Native Phloem and Wood Borers in Australian Mediterranean Forest Trees

Native Mediterranean forests in Australia are dominated by two tree genera, Eucalyptus and Acacia, while Pinus and Eucalyptus dominate plantation forestry. In native forests, there is a high diversity of phloem and wood borers across several families in the Coleoptera and Lepidoptera. In the Coleoptera, cerambycid beetles (Cerambycidae), jewel beetles (Buprestidae), bark, ambrosia and pinhole beetles (Curculionidae) and pinworms (Lymexelidae) are some of the most commonly found beetles attacking eucalypts and acacias. In the Lepidoptera, wood moths (Cossidae), ghost moths (Hepialidae) and borers in the Xyloryctidae (subfamily Xyloryctinae) are most common. In contrast to native forests, there is a much more limited range of native insects present in Australian plantations, particularly in exotic Pinus spp. plantations, although eucalypt plantations do share some borers in common with native forests. This chapter reviews the importance of these borers in Australian forests primarily from an economic perspective (i.e. those species that cause damage to commercial tree species) and highlights a paucity of native forest species that commonly kill trees relative to the large scales regularly seen in North America and Europe.

Reviving extinct Mediterranean forest communities may improve ecosystem potential in a warmer future

The Mediterranean Basin is the region of Europe most vulnerable to negative climate-change impacts, including forest decline, increased wildfire, and biodiversity loss. Because humans have affected Mediterranean ecosystems for millennia, it is unclear whether the region's native ecosystems were more resilient to climate change than current ecosystems, and whether they would provide sustainable management options if restored. We simulated vegetation with the LandClim model, using present-day climate as well as future climate-change scenarios, in three representative areas that encompass a broad range of Mediterranean conditions and vegetation types. Sedimentary pollen records that document now-extinct forests help to validate the simulations. Forests modeled under present climate closely resemble the extinct forests when human disturbance is limited; under future scenarios, characterized by increased temperatures and decreased precipitation, extinct forests are projected to re-emerge. When combined with modeling, paleoecological evidence reveals the potential of native vegetation to re-establish under current and future climate conditions, and provides a template for novel management strategies to maintain forest productivity and biodiversity in a warmer and drier future.

Somatic embryogenesis as a regeneration method for clonal propagation of Mediterranean forest species

La mejora y conservación de recursos genéticos en especies forestales lleva siglos de retraso con respecto a las especies agrícolas. Los recursos forestales se han considerado tradicionalmente como recursos �mineros�, en los que primaba la mera extracción dejando exclusivamente a la regeneración natural la labor de sostenibilidad en los montes y dehesas o montados. Hoy en día, el necesario desarrollo del medio rural obliga a la explotación racional de los recursos como medio de garantizar su sostenibilidad. Por ello se está empezando a extender el criterio de que las especies forestales se pueden y deben �cultivar� en determinados espacios. Las características biológicas de las especies forestales las hacen, a menudo, recalcitrantes a las técnicas de mejora y conservación de recursos genéticos tradicionalmente aplicadas a especies agrícolas. En particular, la propagación vegetativa se ha utilizado ampliamente en muchos cultivos leñosos como una herramienta muy poderosa para capturar todo el potencial genético de combinaciones genéticas valiosas. En especies forestales, en particular en las mediterráneas, esta posibilidad raramente se ha podido aplicar debido a la baja capacidad morfogénica de estas especies y la fuerte influencia de la maduración o cambio de fase. En los últimos años la biotecnología forestal ha tenido un desarrollo espectacular. En particular las técnicas de regeneración clonal de plantas basadas en técnicas de cultivo in vitro, fundamentalmente vía embriogénesis somática, se están ya aplicando por muchas empresas privadas e instituciones públicas a nivel semi-operativo con diversas especies, para la conservación de material selecto y el establecimiento de ensayos clonales. Nuestros grupos de trabajo están desarrollando protocolos de regeneración por embriogénesis somática en distintas especies forestales. En esta comunicación se presenta el estado actual de los conocimientos en dos especies típicamente mediterráneas, el alcornoque (Quercus suber L.) y el pino piñonero (Pinus pinea L.), destacando los principales cuellos de botella para su aplicación a gran escala.

