Distribution and Pathogenicity of Colletotrichum Species Associated With Mango Anthracnose in Mexico.

Mango anthracnose, caused by Colletotrichum spp., is the most significant disease of mango (Mangifera indica L.) in almost all production areas around the world. In Mexico, mango anthracnose has only been attributed to C. asianum and C. gloeosporioides. The aims of this study were to identify the Colletotrichum species associated with mango anthracnose symptoms in Mexico by phylogenetic inference using the ApMat marker, to determine the distribution of these species, and to test their pathogenicity and virulence on mango fruits. Surveys were carried out from 2010 to 2012 in 59 commercial orchards in the major mango growing states of Mexico, and a total of 118 isolates were obtained from leaves, twigs, and fruits with typical anthracnose symptoms. All isolates were tentatively identified in the C. gloeosporioides species complex based on morphological and cultural characteristics. The Bayesian inference phylogenetic tree generated with Apn2/MAT intergenic spacer sequences of 59 isolates (one per orchard) revealed that C. alienum, C. asianum, C. fructicola, C. siamense, and C. tropicale were associated with symptoms of mango anthracnose. In this study, C. alienum, C. fructicola, C. siamense, and C. tropicale are reported for the first time in association with mango tissues in Mexico. This study represents the first report of C. alienum causing mango anthracnose worldwide. The distribution of Colletotrichum species varied among the mango growing states from Mexico. Chiapas was the only state in which all five species were found. Pathogenicity tests on mango fruit cultivar Manila showed that all Colletotrichum species from this study could induce anthracnose lesions. However, differences in virulence were evident among species. C. siamense and C. asianum were the most virulent, whereas C. alienum and C. fructicola were considered the least virulent species.

Adolescent parenthood associated with adverse socio-economic outcomes at age 30 years in women and men of the Pelotas, Brazil: 1982 Birth Cohort Study.

OBJECTIVE: To investigate the potential long-term effects of adolescent parenthood on completed education and income. DESIGN: Population-based birth cohort study. SETTING: All live births in 1982, whose mothers lived in the urban area of Pelotas, southern Brazil. SAMPLE: A total of 3701 participants: 1914 women and 1787 men at age 30 years. METHODS: Questionnaires were completed by the mothers in the early phases of this study, and by the cohort members in adolescence and adulthood. Linear regression models and G-computation were used in the analyses. MAIN OUTCOME MEASURES: Educational attainment and income at age 30 years. RESULTS: In women, adolescent parenthood was associated with lower attained education compared with women without adolescent maternity: by -2.8 years [95% confidence interval (CI) -3.2 to -2.3] if their first birth was at age 16-19, and by -4.4 years (-5.5 to -3.3) at age 11-15. These effects were greater among women who had three or more children. Women with adolescent parenthood also had 49 or 33% lower income at age 30 if their first child was born when aged 16-19 or 11-15, respectively. In men, the adverse effect of adolescent parenthood on education appeared to be mediated by a higher number of children and there was no effect of adolescent paternity on income at age 30 years. CONCLUSION: These findings suggest lasting socio-economic disadvantages of adolescent parenthood, with larger effects being apparent in women than in men. TWEETABLE ABSTRACT: Adolescent parenthood has an adverse effect on educational attainment later in life, and on household income among women.

Pre- and Postharvest Strategies to Minimize Mycotoxin Contamination in the Rice Food Chain.

Rice is part of many people's diet around the world, being the main energy source in some regions. Although fewer reports exist on the occurrence of mycotoxins in rice compared to other cereals, fungal contamination and the associated production of toxic metabolites, even at lower occurrence levels compared to other crops, are of concern because of the high consumption of rice in many countries. Due to the diversity of fungi that may contaminate the rice food chain, the co-occurrence of mycotoxins is frequent. Specific strategies to overcome these problems may be applied at the preharvest part of the crop chain, while assuring good practices at harvest and postharvest stages, since different fungi may find suitable conditions to grow at the various stages of the production chain. Therefore, the aim of this review is to present the state-of-the-art knowledge on such strategies in an integrated way, from the field to the final products, to reduce mycotoxin contamination in rice.

Toxic reagents and expensive equipment: are they really necessary for the extraction of good quality fungal DNA?

