Evolutionary drivers of reproductive fitness in two endangered forest trees.

Population genetics theory predicts a relationship between fitness, genetic diversity (H0) and effective population size (Ne), which is often tested through heterozygosity-fitness correlations (HFCs). We tested whether population and individual fertility and heterozygosity are correlated in two endangered Mexican spruces (Picea martinezii and Picea mexicana) by combining genomic, demographic and reproductive data (seed development and germination traits). For both species, there was a positive correlation between population size and seed development traits, but not germination rate. Individual genome-wide heterozygosity and seed traits were only correlated in P. martinezii (general-effects HFC), and none of the candidate single nucleotide polymorphisms (SNPs) associated with individual fertility showed heterozygote advantage in any species (no local-effects HFC). We observed a single and recent (c. 30 thousand years ago (ka)) population decline for P. martinezii; the collapse of P. mexicana occurred in two phases separated by a long period of stability (c. 800 ka). Recruitment always contributed more to total population census than adult trees in P. mexicana, while this was only the case in the largest populations of P. martinezii. Equating fitness to either H0 or Ne, as traditionally proposed in conservation biology, might not always be adequate, as species-specific evolutionary factors can decouple the expected correlation between these parameters.

Natural durability of timber species exposed to xylophagous fungi in southern Durango, Mexico.

Introduction: Wood is a natural resource used for construction and the manufacture of many products. This material is exposed to damage due to biotic and abiotic factors. An important biotic factor is wood-degrading fungi that generate large economic losses. The objectives of this study were to determine the effect of xylophagous fungi (Coniophora puteana and Trametes versicolor) on the natural durability of six timber species in southern Durango, Mexico, and to establish differences between fungal effects on each tree species. Materials and Methods: Samples of Pinus durangensis, P. cooperi, P. strobiformis, Juniperus deppeana, Quercus sideroxyla, and Alnus acuminata were exposed to fungi for 4 months under laboratory conditions according to European Standard EN350-1. Samples of Fagus sylvatica were used as control. Durability was determined as the percentage of wood mass loss for each species. Welch ANOVA tests were performed to establish differences among tree species. Welch t-tests were used to prove loss mass differences between fungi for each tree species. Results: The most resistant species to C. puteana were P. durangensis, J. deppeana, P. cooperi and P. strobiformis, showing mean mass losses lower than 8.08%. The most resistant species to T. versicolor were J. deppeana, P. strobiformis and P. durangensis (mean mass losses lower than 7.39%). Pinus strobiformis and Q. sideroxyla were more susceptible to C. puteana effect; in contrast, P. durangensis and P. cooperi showed more damage due to T. versicolor degradation. Conclusions: Woods of P. durangensis, P. cooperi, P. strobiformis and Juniperus deppeana are well adapted to infection by these xylophagous fungi and are therefore highly recommended for commercial use in southern Durango, Mexico.

Assisted migration and the rare endemic plant species: the case of two endangered Mexican spruces.

Background: In the projected climate change scenarios, assisted migration might play an important role in the ex situ conservation of the threatened plant species, by translocate them to similar suitable habitats outside their native distributions. However, it is unclear if such habitats will be available for the Rare Endemic Plant Species (REPS), because of their very restricted habitats. The aims of this study were to perform a population size assessment for the REPS Picea martinezii Patterson and Picea mexicana Martínez, and to evaluate the potential species distributions and their possibilities for assisted migration inside México and worldwide. Methods: We performed demographic censuses, field surveys in search for new stands, and developed distribution models for Last Glacial Maximum (22,000 years ago), Middle Holocene (6,000 years ago), current (1961-1990) and future (2050 and 2070) periods, for the whole Mexican territory (considering climatic, soil, geologic and topographic variables) and for all global land areas (based only on climate). Results: Our censuses showed populations of 89,266 and 39,059 individuals for P. martinezii and P. mexicana, respectively, including known populations and new stands. Projections for México indicated somewhat larger suitable areas in the past, now restricted to the known populations and new stands, where they will disappear by 2050 in a pessimistic climatic scenario, and scarce marginal areas (p = 0.5-0.79) remaining only for P. martinezii by 2070. Worldwide projections (based only on climate variables) revealed few marginal areas in 2050 only in México for P. martinezii, and several large areas (p ≥ 0.5) for P. mexicana around the world (all outside México), especially on the Himalayas in India and the Chungyang mountains in Taiwan with highly suitable (p ≥ 0.8) climate habitats in current and future (2050) conditions. However, those suitable areas are currently inhabited by other endemic spruces: Picea smithiana (Wall.) Boiss and Picea morrisonicola Hayata, respectively. Conclusions: Assisted migration would only be an option for P. martinezii on scarce marginal sites in México, and the possibilities for P. mexicana would be continental and transcontinental translocations. This rises two possible issues for future ex situ conservation programs: the first is related to whether or not consider assisted migration to marginal sites which do not cover the main habitat requirements for the species; the second is related to which species (the local or the foreign) should be prioritized for conservation when suitable habitat is found elsewhere but is inhabited by other endemic species. This highlights the necessity to discuss new policies, guidelines and mechanisms of international cooperation to deal with the expected high species extinction rates, linked to projected climate change.

