Estudio de la regeneración natural de Pinus nigra y medidas de favorecimiento por siembra y plantación en la reforestación de zonas afectadas por grandes incendios forestales

Los objetivos que se han planteado en el presente proyecto son los siguientes: - Objetivo 1. Evaluación del estado actual de la regeneración natural en el incendio de Bages-Berguedà de 1994. - Objetivo 2. Estudio experimental de la regeneración de Pinus nigra después de grandes incendios forestales. - Objetivo 3. Ensayo de medidas de siembra y plantación, con el objeto de complementar la regeneración natural del pino laricio.

Optimización de la calidad anatómicofisiológica de pinus halepensis y pinus pinea producidos en contenedor y relación con la supervivencia y crecimiento en plantación. Evaluación de las causas de mortalidad en plantaciones establecidas en tierras agrarias

El presente proyecto se plantea por la necesidad de estudiar la problemática derivada de la aplicación del Real Decreto 378/93 que establece un plan de forestación de superficies agrarias adoptando la normativa comunitaria reflejada en el Reglamento 2080/92. El objetivo de la legislación es la forestación de tierras agrícolas de baja rentabilidad, conservando el medio natural y obteniendo otros beneficios de índole diversa. Esta nueva situación genera una problemática que requiere la participación de investigadores, técnicos, gestores, productores y distribuidores para evitar errores que, a largo plazo, pueden provocar un efecto contrario al que pretendía la legislación. La aplicación del RD 378/93 desde el año 1993 afectaba 150.000 ha en el año 1996 con dispar distribución territorial. El éxito en el establecimiento de las plantaciones fue muy heterogéneo, como cabía esperar de la gran diversidad de estaciones forestadas. El objetivo general de la investigación propuesta en el Proyecto era disminuir los índices de mortalidad en plantaciones establecidas en tierras agrarias. La hipótesis de trabajo que se estableció era que la mortalidad elevada se relaciona con la calidad de la planta y que las particulares condiciones de reforestación incrementan la necesidad de utilizar planta con una calidad anatómica y fisiológica óptima. El establecimiento de un sistema de retroalimentación entre los distintos sistemas de producción de planta y los resultados de la plantación permitían diseñar métodos para la evaluación de la calidad de planta de vivero mediante indicadores fisiológicos. La participación en el proyecto de equipos investigadores con experiencia en la caracterización fisiológica de planta forestal producida en vivero (Subproyecto Escuela Técnica Superior de Ingenieros de Montes, en adelante ETSIM) y en la influencia de la calidad del sistema radical en la supervivencia y crecimiento en plantación (Subproyecto Institut de Recerca i Tecnologia Agroalimentaries, en adelante IRTA) nos permitía abordar el objetivo general propuesto. La participación de empresas viverísticas e instituciones colaboradoras (Forestal Catalana S.A., Genforsa, Generalitat de Catalunya) nos garantizaba la disponibilidad de material vegetal, instalaciones y parcelas experimentales. Con este planteamiento, se estableció un Plan de Trabajo con los siguientes objetivos parciales (entre paréntesis los equipos implicados en cada tarea).

Environment-dependent microevolution in a Mediterranean pine (Pinus pinaster Aiton).

BACKGROUND: A central question for understanding the evolutionary responses of plant species to rapidly changing environments is the assessment of their potential for short-term (in one or a few generations) genetic change. In our study, we consider the case of Pinus pinaster Aiton (maritime pine), a widespread Mediterranean tree, and (i) test, under different experimental conditions (growth chamber and semi-natural), whether higher recruitment in the wild from the most successful mothers is due to better performance of their offspring; and (ii) evaluate genetic change in quantitative traits across generations at two different life stages (mature trees and seedlings) that are known to be under strong selection pressure in forest trees. RESULTS: Genetic control was high for most traits (h2 = 0.137-0.876) under the milder conditions of the growth chamber, but only for ontogenetic change (0.276), total height (0.415) and survival (0.719) under the more stressful semi-natural conditions. Significant phenotypic selection gradients were found in mature trees for traits related to seed quality (germination rate and number of empty seeds). Moreover, female relative reproductive success was significantly correlated with offspring performance for specific leaf area (SLA) in the growth chamber experiment, and stem mass fraction (SMF) in the experiment under semi-natural conditions, two adaptive traits related to abiotic stress-response in pines. Selection gradients based on genetic covariance of seedling traits and responses to selection at this stage involved traits related to biomass allocation (SMF) and growth (as decomposed by a Gompertz model) or delayed ontogenetic change, depending also on the testing environment. CONCLUSIONS: Despite the evidence of microevolutionary change in adaptive traits in maritime pine, directional or disruptive changes are difficult to predict due to variable selection at different life stages and environments. At mature-tree stages, higher female effective reproductive success can be explained by differences in their production of offspring (due to seed quality) and, to a lesser extent, by seemingly better adapted seedlings. Selection gradients and responses to selection for seedlings also differed across experimental conditions. The distinct processes involved at the two life stages (mature trees or seedlings) together with environment-specific responses advice caution when predicting likely evolutionary responses to environmental change in Mediterranean forest trees.