The aim of this work was to evaluate a fungal DNA extraction procedure with the lowest inputs in terms of time as well as of expensive and toxic chemicals, but able to consistently produce genomic DNA of good quality for PCR purposes. Two types of fungal biological material were tested - mycelium and conidia - combined with two protocols for DNA extraction using Sodium Dodecyl Sulphate (SDS) and Cetyl Trimethyl Ammonium Bromide as extraction buffers and glass beads for mechanical disruption of cell walls. Our results showed that conidia and SDS buffer was the combination that lead to the best DNA quality and yield, with the lowest variation between samples. This study clearly demonstrates that it is possible to obtain high yield and pure DNA from pigmented conidia without the use of strong cell disrupting procedures and of toxic reagents. SIGNIFICANCE AND IMPACT OF THE STUDY: There are numerous methods for DNA extraction from fungi. Some rely on expensive commercial kits and/or equipments, unavailable for many laboratories, or make use of toxic chemicals such as chloroform, phenol and mercaptoethanol. This study clearly demonstrates that it is possible to obtain high yields of pure DNA from pigmented conidia without the use of strong and expensive cell disrupting procedures and of toxic reagents. The method herein described is simultaneously inexpensive and adequate to DNA extraction from several different types of fungi.

Aberration control in adaptive optics: a numerical study of arbitrarily deformable liquid lenses.

By means of numerical simulations, using a computational fluid dynamics software together with an optical ray tracing analysis platform, we show that we can tune various optical aberrations by electrically manipulating the shape of liquid lenses using one hundred individually addressable electrodes. To demonstrate the flexibility of our design, we define electrode patterns based on specific Zernike modes and show that aspherical, cylindrical and decentered shapes of liquid lenses can be produced. Using different voltages, we evaluate the tuning range of spherical aberration (Z11), astigmatism (Z5 and Z6) and coma (Z7), while a hydrostatic pressure is applied to control the average curvature of a microlens with a diameter of 1mm. Upon activating all electrodes simultaneously spherical aberrations of 0.15 waves at a pressure of 30Pa can be suppressed almost completely for the highest voltages applied. For astigmatic and comatic patterns, the values of Z5, Z6 and Z7 increase monotonically with the voltage reaching values up to 0.06, 0.06 and 0.2 waves, respectively. Spot diagrams, wavefront maps and modulation transfer function are reported to quantify the optical performance of each lens. Crosstalk and independence of tunability are discussed in the context of possible applications of the approach for general wavefront shaping.

Internal amplification control of PCR for the Glu1-Dx5 allele in wheat.

One of the limiting factors in using dominant markers is the unique amplification of the target fragment. Therefore, failures in polymerase chain reaction (PCR) or non-amplifications can be interpreted as an absence of the allele. The possibility of false negatives implies in reduced efficiency in the selection process in genetic breeding programs besides the loss of valuable genetic material. Thus, this study aimed to evaluate the viability of a microsatellite marker as an internal amplification control with a dominant marker for the wheat Glu1-Dx5 gene. A population of 77 wheat cultivars/breeding lines was analyzed. Fourteen microsatellite markers were analyzed in silico regarding the formation of dimers and clamps. The biplex reaction conditions were optimized, and the Xbarc117 marker was selected as the internal amplification control with a Glu1-Dx5 marker in wheat. It was concluded that the Xbarc117 microsatellite marker was effective in the simultaneous amplification with a dominant Glu1-Dx5 marker, making biplex PCR viable in wheat for the studied markers.

Numerical simulation of astigmatic liquid lenses tuned by a stripe electrode.

We propose a new design for tuning the astigmatism of liquid micro-lenses using electric field and hydrostatic pressure as control parameters. We explore the feasibility and operating range of the lens with a self-consistent numerical calculation of the electric field distribution and the shape of the two-phase interface. Equilibrium shapes, including surface profiles parallel and perpendicular to a stripe electrode, are extracted to determine the astigmatism. The wavefronts are decomposed into Zernike polynomials under zero defocus conditions using a commercial ray-tracing software. We observe that the global curvature of the lens is primarily controlled by the hydrostatic pressure, while asphericity and astigmatism are controlled by the electric field. For optimized electrode geometries and simultaneous control of pressure and electric fields the astigmatism can be tuned from Z6 = 0…0.38 µm with minor changes in the focal length.

Resistance to vertical root fracture of root filled teeth using different conceptual approaches to canal preparation.

