The plant nursery industry has become an ideal reservoir for Phytophthora species and other soilborne pathogens. In this context, isolation from tissues and soil of ornamental and forest plants from nurseries in four regions of Spain was carried out. A high diversity of Phytophthora species was confirmed. Fourteen Phytophthora phylotypes (P. cactorum, P. cambivora, P. cinnamomi, P. citrophthora, P. crassamura, P. gonapodyides, P. hedraiandra, P. nicotianae, P. niederhauserii, P. palmivora, P. plurivora, P. pseudocryptogea, P. sansomeana, and Phytophthora sp. tropicalis-like 2) were isolated from over 500 plant samples of 22 species in 19 plant genera. Nine species were detected in water sources, two of them (P. bilorbang and P. lacustris) exclusively from water samples. P. crassamura was detected for the first time in Spain. This is the first time P. pseudocryptogea is isolated from Chamaecyparis lawsoniana and Yucca rostrata in Spain.

Oomycetes of the genus Phytophthora encompass several of the most successful plant pathogens described to date. The success of infection by Phytophthora species is attributed to the pathogens\' ability to secrete effector proteins that alter the host\'s physiological processes. Structural analyses of effector proteins mainly from bacterial and viral pathogens have revealed the presence of intrinsically disordered regions that host short linear motifs (SLiMs). These motifs play important biological roles by facilitating protein-protein interactions as well as protein translocation. Nonetheless, SLiMs in Phytophthora species RxLR effectors have not been investigated previously and their roles remain unknown. Using a bioinformatics pipeline, we identified 333 candidate RxLR effectors in the strain INRA 310 of Phytophthora parasitica. Of these, 71 (21%) were also found to be present in 10 other genomes of P. parasitica, and hence, these were designated core RxLR effectors (CREs). Within the CRE sequences, the N terminus exhibited enrichment in intrinsically disordered regions compared to the C terminus, suggesting a potential role of disorder in effector translocation. Although the disorder content was reduced in the C-terminal regions, it is important to mention that most SLiMs were in this terminus. PpRxLR1 is one of the 71 CREs identified in this study, and its genes encode a 6-amino acid (aa)-long SLiM at the C terminus. We showed that PpRxLR1 interacts with several host proteins that are implicated in defense. Structural analysis of this effector using homology modeling revealed the presence of potential ligand-binding sites. Among key residues that were predicted to be crucial for ligand binding, L102 and Y106 were of interest since they form part of the 6-aa-long PpRxLR1 SLiM. In silico substitution of these two residues to alanine was predicted to have a significant effect on both the function and the structure of PpRxLR1 effector. Molecular docking simulations revealed possible interactions between PpRxLR1 effector and ubiquitin-associated proteins. The ubiquitin-like SLiM carried in this effector was shown to be a potential mediator of these interactions. Further studies are required to validate and elucidate the underlying molecular mechanism of action. IMPORTANCE The continuous gain and loss of RxLR effectors makes the control of Phytophthora spp. difficult. Therefore, in this study, we endeavored to identify RxLR effectors that are highly conserved among species, also known as \"core\" RxLR effectors (CREs). We reason that these highly conserved effectors target conserved proteins or processes; thus, they can be harnessed in breeding for durable resistance in plants. To further understand the mechanisms of action of CREs, structural dissection of these proteins is crucial. Intrinsically disordered regions (IDRs) that do not adopt a fixed, three-dimensional fold carry short linear motifs (SLiMs) that mediate biological functions of proteins. The presence and potential role of these SLiMs in CREs of Phytophthora spp. have been overlooked. To our knowledge, we have effectively identified CREs as well as SLiMs with the potential of promoting effector virulence. Together, this work has advanced our comprehension of Phytophthora RxLR effector function and may facilitate the development of innovative and effective control strategies.

