Diversity of ectomycorrhizal Thelephoraceae in Tuber melanosporum-cultivated orchards of Northern Spain.

Truffles are edible hypogeous ascomycetes highly appreciated worldwide, especially the black truffle (Tuber melanosporum Vittad.). In recent decades, the cultivation of the black truffle has expanded across the Mediterranean climate regions in and outside its native range. Members of the Thelephoraceae (Thelephorales, Agaricomycetes, Basidiomycota) are commonly found in truffle plantations, but their co-occurrence with Tuber species and other members of the fungal community has been scarcely reported. Thelephoraceae is one of the most represented families of the ectomycorrhizal fungal community in boreal and Mediterranean forests. To reveal the diversity of these fungi in T. melanosporum-cultivated plantations, ten orchards located in the Navarra region (Northern Spain) were surveyed for 2 years. Morphological and molecular approaches were used to detect and identify the Thelephoraceae ectomycorrhizas present in those plantations. Ten different mycorrhizal types were detected and described. Four of them were morphologically identified as Tomentella galzinii, Quercirhiza cumulosa, Q. squamosa, and T39 Thelephoraceae type. Molecular analyses revealed 4-6 operational taxonomic units (OTUs), depending on the nucleotide database used, but similarities remained under 95 % and no clear species assignments could be done. The results confirm the diversity and abundance of this fungal family in the ectomycorrhizal community of black truffle plantations, generally established in Mediterranean areas. The occurrence and relative abundance of Thelephoraceae ectomycorrhizas is discussed in relation to their possible influence on truffle production.

Increased evapotranspiration demand in a Mediterranean climate might cause a decline in fungal yields under global warming.

Wild fungi play a critical role in forest ecosystems, and its recollection is a relevant economic activity. Understanding fungal response to climate is necessary in order to predict future fungal production in Mediterranean forests under climate change scenarios. We used a 15-year data set to model the relationship between climate and epigeous fungal abundance and productivity, for mycorrhizal and saprotrophic guilds in a Mediterranean pine forest. The obtained models were used to predict fungal productivity for the 2021-2080 period by means of regional climate change models. Simple models based on early spring temperature and summer-autumn rainfall could provide accurate estimates for fungal abundance and productivity. Models including rainfall and climatic water balance showed similar results and explanatory power for the analyzed 15-year period. However, their predictions for the 2021-2080 period diverged. Rainfall-based models predicted a maintenance of fungal yield, whereas water balance-based models predicted a steady decrease of fungal productivity under a global warming scenario. Under Mediterranean conditions fungi responded to weather conditions in two distinct periods: early spring and late summer-autumn, suggesting a bimodal pattern of growth. Saprotrophic and mycorrhizal fungi showed differences in the climatic control. Increased atmospheric evaporative demand due to global warming might lead to a drop in fungal yields during the 21st century.

Persistence and detection of black truffle ectomycorrhizas in plantations: comparison between two field detection methods.

Owners of black truffle (Tuber melanosporum) plantations are concerned about the persistence of its mycorrhizas and mycelium in the soil, especially until the appearance of the "truffle burn" areas and the triggering of sporocarp production, at least 5-7 years after planting truffle-inoculated seedlings. During this period, the farmer does not know whether his management is promoting black truffle development. To study the presence and abundance of T. melanosporum ectomycorrhizas in plantations, two sampling methods, direct sampling of root tips and soil core collection, are compared by analyzing 48 evergreen oak trees (Quercus ilex) inoculated with truffle. Those trees are grouped by age (<6, 6-9, >9 years old) and presence or absence of truffle production. T. melanosporum was present in 46 out of the 48 studied trees, and its ectomycorrhizas appeared in 65% of the ectomycorrhizal tips. Its abundance is significantly higher with productive trees and young trees. Direct sampling of root tips and soil core collection were equally effective in detecting this species, although soil core collection proved a better method to also evaluate ectomycorrhizal fungal diversity. To detect the presence of T. melanosporum in a given plantation, three samples suffice, with a single sample per random tree. Although the presence of mycorrhizas is not a sure sign of the future success of a black truffle plantation, its absence influences managers as to whether to continue culturing truffles in a plantation.

Soil physical properties influence "black truffle" fructification in plantations.