Stem CO2 efflux and its contribution to ecosystem CO2 efflux decrease with drought in a Mediterranean forest stand

tThe rate of metabolic processes demanding energy in tree stems changes in relation with prevailing cli-matic conditions. Tree water availability can affect stem respiration through impacts on growth, phloemtransport or maintenance of diverse cellular processes, but little is known on this topic. Here we moni-tored seasonal changes in stem CO2efflux (Fs), radial growth, sap flow and non-structural carbohydrates intrees of Quercus ilex in a Mediterranean forest stand subjected since 2003 to either partial (33%) through-fall exclusion (E) or unchanged throughfall (C). Fsincreased exponentially during the day by an effectof temperature, although sap flow attenuated the increase in Fsduring the day time. Over the year, Fsalso increased exponentially with increasing temperatures, but Fscomputed at a standard temperatureof 15?C (F15s) varied by almost 4-fold among dates. F15swas the highest after periods of stem growth anddecreased as tree water availability decreased, similarly in C and E treatments. The decline in F15swas notlinked to a depletion of soluble sugars, which increased when water stress was higher. The proportionof ecosystem respiration attributed to the stems was highest following stem growth (23.3%) and lowestduring the peak of drought (6.5%). High within-year variability in F15smakes unadvisable to pool annualdata of Fsvs. temperature to model Fsat short time scales (hours to months) in Mediterranean-type for-est ecosystems. We demonstrate that water availability is an important factor governing stem CO2effluxand suggest that trees in Mediterranean environments acclimate to seasonal drought by reducing stemrespiration. Stem respiratory rates do not seem to change after a long-term increase in drought intensity,however.

Facilitation Among Saproxylic Insects Inhabiting Tree Hollows in a Mediterranean Forest: The Case of Cetonids (Coleoptera: Cetoniidae) and Syrphids (Diptera: Syrphidae)

Tree hollows offer an ideal niche for saproxylic insects in mature Mediterranean forests, where Diptera and Coleoptera are the richest groups. Co-occurrence is frequently observed among many species of both groups in these microhabitats, and some of these species have been considered to facilitate the presence of other species by acting as ecosystem engineers. One of the systems that is found in Mediterranean tree hollows is formed by cetonid (Coleoptera: Cetoniidae) and syrphid (Diptera: Syrphidae) larvae. Here, cetonid larvae feed on wood and litter and produce a substrate that is easier to decompose. To assess the possible role of these larvae as facilitating agents for the saproxylic guild, we studied whether the presence of saprophagous Syrphidae inside tree hollows is associated with the activity of cetonid larvae. Furthermore, in laboratory conditions, we tested whether cetonid larvae activity can improve the development and fitness of the saprophagous syrphid species. Our results show that “cetonid activity” was the variable that best explained the presence of saprophagous syrphid species in natural conditions. Myathropa florea (L., 1758) was one of the species most influenced by this activity. The laboratory experiment gave similar results, demonstrating that an enriched substrate with Cetonia aurataeformis Curti, 1913 larval feces improves syrphid larval growth rate and fitness of adults (measured as longer wing length) of M. florea.

Problems of dynamic in mediterranean forests

The mediterranean forest ecosystems constitute a complex unit which is marked off the Northern and middle European ones by its climate, ecology and management.The ecosystems are related to a climate with estival droughtlasting from 1 to 6 months and the peculiar species which constitute them are generally not worth much.The anthropic pressure exerts especially by grazing and moreover they have been stricken by dramatic fires for many decades. Their large variety related to the bioclimatic and edaphic heterogeneity of the region can explain why the data concerning their functioning and dynamics are still fragmentary. Besides, their ecological and pedological characteristics make uneasy the use of schemes and patterns designed for middle european forests.