AIM: To evaluate the resistance to vertical root fracture of root filled teeth using different root canal preparation concepts: apical stop and continuously tapering preparation, with different foraminal enlargement. In addition, the effect of filling associated with these different concepts was assessed. METHODOLOGY: Ninety single-rooted premolars were used. The crowns were removed to standardize the root length at 11 mm. Ten roots were not instrumented and not filled (control group). The other roots were divided into four groups (n = 20): apical stop to finishing file size 25, 0.08 taper; apical stop to finishing file size 45, 0.02 taper; continuously tapering preparation to finishing file size 25, 0.08 taper; and continuously tapering preparation to finishing file size 45, 0.02 taper. Afterwards, 10 roots of each group were root filled using Gutta-percha and AH Plus. A continuous wave of condensation was used as the filling technique. The roots were evaluated as regards resistance to vertical fracture using a universal testing machine. The data were evaluated using the Kruskal-Wallis and Mann-Whitney tests. RESULTS: No significant difference in performance was observed between continuously tapering preparation size 25, 0.08 taper and apical stop size 45, 0.02 taper groups. Lower resistance values were found in continuously tapering preparation size 45, 0.02 taper group and the highest in apical stop size 25, 0.08 taper group. After filling, a significant increase in resistance values was observed in all groups. In addition, the continuously tapering preparation size 45, 0.02 taper group had values comparable with those of the control. CONCLUSION: The different canal preparation techniques reduced resistance to fracture when compared with the control group; however, after root filling, there was a significant increase in resistance.

Resveratrol prevents hyperleptinemia and central leptin resistance in adult rats programmed by early weaning.

We have previously shown that early weaning in rats increases the risk of obesity and insulin resistance at adulthood, and leptin resistance can be a prime factor leading to these changes. Resveratrol is reported to decrease oxidative stress, insulin resistance, and cardiovascular risk. However, there is no report about its effect on leptin resistance. Thus, in this study we have evaluated resveratrol-preventing effect on the development of visceral obesity, insulin, and leptin resistance in rats programmed by early weaning. To induce early weaning, lactating dams were separated into 2 groups: early weaning (EW)--dams were wrapped with a bandage to interrupt lactation in the last 3 days of lactation and control (C)--dams whose pups had free access to milk during throughout lactation period (21 days). At 150 days-old, EW offspring were subdivided into 2 groups: EW+res--treated with resveratrol solution (30 mg/kg BW/day) or EW--receiving equal volume of vehicle solution, both given by gavage during 30 days. Control group received vehicle solution. Resveratrol prevented the higher body weight, hyperphagia, visceral obesity, hyperleptinemia, hyperglycemia, insulin resistance, and hypoadiponectinemia at adulthood in animals that were early weaned. Leptin resistance, associated with lower JAK2 and pSTAT3 and higher NPY in hypothalamus of EW rats were also normalized by resveratrol. The present results suggest that resveratrol is useful as therapeutic tool in treating obesity, mainly because it prevents the development of central leptin resistance.

Synthesis, characterization and antifungal activity of chemically and fungal-produced silver nanoparticles against Trichophyton rubrum.

AIMS: To characterize and explore the potential in extracellular biosynthesis of silver nanoparticles (AgNPs) by Penicillium chrysogenum and Aspergillus oryzae and to investigate the antifungal effect of chemically vs biologically synthesized AgNPs comparing with conventional antifungal drugs against Trichophyton rubrum. METHODS AND RESULTS: Chemically synthesized AgNPs (Chem-AgNPs) coated with polyvinylpyrrolidone (PVP) were synthesized by chemical reduction method with glucose in PVP aqueous solution. Biologically synthesized AgNPs (Bio-AgNPs) were produced from the extracellular cell-free filtrate of P. chrysogenum MUM 03.22 and A. oryzae MUM 97.19. Among the commercial antifungal drugs, terbinafine exhibited the lower minimal inhibitory concentration (MIC) range values of 0·063-0·25 µg ml(-1) for the clinical strains. Chem-AgNPs exhibited antifungal activity against all T. rubrum strains. Bio-AgNPs produced by the fungal cell-free filtrate of P. chrysogenum showed an antifungal activity higher than fluconazole but less than terbinafine, itraconazole and Chem-AgNPs. CONCLUSION: The synthesis parameters in future works should be carefully studied to take full advantage of all the potential of filamentous fungi in the synthesis of AgNPs. SIGNIFICANCE AND IMPACT OF THE STUDY: Bio-AgNPs could be used as antifungal agents, namely against dermatophytes.

