Non-host Species Research Articles

Spatial Pattern of Host Tree Size, Rather than of Host Tree Itself, Affects the Infection Likelihood of a Fungal Stem Disease.

The spatial pattern of diseased forest trees is a product of the spatial pattern of host trees and the disease itself. Previous studies have focused on describing the spatial pattern of diseased host trees, and it remains largely unknown whether an antecedent spatial pattern of host/nonhost trees affects the infection pattern of a disease and how large the effect sizes of the spatial pattern of host/nonhost trees and host size are. The results from trivariate random labeling showed that the antecedent pattern of the host ash tree, Fraxinus mandshurica, but not of nonhost tree species, impacted the infection pattern of a stem fungal disease caused by Inonotus hispidus. To investigate the effect size of the spatial pattern of ash trees, we employed the SADIE (Spatial Analysis by Distance IndicEs) aggregation index and clustering index as predictors in the GLMs. Globally, the spatial pattern (vi index) of ash trees did not affect the infection likelihood of the focal tree; however, the spatial pattern of DBH (diameter at breast height) of ash trees significantly affected the infection likelihood of the focal tree. We sampled a series of circular plots with different radii to investigate the spatial pattern effect of host size on the infection likelihood of the focal tree locally. The results showed that the location (patch/gap) of the DBH of the focal tree, rather than that of the focal tree itself, significantly affected its infection likelihood in most plots of the investigated sizes. A meta-analysis was employed to settle the discrepancy between plots of different sizes, which led to results consistent with those of global studies. The results from meta-regression showed that plot size had no significant effects.

Reduction of damage by the exotic invasive conifer seed bug Leptoglossus occidentalis in mixed pine-birch plantations: A case of biotic resistance

The number of invasive non-native pests is increasing rapidly in forests as a result of global change. It is therefore important to prevent their damage in order to preserve the integrity of forest ecosystems and the associated services. According to the biotic resistance hypothesis, species-rich communities are less likely to be invaded. The associational resistance hypothesis states that insect herbivores are more likely to colonise and exploit plants surrounded by conspecific neighbours than heterospecific, non-host plant species. More diverse forests would therefore be less damaged by non-native pests than tree monocultures. We tested these hypotheses by comparing the damage caused to seeds by the western conifer bug Leptoglossus occidentalis, an invasive insect native to North America, in plots of pure maritime pine and mixed plots of maritime pine and birch. These plots were in two tree diversity experiments in Europe. Mixed pine plots differed in terms of pine density (1250 versus 625 pines/ha), proportion of pine and birch (25 %, 50 %, 75 %) and spatial mixing pattern (aggregated versus dispersed). We sampled 635 cones in 37 plots. Overall, the proportion of seeds damaged by the invasive bug was significantly lower in mixed plots than in pure pine plots at both experimental sites. There was no significant effect of pine density or relative proportion in the mixtures on seed damage. Aggregated pines in the mixed plots were significantly more damaged than dispersed pines, suggesting that bugs may have done less damage when they had more difficulty locating their host trees. This is consistent with the hypothesis of lower plant apparency. These results support the view that forest plantations of mixed species, which are known to be less vulnerable to attack by native insects, may also be more resistant to infestation by exotic insects. Row-wise, intimate mixtures of two species represent a promising option for designing tree plantations that are more resistant and easier to manage.

Genomic and culture-based analysis of Cyclaneusma minus in New Zealand provides evidence for multiple morphotypes

Cyclaneusma needle cast, caused by Cyclaneusma minus, affects Pinus species world wide. Previous studies suggested the presence of two distinct morphotypes in New Zealand, ‘verum’ and ‘simile’. Traditional mycological analyses revealed a third morphotype with clear differences in colony morphology and cardinal growth rates at varying temperatures. Genome sequencing of eight C. minus isolates provided further evidence of the existence of a third morphotype, named as ‘novus’ in this study. To further analyse these morphotypes, we predicted candidate effector proteins for all eight isolates, and also characterized a cell-death eliciting effector family, Ecp32, which is present in other pine phytopathogens. In concordance with their distinct classification into three different morphotypes, the number of Ecp32 family members differed, with patterns of pseudogenization in the ‘simile’ morphotype, and some members being found exclusively either in the ‘simile’ or ‘verum’ morphotypes. We also showed that the Ecp32 family proteins trigger cell death in non-host Nicotiana species, and, as previously demonstrated in other plant pathogens, the Ecp32 family proteins in C. minus adopt a β-trefoil fold. These analyses provide further evidence that the three morphotypes might be distinct species that need formal descriptions. Understanding the geographical range of different Cyclaneusma species and variations in virulence and pathogenicity will provide a better understanding of pine needle diseases and enable the development of more durable methods to control this disease.

