulo metropolitan area (Brazil): species and numeric concentrations

The aim of this study was to estimate the indoor and outdoor concentrations of fungal spores in the Metropolitan Area of Sao Paulo (MASP), collected at different sites in winter/spring and summer seasons. The techniques adopted included cultivation (samples collected with impactors) and microscopic enumeration (samples collected with impingers). The overall results showed total concentrations of fungal spores as high as 36,000 per cubic meter, with a large proportion of non culturable spores (around 91 per cent of the total). Penicillium sp. and Aspergillus sp. were the dominant species both indoors and outdoors, in all seasons tested, occurring in more than 30 per cent of homes at very high concentrations of culturable airborne fungi [colony forming units(CFU) m−3]. There was no significant difference between indoor and outdoor concentrations. The total fungal spore concentration found in winter was 19 per cent higher than that in summer. Heat and humidity were the main factors affecting fungal growth; however, a non-linear response to these factors was found. Thus, temperatures below 16°C and above 25°C caused a reduction in the concentration (CFU m−3) of airborne fungi, which fits with MASP climatalogy. The same pattern was observed for humidity, although not as clearly as with temperature given the usual high relative humidity (above 70 per cent) in the study area. These results are relevant for public health interventions that aim to reduce respiratory morbidity among susceptible populations

Sections of some Carboniferous dispersed spores

Sections of microspores, some cingulate, one zonate and one saccate, are discussed and illustrated. It is shown that sections aid the elucidation of wall structures; thus diagnoses can be more precisely written and this may eventually remove some classificatory difficulties. A sectioning technique is described.

Sections of some spores from the Lower Carboniferous of Spitsbergen

Sections of four lower Carboniferous dispersed-spore species, Labiadensites fimbriatus (Waltz) Hacquebard and Barss, Cristatisporites echinatus Playford, Camptozonotriletes velatus (Waltz) Playford, and Spinozonotriletes uncatus Hacquebard, are discussed and illustrated.

Heat resistance of Bacillus spores when adhered to stainless steel and its relationship to spore hydrophobicity

Twenty-one strains of Bacillus (10 B. stearothermophilus, 3 B. cereus, and 8 B. licheniformis strains) were assayed for spore surface hydrophobicity on the basis of three measures: contact angle measurement (CAM), microbial adhesion to hydrocarbons (MATH), and hydrophobic interaction chromatography (HIC). On the basis of the spore surface characteristics obtained from these assays, along with data on the heat resistance of these spores in water, eight strains of Bacillus (three B. stearothermophilus, three B. cereus, and two B. licheniformis strains) either suspended in water or adhering to stainless steel were exposed to sublethal heat treatments at 90 to 110degreesC to determine heat resistance (D-value). Significant increases in heat resistance (ranging from 3 to 400%) were observed for the eight strains adhering to stainless steel. No significant correlation was found between these heat resistance increases and spore surface characteristics as determined by the three hydrophobicity assays. There was a significant positive correlation between the hydrophobicity data obtained by the MATH assay and those obtained by the HIC assay, but these data did not correlate with those obtained by the CAM assay.

Cytotoxicity Induced by Extracts of Pisolithus tinctorius Spores on Human Cancer and Normal Cell Lines—Evaluation of the Anticancer Potential

Fungi have been considered a potential source of natural anticancer drugs. However, studies on these organisms have mainly focused on compounds present in the sporocarp and mycelium. The aim of this study was to assess the anticancer potential of fungal spores using a bioassay-guided fractionation with cancer and normal cell lines. Crude extracts from spores of the basidiomycetous fungus Pisolithus tinctorius were prepared using five solvents/solvent mixtures in order to select the most effective crude extraction procedure. A dichloromethane/methanol (DCM/MeOH) mixture was found to produce the highest extraction yield, and this extract was fractionated into 11 fractions. Crude extracts and fractions were assayed for cytotoxicity in the human osteocarcinoma cell line MG63, the human breast carcinoma cell line T47D, the human colon adenocarcinoma cell line RKO, and the normal human brain capillary endothelial cell line hCMEC/D3. Cytotoxicity was assessed by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) reduction assay. The results showed a reduction in cancer cell viability of approximately 95% with 4 of 11 fractions without a significant reduction in viability of hCMEC/D3 cells. Data demonstrated that spores of P. tinctorius might serve as an interesting source of compounds with potential anticancer properties.

Chitinolytic activity in viable spores of encephalitozoon species

By employing 4-methylumbelliferyl-beta-D-NN',N"-triacetylchitotriose substrate in a semi quantitative assay, chitinolytic activity in viable spores of Encephalitozoon cuniculi and E. intestinalis was detected and dependence on reaction time, spore concentration, concentration of substrate and temperature were demonstrated. It was possible to block the chitinolytic activity by chitin hydrolysate. By incubation at 80°C for 10 min or at 55°C for 20 min the spores were loosing the chitinolytic activity. Incubation of the spores in trypsin reduced the chitinolytic activity. Cellulase activity could not be detected.