Influence of road noise disturbances on black vulture nesting area: Case of "Cabeza de Hierro" state in Spanish Mediterranean Central Mountain

The real and potential road influence on soundscape is considered a relevant management aspect that can assess the negative effects of massive visitants on such sensitive species -that have been living for centuries in the area-. As a first approach to the study of human disturbances sound impact, acoustic engineering tools allow us to model noise pollution caused by the main road that crosses the state ?Cabeza de Hierro? (M-604). For these preliminary results we use the French method XPS 31-133, recommend at EU level. Noise emission levels in black vulture nesting area are analyzed to understand the influence of human activities on rural areas and road management on biodiversity conservation. This approach develops a useful tool to make compatible the public enjoyment of forest services such as recreation or landscape scenary, the conservation of biodiversity as well as a suitable social and economic activity level ?timber and firewood harvesting, industry?- at the region.

A PCR based method to detect Russula spp. in soil samples and Limodorum abortivum roots in Mediterranean environments

Aim of study. Orchidaceae has the largest number of species of any family in the plant kingdom. This family is subject to a high risk of extinction in natural environments, such as natural parks and protected areas. Recent studies have shown the prevalence of many species of orchids to be linked to fungal soil diversity, due to their myco-heterotrophic behaviour. Plant communities determine fungal soil diversity, and both generate optimal conditions for orchid development. Area of study. The work was carried out in n the two most important natural parks in Alicante (Font Roja and Sierra Mariola), in South-eastern of Spain. Material and Methods. We designed a molecular tool to monitor the presence of Russula spp. in soil and orchids roots, combined with phytosociological methods. Main results. Using a PCR-based method, we detected the presence in the soil and Limodorum abortivum orchid roots of the mycorrhizal fungi Russula spp. The species with highest coverage was Quercus rotundifolia in areas where the orchid was present. Research highlights. We present a useful tool based on PCR to detect the presence of Russula spp. in a natural environment. These results are consistent with those obtained in different studies that linked the presence of the mycorrhizal fungi Russula spp. in roots of the species Limodorum and the interaction between these fungal species and Quercus ilex trees in Mediterranean forest environments.

An Ecological Model for Predicting Behaviour of Mediterranean Shrublands

In order to build dynamic models for prediction and management of degraded Mediterranean forest areas was necessary to build MARIOLA model, which is a calculation computer program. This model includes the following subprograms. 1) bioshrub program, which calculates total, green and woody shrubs biomass and it establishes the time differences to calculate the growth. 2) selego program, which builds the flow equations from the experimental data. It is based on advanced procedures of statistical multiple regression. 3) VEGETATION program, which solves the state equations with Euler or Runge-Kutta integration methods. Each one of these subprograms can act as independent or as linked programs.

Age determination and Heinrich Events in sediment cores SU90-08 and KET80-03

The Mediterranean Sea hydrology at the time of the Heinrich formation in the North Atlantic Ocean was analyzed by comparing sea surface temperatures (SSTs) and oxygen isotope composition of seawater (dw) changes during the past 75 kyr in two marine cores. These were compared to the palynological variations derived in the Mediterranean Sea core. During the last glacial the two oceanic SST records show similar and synchronous patterns, with several long-term cooling periods, ending by abrupt SST increases. At the time of the Heinrich events, cold SSTs and low salinity prevailed in the Mediterranean Sea. The freshwater budget was similar to the modern one, permitting the presence of a mixed forest on the Mediterranean borderlands. The post-Heinrich periods are marked by a freshwater budget decrease, limiting oak and fir tree growth in the Mediterranean region. Increase of precipitation or reduction of evaporation is observed before the Heinrich episode, and is associated with a well-developed mixed Mediterranean forest.