Herbivory by Leaf-Cutting Ants: Exploring the Jasmonate Response in Host and Non-Host Plants.

Leaf-cutting ants (Formicidae; Atta spp., Acromyrmex spp.) cut off pieces of leaves and other plant tissue and feed it to their symbiotic fungi. As this foraging behavior poses an imminent threat to agriculture, leaf-cutting ants are considered as pests of huge ecologically and economically importance. Consequently, research on leaf-cutting ants focused on their foraging decisions and interactions with their cultivated symbiotic fungi, whereas their effect on the attacked plants, apart from the loss of plant tissue, remains largely unknown. In this study, we investigated the consequences of an attack by leaf-cutting ants and analyzed the plants' defense responses in comparison to chewing caterpillars and mechanical damage. We found that an attack by leaf-cutting ants induces the production of jasmonates in several host and non-host plant species (Arabidopsis thaliana, Vicia faba, Phaseolus lunatus, Tococa quadrialata). Additionally, we showed in the natural host plant lima bean (P. lunatus) that leaf-cutting ant damage immediately leads to the emission of typical herbivory-induced plant volatiles, including green leaf volatiles and terpenoids. Further data exploration revealed clear differences in the defense-related phytohormone profile in plant species of Neotropical and Eurasian origin. Taken together, we show that leaf-cutting ant infestation and their way of clipping the plants' tissues induce jasmonate and jasmonates-mediated responses and do not differ from those to mechanical injury or larval feeding.

The Effects of Infectious Diseases on the Consequences of Interspecific Competition in Grassland Communities

As infectious diseases have a severe impact on the individual survival and competitive ability of host species in grassland communities, competition between relevant species may have different consequences, potentially influencing the species composition of grassland communities and the functioning of grassland ecosystems. Understanding the impact of infectious diseases on competition is thus crucial for the health management of grassland ecosystems. How do infectious diseases affect the competitive coexistence of grassland plant species? In this study, by drawing on Tilman’s multispecies competition model and incorporating the spread characteristics of grassland plant diseases, we built three spatial competition models of two grass species subject to infectious diseases, and we analyzed the invasion conditions of infectious diseases and how they impact the population dynamics and competitive consequences of the species. Our model revealed the following: (1) Diseases with high transmission rates and low virulence are more likely to invade populations, while the presence of non-host species is detrimental to disease invasion. (2) Disease promotes the coexistence of competing species, breaking down the original competition–compromise trade-off mechanism for species coexistence and greatly expanding the range of parameters within which the two species can coexist. (3) Pathogen infections indirectly weaken the suppression of inferior species by dominant species, which is similar to the cascade effect seen in trophic interactions. Our findings highlight the importance of disease in species coexistence and grassland biodiversity maintenance.

Arbuscular mycorrhizal fungi induce Zn uptake and antioxidant efficiency in broccoli (Brassica oleracea L.) exposed to Zn deficiency