Although the important effects of pH and carbonate content of soils on "black truffle" (Tuber melanosporum) production are well known, we poorly understand the influence of soil physical properties. This study focuses on physical soil characteristics that drive successful production of black truffles in plantations. Seventy-eight Quercus ilex ssp. ballota plantations older than 10 years were studied in the province of Teruel (eastern Spain). Soil samples were analyzed for various edaphic characteristics and to locate T. melanosporum ectomycorrhizae. The influence of cultivation practices, climatic features, and soil properties on sporocarp production was assessed using multivariate analyses. Low contents of fine earth and silt and high levels of bulk density, clay content, and water-holding capacity appear to promote fructification. Watering is also highly positive for truffle fructification. We develop and discuss a logistic model to predict the probability of truffle fructification in field sites under consideration for truffle plantation establishment. The balance between water availability and aeration plays a crucial role in achieving success in black truffle plantations.

Ectomycorrhizal fungus diversity and community structure with natural and cultivated truffle hosts: applying lessons learned to future truffle culture.

Since the first truffle plantations were established in France, Italy and other parts in the world, many studies have been carried out to improve their productivity and sustainability. Success of plantations is clearly related to the mycorrhizal status of the host trees over the years, from inoculated seedlings to truffle-producing trees. The experience gained in monitoring the ectomycorrhizal fungus status in cultivated truffle grounds has allowed us to develop an extensive catalogue of the ectomycorrhizal fungi present in truffle plantations. Herein, we summarize fungal community data from 85 references that represent different truffle studies in natural habitats and plantations. Approximately 25% of the ectomycorrhizae reported in the 85 references are common to most of the studies. In general, more fungal species are detected in productive plantations than in the non-productive ones. Truffle plantations display a diverse ectomycorrhizal fungal community, in which species of the genus Tuber are well represented. Tuber rufum and some members of Boletales are typically restricted to productive truffle plots. On the other hand, Hebeloma, Laccaria and Russula species are mostly associated with unproductive plots. Ectomycorrhizae belonging to Thelephoraceae are frequently found in mature truffle orchards but do not seem to affect sporocarp production. Several biotic and abiotic factors affect the ectomycorrhizal fungus communities associated with truffle orchards. Among them are plantation age, host species and its growth, the surrounding environment (particularly the presence of other ectomycorrhizal hosts), and plantation management. Understanding the ectomycorrhizal fungal communities inhabiting different plantations may give us clues about the dynamics of the targeted truffles and the possibility of identifying mycorrhizal fungal species that are good indicators of successful truffle plantations.

Age class influence on the yield of edible fungi in a managed Mediterranean forest.

Lack of information and difficulty in predicting wild edible sporocarp yields is blocking its integration in forest management. In the Mediterranean area, this nontimber forest product has increased its market value, consumption demand, and interest over the last decade. In this work, sampling year and stand age effects are analyzed in order to advance knowledge of edible fungi community structure, dynamics, and production. Weekly autumnal sporocarp monitoring was performed from 1997 to 2011 in a Pinus pinaster managed forest in central Spain. After applying a random stand age-stratified survey, 21 plots of 150 m(2) have been set with three per stand age class. The forest age classes have been defined as follows: 0-10 years, mixture of parent and regenerated trees, 11-20, 21-40, 41-60, 61-90, and over 90 years. A total of 153 species belonging to 56 genera were recorded, 55 of which are edible. The production of edible sporocarps was 19.8 kg ha(-1), representing 31 % of total production. Sporocarp production presents a sharp interannual variability with autumns 62 times more productive than others. The most abundant edible species in terms of fresh weight per hectare has been Lactarius deliciosus with 7.0 kg ha(-1). Edible fungi yields registered a significant decline in 10 years following regenerative cutting. The presence of parent trees significantly increases production with regard to the first class. The highest production of edible species occurs in the middle age, 41-60 years, and in the following classes, a decrease is produced. L. deliciosus production registered differences with age, manifesting in a high yield in young stands (11-20 years) and significant recovery in woodlands near to the cutting.

Seasonal dynamics of Boletus edulis and Lactarius deliciosus extraradical mycelium in pine forests of central Spain.

