Rafael Zas

Foliar nutrient status and nutritional relationships of young Pinus radiata D. Don plantations in northwest Spain

Abstract Fifty-four plots of young Pinus radiata D. Don plantations on abandoned agricultural land in Galicia (northwest Spain) were selected for nutritional diagnosis. Nutritional status was assessed by foliar analysis using the critical levels method. The relationships between foliar nutrient concentrations and the plantation’s growth and survival and the topsoil physical and chemical properties were also analyzed. The most common deficiencies among plots were P and Mg. Foliar N concentrations were relatively high and generally they were above the critical level. Foliar N:P, N:K and N:Mg ratios were significantly unbalanced in 31, 44 and 17 out of 54 studied plots. Foliar P levels were significantly higher on sites with previous herbaceous land use and on soils with a finer texture, whereas foliar Mg levels were higher at low altitudes. Foliar nutrient levels were not significantly correlated with their respective soil levels except in the case of K. Foliar Ca and Mg concentrations correlated positively with topsoil pH. Growth correlated positively with foliar K and Mg concentrations and negatively with foliar N:P, N:K and N:Mg ratios. Percentage of dead and poor state plants were negatively correlated with foliar P, K and Ca levels and positively correlated with foliar N:P and N:Mg ratios. These results suggested that nitrogen may be in excess and may condition the uptake of other nutrients. Foliar N level was considered as an unfavorable parameter, whereas foliar P, K, Ca and Mg concentrations were considered as favorable parameters for P. radiata establishment in Galicia.

Iterative kriging for removing spatial autocorrelation in analysis of forest genetic trials

Conventional analysis of spatially correlated data in inadequately blocked field genetic trials may give erroneous results that would seriously affect breeding decisions. Forest genetic trials are commonly very large and strongly heterogeneous, so adjustments for micro-environmental heterogeneity become indispensable. This study explores the use of geostatistics to account for the spatial autocorrelation in four Pinus pinaster Ait. progeny trials established on hilly and irregular terrains with a randomized complete block design and large blocks. Data of five different traits assessed at age 8 were adjusted using an iterative method based on semivariograms and kriging, and the effects on estimates of variance components, heritability, and family effects were evaluated in relation to conventional analysis. Almost all studied traits showed nonrandom spatial structures. Therefore, after the adjustments for spatial autocorrelation, the block and family × block variance components, which were extremely high in the conventional analysis, almost disappeared. The reduction of the interaction variance was recovered by the family variance component, resulting in higher heritability estimates. The removal of the spatial autocorrelation also affected the estimation of family effects, resulting in important changes in family ranks after the spatial adjustments. Comparison among families was also greatly improved due to higher accuracy of the family effect estimations. The analysis improvement was larger for growth traits, which showed the strongest spatial heterogeneity, but was also evident for other traits such as straightness or number of whorls. The present paper demonstrates how spatial autocorrelation can drastically affect the analysis of forest genetic trials with large blocks. The iterative kriging procedure presented in this paper is a promising tool to account for this spatial heterogeneity.

Climatic control of intra-annual wood density fluctuations of Pinus pinaster in NW Spain

Tree rings of Pinus pinaster often contain intra-annual density fluctuations (IADFs), which have been attributed to the succession of dry and rainy periods typical of Mediterranean climate, but their formation has not been studied yet under Atlantic climate. We analyzed the occurrence and climatic significance of replicated IADFs in ten monospecific stands in NW Spain. The frequency of IADFs was higher than previously reported for this species under Mediterranean conditions and consistently decreased with increasing elevation. The formation of bands of latewood-like tracheids within the earlywood was favored by dry previous August, cold previous winter and dry April. Bands of earlywood-like tracheids within the early latewood were also favored by low winter temperatures. However, their occurrence was geographically heterogeneous, with two groups of stands being defined by their distances to the shoreline. In coastal stands, cold May–August triggered IADFs formation, while in inland stands their formation was favored by dry May–July. Regional winter temperatures and April water balance were strongly related to the East Atlantic (EA) pattern, which greatly conditioned the occurrence of IADFs in the earlywood and the early latewood. By contrast, the presence of bands of earlywood-like tracheids in the late latewood was independent of the EA pattern, being strongly related to warm conditions in spring and especially to a wet October. The link between regional climate and the EA pattern strongly controlled the physiological processes that determine intra-annual growth dynamics and short-term cell enlargement of P. pinaster in NW Spain.

Vermicompost enhances germination of the maritime pine (Pinus pinaster Ait.)