Zinc (Zn) is an essential micronutrient for plant growth and development, and its deficiency in soil can be a significant problem for broccoli (Brassica oleracea L.) production. While the genetic efforts to breed Zn-efficient broccoli are complex, harnessing microbiomes is an emerging way to induce mineral efficiency in crops. In this study, broccoli, a non-host mycorrhizal species, was cultivated with or without zinc deficiency in the presence or absence of arbuscular mycorrhizal fungi (AMF), confirmed by the colonization efficiency in roots. Zn deficiency caused a significant decrease in morphological parameters and photosynthetic attributes being consistent with the decreased Zn levels in root and shoot relative to Zn-sufficient plants. However, the broccoli plants inoculated with AMF showed a substantial improvement in morphological and photosynthetic parameters in Zn-deficient conditions due to the significant increase in tissue Zn levels. In addition, AMF-inoculated plants under Zn deficiency showed a significant decrease in cell death (%), electrolyte leakage, and H2O2 levels compared to Zn-deficient plants, which further suggests the improvements in cellular status due to the AMF colonization. The real-time PCR experiments showed a significant induction in the expression of BoZIP1 and BoNRAMP1 genes in the roots of Zn-deficient broccoli inoculated with AMF, suggesting that AMF may be associated with the induction of these genes responsible for Zn-uptake and transport. We further observed the induction of POD (peroxidase), APX (ascorbate peroxidase) and SOD (superoxide dismutase), and S-metabolites (cysteine and glutathione) predominantly in the roots of Zn-starved broccoli inoculated with AMF which may confer tolerance to Zn-deficiency induced oxidative damage. This is the first report on the role of AMF in mitigating Zn-deficiency in broccoli which may promote the microbiome-aided improvement in plant health suffering from Zn-deficiency in broccoli and other vegetable crops.

Seeds of nonhost species as sources of toxic compounds for the cowpea weevil Callosobruchus maculatus (F.)

Cowpea weevil, Callosobruchus maculatus, is the primary pest of stored cowpea seeds. The management of this infestation currently relies on insecticides, resulting in environmental pollution and selection of insecticide-resistant pests. Consequently, research efforts are being devoted to identify natural insecticides as sustainable and environment friendly alternatives for the control of C. maculatus. In this study, we explore the toxic effects of the nonhost seeds Parkia multijuga, Copaifera langsdorffii, Ormosia arborea, Amburana cearensis, Lonchocarpus guilleminianus, Sapindus saponaria, and Myroxylon peruiferum, on the cowpea weevil C. maculatus. Notably, all nonhost seeds led to reductions between 60 and 100% in oviposition by C. maculatus females. Additionally, the larvae were unable to penetrate the nonhost seeds. Artificial seeds containing 0.05% to 10% of cotyledon flour were toxic to C. maculatus larvae. Approximately 40% of larvae that consumed seeds containing 0.05% of O. arborea failed to develop, in contrast to control larvae. Proteomic analysis of A. cearensis and O. arborea seeds identify revealed a total of 371 proteins. From those, 237 are present in both seeds, 91 were exclusive to O. arborea seeds, and 43 were specific to A. cearensis seeds. Some of these proteins are related to defense, such as proteins containing the cupin domain and 11S seed storage protein. The in silico docking of cupin domain-containing proteins and 11S storage protein with N-acetylglucosamine (NAG)4 showed negative values of affinity energy, indicating spontaneous binding. These results showed that nonhost seeds have natural insecticide compounds with potential to control C. maculatus infestation.

Exploring the xylem-sap to unravel biological features of Xylella fastidiosa subspecies pauca ST53 in immune, resistant and susceptible crop species through metabolomics and in vitro studies.

Xylella fastidiosa subsp. pauca ST53 (Xfp) is a pathogenic bacterium causing one of the most severe plant diseases currently threatening the olive-growing areas of the Mediterranean, the Olive Quick Decline Syndrome (OQDS). The majority of the olive cultivars upon infections more or less rapidly develop severe desiccation phenomena, while few are resistant (e.g. Leccino and FS17), being less impacted by the infections. The present study contributes to elucidating the basis of the resistance phenomenon by investigating the influence of the composition of the xylem sap of plant species on the rate of bacterial multiplication. Xylem saps from Xfp host and non-host species were used for growing the bacterium in vitro, monitoring bacterial growth, biofilm formation, and the expression of specific genes. Moreover, species-specific metabolites, such as mannitol, quinic acid, tartaric acid, and choline were identified by non-targeted NMR-based metabolomic analysis in olive, grapevine, and citrus. In general, the xylem saps of immune species, including grapevine and citrus, were richer in amino acids, organic acids, and glucose. The results showed greater bacterial growth in the olive cultivar notoriously susceptible to Xfp (Cellina di Nardò), compared to that recorded in the resistant cultivar Leccino. Conversely, higher biofilm formation occurred in Leccino compared to Cellina di Nardò. Using the xylem saps of two Xfp-immune species (citrus and grapevine), a divergent bacterial behavior was recorded: low planktonic growth and biofilm production were detected in citrus compared to the grapevine. A parallel evaluation of the expression of 15 genes showed that Xfp directs its molecular functions mainly to virulence. Overall, the results gained through this multidisciplinary study contribute to extending the knowledge on the host-pathogen interaction, while confirming that the host response and resistance mechanism have a multifactorial basis, most likely with a cumulative effect on the phenotype.

