Forests are essential resources on a global scale, not only for the ecological benefits, but also for economical and landscape purposes. However, in recent years, a large number of forest species have suffered a serious decline, with maritime pine being one of the most affected. In Portugal, the maritime pine forest has been devastated by the pine wood nematode (PWN), the causal agent of pine wilt disease. In this study, RNA-Seq data was used to characterize the maritime pine response to infection with PWN, by determining the differentially expressed genes and identifying the regulatory networks and pathways associated. The analyses showed clear differences between an early response that occurs immediately after inoculation and a late response that is observed seven days after inoculation. Moreover, differentially expressed genes related to secondary metabolism, oxidative stress and defense against pathogen infection were identified over different time points. These results provide new insights about the molecular mechanisms and metabolic pathways involved in the response of Pinus pinaster against PWN infection, which will be a useful resource in follow-up studies and for future breeding programs to select plants with lower susceptibility to this disease.

Pine wilt disease (PWD) caused by pine wood nematode (PWN), Bursaphelenchus xylophilus, is a serious threat to global forest populations of conifers, in particular Pinus spp. Recently, the presence of PWN was reported in dead Yunnan pine (Pinus yunnanensis) trees under natural conditions. To further understand the potential impact caused by PWN in Yunnan pine populations, a transcriptional profiling analysis was performed over different time points (0 hours (h), 6 h, 24 h, 48 h, and 7 days) after PWN inoculation. A total of 9961 differentially expressed genes were identified after inoculation, which suggested a dynamic response against the pathogen, with a more intense pattern at 48 h after inoculation. The results also highlighted a set of biological mechanisms triggered after inoculation that provide valuable information regarding the response of Yunnan pine to PWN infection. When compared with maritime pine (Pinus pinaster), the Yunnan pine response was less complex and involved a smaller number of differentially expressed genes, which may be associated with the increased degree of resistance to PWN displayed by Yunnan pine. These results revealed different strategies to cope with PWN infection by these two pine species, which display contrasting degrees of susceptibility, especially in the timely perception of the infection and response magnitude.

The ambrosia beetle Platypus cylindrus Fab. (Coleoptera: Curculionidae) is a major cork oak pest in Portugal. Female and male beetles have different roles in host tree colonization and are both equipped with prothoracic mycangia for fungal transportation. Despite a known beneficial role of bacteria in ambrosia beetles, information on bacterial composition associated with prothoracic mycangia structures is scarce. Bacterial community from mycangia of P. cylindrus male and female beetles collected from cork oak galleries was investigated by means of 16S metagenomics. Mycangia anatomical structure was also explored with histological techniques and X-ray computed microtomography to highlight evidence supporting biological sexual dimorphism. A bacterial community with highly diverse bacterial taxa with low abundances at the genus level was revealed. Lactobacillales, Leptotrichia, Neisseria, Rothia, and Sphingomonadaceae were significantly more abundant in males, while Acinetobacter, Chitinophagaceae, Enterobacteriaceae, Erwiniaceae, Microbacteriaceae, and Pseudoclavibacter were more abundant in females. Additionally, a core bacteriome of five genera was shared by both sexes. Histological examination revealed visible connections linking external and internal tissues in females, but none in males. Overall, these results provide the first insights into sexual differentiation for bacteria in a Platypodinae beetle species, identifying key patterns of bacteria distribution in the context of beetle ecology and functional behavior.

Pine wilt disease (PWD), caused by the pinewood nematode (PWN) Bursaphelenchus xylophilus, is one of the most destructive diseases in trees of the genus Pinus and is responsible for significant environmental and economic losses in North America, Eastern Asia, and Western Europe. However, pine species are not equally affected, with some being tolerant/resistant while others are susceptible to nematode infection for reasons still unclear. The present study aims to investigate differential chemical responses of susceptible and tolerant/resistant pine species shortly after nematode infection by characterizing the phenolic profiles of adult Pinus sylvestris, Pinus pinaster, Pinus pinea, and Pinus halepensis. HPLC and LC-MS were used to identify and quantify the pine´s phenolic compounds: gallic acid, ferulic acid, taxifolin, rutin, resveratrol, (+)-secoisolariciresinol, (−)-epicatechin, protocatechuic acid hexoside, gallic acid hexoside, ferulic acid glucoside, quercetin hexoside, and two unidentified compounds (#A and #B). Prior to infection, we could not differentiate between nematode-tolerant/resistant and susceptible adult pine species based on their constitutive phenolic compounds. In the presence of the PWN, the phenolic profile allowed for a noticeable separation of the PWN-tolerant/resistant P. halepensis from the susceptible P. sylvestris, contrasting with a more homogenous response from P. pinea and P. pinaster. Observations on P. halepensis suggest that taxifolin, resveratrol, and rutin may have an active role in protecting against B. xylophilus, possibly in conjugation with other biochemical and anatomical characters. We emphasize the importance of studying pine tolerant/resistance on adult trees, and not on excised branches, saplings, or seedlings to accurately simulate the nematode–pine host interactions occurring under natural conditions.

The wild dioecious plant Corema album, endemic to the Atlantic coast of the Iberian Peninsula, has potential to become a new crop and to integrate the berry fruit market. Due to its potential, the aim of this study was to propose a BBCH (Biologische Bundesanstalt, Bundessortenamt and CHemical Industry) scale for the species. Based on field observation, the principal growth stages were described and sequentially established. Photographs and stereomicroscope acquisition software were used to illustrate the phenological stages of male and female plants. The scale includes seven principal stages starting with bud development (stage 0), followed by leaf development (Stage 1), main stem elongation (Stage 3), inflorescence emergence (Stage 5), flowering (Stage 6), fruit development (Stage 7) and ending with fruit maturation (stage 8), with mesostages for male and female inflorescence (stage 5) and flower development (stage 6). Through field observations carried out in plants in different growth conditions, the proposition of this extended BBCH scale was made, to have a standardized system for Corema album phenological stages. This study will have an application especially in fields of agronomy and plant breeding and will also be a stepping stone for the inclusion of this species as a novelty crop in a nearby future.