Three large saproxylic cerambycids with different pest/legal status co-occur in the Iberian oak woodlands, Cerambyx welensii (Cw), Cerambyx cerdo (Cc) and Prinobius myardi (Pm): Cw is an emerging pest, Cc is a protected but sometimes harmful species and Pm is a secondary/minor pest. A precise taxonomic diagnosis is necessary for research, management or protection purposes, but may be problematic mainly because Cw and Cc larvae are morphologically indistinguishable. To resolve this constraint, we genotyped adults, larvae and eggs collected over a wide geographical range using the mitochondrial barcoding of the cytochrome c oxidase subunit I (COI). A Neighbour-Joining tree phylogram revealed three distinct clusters corresponding to Cw, Cc and Pm. We further first sequenced for Cw and Cc two mitochondrial (12S rRNA and 16S rRNA) and one nuclear (28S rRNA) gene fragments. For the first two genes, interspecific divergence was lower than in COI, and for the 28S (lower mutation rate), the two species shared identical haplotypes. Two approaches for species delimitation (General Mixed Yule Coalescent (GMYC), Barcode Index Number (BIN)) confirmed the species distinctiveness of Cc and Cw. The Bayesian COI gene tree showed a remarkable genetic divergence between Cc populations from Iberia and the rest of Europe. Such divergence has relevant taxonomic connotations and stresses the importance of a wide geographical scale sampling for accurate DNA barcoding species identification. Incongruities between morphology/lineage and COI barcodes in some individuals revealed natural hybridization between Cw and Cc. Natural hybridization is important from a phylogenetic/evolutionary perspective in these cerambycids, but the prevalence of (and the behavioural/ecological factors involved in) interspecific cross-breeding remain to be investigated.

The transfer of the imazethapyr-resistant gene from Clearfield™ (CL) rice to red rice is an ecological risk. Flowering synchronization and genetic compatibility between cultivated rice and red rice could influence gene transfer. We examined the (1) variability in maximum outcrossing rate between 12 red rice biotypes and ‘CL161’ rice during their peak flowering overlap in the field and (2) genetic compatibility of red rice biotypes with CL161 rice. Experiments were conducted at Stuttgart, AR, and Fayetteville, AR, from 2005 to 2007. To evaluate the flowering synchrony of red rice and CL161 rice as well as its impact on outcrossing rate, field experiments were conducted at four planting times from early April to late May. The red rice biotypes were planted in the middle row of nine-row CL161 plots and flowering was monitored. Outcrosses were evaluated in subsequent years by herbicide response and simple-sequence-repeat marker assays. To determine compatibility, manual crosses were performed between 12 red rice biotypes and CL161 rice in the greenhouse. The flowering duration of all red rice types ranged from 5 to 16 d after the onset of flowering in contrast to 6 d in CL161 rice. Ten of the twelve types of red rice had ≥ 70% overlap in flowering time with CL161 rice in at least one planting date. The maximum field outcrossing rate between red rice biotypes and CL161 ranged from 0.03 to 0.25%. The field outcrossing rate between red rice biotypes differed (P < 0.01), but flowering synchronization was not directly related to outcrossing rate. Manual crosses resulted in seed sets of 49 to 94%. The majority of red rice biotypes had similar compatibility with CL161 rice. Thus, other factors must contribute to hybridization rates in the field. Follow-up experiments should investigate other plant factors and environmental influence on hybridization rate.