Small mammal community composition impacts bank vole (Clethrionomys glareolus) population dynamics and associated seroprevalence of Puumala orthohantavirus.

Rodents are important reservoirs for zoonotic pathogens that cause diseases in humans. Biodiversity is hypothesized to be closely related to pathogen prevalence through multiple direct and indirect pathways. For example, the presence of non-host species can reduce contact rates of the main reservoir host and thus reduce the risk of transmission ("dilution effect"). In addition, an overlap in ecological niches between two species could lead to increased interspecific competition, potentially limiting host densities and reducing density-dependent pathogen transmission processes. In this study, we investigated the relative impact of population-level regulation of direct and indirect drivers of the prevalence of Puumala orthohantavirus (PUUV) in bank voles (Clethrionomys glareolus) during years with high abundance. We compiled data on small mammal community composition from four regions in Germany between 2010 and 2013. Structural equation modeling revealed a strong seasonality in PUUV control mechanisms in bank voles. The abundance of shrews tended to have a negative relationship with host abundance, and host abundance positively influenced PUUV seroprevalence, while at the same time increasing the abundance of competing non-hosts like the wood mouse (Apodemus sylvaticus) and the yellow-necked field mouse (Apodemus flavicollis) were associated with reduced PUUV seroprevalence in the host. These results indicate that for PUUV in bank voles, dilution is associated with increased interspecific competition. Anthropogenic pressures leading to the decline of Apodemus spp. in a specific habitat could lead to the amplification of mechanisms promoting PUUV transmission within the host populations.

Parasitoids as taxonomists: how does the parasitoid Chouioia cunea distinguish between a host and a non-host?

How parasitoids distinguish hosts from non-hosts remains an unknown question. Chouioia cunea Yang (Eulophidae) is an important fall webworm parasitoid that attacks many forest and agricultural pests. To study the differences in the chemical clues used by C. cunea to distinguish host and non-host plants, we used gas chromatography-mass spectrometry (GC-MS) to identify volatile compounds of two C. cunea hosts (Hyphantria cunea and Helicoverpa armigera) and two non-hosts (Spodoptera exigua and Spodoptera frugiperda). Additionally, we used behavioral assays to compare the attraction of C. cunea to various compounds. The two natural host species were more attractive than the two non-host species, in the following order: Hyphantria cunea > Helicoverpa armigera > S. exigua = S. frugiperda. The pupae of the natural hosts contained 1-dodecene, which was not produced by the two natural non-hosts. When the 'attractants' based on the difference between the species-specific blend emitted by pupae and the optimal blend were sprayed onto the natural non-host pupae, they significantly improved the attraction of C. cunea to the non-host pupae. These results revealed that specific host-produced volatile compounds guide C. cunea to distinguish between natural hosts and non-hosts. Overall, this study provides a foundation for developing a behavior-modifying strategy to re-direct C. cunea attacks to control important non-host pests. © 2023 Society of Chemical Industry.

The attractive host volatiles can enhance oviposition of Anoplophora glabripennis on a non-host tree.

