When a plant invader meets its old enemy abroad: what can be learnt from accidental introductions of biological control agents.

Accidental introductions of biological weed control (BWC) agents (i) offer opportunities to assess host use of agents with a potentially broader fundamental host-range than those approved for field release directly in target areas; (ii) urge national authorities to rapidly respond as they may threaten native species or crops, and by this (iii) help advancing post-release studies, a neglected aspect of BWC. Through detailed insights gained from studying the recent accidental introduction of the ragweed leaf beetle Ophraella communa into Europe, we derive suggestions for overcoming barriers to adoption of BWC by re-evaluating the predictive power of pre-release studies and, thus, the presently strict criteria for deciding upon their release that might exclude safe and efficient agents. By using the allergenic weed Ambrosia artemisiifolia and the accidentally introduced BWC agent O. communa as study system, we also hope to raise the awareness of authorities to consider biological control more prominently as a key approach for pest management in the 'One Health' concept, which aims to sustainably balance and optimize the health of people, animals, plants and ecosystems. © 2023 Society of Chemical Industry.

Synergistic effects of grass competition and insect herbivory on the weed Rumex obtusifolius in an inundative biocontrol approach.

Outcomes of weed biological control projects are highly variable, but a mechanistic understanding of how top-down and bottom-up factors influence the success of weed biological control is often lacking. We grew Rumex obtusifolius, the most prominent native weed in European grasslands, in the presence and absence of competition from the grass Lolium perenne and subjected it to herbivory through targeted inoculation with root-boring Pyropteron spp. To explore whether the interactive effects of competition and inundative biological control were size-dependent, R. obtusifolius was planted covering a large range of plant sizes found in managed grasslands. Overall, competition from the grass sward reduced aboveground biomass and final root mass of R. obtusifolius about 62- and 7.5-fold, respectively, and increased root decay of R. obtusifolius from 14 to 58%. Herbivory alone increased only root decay. However, grass competition significantly enhanced infestation by Pyropteron spp. and, as a consequence, enhanced the impact of herbivory on aboveground biomass and final root mass. The synergistic effect was so strong that R. obtusifolius plants grown from initially smaller roots did no longer develop. Inoculating R. obtusifolius with Pyropteron species in grasslands should be further pursued as a promising inundative biological control strategy in the weed's native range.

A temporally and spatially explicit, data-driven estimation of airborne ragweed pollen concentrations across Europe.

Ongoing and future climate change driven expansion of aeroallergen-producing plant species comprise a major human health problem across Europe and elsewhere. There is an urgent need to produce accurate, temporally dynamic maps at the continental level, especially in the context of climate uncertainty. This study aimed to restore missing daily ragweed pollen data sets for Europe, to produce phenological maps of ragweed pollen, resulting in the most complete and detailed high-resolution ragweed pollen concentration maps to date. To achieve this, we have developed two statistical procedures, a Gaussian method (GM) and deep learning (DL) for restoring missing daily ragweed pollen data sets, based on the plant's reproductive and growth (phenological, pollen production and frost-related) characteristics. DL model performances were consistently better for estimating seasonal pollen integrals than those of the GM approach. These are the first published modelled maps using altitude correction and flowering phenology to recover missing pollen information. We created a web page (http://euragweedpollen.gmf.u-szeged.hu/), including daily ragweed pollen concentration data sets of the stations examined and their restored daily data, allowing one to upload newly measured or recovered daily data. Generation of these maps provides a means to track pollen impacts in the context of climatic shifts, identify geographical regions with high pollen exposure, determine areas of future vulnerability, apply spatially-explicit mitigation measures and prioritize management interventions.

The novel expression of clonality following whole-genome multiplication compensates for reduced fertility in natural autopolyploids.

Exploring the fitness consequences of whole-genome multiplication (WGM) is essential for understanding the establishment of autopolyploids in diploid parental populations, but suitable model systems are rare. We examined the impact of WGM on reproductive traits in three major cytotypes (2x, 3x, 4x) of Pilosella rhodopea, a species with recurrent formation of neo-autopolyploids in mixed-ploidy populations. We found that diploids had normal female sporogenesis and gametogenesis, high fertility, and produced predominantly euploid seed progeny. By contrast, autopolyploids had highly disturbed developmental programs that resulted in significantly lower seed set and a high frequency of aneuploid progeny. All cytotypes, but particularly triploids, produced gametes of varying ploidy, including unreduced ones, that participated in frequent intercytotype mating. Noteworthy, the reduced investment in sexual reproduction in autopolyploids was compensated by increased production of axillary rosettes and the novel expression of two clonal traits: adventitious rosettes on roots (root-sprouting), and aposporous initial cells in ovules which, however, do not result in functional apomixis. The combination of increased vegetative clonal growth in autopolyploids and frequent intercytotype mating are key mechanisms involved in the formation and maintenance of the largest diploid-autopolyploid primary contact zone ever recorded in angiosperms.