Combined swimming and soybean intake reduce early atherosclerotic lesions and serum oxidative stress in mice.

AIM: Physical exercise and soybean intake reduced oxidative stress and atherosclerosis. However, the associated effects of both interventions have not been yet investigated. Thus, we aimed to evaluate the combined effects of swimming and soybean intake on lipid profile, oxidative stress and atherogenesis. METHODS: Ten-week-old male Low-Density Lipoprotein Receptor Knockout mice were divided into 4 groups (N.=8 for each group): control diet without swimming; control diet with swimming; soybean rich diet without swimming and soybean rich diet with swimming. Diets were based on American Institute of Nutrition 93 Growth. The diet of soybean groups was made by soybean extract contained isoflavones. The animals in the exercise groups underwent a 6-week swimming program five times per week. Plasma lipid profile was determined using enzymatic kits. Oxidative stress was measured by thiobarbituric acid reactive substances, hydroperoxide and the lipid oxidation resistance determinations. Atherosclerotic lesions were calculated by morphometry. RESULTS: Soybean intake increased high density lipoprotein cholesterol. Moreover, soybean and exercise individually reduced hepatic oxidative stress and atherogenesis in aortic valve. No additional effect was seen in soybean+exercise group. However, the association of soybean and exercise reduced the percentage of lesion area in arch, thoracic and abdominal aorta and increased serum antioxidant potential. CONCLUSION: Soybean intake and swimming are beneficial in reducing atherosclerosis besides improving lipid profile and reducing lipid peroxidation. The association of soybean and swimming aggregates beneficial effects in serum antioxidant potential and in aorta lesion.

Epidemiological profile of individuals with American cutaneous leishmaniasis in Jequitinhonha Valley, Brazil.

This study aimed to characterize the epidemiological profile of patients with American cutaneous leishmaniasis (ACL) in 34 municipalities in Jequitinhonha Valley, Minas Gerais, Brazil, registered from 2005 to 2010 with the Regional Health Superintendence, using data from the Brazilian Disease Notification Information System. Among 281 cases, 156 (55.5%) were included in the study. A statistically significant difference was found in ACL among men between 21 and 60 years of age (P = 0.034) in relation to women and other age groups. Analysing the male sample, a significant association was found between ACL and occupation (P = 0.04). The cutaneous form occurred in 87.3% of cases. The medication most often employed was pentavalent antimony (87.3%), with cure achieved in 91.1% of cases. The epidemiological profile of ACL in the Jequitinhonha Valley is similar to that reported for other regions of Brazil, exhibiting a rural occupational nature and affecting men with low levels of schooling within the productive age.

Two models of early weaning decreases bone structure by different changes in hormonal regulation of bone metabolism in neonate rat.

During the last decade a great concern has developed for determining what factors influence bone mineral accretion in healthy children. Mother's milk represents the primary source of calcium and other nutrients in the neonate. The development of bone and adipose tissue has common origins. Since early weaning decreases adipogenesis in neonate, our aim was to evaluate bone metabolism in 2 models of early weaning (EW) in neonate rats. Lactating rats were separated into 3 groups: control: pups had free access to milk; MEW: dams were involved with a bandage mechanically (M) interrupting lactation in the last 3 days; and PEW: dams were pharmacologically (P) treated to block prolactin (0.5 mg bromocryptine/twice a day) 3 days before standard weaning. Significant difference had p<0.05. At weaning, MEW and PEW pups presented lower body weight (-18% and -15%), total body fat (-26% and -27%), total bone mineral density (-7% and -6%), total bone mineral content (-30% and -32%), bone area (-28% and -30%), serum osteocalcin (-20% and -55%), and higher C-terminal cross-linked telopeptide of type I collagen (CTX-I) (1.3 and 1.1-fold increase). However, serum ionized calcium was lower only in MEW pups (-34%), 25-hydroxyvitamin D was higher (1.4-fold increase), and PTH was lower (-26%) only in PEW group. The present study shows that both early weaning models leads to an impairment of osteogenesis associated with lower adipogenesis by different mechanisms, involving mainly changes in vitamin D and PTH.