The Asian longhorned beetle (ALB), Anoplophora glabripennis, is a serious wood borer of hardwood trees. Populus deltoides 'Shalinyang' (PdS) is attractive to ALB adults for oviposition but highly resistant to their offspring. Investigation of the chemicals regulating ALB oviposition is scarce in previous studies until now. To determine which chemicals emitted by PdS were attractive and induced oviposition behavior by ALB on non-host poplar tree species, we first: collected and identified the bio-active volatiles produced by PdS using coupled gas chromatography-mass spectrometry (GC-MS) and coupled gas chromatography-electroantennographic detector (GC-EAD); then evaluated which chemicals were attractive in a Y-tube olfactometer bioassay; and finally screened key compounds affecting ALB oviposition using a 'chemical-stimulated oviposition on non-host tree' bioassay. (E)-2-Hexenal, hexyl acetate, (Z)-3-hexenol acetate, 1-hexanol, (Z)-3-hexenol, β-caryophyllene, and salicylaldehyde emitted from PdS were attractive to ALB. When (E)-2-hexenal, 1-hexanol, (Z)-3-hexenol acetate, and (Z)-3-hexenol were applied to the non-host tree Populus tomentosa, oviposition by ALB females was significantly increased. Furthermore, the mean number of oviposition pits increased as the (Z)-3-hexenol concentrations increased. Further tests on synergy between pairs of chemicals showed that (Z)-3-hexenol stimulated production of the most oviposition pits, but that the percentage of effective oviposition pits (those containing an egg and larva and not empty) decreased. (Z)-3-Hexenol is the main chemical component inducing ALB oviposition. These results increase understanding about the oviposition behavior of ALB and could help improve management strategies that regulate ALB behavior by planting mixed-species forests resistant to ALB. © 2023 Society of Chemical Industry.

Tree canopy patterns in a temperate forest in Oaxaca, Mexico after bark beetle outbreaks: Implications for ecological resistance and resilience

Bark beetles (BB) are an insect that can become a forest pest and that principally affects temperate regions globally, massively killing tree hosts, mainly coniferous species. When tree hosts are the dominant species in the canopy, BB outbreaks may impose important changes in the structure, composition and diversity of forests, from the stand to the regional scale. In Mexico, during the current century, BB pests have become the main biotic threat in forests where Pinus species are dominant. Questions addressed in this study are: What is the structure, composition and diversity of the forest canopy after BB disturbances? and “What are the implications for ecological resistance and resilience of the forest”? The study site was located in the Sierra Norte region in Oaxaca, Mexico, where more a decade ago BB outbreaks affected the forest. In ninety plots of 500 m2 (4.5 ha) all live trees with ≥ 5 cm diameter at breast height (DBH) were located, their DBH was measured, and the species was identified. As well, all stumps resulting from previous sanitation logging and timber extraction (TE) were located and for each condition, basal diameter was estimated and the genera identified. For the post-disturbance canopy, density basal area, species composition and diversity (alpha and beta) were estimated. This data plus the analysis of the stumps was used to estimate the tree canopy structure before the BB outbreaks. Statistical comparisons among BB, TE disturbances and condition without disturbance were conducted. Based on interviews with local experts, a rank of risk to BB among Pinus species was established. A total of 4,053 live trees and 547 stumps were found. The live trees corresponded to 24 species from 8 genera. Pinus genus and, particularly P. patula predominated. Seventy-five percent of the individuals had < 20 cm DBH and a J-inverted diametric distribution resulted. Individuals with DBH ≥ 100 cm were found only in the condition without disturbance. Three of the nine Pinus species, were BB main-host species, and the least preferred host was P. ayacahuite. The three BB main-host species dominated the post-disturbance canopy (including regeneration). In contrast, P. ayacahuite and other non-host tree species dominated in the conditions without disturbance, and the three main-host species showed low density. As well, high species richness occurred in areas affected by BB. Since climate change will likely continue to exacerbate BB outbreaks, it is proposed that to increase ecological resistance and resilience forest management practices should focus on reducing density, promoting a mix of host and non-host species, increasing tree diversity and avoiding stands with monodominance of main-host species.

Isolation characterization and pathogenicity of Helicoverpa armigera single nucleopolyhedrovirus isolate from Türkiye

The cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), is a major pest of agricultural crops, such as cotton, horticultural plants, a range of vegetable crops and forest trees. Many dead and live larvae and pupa samples of H. armigera, an extremely polyphagous pest, were collected from Adana in Türkiye. It was observed that cadaveric integuments were fragile, and their body tissues were also liquefied. Studies with phase-contrast microscopy revealed a very intense baculovirus infection in cadavers. Transmission electron microscopy exhibited that the new baculovirus had one nucleocapsid in an envelope in the occlusion bodies. Therefore, the isolate was named Helicoverpa armigera single nucleopolyhedrovirus-D (HearSNPV-D). The phylogenetic analysis according to the polyhedrin and late expression factor-8 genes appeared that the new isolate was in close relationship with HearSNPV China and Türkiye isolate. The new isolate was tested on six different insects including its own host Helicoverpa armigera, Hyphantria cunea, Malacosoma neustria, Lymantria dispar, Thaumetopea pityocampa and Spodoptera exigua larvae. Bioassay studies showed that HearSNPV-D was virulent to H. armigera and had a narrow host range. The LC50 value was determined as 0.7 × 105 and 1.9 × 107 OBs/ml for H. armigera and S. exigua, respectively. Non-significant mortality was observed in non-host species by HearSNPV-D, which may reveal the highly specific nature of HearSNPV-D. Therefore, HearSNPV-D can be used as a specific, environmentally friendly control agent for H. armigera.

