Alejandro Solla

Biogeographical patterns and determinants of invasion by forest pathogens in Europe

A large database of invasive forest pathogens (IFPs) was developed to investigate the patterns and determinants of invasion in Europe. Detailed taxonomic and biological information on the invasive species was combined with country-specific data on land use, climate, and the time since invasion to identify the determinants of invasiveness, and to differentiate the class of environments which share territorial and climate features associated with a susceptibility to invasion. IFPs increased exponentially in the last four decades. Until 1919, IFPs already present moved across Europe. Then, new IFPs were introduced mainly from North America, and recently from Asia. Hybrid pathogens also appeared. Countries with a wider range of environments, higher human impact or international trade hosted more IFPs. Rainfall influenced the diffusion rates. Environmental conditions of the new and original ranges and systematic and ecological attributes affected invasiveness. Further spread of established IFPs is expected in countries that have experienced commercial isolation in the recent past. Densely populated countries with high environmental diversity may be the weakest links in attempts to prevent new arrivals. Tight coordination of actions against new arrivals is needed. Eradication seems impossible, and prevention seems the only reliable measure, although this will be difficult in the face of global mobility.

Influence of Plant Age on Symptom Development in Ulmus minor Following Inoculation by Ophiostoma novo-ulmi

In American and European breeding programs, numerous elm trees from many species (Ulmus spp.) and hybrids have been inoculated annually with the fungus Ophiostoma novo-ulmi (the Dutch elm disease pathogen) in screening tests for resistance. Because trees were inoculated at different ages, it appeared necessary to study the influence of host age on the symptoms shown. Four Ulmus minor trees and one U. minor × U. pumila tree were cloned annually from 1994 to 1999. The replicates obtained (usually n = 6) were inoculated on 17 May 2000 with an O. novoulmi strain. At the end of the season, 2-year-old U. minor trees showed average wilting of 8 ± 3% (ES), significantly less than that of 3-year-old trees (34 ± 5%) (P ≤ 0.01). Elms that were 4, 5, 6, and 7 years old showed wilting values of around 50%. A positive relationship between the symptoms shown and the diameter of the elm xylem vessels was observed, and the implications for elm resistance and breeding are discussed. Breeders and pathologists should use trees of the same age and physiological phase when determining the relative resistance among elm clones. It is concluded that under the growing conditions of this experiment, the optimal age for U. minor screening was 4-year-old plants.

Antifungal effect and reduction of Ulmus minor symptoms to Ophiostoma novo-ulmi by carvacrol and salicylic acid

There are still no effective means to control Dutch elm disease (DED), caused by the vascular fungi Ophiostoma ulmi and O. novo-ulmi. Plant phenolics may provide a new strategy for DED control, given their known antifungal activity against pathogens and their involvement in plant defence mechanisms. The in vitro antifungal activity of salicylic acid, carvacrol, thymol, phenol, o-cresol, m-cresol, p-cresol, and 2,5-xylenol against the DED pathogens was tested. Also, the protective effect of watering Ulmus minor seedlings with these compounds was tested against O. novo-ulmi. Salicylic acid, carvacrol, and thymol showed the strongest antifungal in vitro activity, while carvacrol and salicylic acid provided the strongest in vivo protection against O. novo-ulmi (63 and 46% reduction of leaf wilting symptoms with respect to controls, respectively). The effect of the treatments on tree phenology was low, and a significant negative relation was observed between the number of days to bud burst and the leaf wilting symptoms after inoculation, probably determined by genetic differences among the elm tree progenies used. The treatments with salicylic acid, carvacrol and thymol induced the highest shift in phenolic metabolite profile with respect to control trees. The protective effect of carvacrol and salicylic acid is discussed in terms of their combined activity as antifungal compounds and as inductors of tree defence responses.

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.

Environmental Maternal Effects Mediate the Resistance of Maritime Pine to Biotic Stress

The resistance to abiotic stress is increasingly recognised as being impacted by maternal effects, given that environmental conditions experienced by parent (mother) trees affect stress tolerance in offspring. We hypothesised that abiotic environmental maternal effects may also mediate the resistance of trees to biotic stress. The influence of maternal environment and maternal genotype and the interaction of these two factors on early resistance of Pinus pinaster half-sibs to the Fusarium circinatum pathogen was studied using 10 mother genotypes clonally replicated in two contrasting environments. Necrosis length of infected seedlings was 16% shorter in seedlings grown from favourable maternal environment seeds than in seedlings grown from unfavourable maternal environment seeds. Damage caused by F. circinatum was mediated by maternal environment and maternal genotype, but not by seed mass. Mechanisms unrelated to seed provisioning, perhaps of epigenetic nature, were probably involved in the transgenerational plasticity of P. pinaster, mediating its resistance to biotic stress. Our findings suggest that the transgenerational resistance of pines due to an abiotic stress may interact with the defensive response of pines to a biotic stress.

Alterations of the resin canal system of Pinus pinaster seedlings after fertilization of a healthy and of a Hylobius abietis attacked stand

Changes in resource availability and biotic and abiotic stress may alter the defensive mechanisms of pine trees. The effect of fertilisation on the resin canal structure of Pinus pinaster seedlings established in two trials in NW Spain, one attacked by Hylobius abietis and the other non-attacked, was studied. The leaders of 50 plants were destructively sampled and the resin canal density, the canal area and its relative conductive area in the phloem and xylem were assessed. Experimentally increased nutrient availability significantly decreased resin canal density in the phloem of the seedlings in the two analysed trials, where unfertilised seedlings presented up to 30% more resin canal density than the fertilised seedlings (mean value ± SEM = 0.32 ± 0.02 resin canals mm−2 in the fertilised plants versus 0.45 ± 0.04 resin canals mm−2 in the control plants). Fertilisation had no effect on the resin canal system in the xylem, but significantly increased tracheid size. Significant differences of resin canals among sites were observed mainly in the xylem; the resin canal density was 1.7-fold greater in the attacked site than in the non-attacked site. The similar structure of phloem resin canals in both sites supports that phloem resin canals are constitutive mechanisms of defence in P. pinaster, whereas xylem resin canals would be constitutive mechanisms but also inducible mechanisms of resistance following the attack of pine weevils or bark beetles.

