DNA Barcoding and geographical scale effect: The problems of undersampling genetic diversity hotspots.

DNA barcoding identification needs a good characterization of intraspecific genetic divergence to establish the limits between species. Yet, the number of barcodes per species is many times low and geographically restricted. A poor coverage of the species distribution range may hamper identification, especially when undersampled areas host genetically distinct lineages. If so, the genetic distance between some query sequences and reference barcodes may exceed the maximum intraspecific threshold for unequivocal species assignation. Taking a group of Quercus herbivores (moths) in Europe as model system, we found that the number of DNA barcodes from southern Europe is proportionally very low in the Barcoding of Life Data Systems. This geographical bias complicates the identification of southern query sequences, due to their high intraspecific genetic distance with respect to barcodes from higher latitudes. Pairwise intraspecific genetic divergence increased along with spatial distance, but was higher when at least one of the sampling sites was in southern Europe. Accordingly, GMYC (General Mixed Yule Coalescent) single-threshold model retrieved clusters constituted exclusively by Iberian haplotypes, some of which could correspond to cryptic species. The number of putative species retrieved was more reliable than that of multiple-threshold GMYC but very similar to results from ABGD and jMOTU. Our results support GMYC as a key resource for species delimitation within poorly inventoried biogeographic regions in Europe, where historical factors (e.g., glaciations) have promoted genetic diversity and singularity. Future European DNA barcoding initiatives should be preferentially performed along latitudinal gradients, with special focus on southern peninsulas.

Seed loss before seed predation: experimental evidence of the negative effects of leaf feeding insects on acorn production.

Insect herbivory decreases plant fitness by constraining plant growth, survival and reproductive output. Most studies on the effects of herbivory in trees rely on correlational inter-individual comparisons and could, thus, be affected by confounding factors linked to both herbivory and plant performance. Using the Mediterranean Holm oak (Quercus ilex) as a study model, we followed an experimental approach in which leaf-feeding insects (mainly Lepidoptera caterpillars) were excluded from some shoots in all study trees. Shoots subjected to herbivore exclusion exhibited lower defoliation rates and produced more acorns than control shoots. Defoliation constrained shoot growth throughout the study period, but had no effect on the number of female flowers produced per shoot. Acorn production was, however, lower in control shoots due to their higher abortion rates, and also to their greater mortality risk during summer drought, as shoots with fewer leaves were less likely to survive. Plant reaction to herbivory inhibits certain physiological pathways involved in plant growth, which, together with the effects of physical damage, reduces the amount and efficiency of the photosynthetic tissue. This increases their vulnerability to environmental stresses, such as water deficit, which limit resource assimilation. Defoliation is likely a key factor affecting oak regeneration, as it may be a significant source of seed loss prior to pre-dispersal acorn predation. Further experimental studies could help to elucidate its effects in contrasting environments. In Mediterranean regions, the harsher droughts predicted by climate change models could worsen the effects of insect herbivory on oak reproductive output.