Graft survival of Pinus engelmannii Carr. in relation to two grafting techniques with dormant and sprouting buds.

Developing methods for successfully grafting forest species will be helpful for establishing asexual seed orchards and increasing the success of forest genetic improvement programs in Mexico. In this study we investigated the effects of two grafting techniques (side veneer and top cleft) and two phenological stages of the scion buds (end of latency and beginning of sprouting), in combination with other seven grafting variables, on the sprouting and survival of 120 intraspecific grafts of Pinus engelmannii Carr. The scions used for grafting were taken from a 5.5-year-old commercial forest plantation. The first grafting was performed on January 18 (buds at the end of dormancy) and the second on February 21 (buds at the beginning of sprouting). The data were examined by analysis of variance and a test of means and were fitted to two survival models (the Weibull's accelerated failure time and the Cox's proportional hazards model) and the respective hazard ratios were calculated. Survival was higher in the top cleft grafts made with buds at the end of latency, with 80% sprouting and an estimated average survival time of between 164 and 457 days after the end of the 6-month evaluation period. Four variables (grafting technique, phenological stage of the scion buds, scion diameter and rootstock height) significantly affected the risk of graft death in both survival models. Use of top cleft grafts with buds at the end of the latency stage, combined with scion diameters smaller than 11.4 mm and rootstock heights greater than 58.5 cm, was associated with a lower risk of death.

Improving the physical, mechanical and energetic properties of Quercus spp. wood pellets by adding pine sawdust.

BACKGROUND: Biomass usage for energy purposes has emerged in response to global energy demands and environmental problems. The large amounts of by-products generated during logging are rarely utilized. In addition, some species (e.g., Quercus spp.) are considered less valuable and are left in the cutting areas. Production of pellets from this alternative source of biomass may be possible for power generation. Although the pellets may be of lower quality than other types of wood pellets, because of their physical and technological properties, the addition of different raw materials may improve the characteristics of the oak pellets. METHODS: Sawdust from the oak species Quercus sideroxyla, Q. rugosa, Q. laeta and Q. conzattii was mixed with sawdust from the pine Pinus durangensis in different ratios of oak to pine (100:0, 80:20, 60:40, 40:60 and 20:80). Physical and mechanical properties of the pellets were determined, and calorific value tests were carried out. For each variable, Kolmogorov-Smirnov normality and Kruskal-Wallis tests were performed and Pearson's correlation coefficients were determined (considering a significance level of p < 0.05). RESULTS: The moisture content and fixed carbon content differed significantly (p < 0.05) between the groups of pellets (i.e., pellets made with different sawdust mixtures). The moisture content of all pellets was less than 10%. However, volatile matter and ash content did not differ significantly between groups (p ≥ 0.05). The ash content was less than 0.7% in all mixtures. The addition of P. durangensis sawdust to the mixtures improved the bulk density of the pellets by 18%. Significant differences (p < 0.05) in particle density were observed between species, mixtures and for the species × mixture interaction. The particle density was highest in the 80:20 and 60:40 mixtures, with values ranging from 1,245 to 1,349 kg m-3. Bulk density and particle density of the pellets were positively correlated with the amount of P. durangensis sawdust included. The mechanical hardness and impact resistance index (IRI) differed significantly (p < 0.05) between groups. The addition of pine sawdust decreased the mechanical hardness of the pellets, up to 24%. The IRI was highest (138) in the Q. sideroxyla pellets (100:0). The mechanical hardness and IRI of the pellets were negatively correlated with the amount of P. durangensis sawdust added. The bulk density of the pellets was negatively correlated with mechanical hardness and IRI. The calorific value of mixtures and the species × mixture interaction differed significantly between groups. Finally, the mean calorific value was highest (19.8 MJ kg-1) in the 20:80 mixture. The calorific value was positively related to the addition of P. durangensis sawdust.

Survival of side grafts with scions from pure species Pinus engelmannii Carr. and the P. engelmannii × P. arizonica Engelm. var. arizonica hybrid.