The genome of Lactuca saligna, a wild relative of lettuce, provides insight into non-host resistance to the downy mildew Bremia lactucae.

Lactuca saligna L. is a wild relative of cultivated lettuce (Lactuca sativa L.), with which it is partially interfertile. Hybrid progeny suffer from hybrid incompatibility (HI), resulting in reduced fertility and distorted transmission ratios. Lactuca saligna displays broad-spectrum resistance against lettuce downy mildew caused by Bremia lactucae Regel and is considered a non-host species. This phenomenon of resistance in L. saligna is called non-host resistance (NHR). One possible mechanism behind this NHR is through the plant-pathogen interaction triggered by pathogen recognition receptors, including nucleotide-binding leucine-rich repeat (NLR) proteins and receptor-like kinases (RLKs). We report a chromosome-level genome assembly of L. saligna (accession CGN05327), leading to the identification of two large paracentric inversions (>50 Mb) between L. saligna and L. sativa. Genome-wide searches delineated the major resistance clusters as regions enriched in NLRs and RLKs. Three of the enriched regions co-locate with previously identified NHR intervals. RNA-seq analysis of Bremia-infected lettuce identified several differentially expressed RLKs in NHR regions. Three tandem wall-associated kinase-encoding genes (WAKs) in the NHR8 interval display particularly high expression changes at an early stage of infection. We propose RLKs as strong candidates for determinants of the NHR phenotype of L. saligna.

When tree diversity does not work: The interacting effects of tree diversity, altitude and amount of spruce on European spruce bark beetle outbreaks

The frequency of large-scale insect outbreaks is expected to increase with climate change, and the European spruce bark beetle (Ips typographus) is a major forest pest in Europe that has caused more extensive and widespread outbreaks in recent years. It is therefore important to evaluate forest management strategies for controlling bark beetle populations. While tree species diversity is often promoted as a way to improve forest resistance to specialist insects, the relationship between tree species diversity and bark beetle outbreaks is not well understood, partly due to the complex interactions with other factors. The aim of this study was to assess the impact of tree species diversity on European spruce bark beetle outbreaks, while controlling for host availability across different altitudes. We analysed data from a systematic 1 × 1 km grid in Slovenia. For each grid cell, we extracted data on the sanitary felling of Norway spruce that followed bark beetle outbreaks from a national data base covering the period 1996–2017. Additionally, we extracted data on resource availability (i.e., the percentage of Norway spruce in the grid cell), non-host tree species diversity and average altitude. We analysed the effect of altitude, forest cover, percentage of spruce and non-host tree species diversity during outbreak and non-outbreak periods on the probability of sanitary felling and on the amount of sanitary felling, separately. The probability of sanitary felling was higher at low altitudes and increased with an increasing proportion of spruce in the growing stock in the grid cell, but it was not affected by non-host tree species diversity. In contrast, the amount of sanitary felling decreased with increasing non-host tree species diversity, particularly at low altitudes. However, the effect of non-host tree species diversity on the amount of sanitary felling diminished as the percentage of Norway spruce increased. Thus, non-host tree species diversity per se is not the main driver of outbreak risk but strengthens biotic resistance with lower host availability at low altitudes where abiotic conditions are the least favourable to Norway spruce.

Competitive interactions in two different plant species: Do grassland mycorrhizal communities and nitrogen addition play the same game?