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Predicting whether naïve tree populations have the potential to adapt to exotic pathogens is necessary owing to the increasing rate of invasions. Adaptation may occur as a result of natural selection when heritable variation in terms of susceptibility exists in the naïve population. We searched for signs of selection on black alder (Alnus glutinosa) stands growing on riverbanks invaded by two pathogens differing in aggressiveness, namely, Phytophthora uniformis (PU) and Phytophthora × alni (PA). We compared the survival and heritability measures from 72 families originating from six invaded and uninvaded (naïve) sites by performing in vitro inoculations. The results from the inoculations were used to assess the relative contribution of host genetic variation on natural selection. We found putative signs of natural selection on alder exerted by PU but not by PA. For PU, we found a higher survival in families originating from invaded sites compared with uninvaded sites. The narrow sense heritability of susceptibility to PU of uninvaded populations was significantly higher than to PA. Simulated data supported the role of heritable genetic variation on natural selection and discarded a high aggressiveness of PA decreasing the transmission rate as an alternative hypothesis for a slow natural selection. Our findings expand on previous attempts of using heritability as a predictor for the likelihood of natural adaptation of naïve tree populations to invasive pathogens. Measures of genetic variation can be useful for risk assessment purposes or when managing Phytophthora invasions.

Phytophthora cinnamomi is one of the world\'s most invasive plant pathogens affecting ornamental plants, horticultural crops and natural ecosystems. Accurate diagnosis is very important to determine the presence or absence of this pathogen in diseased and asymptomatic plants. In previous studies, P. cinnamomi species-specific primers were designed and tested using various polymerase chain reaction (PCR) techniques including conventional PCR, nested PCR and quantitative real-time PCR. In all cases, the primers were stated to be highly specific and sensitive to P. cinnamomi. However, few of these studies tested their primers against closely related Phytophthora species (Phytophthora clade 7). In this study, we tested these purported P. cinnamomi-specific primer sets against 11 other species from clade 7 and determined their specificity; of the eight tested primer sets only three were specific to P. cinnamomi. This study demonstrated the importance of testing primers against closely related species within the same clade, and not just other species within the same genus. The findings of this study are relevant to all species-specific microbial diagnosis.

A major aim of invasion ecology is to identify characteristics of successful invaders. However, most plant groups studied in detail (e.g. pines and acacias) have a high percentage of invasive taxa. Here we examine the global introduction history and invasion ecology of Proteaceae--a large plant family with many taxa that have been widely disseminated by humans, but with few known invaders. To do this we compiled a global list of species and used boosted regression tree models to assess which factors are important in determining the status of a species (not introduced, introduced, naturalized or invasive). At least 402 of 1674 known species (24%) have been moved by humans out of their native ranges, 58 species (14%) have become naturalized but not invasive, and 8 species (2%) are invasive. The probability of naturalization was greatest for species with large native ranges, low susceptibility to Phytophthora root-rot fungus, large mammal-dispersed seeds, and with the capacity to resprout. The probability of naturalized species becoming invasive was greatest for species with large native ranges, those used as barrier plants, tall species, species with small seeds, and serotinous species. The traits driving invasiveness of Proteaceae were similar to those for acacias and pines. However, while some traits showed a consistent influence at introduction, naturalization and invasion, others appear to be influential at one stage only, and some have contrasting effects at different stages. Trait-based analyses therefore need to consider different invasion stages separately. On their own, these observations provide little predictive power for risk assessment, but when the causative mechanisms are understood (e.g. Phytophthora susceptibility) they provide valuable insights. As such there is considerable value in seeking the correlates and mechanisms underlying invasions for particular taxonomic or functional groups.

Phytophthora infestans is the most important fungal pathogen in the cultivated potato (Solanum tuberosum). Dominant, race-specific resistance alleles and quantitative resistance--the latter being more important for potato breeding--are found in the germplasm of cultivated and wild potato species. Quantitative trait loci (QTLs) for resistance to two races of P. infestans have been mapped in an F1 progeny of a cross between non-inbred diploid potato parents with multiple alleles. Interval mapping methods based on highly informative restriction fragment length polymorphism markers revealed 11 chromosome segments on 9 potato chromosomes showing significant contrasts between marker genotypic classes. Whereas phenotypically no difference in quantitative resistance response was observed between the two fungal races, QTL mapping identified at least one race specific QT locus. Two QT regions coincided with two small segments on chromosomes V and XII to which the dominant alleles R1, conferring race specific resistance to P. infestans, Rx1 and Rx2, both inducing extreme resistance to potato virus X, have been allocated in independent mapping experiments. Some minor QTLs were correlated with genetic loci for specific proteins related to pathogenesis, the expression of which is induced after infection with P. infestans.