Developmental plasticity of endocrine disorders in obesity model primed by early weaning in dams.

Early weaning is associated with changes in the developmental plasticity. Here, we studied the adipocytes morphology, adipokines expression or content in adipose tissue as well as adrenal and thyroid function of neonate and adult offspring primed by early weaning. After birth, lactating rats were divided into 2 groups: EW (early weaning)--dams were wrapped with a bandage to block access to milk during the last 3 days of lactation, and Control--dams whose pups had free access to milk throughout lactation (21 days). At postnatal day (PN) 21, EW pups had lower visceral and subcutaneous adipocyte area (-67.7% and -62%, respectively), body fat mass (-26%), and leptin expression in visceral adipocyte (-64%) but higher leptin expression in subcutaneous adipocyte (2.9-fold increase). Adrenal evaluations were normal, but neonate EW pups presented lower serum T3 (-55%) and TSH (-44%). At PN 180, EW offspring showed higher food intake, higher body fat mass (+21.6%), visceral and subcutaneous adipocyte area (both 3-fold increase), higher leptin (+95%) and ADRß3 (2-fold increase) content in visceral adipose tissue, and higher adiponectin expression in subcutaneous adipose tissue (+47%) but lower in visceral adipose tissue (-40%). Adult EW offspring presented higher adrenal catecholamine content (+31%), but no changes in serum corticosterone or thyroid status. Thus, early weaning primed for hypothyroidism at weaning, which can be associated with the adipocyte hypertrophy at adulthood. The marked changes in catecholamine adrenal content and visceral adipocyte ADRB3 are generally found in obesity, contributing to the development of other cardiovascular and metabolic disturbances.

Biofilms from a Brazilian water distribution system include filamentous fungi.

Filamentous fungi in drinking water can block water pipes, can cause organoleptic biodeterioration, and are a source of pathogens. There are increasing reports of the involvement of the organisms in biofilms. This present study describes a sampling device that can be inserted directly into pipes within water distribution systems, allowing biofilm formation in situ. Calcofluor White M2R staining and fluorescent in situ hybridization with morphological analyses using epifluorescent microscopy were used to analyse biofilms for filamentous fungi, permitting direct observation of the fungi. DAPI (4',6-diamidino-2-phenylindole) was applied to detect bacteria. Filamentous fungi were detected in biofilms after 6 months on coupons exposed to raw water, decanted water and at the entrance of the water distribution system. Algae, yeast, and bacteria were also observed. The role of filamentous fungi requires further investigations.

Fatigue is mediated by cholinoceptors within the ventromedial hypothalamus independent of changes in core temperature.

We investigated brain mechanisms modulating fatigue during prolonged physical exercise in cold environments. In a first set of studies, each rat was subjected to three running trials in different ambient temperatures (T(a)). At 8 °C and 15 °C, core body temperature (T(core)) decreased and increased, respectively, whereas at 12 °C, the T(core) did not change throughout the exercise. In another set of experiments, rats were randomly assigned to receive bilateral 0.2 µL injections of 2.5 × 10(-2) M methylatropine or 0.15 M NaCl solution into the ventromedial hypothalamic nuclei (VMH). Immediately after the injections, treadmill exercise was started. Each animal was subjected to two experimental trials at one of the following T(a) : 5 °C, 12 °C or 15 °C. Muscarinic blockade of the VMH reduced the time to fatigue (TF) in cold environments by 35-37%. In all T(a) studied, methylatropine-treated rats did not present alterations in T(core) and tail skin temperature compared with controls. These results indicate that, below the zone of thermoneutrality, muscarinic blockade of the VMH decreases the TF, independent of changes in T(core). In conclusion, our data suggest that VMH muscarinic transmission modulates physical performance, even when the effects of thermoregulatory adjustments on fatigue are minimal.

Interaction with Penicillium expansum enhances Botrytis cinerea growth in grape juice medium and prevents patulin accumulation in vitro.