Identification and isolation of two ascomycete fungi from spores of the arbuscular mycorrhizal fungus Scutellospora castanea.

Two filamentous fungi with different phenotypes were isolated from crushed healthy spores or perforated dead spores of the arbuscular mycorrhizal fungus (AMF) Scutellospora castanea. Based on comparative sequence analysis of 5.8S ribosomal DNA and internal transcribed spacer fragments, one isolate, obtained from perforated dead spores only, was assigned to the genus Nectria, and the second, obtained from both healthy and dead spores, was assigned to Leptosphaeria, a genus that also contains pathogens of plants in the Brassicaceae. PCR and randomly amplified polymorphic DNA-PCR analyses, however, did not indicate similarities between pathogens and the isolate. The presence of the two isolates in both healthy spores and perforated dead spores of S. castanea was finally confirmed by transmission electron microscopy by using distinctive characteristics of the isolates and S. castanea. The role of this fungus in S. castanea spores remains unclear, but the results serve as a strong warning that sequences obtained from apparently healthy AMF spores cannot be presumed to be of glomalean origin and that this could present problems for studies on AMF genes.

Reserve substances and storage of Cyathea delgadii Sternb. spores

The spores used were collected from a population of trees of Cyathea delgadii Sternb. growing in a gallery forest at the Reserva Biológica e Estação Experimental de Moji Guaçu, São Paulo state, Brazil (22°18’S and 47°11’W). The germination of spores of Cyathea delgadii decreases with time when kept in closed bottles under storage at 4°C in darkness. Germination is still very high after storage for one year. Spores stored for three years do not germinate. The results also show a decrease in soluble proteins and an increase in starch after several months storage.

Light and storage on the germination of spores of Dicksonia sellowiana (Presl.) Hook., Dicksoniaceae

Spores of Dicksonia sellowiana are positively photoblastic and reach the maximum percentage of germination at 23 ± 2°C in white light after seven days of imbibition. The pre-induction phase for spores induced by white or red light for 24 hours was 72 hours. Gametophytes grown in white light were plane and bidimensional, while those grown under red light were filamentous. The higher the number of hours of light applied per day during 10 days, the higher the percentage of germination. Germination was higher for long white light treatments applied on a daily basis. The effect of different light intensities on germination was also investigated here. The lower percentages of germination were observed for spores kept under 43% and 2% of full sunlight, while those kept under 26, 19 and 4% presented higher percentages. Spores presented circa 82% of germination after 731 days of storage under refrigeration at aproximately 10°C.

The survival and establishment potential of spores of Cyathea delgadii Sternb. in soils from Itirapina and Moji Guaçu (SP), Brazil

Germination of spores of Cyathea delgadii was carried out in soils collected in March 1997 (rainy season) at three depths (0-5, 5-10 and 10-15 cm) from the cerrado, open-cerrado, gallery forest and marsh at Moji Guaçu and cerradão, cerrado and gallery forest at Pedregulho, in Itirapina, both in the state of São Paulo, Brazil. Viability of spores mixed with soil and buried in the cerrado at the Reserva in Moji Guaçu were conducted for up to 10 months. The spores germinated in all soils. Germination in the soils was significantly lower than in distilled water. Germination was higher in soils from the gallery forest and cerrado than from open cerrado, cerradão and marsh. The germination was the same in the soil samples from the three depths of gallery forest, cerrado and cerradão. Spores of C. delgadii maintained viability longer when buried in cerrado soil than when dry stored at 4°C. After 10 months germination was 50.5% in buried spores against 3.8% in spores of the same age and harvest, stored at 4°C.

High irradiance and temperature inhibit the germination of spores of the fern Rumohra adiantiformis (Forst.) Ching (Dryopteridaceae)

Rumohra adiantiformis (Forst.) Ching is a fern (Dryopteridaceae) which is used to compose floral arrangements. Fertile fronds were harvested in the "Permanently Protected Area" of Ilha Comprida, São Paulo, Brazil. Sterilized spores were germinated in Mohr liquid medium modified by Dyer. The effect of 72%, 54%, 17% and 9% of total irradiance on germination under field conditions, was analyzed. Experiments were carried out in March (I), April (II) and August of 2000 (III). Under 54% and 72% of total irradiance in Experiment I (March) the germination was completely inhibited and partially inhibited under 72% of total irradiance in Experiment II (April). The lowest mean germination time () was observed under 9% of total irradiance in Experiments II (11.62 days) and III (8.80 days) respectively, followed by 17% in Experiment III (10.12 days) and 9% of total irradiance in the Experiment I (11.62 days ). The effect of temperatures of 15 ± 1, 20 ± 1, 25 ± 1 and 30 ± 1 ºC on germination was also analyzed. The lowest mean germination time (7.93 days) was observed at 25 ± 1 °C followed by 20 ± 1 °C. The highest mean germination time was observed at 15 ± 1 °C (12.10 days) followed by 30 ± 1 °C (10.63 days), which inhibited germination. The germination of R. adiantiformis was photoinhibited by high irradiance and partially inhibited by the highest temperature tested.