First insights into the transcriptome and development of new genomic tools of a widespread circum-Mediterranean tree species, Pinus halepensis Mill.

Aleppo pine (Pinus halepensis Mill.) is a relevant conifer species for studying adaptive responses to drought and fire regimes in the Mediterranean region. In this study, we performed Illumina next-generation sequencing of two phenotypically divergent Aleppo pine accessions with the aims of (i) characterizing the transcriptome through Illumina RNA-Seq on trees phenotypically divergent for adaptive traits linked to fire adaptation and drought, (ii) performing a functional annotation of the assembled transcriptome, (iii) identifying genes with accelerated evolutionary rates, (iv) studying the expression levels of the annotated genes and (v) developing gene-based markers for population genomic and association genetic studies. The assembled transcriptome consisted of 48,629 contigs and covered about 54.6 Mbp. The comparison of Aleppo pine transcripts to Picea sitchensis protein-coding sequences resulted in the detection of 34,014 SNPs across species, with a Ka /Ks average value of 0.216, suggesting that the majority of the assembled genes are under negative selection. Several genes were differentially expressed across the two pine accessions with contrasted phenotypes, including a glutathione-s-transferase, a cellulose synthase and a cobra-like protein. A large number of new markers (3334 amplifiable SSRs and 28,236 SNPs) have been identified which should facilitate future population genomics and association genetics in this species. A 384-SNP Oligo Pool Assay for genotyping with the Illumina VeraCode technology has been designed which showed an high overall SNP conversion rate (76.6%). Our results showed that Illumina next-generation sequencing is a valuable technology to obtain an extensive overview on whole transcriptomes of nonmodel species with large genomes.

Ocorrência e flutuação populacional de Chrysopidae (Neuroptera) em áreas de plantio de Pinus taeda (L.) (Pinaceae) no sul do Paraná

The lacewings are very voracious predators of aphids. The objective of this research was to evaluate the occurrence of adult chrysopids in areas of Pinus reforestation where the giant conifer aphid Cinara spp. (Hemiptera: Aphididae) is causing severe damages. A total of 47 specimens were collected during one year and identified as: Chrysoperla externa (Hagen, 1861), Leucochrysa (Nodita) intermedia (Scheneir, 1851) and Leucochrysa (Nodita) vieirana (Navás, 1913). The captures in the area where the plants were one year old represented about 75% of the adults probably due to the high Cinara infestation on the trees in this area. The chrysopids were recorded mostly during the summer, possibly influenced by temperature.

Occurrence of Cinara spp. (Hemiptera, Aphididae) on Pinus spp. (Pinaceae), in the county of Lages-SC, Brazil

A population survey of the giant conifer aphid Cinara spp. was carried out in two areas of Pinus elliottii using yellow pan traps, during two years, from July 1997 to June 1999. During the collection period, the predominant species was Cinara pinivora (Wilson, 1919), representing 99% of the collected species. A few specimens of Cinara maritimae (Dufour, 1833) and only one winged female of Cinara fresai Blanchard, 1939 were collected. The population of C. pinivora was statistically higher in the area with 4.5 year-old trees than in the 1.5 year-old ones. The highest population peaks were registered in July 1997 and in August 1998. An unexpected high number of winged parthenogenetic females was collected in December 1998. C. fresai Blanchard, 1939 is recorded for the first time for Brazil.

Ciclo biológico e predação de três espécies de coccinelídeos (Coleoptera, Coccinellidae) sobre o pulgão-gigante-do-pinus Cinara atlantica (Wilson) (Hemiptera, Aphididae)

Afídeos do gênero Cinara Curtis são importantes pragas de coníferas em vários países. No Brasil Cinara atlantica (Wilson, 1919) tem causado danos em plantações comerciais de Pinus spp. e o controle biológico com predadores pode ser uma opção melhor que o controle químico. Este trabalho teve por objetivo estudar a biologia e a capacidade de predação dos coccinelídeos Hippodamia convergens Guérin-Méneville, 1842, Cycloneda sanguinea (Linnaeus, 1763) e Eriopis connexa (German, 1824) (Coleoptera, Coccinellidae) sobre ninfas de C. atlantica. O estudo foi conduzido sob condições controladas (temperatura: 23 ± 1ºC, UR: 70 ± 10% e fotofase: 14 h.). Foi verificado que ninfas de C. atlantica são adequadas como alimento para as três espécies de coccinelídeos, assegurando seu desenvolvimento e reprodução. H. convergens e C. sanguinea apresentaram maior longevidade e capacidade de reprodução e também maior capacidade de predação (3832 e 3633 ninfas de C. atlantica em comparação a 2735 ninfas consumidas por E. connexa durante o ciclo completo, respectivamente para as espécies). Estas espécies de predadores podem contribuir para a redução da população de Cinara no campo.