In the Tibetan Plateau grassland ecosystems, nitrogen (N) availability is rising dramatically; however, the influence of higher N on the arbuscular mycorrhizal fungi (AMF) might impact on plant competitive interactions. Therefore, understanding the part played by AMF in the competition between Vicia faba and Brassica napus and its dependence on the N-addition status is necessary. To address this, a glasshouse experiment was conducted to examine whether the grassland AMF community's inocula (AMF and NAMF) and N-addition levels (N-0 and N-15) alter plant competition between V. faba and B. napus. Two harvests took day 45 (1st harvest) and day 90 (2nd harvest), respectively. The findings showed that compared to B. napus, AMF inoculation significantly improved the competitive potential of the V. faba. In the occurrence of AMF, V. faba was the strongest competitor being facilitated by B. napus in both harvests. While under N-15, AMF significantly enhanced tissue N:P ratio in B. napus mixed-culture at 1st harvest, the opposite trend was observed in 2nd harvest. The mycorrhizal growth dependency slightly negatively affected mixed-culture compared to monoculture under both N-addition treatments. The aggressivity index of AMF plants was higher than NAMF plants with both N-addition and harvests. Our observation highlights that mycorrhizal associations might facilitate host plant species in mixed-culture with non-host plant species. Additionally, interacting with N-addition, AMF could impact the competitive ability of the host plant not only directly but also indirectly, thereby changing the growth and nutrient uptake of competing plant species.

Factors affecting pine processionary moth (Thaumetopoea pityocampa) incidence in Mediterranean pine stands: A multiscale approach

The occurrence of the pine processionary moth (Thaumetopoea pityocampa, hereafter PPM) in Mediterranean pine forests and plantations is of increasing concern as processes such as global warming and forest cover expansion have amplified the frequency and virulence of outbreaks. This study attempts to provide as complete an overview as possible of the different factors involved in pine forest susceptibility to PPM in 98 pine stands in Central Spain. A large study area (∼3,000 km2) was considered, including three different spatial scales (<1 ha, 1 ha, 10 ha) and four different pine tree species (Pinus pinaster, P. nigra, P. sylvestris, P. pinea), monitored over a 4-year period. Models were fitted for each of the three spatial scales, as well as for a combination of them (multiscale model). Pine species had a strong influence on vulnerability to PPM, with higher incidences in stands dominated by P. pinaster and P. nigra and lower for those dominated by P. sylvestris and P. pinea. Grazing pressure, a factor thus far disregarded, was also determining, with a prominent role at both the multiscale and the stand-scale levels. Other variables were highly influential at some of the spatial scales, showing a greater resistance in pine forests or plantations with a higher diversity of pine sizes, greater tree density, and a larger cover of non-host tree species. Open habitats also played a role in PPM incidence: we detected that, both at the locality and the plot scale, the presence of clearings occupied by bushes or rocky outcrops reduced vulnerability to PPM, while the clearings with a more simplified structure produced the opposite effect. We conclude that PPM incidence is greater in structurally simple pine forests, with strong grazing pressure and abundant open spaces without woody vegetation that may favour PPM pupation. To reduce damage by PPM, pine forest and plantations should be managed to avoid oversimplified forest structures. Patches of non-host tree species should be promoted, as well as some presence of shrubs in open areas. This implies avoiding overgrazing and other management models leading to an excessive increase in bare soil. Nevertheless, it should be noted that the implementation of these recommendations should consider the remaining challenges that need to be addressed in the current context of global change, such as the increased risk of wildfires and the emergence of other pests and diseases.

What Was the Reason for the Durable Effect of Sr31 against Wheat Stem Rust?

Common wheat cultivars have been protected from stem rust for several decades worldwide by the Sr31 resistance gene transferred from Secale cereale L. (cv. Petkus). In 1998, Sr31 was overcome in Uganda by the Ug99 race of Puccinia graminis f. sp. tritici Eriks. &amp; Henn. (Pgt). The Ug99 race and its derivatives have spread widely in Africa, neighboring regions and Europe. However, Sr31 remains effective in other areas of the world, including Russia. To breed wheat with durable resistance, it is promising to research the resistance mechanisms of nonhost species and introgressive cultivars. The aim of the research was to estimate the resistance of S. cereale and Triticum aestivum cultivars with Sr31 to stem rust and to study the mechanisms of incompatibility of Pgt with plants at the cellular level. The research was carried out in Western Siberia (Russia, Omsk region) in 2018–2022. Rye and wheat with Sr31 (cvs. Kavkaz, Seri 82, Bacanora (=Kauz’s’), NIL Thatcher TcLr26/Sr31) were resistant at the stages of seedling and adult plant, and cv. PWB343 was more susceptible to disease. Cytological studies have shown that Pgt died on the rye plants on the surface, and cv. Petkus intensively suppressed the development of the appressoria necessary to penetrate into tissues. Wheat cultivars inhibited the Pgt development mainly on the surface and while it attempted to penetrate into the stomata (pre-haustorial resistance). It has been demonstrated that Pgt has to adapt step-by-step to the surface and tissue properties for compatible interaction, which may be the reason for the durable effectiveness of Sr31.