The annual belowground dynamics of extraradical soil mycelium and sporocarp production of two ectomycorrhizal fungi, Boletus edulis and Lactarius deliciosus, have been studied in two different pine forests (Pinar Grande and Pinares Llanos, respectively) in Soria (central Spain). Soil samples (five per plot) were taken monthly (from September 2009 to August 2010 in Pinar Grande and from September 2010 to September 2011 in Pinares Llanos) in eight permanent plots (four for each site). B. edulis and L. deliciosus extraradical soil mycelium was quantified by real-time polymerase chain reaction, with DNA extracted from soil samples, using specific primers and TaqMan® probes. The quantities of B. edulis soil mycelium did not differ significantly between plots, but there was a significant difference over time with a maximum in February (0.1576 mg mycelium/g soil) and a minimum in October (0.0170 mg mycelium/g soil). For L. deliciosus, significant differences were detected between plots and over time. The highest amount of mycelium was found in December (1.84 mg mycelium/g soil) and the minimum in February (0.0332 mg mycelium/g soil). B. edulis mycelium quantities were positively correlated with precipitation of the current month and negatively correlated with the mean temperature of the previous month. Mycelium biomass of L. deliciosus was positively correlated with relative humidity and negatively correlated with mean temperature and radiation. No significant correlation between productivity of the plots with the soil mycelium biomass was observed for any of the two species. No correlations were found between B. edulis sporocarp production and weather parameters. Sporocarp production of L. deliciosus was positively correlated with precipitation and relative humidity and negatively correlated with maximum and minimum temperatures. Both species have similar distribution over time, presenting an annual dynamics characterized by a seasonal variability, with a clear increase on the amounts of biomass during the coldest months of the year. Soil mycelial dynamics of both species are strongly dependent on the weather.

Edible sporocarp production by age class in a Scots pine stand in Northern Spain.

With the aim of increasing knowledge of community structure, dynamics and production of ectomycorrhizal fungi, edible sporocarp yields were monitored between 1995 and 2004 in a Pinus sylvestris stand in the northeast zone of the Iberian Peninsula. A random sampling design was performed by stand age class according to the forest management plan: 0-15, 16-30, 31-50, 51-70 and over 71-years-old. Eighteen 150 m plots were established and sampled weekly every year from September to December. One hundred and nineteen taxa belonging to 51 genera were collected, 40 of which were edible and represented 74% of the total biomass. Boletus edulis, Lactarius deliciosus, Cantharellus cibarius and Tricholoma portentosum sporocarps, which are considered to be of high commercial value, represented 34% of the total production. B. edulis and L. deliciosus were the most remarkable and abundant species, and both were collected in more than 60% of the samplings. B. edulis fructified every year of the experiment; its mean production was 40 kg/ha and year and its maximum productivity was more than 94 kg/ha in 1998. The age class with the largest production of this taxa was the fourth (51-70 years), with 70 kg/ha. L. deliciosus only failed to fructify one autumn (2000); its mean production was almost 10 kg/ha and its maximum productivity close to 30 kg/ha in 1997. The maximum productivity of this species was found in the second (16-30 years) and fifth (71-90 years) stand age classes, with 18 and 16 kg/ha, respectively. Advances in this field can certainly offer new insights into factors affecting sporocarp production.

Quantification of extraradical soil mycelium and ectomycorrhizas of Boletus edulis in a Scots pine forest with variable sporocarp productivity.

The availability of most edible ectomycorrhizal mushrooms depends on their natural fructification. Sporocarp formation of these fungi is linked to habitat characteristics and climate conditions, but these data alone do not explain all the trends of fungal fruiting and dynamics. It could be hypothesized that the amount of soil mycelia could also be related to the production of carpophores. Soil samples (five cylinders of 250 cm(3) per plot) were taken monthly, from September to November, in five fenced permanent plots (5 × 5 m) in Pinar Grande (Soria, Spain), a Pinus sylvestris stand situated in the north of the Sistema Ibérico mountain range. Plots were chosen to establish a gradient of Boletus edulis productivity from 0 to 38.5 kg/ha year, according to the mean fresh weight of sporocarps collected during the last 10 years. B. edulis ectomycorrhizal root tips were identified in each soil sample according to its morphology and counted. DNA extractions were performed with the PowerSoil(TM) DNA Isolation Kit and quantification of extraradical soil mycelium by real-time polymerase chain reaction using specific primers and a TaqMan® probe. The concentration of soil mycelium of B. edulis (mg mycelium/g soil) did not differ significantly between plots (p = 0.1397), and sampling time (p = 0.7643) within the fructification period. The number of mycorrhizal short roots per soil volume showed significant differences between the plots (p = 0.0050) and the three sampling times (p < 0.0001). No significant correlation between the number of mycorrhizas and the productivity of the plot (kg of B. edulis/ha year) was detected (p = 0.615). A statistically significant positive correlation (p = 0.0481) was detected between the concentration of mycelia of B. edulis in the soil samples and the presence of short roots mycorrhizal with B. edulis in these samples. The productivity of the plots, in terms of sporocarps produced during the last 10 years, was not correlated either with the concentration of soil mycelium or with the presence or abundance of ectomycorrhizas.