Clonal functional traits favor the invasive success of alien plants into native communities.

Functional traits are frequently proposed to determine the invasiveness of alien species. However, few empirical studies have directly manipulated functional traits and tested their importance in the invasion success of alien species into native plant communities, particularly under global change. We manipulated clonal integration (a key clonal functional trait) of four alien clonal plants by severing inter-ramet connections or keeping them intact and simulated their invasion into native plant communities with two levels of species diversity, population density and nutrient availability. High community diversity and density impeded the invasion success of the alien clonal plants. Clonal integration of the alien plants promoted their invasion success, particularly in the low-density communities associated with low species diversity or nutrient addition, which resulted in a negative correlation between the performance of alien plants and native communities, as expected under global change. Thus, clonal integration can favor the invasion success of alien clonal plants into degraded resident communities with a high degree of disturbance and eutrophication. Our findings confirm the role of clonal functional traits in facilitating alien plant invasions into native plant communities and suggest that clonal functional traits should be considered to efficiently restore degraded communities heavily invaded by alien clonal plants.

Climate warming can reduce biocontrol efficacy and promote plant invasion due to both genetic and transient metabolomic changes.

Climate change may affect plant-herbivore interactions and their associated ecosystem functions. In an experimental evolution approach, we subjected replicated populations of the invasive Ambrosia artemisiifolia to a combination of simulated warming and herbivory by a potential biocontrol beetle. We tracked genomic and metabolomic changes across generations in field populations and assessed plant offspring phenotypes in a common environment. Using an integrated Bayesian model, we show that increased offspring biomass in response to warming arose through changes in the genetic composition of populations. In contrast, increased resistance to herbivory arose through a shift in plant metabolomic profiles without genetic changes, most likely by transgenerational induction of defences. Importantly, while increased resistance was costly at ambient temperatures, warming removed this constraint and favoured both vigorous and better defended plants under biocontrol. Climate warming may thus decrease biocontrol efficiency and promote Ambrosia invasion, with potentially serious economic and health consequences.

High Phenotypic Plasticity in a Prominent Plant Invader along Altitudinal and Temperature Gradients.

Studies on plant growth and trait variation along environmental gradients can provide important information for identifying drivers of plant invasions and for deriving management strategies. We used seeds of the annual plant invader Ambrosia artemisiifolia L. (common ragweed) collected from an agricultural site in Northern Italy (226 m. a.s.l; Mean Annual Air Temperature: 12.9 °C; precipitations: 930 mm) to determine variation in growth trajectories and plant traits when grown along a 1000-m altitudinal gradient in Northern Italy, and under different temperature conditions in the growth chamber (from 14/18 °C to 26/30 °C, night/day), using a non-liner modeling approach. Under field conditions, traits related to plant height (maximum height, stem height, number of internodes) followed a three-parameter logistic curve. In contrast, leaf traits (lateral spread, number of leaves, leaf length and width) followed non-monotonic double-Richards curves that captured the decline patterns evident in the data. Plants grew faster, reaching a higher maximum plant height, and produced more biomass when grown at intermediate elevations. Under laboratory conditions, plants exhibited the same general growth trajectory of field conditions. However, leaf width did not show the recession after the maximum value shown by plants grown in the field, although the growth trajectories of some individuals, particularly those grown at 18 °C, showed a decline at late times. In addition, the plants grown at lower temperatures exhibited the highest value of biomass and preserved reproductive performances (e.g., amount of male inflorescence, pollen weight). From our findings, common ragweed exhibits a high phenotypic plasticity of vegetative and reproductive traits in response to different altitudes and temperature conditions. Under climate warming, this plasticity may facilitate the shift of the species towards higher elevation, but also the in situ resistance and (pre)adaptation of populations currently abundant at low elevations in the invasive European range. Such results may be also relevant for projecting the species management such as the impact by possible biocontrol agents.

Rapid genomic and phenotypic change in response to climate warming in a widespread plant invader.