Ocorrência e recomendações para o manejo de Sirex noctilio Fabricius (Hymenoptera, Siricidae) em plantios de Pinus patula (Pinaceae) em Minas Gerais, Brasil

A vespa-da-madeira Sirex noctilio Fabricius, 1793 (Hymenoptera, Siricidae) é a praga mais importante das florestas cultivadas com Pinus spp. no Brasil. Foi introduzida no Rio Grande do Sul em 1988 e sua dispersão ficou restrita aos estados do sul do país até 2004, quando foi detectada em São Paulo. Neste trabalho é relatada a ocorrência de S. noctilio em plantios de Pinus patula em Minas Gerais, em janeiro de 2005. São discutidas algumas medidas que podem ser adotadas para restringir a dispersão de S. noctilio na região Sudeste.

Efeito do monocultivo de Pinus e da queima do campo nativo em atributos biológicos do solo no Planalto Sul Catarinense

O presente estudo objetivou avaliar o impacto da queima tradicional de campo nativo e do monocultivo de Pinus sp. em Lages (SC), no CO total do solo (COT), carbono da biomassa microbiana (CBM), respiração basal (C-CO2), quociente metabólico microbiano (qCO2) e relação CBM:COT. Foram selecionadas quatro áreas representativas da região, de 0,5 ha cada, sendo: (a) Campo nativo, sem queima nos últimos 50 anos (CN); (b) Campo nativo submetido à queima tradicional (CNQ), ambas, pastagens naturais; (c) Mata natural com predominância de Araucaria angustifolia (MATA); e (d) Reflorestamento de Pinus taeda com oito anos (PINUS). Para tanto, coletaram-se seis amostras, compostas de nove subamostras de solo em cada área, na profundidade de 0-5 cm, em dezembro de 2002. Para avaliar o CBM, foi utilizado o método da fumigação-extração. O C-CO2 foi determinado em laboratório. Os maiores valores de liberação de C-CO2 foram encontrados na MATA e no PINUS, seguidos de CNQ e CN, respectivamente. Os maiores valores de CBM, COT e relação CBM:COT foram encontrados na MATA, não tendo as demais áreas estudadas apresentado diferenças entre si. O qCO2 foi maior no PINUS, seguido de CNQ, em comparação com MATA e CN. A análise multivariada mostrou ser uma ferramenta auxiliar importante ao discriminar o CBM como sendo o atributo que mais contribuiu na separação entre as áreas estudadas.

The role of population origin and microenvironment in seedling emergence and early survival in Mediterranean maritime pine (Pinus pinaster Aiton).

Understanding tree recruitment is needed to forecast future forest distribution. Many studies have reported the relevant ecological factors that affect recruitment success in trees, but the potential for genetic-based differences in recruitment has often been neglected. In this study, we established a semi-natural reciprocal sowing experiment to test for local adaptation and microenvironment effects (evaluated here by canopy cover) in the emergence and early survival of maritime pine (Pinus pinaster Aiton), an emblematic Mediterranean forest tree. A novel application of molecular markers was also developed to test for family selection and, thus, for potential genetic change over generations. Overall, we did not find evidence to support local adaptation at the recruitment stage in our semi-natural experiment. Moreover, only weak family selection (if any) was found, suggesting that in stressful environments with low survival, stochastic processes and among-year climate variability may drive recruitment. Nevertheless, our study revealed that, at early stages of recruitment, microenvironments may favor the population with the best adapted life strategy, irrespectively of its (local or non-local) origin. We also found that emergence time is a key factor for seedling survival in stressful Mediterranean environments. Our study highlights the complexity of the factors influencing the early stages of establishment of maritime pine and provides insights into possible management actions aimed at environmental change impact mitigation. In particular, we found that the high stochasticity of the recruitment process in stressful environments and the differences in population-specific adaptive strategies may difficult assisted migration schemes.

Molecular Proxies for Climate Maladaptation in a Long-Lived Tree (Pinus pinaster Aiton, Pinaceae).

Understanding adaptive genetic responses to climate change is a main challenge for preserving biological diversity. Successful predictive models for climate-driven range shifts of species depend on the integration of information on adaptation, including that derived from genomic studies. Long-lived forest trees can experience substantial environmental change across generations, which results in a much more prominent adaptation lag than in annual species. Here, we show that candidate-gene SNPs (single nucleotide polymorphisms) can be used as predictors of maladaptation to climate in maritime pine (Pinus pinaster Aiton), an outcrossing long-lived keystone tree. A set of 18 SNPs potentially associated with climate, 5 of them involving amino acid-changing variants, were retained after performing logistic regression, latent factor mixed models, and Bayesian analyses of SNP-climate correlations. These relationships identified temperature as an important adaptive driver in maritime pine and highlighted that selective forces are operating differentially in geographically discrete gene pools. The frequency of the locally advantageous alleles at these selected loci was strongly correlated with survival in a common garden under extreme (hot and dry) climate conditions, which suggests that candidate-gene SNPs can be used to forecast the likely destiny of natural forest ecosystems under climate change scenarios. Differential levels of forest decline are anticipated for distinct maritime pine gene pools. Geographically defined molecular proxies for climate adaptation will thus critically enhance the predictive power of range-shift models and help establish mitigation measures for long-lived keystone forest trees in the face of impending climate change.