O ensino de pediatria na atenção básica em saúde entre as fronteiras do modelo biomédico e a perspectiva da integralidade do cuidado: a visão dos médicos supervisores / Pediatrics teaching in primary heath care between the boundaries of the biomedical model and the perspective of integrality of care: the view of the supervisor doctors / La enseñanza de la pediatría en atención básica en salud; los límites entre el modelo biomédico y la perspectiva de la atención integral: una visión de los supervisores médicos

Trata-se de estudo sobre educação médica, focalizado no ensino de pediatria na atenção básica, com o objetivo de analisar a contribuição da disciplina de Pediatria Social e Comunitária no 4º ano de graduação em medicina da Faculdade de Medicina de Botucatu, Universidade Estadual Paulista.Utilizou-se metodologia qualitativa de pesquisa, tendo como estratégia o estudo de caso. A análise foi fundamentada na abordagem sócio-histórica, subsidiada pelos estudos de Vigotski. Identificou-se o que os supervisores consideraram como aprendizados relevantes a sua contribuição no processo de mediação para que os estudantes aprendam sobre o processo de trabalho na atenção básica, o que não poderiam alcançar sozinhos. As entrevistas transcritas revelaram núcleos de significação comuns: importância da diversificação de cenários de ensino, aprendizado dos principais problemas de saúde, integralidade no atendimento da criança, com ênfase no vínculo e oportunidade do estudante aprender sobre promoção da saúde.

This is a study on medical education that focuses on pediatrics teaching in primary care. The aim is to analyze the contribution of the discipline of Social and Community Pediatrics in the 4th year of the Medicine undergraduate course at the Botucatu Medical School, Universidade Estadual Paulista. This qualitative research was based on a case study. The analysis was grounded on the social-historical approach, according to Vygotsky's studies. We found out what the supervisors considered to be relevant learning to their contribution to the mediation process so that students can learn about work at primary health care, something that they would not be able to reach by themselves. The interviews with the doctors revealed signification nuclei which are common to all these actors: the importance of the diversification of teaching scenarios, learning the main health problems, integral childcare with emphasis on the bond, and the student's opportunity to learn about health promotion.

El estudio presenta una contribución de la pediatría social y comunitaria en el 4º año los de graduación en medicina de la Facultad de Medicina de Botucatu, Universidade Estadual Paulista, Brasil. Se utilizó una metodología de investigación cualitativa, con una estrategia de estudio de caso. El análisis se basó en el enfoque histórico-social, apoyado por los estudios de Vigotsky. Se encontró lo que los supervisores de disciplina consideran como aprendizaje relevante a su contribución en el proceso de mediación para que los estudiantes aprendan sobre el proceso de trabajo en los cuidados básicos, lo que no podrían lograr por sí solos.Las entrevistas con los médicos revelan núcleos de significación que son comune: la importancia de los diferentes entornos de enseñanza, el aprendizaje de los principales problemas de salud, integración en el cuidado del niño, con énfasis en el vínculo y oportunidad del estudiante para aprender sobre promoción de la salud.

Tree age influences on the development of edible ectomycorrhizal fungi sporocarps in Pinus sylvestris stands.