The relevance of seed size in modulating leaf physiology and early plant performance in two tree species

he size of seeds and the microsite of seed dispersal may affect the early establishment of seedlings through different physiological processes. Here, we examined the effects of seed size and light availability on seedling growth and survival, and whether such effects were mediated by water use efficiency. Acorns of Quercus petraea and the more drought-tolerant Quercus pyrenaica were sowed within and around a tree canopy gap in a sub-Mediterranean forest stand. We monitored seedling emergence and measured predawn leaf water potential (Ψpd), leaf nitrogen per unit area (Na), leaf mass per area, leaf carbon isotope composition (δ13C) and plant growth at the end of the first summer. Survival was measured on the next year. Path analysis revealed a consistent pattern in both species of higher δ13C as Ψpd decreased and higher δ13C as seedlings emerged later in the season, indicating an increase in 13C as the growing season is shorter and drier. There was a direct positive effect of seed size on δ13C in Q. petraea that was absent in Q. pyrenaica. Leaf δ13C had no effect on growth but the probability of surviving until the second year was higher for those seedlings of Q. pyrenaica that had lower δ13C on the first year. In conclusion, leaf δ13C is affected by seed size, seedling emergence time and the availability of light and water, however, leaf δ13C is irrelevant for first year growth, which is directly dependent on the amount of seed reserves.

Development of somatic embryogenesis for cloning and conservation of mature holm oak trees (Quercus ilex L.)