We investigated the effect of vermicompost on the germination and early development of six different progenies of the maritime pine (Pinus pinaster Ait.). We compared the effects of incorporating solid vermicompost into the potting media to those of vermicompost water extract. The incorporation of vermicompost in the growing media of maritime pine increased germination by 16%, and particularly, addition of vermicompost water extract produced the best results. Plants germinated with vermicompost showed higher N content as compared to control plants, and this could have determined the faster maturation of the treated seedlings. Since the best effects on pine germination were observed after application of vermicompost water extract, other mechanisms, rather than the physical amelioration of the substrate, such as the presence of water soluble nutrients and organic compounds (i.e. humic acids and plant growth regulating substances) in the vermicompost, might be involved in the promotion of germination. We also report variation in the response of the different pine progenies to vermicompost application thereby confirming the necessity of taking into account genetic variability in order to study the potential of vermicompost and other biologically-active organic materials as a potting amendments.

Bottom-up effects of host-plant species diversity and top-down effects of ants interactively increase plant performance

While plant diversity is well known to increase primary productivity, whether these bottom-up effects are enhanced by reciprocal top-down effects from the third trophic level is unknown. We studied whether pine tree species diversity, aphid-tending ants and their interaction determined plant performance and arthropod community structure. Plant diversity had a positive effect on aphids, but only in the presence of mutualistic ants, leading to a threefold greater number of both groups in the tri-specific cultures than in monocultures. Plant diversity increased ant abundance not only by increasing aphid number, but also by increasing ant recruitment per aphid. The positive effect of diversity on ants in turn cascaded down to increase plant performance; diversity increased plant growth (but not biomass), and this effect was stronger in the presence of ants. Consequently, bottom-up effects of diversity within the same genus and guild of plants, and top-down effects from the third trophic level (predatory ants), interactively increased plant performance.

Genetics, phosphorus availability, and herbivore-derived induction as sources of phenotypic variation of leaf volatile terpenes in a pine species

Oleoresin produced and stored in pine tree leaves provides direct resistance to herbivores, while leaf volatile terpenes (LVT) in the resin are also powerful airborne infochemicals. Resin concentration and profile show considerable spatial and temporal phenotypic variation within and among pine populations. LVT biochemistry is known to be under genetic control, and although LVT should be plastic to diverse abiotic and biotic environmental factors such as nutrient availability and herbivore attack, little is known about their relative contributions and interactive effects. The aim of this paper was to clarify whether reduced phosphorus availability could increase the LVT concentration and affect the expression of herbivore-derived induced defences, and how plasticity would contribute to the phenotypic variation of LVT. The constitutive and methyl-jasmonate (MeJa) induced LVT concentration and profile were analysed in 17 half-sib Pinus pinaster families growing under two levels of P-availability (complete and P-limited fertilization). Individual terpene concentrations showed large additive genetic variation, which was more pronounced in the control than in MeJa-induced pines. MeJa application did not affect the LVT concentration, but significantly modified the LVT profile by depleting the α-pinene content and reducing the sesquiterpene fraction. Low P-availability strongly reduced plant growth and foliar nutrient concentrations, but did not affect LVT concentration and profile, and did not interact with MeJa-induction. Results indicate a strong homeostasis of LVT concentration to P-availability, and minor changes in the LVT profile due to MeJa-induction. Genetic variation appears to be the main source of phenotypic variation affecting the LVT concentration in this pine species.

Differential Allocation of Constitutive and Induced Chemical Defenses in Pine Tree Juveniles: A Test of the Optimal Defense Theory

Optimal defense theory (ODT) predicts that the within-plant quantitative allocation of defenses is not random, but driven by the potential relative contribution of particular plant tissues to overall fitness. These predictions have been poorly tested on long-lived woody plants. We explored the allocation of constitutive and methyl-jasmonate (MJ) inducible chemical defenses in six half-sib families of Pinus radiata juveniles. Specifically, we studied the quantitative allocation of resin and polyphenolics (the two major secondary chemicals in pine trees) to tissues with contrasting fitness value (stem phloem, stem xylem and needles) across three parts of the plants (basal, middle and apical upper part), using nitrogen concentration as a proxy of tissue value. Concentration of nitrogen in the phloem, xylem and needles was found to be greater higher up the plant. As predicted by the ODT, the same pattern was found for the concentration of non-volatile resin in the stem. However, in leaf tissues the concentrations of both resin and total phenolics were greater towards the base of the plant. Two weeks after MJ application, the concentrations of nitrogen in the phloem, resin in the stem and total phenolics in the needles increased by roughly 25% compared with the control plants, inducibility was similar across all plant parts, and families differed in the inducibility of resin compounds in the stem. In contrast, no significant changes were observed either for phenolics in the stems, or for resin in the needles after MJ application. Concentration of resin in the phloem was double that in the xylem and MJ-inducible, with inducibility being greater towards the base of the stem. In contrast, resin in the xylem was not MJ-inducible and increased in concentration higher up the plant. The pattern of inducibility by MJ-signaling in juvenile P. radiata is tissue, chemical-defense and plant-part specific, and is genetically variable.