Grafting is one of the most widely used methods for vegetative propagation, particularly for multiplying trees considered important, but there has been little research done on the effect of hybridization on grafts from the genus Pinus. Sometimes hybrids show the ability to reproduce and adapt efficiently to the environment. However, they reduce the genetic gain of seed orchards. The objective in this research was to evaluate the effect of scion grafts from pure species donor Pinus engelmannii Carr. and from putative hybrid trees P. engelmannii × P. arizonica Engelm., grafted on rootstocks of pure species P. engelmannii, along with the effect of the position of the scion in the donor tree crown (upper third and middle third). The scions were collected from three trees of the pure species and three hybrids. In each tree, 20 scions were collected from each third of the crown evaluated. 120 side-veneer grafts were made at the beginning of spring (March) 2018. Variance analyses were performed to evaluate the treatments and adjustments of the Logit and Weibull models to obtain the probability of graft survival. Significant differences were found between the origins of scions (p < 0.0083, after Bonferroni correction), showing grafts with hybrid tree scions taking hold better. In addition, the probability of survival at 5 months after grafting with hybrid tree scions was greater (p < 0.0001) than in grafts with scions from trees of the pure species (Logit model), which coincides with the results of the Weibull model, which indicated that the probability of graft death with pure species donor tree scions is greater than for grafts with hybrid scions. There were no significant differences regarding the position of the scion in the donor tree crown.

Discrimination of Picea chihuahuana Martinez populations on the basis of climatic, edaphic, dendrometric, genetic and population traits.

BACKGROUND: Picea chihuahuana, which is endemic to Mexico, is currently listed as "Endangered" on the Red List. Chihuahua spruce is only found in the Sierra Madre Occidental (SMO), Mexico. About 42,600 individuals are distributed in forty populations. These populations are fragmented and can be classified into three geographically distinct clusters in the SMO. The total area covered by P. chihuahuana populations is less than 300 ha. A recent study suggested assisted migration as an alternative to the ex situ conservation of P. chihuahuana, taking into consideration the genetic structure and diversity of the populations and the predictions regarding the future climate of the habitat. However, detailed background information is required to enable development of plans for protecting and conserving species and for successful assisted migration. Thus, it is important to identify differences between populations in relation to environmental conditions. The genetic diversity of populations, which affect vigor, evolution and adaptability of the species, must also be considered. In this study, we examined 14 populations of P. chihuahuana, with the overall aim of discriminating the populations and form clusters of this species. METHODS: Each population was represented by one 50 × 50 m plot established in the center of its respective location. Climate, soil, dasometric, density variables and genetic and species diversities were assessed in these plots for further analyses. The putatively neutral and adaptive AFLP markers were used to calculate genetic diversity. Affinity Propagation (AP) clustering technique and k-means clustering algorithm were used to classify the populations in the optimal number of clusters. Later stepwise binomial logistic regression was applied to test for significant differences in variables of the southern and northern P. chihuahuana populations. Spearman's correlation test was used to analyze the relationships among all variables studied. RESULTS: The binomial logistic regression analysis revealed that seven climate variables, the geographical longitude and sand proportion in the soil separated the southern from northern populations. The northern populations grow in more arid and continental conditions and on soils with lower sand proportion. The mean genetic diversity using all AFLP studied of P. chihuahuana was significantly correlated with the mean temperature in the warmest month, where warmer temperatures are associated to larger genetic diversity. Genetic diversity of P. chihuahuana calculated with putatively adaptive AFLP was not statistically significantly correlated with any environmental factor. DISCUSSION: Future reforestation programs should take into account that at least two different groups (the northern and southern cluster) of P. chihuahuana exist, as local adaptation takes place because of different environmental conditions.

Degree of Hybridization in Seed Stands of Pinus engelmannii Carr. In the Sierra Madre Occidental, Durango, Mexico.

Hybridization is an important evolutionary force, because interspecific gene transfer can introduce more new genetic material than is directly generated by mutations. Pinus engelmannii Carr. is one of the nine most common pine species in the pine-oak forest ecoregion in the state of Durango, Mexico. This species is widely harvested for lumber and is also used in reforestation programmes. Interspecific hybrids between P.engelmannii and Pinus arizonica Engelm. have been detected by morphological analysis. The presence of hybrids in P. engelmannii seed stands may affect seed quality and reforestation success. Therefore, the goals of this research were to identify introgressive hybridization between P. engelmannii and other pine species in eight seed stands of this species in Durango, Mexico, and to examine how hybrid proportion is related to mean genetic dissimilarity between trees in these stands, using Amplified Fragment Length Polymorphism (AFLP) markers and morphological traits. Differences in the average current annual increment of putative hybrids and pure trees were also tested for statistical significance. Morphological and genetic analyses of 280 adult trees were carried out. Putative hybrids were found in all the seed stands studied. The hybrids did not differ from the pure trees in vigour or robustness. All stands with putative P. engelmannii hybrids detected by both AFLPs and morphological traits showed the highest average values of the Tanimoto distance, which indicates: i) more heterogeneous genetic material, ii) higher genetic variation and therefore iii) the higher evolutionary potential of these stands, and iv) that the morphological differentiation (hybrid/not hybrid) is strongly associated with the Tanimoto distance per stand. We conclude that natural pairwise hybrids are very common in the studied stands. Both morphological and molecular approaches are necessary to confirm the genetic identity of forest reproductive material.