Interactions between fungi occur when they grow on the same host plant. This is the case of Botrytis cinerea and Penicillium expansum on grape. P. expansum is also responsible for production of the mycotoxin patulin. In this study, the influence of the interaction between both fungi on fungal growth parameters was studied as well as the effect on the accumulation of patulin by P. expansum. For that purpose, spores of B. cinerea and P. expansum were inoculated together (mixed inoculum), and the parameters growth rate, time for growth and patulin accumulation were assessed. The presence of P. expansum conidia shortened the time for growth of mixed inoculum colonies which, at the end of incubation, were B. cinerea-like. Although some P. expansum growth was observed in mixed inoculum colonies, very low levels of patulin were observed. In assays carried out in patulin-spiked medium, B. cinerea was capable to metabolize the mycotoxin. The capabilities of B. cinerea to shorten time for growth and prevent patulin accumulation are competing abilities that facilitate grape colonization.

First Report of Mango Anthracnose Caused by Colletotrichum karstii in Brazil.

From April to June 2010, mango fruits (Mangifera indica L.) (cv. Tommy Atkins) showing post-harvest anthracnose symptoms were collected during a survey conducted in São Francisco Valley, northeastern Brazil. Fruits affected by anthracnose showed sunken, prominent, dark brown to black decay spots. Small pieces (4 to 5 mm) of necrotic tissues were surface sterilized for 1 min in 1.5% NaOCl, washed twice with sterile distilled water, and plated onto potato dextrose agar (PDA) amended with 0.5 g liter-1 streptomycin sulfate. Plates were incubated at 25°C in the dark for 5 to 7 days and colonies that were morphologically similar to species of Colletotrichum were transferred to PDA (1). Identification was made using morphological characteristics and phylogenetic analysis. Two isolates (CMM 4101 and CMM 4102) presented colonies that had white aerial mycelia and orange conidial mass, varying between colorless and pale orange in reverse. Conidia were hyaline, cylindrical, and aseptate 14.52 (10.40 to 20.20) µm long and 4.90 (3.80 to 6.50) µm wide, length/width ratio = 3.0. Mycelial growth rate was 5.20 mm per day at 25°C. Morphological and cultural characterizations were consistent with the description of Colletotrichum karstii (3). PCR amplification by universal primers (ITS1/ITS4) and DNA sequencing of the internal transcribed spacer (ITS1-5.8S-ITS2 rRNA gene cluster) were conducted to confirm the identifications. Analysis of representative sequences (GenBank Accession Nos. HM585409 and HM585406) suggested that the isolated pathogen was C. karstii. Using published ITS data for C. karstii (3), a phylogenetic analysis was made via Bayesian inference, which shows that the isolated fungi belong to the C. karstii clade. Sequences of the isolates obtained in this study were deposited in GenBank (KC295235 and KC295236), and cultures were deposited in the Culture Collection of Phytopathogenic Fungi of the Universidade Federal Rural de Pernambuco (CMM, Recife, Brazil). Pathogenicity tests were conducted with the C. karstii strains on mango fruits cv. Tommy Atkins. Mycelial plugs taken from the margin of actively growing colonies (PDA) of each isolate were applied in shallow wounds (0.4 cm in diameter) at the medium region of the each fruit. PDA discs without fungal growing were used as controls. Inoculated fruits were placed in plastic containers lined with paper towels wetted in distilled water. The containers were partially sealed with plastic bags to maintain high humidity and incubated at 25°C in the dark. The plastic bags and paper towels were removed after 24 h, and fruits were kept at the same temperature. The experiment was arranged in a completely randomized design with four replicates per treatment (isolate) and four fruits per replicate. Typical anthracnose symptoms were observed after 10 days in mango fruits. C. karstii was successfully reisolated from symptomatic mango fruits to fulfill Koch's postulates. C. karstii was previously described from Orchidaceae in southwest China and the United States (2,3). To our knowledge, this is the first report of C. karstii causing mango anthracnose in Brazil and worldwide. References: (1) U. Damm et al. Stud. Mycol. 73:1, 2012. (2) I. Jadrane et al. Plant Dis. 96:1227, 2012. (3) Yang et al. Cryptogamie Mycol. 32:229, 2011.

How cutting types and shading levels influence the vegetative propagation of Pereskia aculeata?