La encina (Quercus ilex L.) es una de las especies forestales mediterráneas más importantes. Constituye gran parte del estrato arbóreo de dehesas o montados, produce bellota como alimento del ganado y establece simbiosis con hongos micorrizógenos de gran valor económico. La encina está considerada como una especie recalcitrante en términos de conservación de semillas y capacidad morfogénica, lo que dificulta los programas de conservación de recursos genéticos y la mejora de la especie. La propagación vegetativa es una potente herramienta de los programas de mejora, por lo que es preciso desarrollar protocolos de regeneración somática en encina. La embriogénesis somática está considerada como la modalidad más adecuada de regeneración basada en técnicas de cultivo de tejidos vegetales utilizada en biotecnología forestal. Este trabajo se centra en el estudio de determinados aspectos de la embriogénesis somática para la regeneración clonal de encinas adultas. La memoria de esta tesis se ha dividido en capítulos que se corresponden con diferentes aspectos del sistema embriogénico. La embriogénesis somática se indujo en tegumentos maternos de óvulos en desarrollo procedentes de bellotas inmaduras de encinas adultas. A pesar de las bajas frecuencias de inducción, las líneas embriogénicas generadas se amplificaron mediante embriogénesis secundaria observándose cierta pérdida de la capacidad de diferenciación con el tiempo. Tanto el genotipo como la formulación del medio de cultivo influyeron en la respuesta embriogénica, concluyendo que la formulación de macronutrientes de Schenk y Hildebrant del medio sin reguladores de crecimiento fue la combinación más efectiva en la inducción. Los resultados sugirieron la existencia de una ventana en el desarrollo del óvulo más sensible a la inducción. El genotipo in[luyó en la capacidad proliferativa de los cultivos y en la conversión de los embriones somáticos, que se incrementó suplementando el medio con ácido indol-3-butírico y 6-benciladenina. El cultivo en medio líquido de líneas embriogénicas en condiciones de inmersión transitoria incrementó el crecimiento, dependiendo del genotipo, con respecto al cultivo en medio semisólido. Sin embargo, no mejoró la capacidad de diferenciar embriones cotiledonares aislados. Se estableció un protocolo de inicio y mantenimiento de cultivos en suspensión para varias líneas embriogénicas mediante inoculación en alta densidad de agregados embrionarios procedentes del medio semisólido. Para evitar la pérdida de vigor y la capacidad morfogénica debida al cultivo prolongado se desarrolló un protocolo de crioconservación de líneas embriogénicas mediante vitrificación. Al determinar la influencia de los agentes crioprotectores antes y después de su inmersión en nitrógeno líquido se concluyó que las respuestas de capacidad de crecimiento y de diferenciación del material embriogénico son independientes, además de estar bajo influencia del genotipo y el tipo de material crioconservado. La combinación de sacarosa y PVS2 previa a la inmersión en nitrógeno líquido proporcionó la mayor tasa de recuperación. Cuando las líneas fueron crioconservadas 30 días la capacidad de diferenciación se perdió en todas ellas. El análisis de SSR detectó variación somaclonal en el material crioconservado a corto plazo. SSR y RAPD mostraron importantes diferencias genéticas entre los árboles donantes y el material embriogénico que dependieron del genotipo. El grado de detección dependió del marcador empleado. Ambos marcadores revelaron baja inestabilidad intraclonal. Los RAPD revelaron variación genética intra-individuo en las encinas donantes. Se discuten la variación genética pre-existente en encina, su aparición durante las primeras fases de la inducción de embriogénesis, y la presencia de tejidos provenientes de la fertilización en el explanto materno. Esto hace preciso definir la identidad genética del material donante y acometer ensayos de detección precoz de variación somaclonal. ABSTRACT Holm oak (Quercus ilex L.) is one of the most important Mediterranean forest species. It conforms the tree layer of dehesas or montados, it produces acorns to feed the livestock and it establishes symbiosis with profitable mycorrhizal fungi. Holm oak is considered as recalcitrant species in terms of seed conservation and morphogenic capacities, which complicates the development of genetic conservation and improvement programs. Vegetative propagation is one of the mightiest tools for breeding programs therefore; developing protocols for clonal regeneration of holm oak is essential. Somatic embryogenesis is considered the best tissue culture-based way of plant regeneration in forest biotechnology. The present study is focused on the study of certain aspects of somatic embryogenesis for clonal regeneration of mature holm oak. This thesis manuscript is divided into several chapters that match with different aspects of the embryogenic system. Somatic embryogenesis induction was achieved on maternal teguments of developing ovules from immature acorns of adult holm oak trees. Despite the low induction frequencies, the generated embryogenic lines were amplified by secondary embryogenesis. A decline in the differentiation capacity over time was also observed. It was concluded that both genotype and culture media formulation influenced the embryogenic response, being the Schenk and Hildebrandt´s macronutrients formulation from culture medium and the lack of plant growth regulators the most effective combination for the induction of the embryogenic response. It has been suggested the existence of a developmental window in which ovules are prone to induction. Genotype influenced the proliferation capacity and the plant conversion of somatic embryos, which was also favoured by the presence of indol-3-butyric acid and 6-bencyladenine. The use of temporary immersion systems as proliferation in liquid culture of the embryogenic lines increased the growth depending on genotype, when compared to semisolid cultures. However, it did not improve the differentiation of single cotyledonary embryos. A protocol for the initiation and maintenance of embryogenic suspension cultures was established for several embryogenic lines with highly dense inoculi of embryogenic clusters from proliferating semisolid cultures. In order to avoid the loss of vigour and morphogenic ability of embryogenic lines due to prolonged cultures, a cryopreservation protocol for embryogenic lines of holm oak has been developed. During the determination of the influence of cryoprotective agents on the growth and differentiation capacities before and after liquid nitrogen immersion, it was concluded that both responses were independent from each other and also under the influence of genotype and the type of cryopreserved material. The combination of sucrose and PVS2 prior liquid nitrogen immersion provided higher recovery rates. When the same embryogenic lines were cryopreserved for 30 days, none was able to differentiate. The SSRs analysis of the short-term cryopreserved material detected somaclonal variation. Both SSR and RAPD markers showed high sensitivity to detect genetic differences between the donor trees and the generated embryogenic material. Nevertheless, the degree of instability detection depended on the marker. The SSR analysis indicated a relationship between genotype, the studied loci and the located polymorphisms. Also, both markers revealed low intraclonal genetic variation. The RAPD detected genetic variation within the donor trees. The presence of pre-existent genetic variation within mature trees, in addition to its occurrence during the early stages of the embryogenic induction, and the presence of tissues of fertilisation origin within the maternal explants are all discussed. Nonetheless, the determination of the genetic identity of donor material is required, in addition to early detection methods of somaclonal variation.