Western Spruce Budworm Effects on Forest Resilience.

Western spruce budworm (Choristoneura freemani Razowski) is the most destructive defoliator of forests in the western US. Forests in northern New Mexico experienced high levels of WSBW-caused defoliation and subsequent mortality between the 1980s and 2010s. The effects of severe western spruce budworm outbreaks on stand dynamics in the US Southwest are still relatively unknown, but understanding the impacts is important to the management and resilience of these forests. To begin addressing this knowledge gap, we conducted a study along two gradients: an elevational gradient from mixed-conifer to spruce-fir forests and a gradient of WSBW-caused defoliation intensity. We recorded overstory and understory stand conditions (size structure, species composition, damaging agents). Western spruce budworm was the primary damaging agent of host trees in all stands andcaused host tree mortality across all size classes, particularly in spruce-fir stands. Results indicate an unsustainable level of mortality in spruce-fir stands and a transition towards non-host species in mixed-conifer stands. Low levels of regeneration coupled with high overstory mortality rates indicate a potential lack of resilience in spruce-fir stands, whereas resilience to future western spruce budworm defoliation events may have increased in mixed-conifer stands affected by these outbreaks.

Characterization of two conserved cell death elicitor families from the Dothideomycete fungal pathogens Dothistroma septosporum and Fulvia fulva (syn. Cladosporium fulvum).

Dothistroma septosporum (Ds) and Fulvia fulva (Ff; previously called Cladosporium fulvum) are two closely related Dothideomycete fungal species that cause Dothistroma needle blight in pine and leaf mold in tomato, respectively. During host colonization, these pathogens secrete virulence factors termed effectors to promote infection. In the presence of corresponding host immune receptors, however, these effectors activate plant defenses, including a localized cell death response that halts pathogen growth. We identified two apoplastic effector protein families, Ecp20 and Ecp32, which are conserved between the two pathogens. The Ecp20 family has four paralogues in both species, while the Ecp32 family has four paralogues in D. septosporum and five in F. fulva. Both families have members that are highly expressed during host infection. Members of the Ecp20 family have predicted structural similarity to proteins with a β-barrel fold, including the Alt a 1 allergen from Alternaria alternata, while members of the Ecp32 family have predicted structural similarity to proteins with a β-trefoil fold, such as trypsin inhibitors and lectins. Using Agrobacterium tumefaciens-mediated transient transformation assays, each family member was assessed for its ability to trigger cell death in leaves of the non-host species Nicotiana benthamiana and N. tabacum. Using this approach, FfEcp20-2, DsEcp20-3, and FfEcp20-3 from the Ecp20 family, and all members from the Ecp32 family, except for the Ds/FfEcp32-4 pair, triggered cell death in both species. This cell death was dependent on secretion of the effectors to the apoplast. In line with recognition by an extracellular immune receptor, cell death triggered by Ds/FfEcp20-3 and FfEcp32-3 was compromised in N. benthamiana silenced for BAK1 or SOBIR1, which encode extracellular co-receptors involved in transducing defense response signals following apoplastic effector recognition. We then investigated whether DsEcp20-3 and DsEcp20-4 triggered cell death in the host species Pinus radiata by directly infiltrating purified protein into pine needles. Strikingly, as in the non-host species, DsEcp20-3 triggered cell death, while DsEcp20-4 did not. Collectively, our study describes two new candidate effector families with cell death-eliciting activity from D. septosporum and F. fulva and provides evidence that members of these families are recognized by plant immune receptors.