Purification and characterization of a phytoalexin elicitor from spores of the saprobe Mucor ramosissimus

Plants accumulate antimicrobial compounds (phytoalexins) in response to a wide variety of microorganisms. Mucor ramosissimus Samutsevitsch is a saprobe capable of inducing phytoalexin production in soybean cotyledons and in the leaves of tropical Rubiaceae on whose surface it has been found. In the present study, the elicitor from M. ramosissimus was partially purified and the activity compared to that of a glucan elicitor isolated from Phytophthora sojae. Optimal isolation of the elicitor (based on fungal growth, yield of spores and elicitor activity) was achieved by autoclaving spores obtained from nine day-old cultures of the fungus. The elicitor was precipitated with ethanol and purified by chromatography on an anion exchange column, which retained the elicitor, and a Concanavalin A-affinity matrix, to which the elicitor did not bind. The purification resulted in a considerable increase (six-fold) in the specific activity of the elicitor. Neutral sugar composition, analyzed by HPLC, revealed the predominance of mannose, followed by glucose and galactose, whereas colorimetric quantification showed the presence of uronic acids. GC-MS analysis of the elicitor revealed the predominance of glucuronic acid and mannose. These results suggest that fragments of mucoran-type polysaccharides are the phytoalexin elicitors present in the spores of the saprobe M. ramosissimus. Our results also indicate for the first time that soybean cotyledon tissues can recognize fragments of glucuronic-acid heteropolymers as phytoalexin elicitors.

Germination of spores and growth of gametophytes and sporophytes of Rumohra adiantiformis (Forst.) Ching (Dryopteridaceae) after spore cryogenic storage

Rumohra adiantiformis (Forst.) Ching is a fern (Dryopteridaceae) used in floral arrangements. Spores sterilized in 15% (v/v) solution of commercial sodium hypochlorite for 10 minutes and unsterilized spores were plunged in liquid nitrogen and held for 15 minutes and for 90 days. After the cryogenic treatments, spores were taken out of liquid nitrogen and rapidly thawed out in a water bath or slowly at room temperature and were cultured in Mohr's mineral solution as modified by Dyer, kept at 25 ± 2 ºC and a 16-hours photoperiod. Statistical differences were not observed in the germination of unsterilized spores immersed or not immersed in liquid nitrogen, but when the spores were previously sterilized, a severe inhibition of germination was observed in cryopreserved spores. Faster mean germination time was observed for unsterilized spores cryopreserved in liquid nitrogen for 15 minutes. The germination of spores stored in liquid nitrogen for 90 days reached the maximum percentage after 12 days, while control spores reached their maximum percentage after 16 days. Levels of soluble sugars did not vary among treatments in gametophytes cultivated for 10 weeks after spore inoculation. The number of fronds and the length of the longest frond on sporophytes did not differ statistically among treatments. The relative growth rate of sporophytes grown from cryopreserved and control spores were not statistically different among treatments. Spores of R. adiantiformis immersed in liquid nitrogen for 15 minutes apparently produced phenotypically normal plants.

Determination of thermobacteriological parameters and size of Bacillus stearothermophilus ATCC 7953 spores

In order to determine thermobacteriological parameters for B. stearothermophilus spores, they were diluted in a saline solution medium and in ground corn-soybean mix, distributed in TDT tube, and submitted to heat for a specific period of time. The D value (time to reduce 1 log cycle of microbial count under a certain temperature) and z value (variation of temperature to cause 10-fold change in D value) were estimated. To estimate their dimensions, the spores were visualized by using a scanning electron microscope. D121.1 ºC and z values for these spores, as determined in the saline solution, were 8.8 minutes and 12.8 ºC, respectively. D121,1 ºC and z values determined in the corn-soy mix were 14.2 minutes and 23.7 ºC, respectively. The micrographs indicated that the spores have homogeneous shape and size, with length and diameter of 2 and 1 µm, respectively. These results confirm that the spore is highly thermal-resistant, and it is a good biological indicator to evaluate the extrusion process as a feed sterilizer.