Information from Pereskia aculeata Miller (Cactaceae) responses regarding the portion of the stem cuttings branch parental plants and the shading level need to be investigated in order to establish techniques for their producing seedlings. We aimed was to evaluate the effect of cutting types and shading levels in the production of P. aculeata seedlings. We studied three cutting types, collected from different portions of the stem cuttings branch: herbaceous, semi-hardwood, and hardwood, which were arranged under two shading levels: 0% (full sun) or 50% (shade). The selected parent plants had an adequate and vigorous phytosanitary aspect. After 90 days after the cuttings, the seedlings were evaluated regarding survival, growth indicators, biomass production and partitioning, and allometric indices. Seedlings from hardwood cuttings and produced under 0% shading showed higher survival. The largest number of sprouts occurred in seedlings of semi-hardwood and hardwood cuttings. The highest leaf area values ​​occurred in seedlings of semi-hardwood and hardwood cuttings under 0% shading. Seedlings from hardwood cuttings had higher biomass allocation in the roots under 50% shading. Seedlings of herbaceous and semi-hardwood portion allocated values ≥ 70% of the biomass in the aerial part. Seedlings of species are able to adjust to different shading levels as a plasticity mechanism. For the production of P. aculeata seedlings, the hardwood portion of the stem cuttings branch under full sun cultivation is recommended. In addition, semi-hardwood cuttings grown under 50% shading can also be used for seedlings production.

First Report of Mango Dieback Caused by Pseudofusicoccum stromaticum in Brazil.

From September to December 2010, mango (Mangifera indica L.) stems showing dieback symptoms were collected during a survey conducted in São Francisco Valley, northeastern Brazil. Small pieces (4 to 5 mm) of necrotic tissues were surface sterilized for 1 min in 1.5% NaOCl, washed twice with sterile distilled water, and plated onto potato dextrose agar (PDA) amended with 0.5 g liter-1 streptomycin sulfate. Plates were incubated at 25°C in the dark for 14 to 21 days and colonies that were morphologically similar to species of Botryosphaeriaceae were transferred to PDA. Colonies developed a compact mycelium that was initially white, but becoming gray dark after 4 to 6 days of incubation at 25°C in darkness. Identification was made using morphological characteristics and DNA based molecular techniques. Pycnidia were obtained on 2% water agar with sterilized pine needles as substratum after 3 weeks of incubation at 25°C under near-UV light. Pycnidia were large, multilocular, eustromatic, covered with hyphae; locule totally embedded without ostioles, locule walls consisting of a dark brown textura angularis, becoming thinner and hyaline toward the conidiogenous region. Conidia were hyaline, thin to slightly thickened walled, aseptate, with granular contents, bacilliform, straight to slightly curved, apex and base both bluntly rounded or just blunt, 15.6 to 25.0 (20.8) µm long, and 2.7 to 7.9 (5.2) µm wide, length/width = 4.00. According to these morphological characteristics, three isolates (CMM1364, CMM1365, and CMM1450) were identified as Pseudofusicoccum stromaticum (1,3,4). PCR amplification by universal primers (ITS4/ITS5) and DNA sequencing of the internal transcribed spacer (ITS1-5.8S-ITS2 rRNA gene cluster) were conducted to confirm the identifications through BLAST searches in GenBank. The isolates were 100% homologous with P. stromaticum (3) (GenBank Accession Nos. AY693974 and DQ436935). Representative sequences of the isolates were deposited in GenBank (Accession Nos. JF896432, JF966392, and JF966393). Pathogenicity tests were conducted with the P. stromaticum strains on 5-month-old mango seedlings (cv. Tommy Atkins). Mycelial plugs taken from the margin of actively growing colonies (PDA) of each isolate were applied in shallow wounds (0.4 cm in diameter) on the stem (center) of each plant. Inoculation wounds were wrapped with Parafilm. Control seedlings received sterile PDA plugs. Inoculated and control seedlings (five each) were kept in a greenhouse at 25 to 30°C. After 5 weeks, all inoculated seedlings showed leaf wilting, drying out of the branches, and necrotic lesions in the stems. No symptoms were observed in the control plants. P. stromaticum was successfully reisolated from symptomatic plants to fulfill Koch's postulates. P. stromaticum was described from Acacia, Eucalyptus, and Pinus trees in Venezuela (3,4), and there are no reports of this fungus in other hosts (2). To our knowledge, this is the first report of P. stromaticum causing mango dieback in Brazil and worldwide. References: (1) P. W. Crous et al. Stud. Mycol. 55:235, 2006. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , 18 May 2011. (3) S. Mohali et al. Mycol. Res. 110:405, 2006. (4) S. R. Mohali et al. Fungal Divers. 25:103, 2007.