The study of factors influencing the production and development of wild edible mushroom sporocarps is extremely important in the characterization of the fungi life cycle. The main objective of this work is to determine how tree age influences the speed of sporocarp growth of edible ectomycorrhizal fungi Boletus edulis and Lactarius deliciosus in a Pinus sylvestris stand. This study is based on information recorded on a weekly basis every autumn between 1995 and 2008 in a set of permanent plots in Spain. Sporocarps are collected weekly, and as a result, specimens may not have reached their maximum size. The study area is a monospecific P. sylvestris stand. Three age classes were considered: under 30 years, between 31 and 70 years, and over 70 years. Sporocarps of B. edulis and L. deliciosus grow faster in the first age class stands than in the other two, and in the second age class stands, sporocarps are more than 50% smaller. The average weight of the picked B. edulis sporocarps clearly varies in the three age classes considered, with its maximum in the first age class (127 g and 6.8 cm cap diameter), minimum in the second age class (68 g and 4.7 cm cap diameter), and showing a relative maximum in the third (79 and 4.3 cm cap diameter). L. deliciosus sporocarps are on average larger in the first age class (48 g and 7.4 cm cap diameter), decreasing in the second (20 g and 5.8 cm cap diameter) and also in the third (21 g and 5.3 cm cap diameter). The results show the influence of tree age in speed of sporocarp growth for the two ectomycorrhizal species.

Mycorrhizal synthesis between Boletus edulis species complex and rockroses (Cistus sp.).

Ectomycorrhizas of Boletus aereus, Boletus edulis, and Boletus reticulatus were synthesized with Cistus sp. under laboratory conditions using synthesis tubes filled with a mixture of sterilized peat-vermiculite and nutrient solution. The fungal strains isolated from sporocarps were identified by molecular techniques. The inoculated seedlings were grown for 4-5 months. The ectomycorrhizas formed were described based on standard morphological and anatomical characters. The three ectomycorrhizas described were very similar, with white monopodial-pinnate morphology, a three-layered plectenchymatous mantle on plan view and boletoid rhizomorphs.

Characterization and identification of field ectomycorrhizae of Boletus edulis and Cistus ladanifer.

Field ectomycorrhizae sampled under Boletus edulis and Cistus ladanifer have been characterized and described in detail based on standard morphological and anatomical characters. The described ectomycorrhiza has traits typical of Boletales: whitish with three differentiated plectenchymatous layers in the mantle in plan view forming ring-like structures and rhizomorphs with highly differentiated hyphae. The inflated, smooth cystidia-like clavate end cells on the surface of the rhizomorphs and their slightly twisted external hyphae are additional characterizing features. The Hartig net occupies 1 1/2 rows of cortical cells, partly reaching the endodermis. Not all hyphae have clamps. The identification of the fungal symbiont as B. edulis was confirmed by ITS rDNA sequence comparison between mycorrhizas and sporocarps. The singularity of this symbiotic association, as well as its ecological and practical implications, are discussed.

Contribuição à mudança curricular na graduação da Faculdade de Medicina de Botucatu / Contributions to curriculum changes at Botucatu Medical School

Descreve-se a inovação pedagógica de dois casos na graduação médica: uma matéria denominada Saúde Coletiva III, com (Administração, Ciências Sociais, Epidemiologia, Ética e Nutrição em Saúde Publica) e a disciplina de Semiologia Pediátrica. Para descrição e avaliação dos casos, utilizaram-se métodos qualitativos. A Saúde Coletiva III foi organizada por núcleos temáticos: Problemas em Saúde Publica; Nutrição em Saúde Publica; Planejamento em Saúde. O modelo de ensino centrou-se na problematização de situações vivenciadas na prática da Saúde Publica, trabalhando-se em centros, serviços e organizações de saúde. A Semiologia Pediátrica privilegiou a atenção integral à saúde da criança. O modelo de ensino centrou-se na aprendizagem baseada em problemas e no aprendizado prático da semiologia pediátrica em diferentes cenários, enfatizando-se o ensino ambulatorial. Privilegiou-se o trabalho em pequenos grupos, com a orientação docente. A principal missão voltou-se à utilização de estratégias que valorizassem o ensino centrado no estudante e sua capacidade de construir conhecimento com autonomia. No caso da Pediatria, avançou-se rumo a Medicina Integral, com enfoque amplo do modelo de atenção à criança. A Saúde Coletiva aproximou-se da Medicina Comunitária problematizando situações concretas no SUS e na atenção primária. O estudo mostra a possibilidade de inovação no ensino, e podendo contribuir para a mudança institucional.