Mediation of seed provisioning in the transmission of environmental maternal effects in Maritime pine (Pinus pinaster Aiton)

Although maternal environmental effects are increasingly recognized as an important source of phenotypic variation with relevant impacts in evolutionary processes, their relevance in long-lived plants such as pine trees is largely unknown. Here, we used a powerful sample size and a strong quantitative genetic approach to analyse the sources of variation of early seedling performance and to identify seed mass (SM)-dependent and -independent maternal environmental effects in Maritime pine. We measured SM of 8924 individual seeds collected from 10 genotypes clonally replicated in two environments of contrasting quality (favourable and stressful), and we measured seedling growth rate and biomass allocation to roots and shoots. SM was extremely variable (up to 14-fold) and strongly determined by the maternal environment and the genotype of the mother tree. The favourable maternal environment led to larger cones, larger seeds and reduced SM variability. The maternal environment also determined the offspring phenotype, with seedlings coming from the favourable environment being 35% larger and with greater root/shoot ratio. Transgenerational plasticity appears, thus, to be a relevant source of phenotypic variation in the early performance of this pine species. Seed provisioning explained most of the effect of the maternal environment on seedling total biomass. Environmental maternal effects on seedling biomass allocation were, however, determined through SM-independent mechanisms, suggesting that other epigenetic regulation channels may be involved.

Differentiation of persistent anatomical defensive structures is costly and determined by nutrient availability and genetic growth-defence constraints

Conifers exhibit a number of chemical and anatomical mechanisms to defend against pests and pathogens. Theory predicts an increased investment in plant defences under limited nutrient availability, but while this has been demonstrated for chemical defences, it has rarely been shown for anatomical defensive structures. In a long-lived woody plant, we tested the hypothesis that limited nutrient availability may promote an improved differentiation of persistent anatomical defences. We also hypothesized that the costs of differentiation of those long-term anatomical structures may be determined by genetic constraints on early growth potential. Using Pinus pinaster Ait. juveniles, we performed a greenhouse study with 15 half-sib families subjected to experimental manipulation of phosphorus (P) availability and herbivory-related induced responses. When plants were ∼30 cm high, half of the plant material was treated with methyl jasmonate to induce defences, and 2 weeks later plants were harvested and the abundance of resin canals in the cortex and xylem was assessed. Density of constitutive resin canals in the cortex and the total canal system was ∼1.5-fold higher in plants under limited P availability than in fully fertilized plants. Availability of P did not significantly influence the inducibility of resin canal traits. We found negative genetic correlations between plant growth and the density of constitutive canals in the xylem and total canal system, but only under conditions of limited nutrition. These results demonstrate for the first time that differentiation of constitutive anatomical-based defences is affected by P limitation. Moreover, results also evidence the existence of genetic constraints between plant growth and constitutive defensive investment, where lineages with the highest growth potential showed the lowest investment in constitutive resin canals.

Assessment of Plant Growth Promotion by Vermicompost in Different Progenies of Maritime Pine (Pinus pinaster Ait.)

The feasibility of incorporating vermicompost as a plant growth promoter into pine (Pinus pinaster Ait.) nurseries was investigated. Pines were grown in conventional peat-based nursery potting media where the peat was substituted by 0%, 2.5%, 5%, 10% and 25% solid vermicompost. In addition, in order distinguish between possible physical and biochemical mechanisms, the effects of solid vermicompost and vermicompost extracts on pine seedling growth were compared. Five different open-pollinated pine progenies were used in order to evaluate the possible genotype-dependent effects of vermicompost. All pots were provided with adequate mineral fertilization in order to avoid nutrient limitations and arranged in the greenhouse following a bifactorial randomized block design. Twenty weeks after sowing, seedlings were harvested and their aerial and root growth were measured, as well as biomass partitioning and seedling maturity. Amendment with solid vermicompost at 2.5% and 10% significantly stimulated pine seedling height, but not aerial biomass. Vermicompost also produced seedlings with greater shoot:root biomass ratios than the control. Besides, we noticed a significant inhibition of aerial and root biomass with the higher dose of solid vermicompost (25% substrate substitution). No effects, either positive or negative, were detected in plant growth due to the vermicompost extracts. All the reported results were the same for all the different progenies assayed, and therefore no genotype dependent effects were detected.