Chemical changes in Ulmus minor xylem tissue after salicylic acid or carvacrol treatments are associated with enhanced resistance to Ophiostoma novo-ulmi

Application of endogenous plant hormone salicylic acid (SA) or essential oil component carvacrol (CA) in elms enhances tree resistance to the Dutch elm disease pathogen, although the effect of these compounds on tree metabolism is unknown. The chemical changes induced by SA or CA treatments in Ulmus minor were studied through gas chromatography-mass spectrometry (GC-MS) analysis of xylem tissues. Treatments consisted of fortnightly irrigating seedlings with water, SA or CA at 600 mg L⁻¹. The chemical composition of the xylem tissues sampled from treated trees was significantly altered depending on the treatment type. SA treatment induced an accumulation of the sinapyl alcohol, a precursor of lignin and other phenylpropanoid-derived products. CA treatment induced an accumulation of the methyl esters of palmitic, linoleic and stearic acids. Both treatments resulted in early bud burst and SA significantly reduced sapwood radial growth, possibly as a consequence of a trade-off between tree growth and tree defence. The enhanced resistance provided by these treatments is discussed.

Root system of Quercus suber L. seedlings in response to herbaceous competition and different watering and fertilisation regimes

Different management practices are commonly applied to increase pasture yield of Mediterranean open woodlands, but the consequences of increasing competition for soil resources with these practices on tree recruitment are still unknown. In a greenhouse study, belowground competition of Quercus suber L. seedlings growing together with natural (OakNP) or improved pasture (OakIP) was evaluated, and their root systems compared with those of seedlings growing in bare soil (OakBS). Two watering levels and two regimes of P2O5 fertilisation were also tested. Because of competition, the OakIP seedlings had their fine root mass density, coarse root length, and shoot mass reduced by up to 40, 36, and 39%, respectively, when compared to OakNP seedlings. OakNP and OakBS seedlings showed similar average root density parameters and shoot mass values, indicating that Q. suber seedlings growing with natural pasture is a low competitive system. High availability of water and phosphorus did not mitigate the strength of competition between herbaceous plants and oak seedlings, and favoured the pasture to the detriment of the trees. Our findings suggest that P2O5 fertilisation and irrigation practices performed to improve herbaceous productivity will negatively influence recruitment of Q. suber seedlings.

Spanish Program for the Conservation and Breeding of Elms Against DED

The fungus Ophiostoma novo-ulmi, responsible for recent epidemics of Dutch elm disease, was detected in Spain at the end of the 1970s. As a response to high mortality of U. minor (sensu lato) throughout the country, a program was begun in 1986 by the Universidad Politecnica de Madrid. The program’s objectives are to preserve the largest possible variety of native elms, and to obtain resistant individuals by means of crossings with U. pumila, a well adapted species immune to disease. At present, 253 genotypes from throughout the country have been preserved, the majority of which have been characterized in morphology and phenology. In addition, numerous hybridizations have been carried out on a crossing population and the seedlings obtained have been inoculated. Some elms have shown acceptable resistance. The activities of the program have also been focused towards the development of early testing techniques, and in vitro methods and anatomical and physiological parameters have been used.

Susceptibility to the pinewood nematode (PWN) of four pine species involved in potential range expansion across Europe.

The pine wilt disease (PWD), caused by the pinewood nematode (PWN) Bursaphelenchus xylophilus (Steiner et Buhrer) Nickle, is one of the most serious threats to pine forests worldwide. Here we studied several components of susceptibility to PWN infection in a model group of pine species widely distributed in Europe (Pinus pinaster Ait., P. pinea L., P. sylvestris L. and P. radiata D. Don), specifically concerning anatomical and chemical traits putatively related to nematode resistance, whole-plant nematode population after experimental inoculation, and several biochemical and physiological traits indicative of plant performance, damage and defensive responses 60 days post inoculation (dpi) in 3-year-old plants. Pinus pinaster was the most susceptible species to PWN colonization, with a 13-fold increase in nematode population size following inoculation, showing up to 35-fold more nematodes than the other species. Pinus pinea was the most resistant species, with an extremely reduced nematode population 60 dpi. Axial resin canals were significantly wider in P. pinaster than in the other species, which may have facilitated nematode dispersal through the stem and contributed to its high susceptibility; nevertheless, this trait does not seem to fully determinate the susceptible character of a species, as P. sylvestris showed similar nematode migration rates to P. pinaster but narrower axial resin canals. Nematode inoculation significantly affected stem water content and polyphenolic concentration, and leaf chlorophyll and lipid peroxidation in all species. In general, P. pinaster and P. sylvestris showed similar chemical responses after infection, whereas P. radiata, which co-exists with the PWN in its native range, showed some degree of tolerance to the nematode. This work provides evidence that the complex interactions between B. xylophilus and its hosts are species-specific, with P. pinaster showing a strong susceptibility to the pathogen, P. pinea being the most tolerant species, and P. sylvestris and P. radiata having a moderate susceptibility, apparently through distinct coping mechanisms.