Predicting plant distributions under climate change is constrained by our limited understanding of potential rapid adaptive evolution. In an experimental evolution study with the invasive common ragweed (Ambrosia artemisiifolia L.) we subjected replicated populations of the same initial genetic composition to simulated climate warming. Pooled DNA sequencing of parental and offspring populations showed that warming populations experienced greater genetic divergence from their parents, than control populations. In a common environment, offspring from warming populations showed more convergent phenotypes in seven out of nine plant traits, with later flowering and larger biomass, than plants from control populations. For both traits, we also found a significantly higher ratio of phenotypic to genetic differentiation across generations for warming than for control populations, indicating stronger response to selection under warming conditions. As a measure for evolutionary rate, the phenotypic and sequence divergence between generations were assessed using the Haldane metric. Our approach combining comparisons between generations (allochronic) and between treatments (synchronic) in an experimental evolutionary field study, and linking population genomic data with phenotyping analyses provided a powerful test to detect rapid responses to selection. Our findings demonstrate that ragweed populations can rapidly evolve in response to climate change within a single generation. Short-term evolutionary responses to climate change may aggravate the impact of some plant invaders in the future and should be considered when making predictions about future distributions and impacts of plant invaders.

Genome Assembly of the Ragweed Leaf Beetle: A Step Forward to Better Predict Rapid Evolution of a Weed Biocontrol Agent to Environmental Novelties.

Rapid evolution of weed biological control agents (BCAs) to new biotic and abiotic conditions is poorly understood and so far only little considered both in pre-release and post-release studies, despite potential major negative or positive implications for risks of nontargeted attacks or for colonizing yet unsuitable habitats, respectively. Provision of genetic resources, such as assembled and annotated genomes, is essential to assess potential adaptive processes by identifying underlying genetic mechanisms. Here, we provide the first sequenced genome of a phytophagous insect used as a BCA, that is, the leaf beetle Ophraella communa, a promising BCA of common ragweed, recently and accidentally introduced into Europe. A total 33.98 Gb of raw DNA sequences, representing ∼43-fold coverage, were obtained using the PacBio SMRT-Cell sequencing approach. Among the five different assemblers tested, the SMARTdenovo assembly displaying the best scores was then corrected with Illumina short reads. A final genome of 774 Mb containing 7,003 scaffolds was obtained. The reliability of the final assembly was then assessed by benchmarking universal single-copy orthologous genes (>96.0% of the 1,658 expected insect genes) and by remapping tests of Illumina short reads (average of 98.6 ± 0.7% without filtering). The number of protein-coding genes of 75,642, representing 82% of the published antennal transcriptome, and the phylogenetic analyses based on 825 orthologous genes placing O. communa in the monophyletic group of Chrysomelidae, confirm the relevance of our genome assembly. Overall, the genome provides a valuable resource for studying potential risks and benefits of this BCA facing environmental novelties.

How to better predict long-term benefits and risks in weed biocontrol: an evolutionary perspective.

Classical biological control (also called importation biological control) of weeds has a remarkable track record for efficiency and safety, but further improvement is still needed, particularly to account for potential evolutionary changes after release. Here, we discuss the increasing yet limited evidence of post-introduction evolution and describe approaches to predict evolutionary change. Recent advances include using experimental evolution studies over several generations that combine -omics tools with behavioral bioassays. This novel approach in weed biocontrol is well suited to explore the potential for rapid evolutionary change in real-time and thus can be used to estimate more accurately potential benefits and risks of agents before their importation. We outline this approach with a chrysomelid beetle used to control invasive common ragweed.

Biological weed control to relieve millions from Ambrosia allergies in Europe.

Invasive alien species (IAS) can substantially affect ecosystem services and human well-being. However, quantitative assessments of their impact on human health are rare and the benefits of implementing IAS management likely to be underestimated. Here we report the effects of the allergenic plant Ambrosia artemisiifolia on public health in Europe and the potential impact of the accidentally introduced leaf beetle Ophraella communa on the number of patients and healthcare costs. We find that, prior to the establishment of O. communa, some 13.5 million persons suffered from Ambrosia-induced allergies in Europe, causing costs of Euro 7.4 billion annually. Our projections reveal that biological control of A. artemisiifolia will reduce the number of patients by approximately 2.3 million and the health costs by Euro 1.1 billion per year. Our conservative calculations indicate that the currently discussed economic costs of IAS underestimate the real costs and thus also the benefits from biological control.

Predicting impact of a biocontrol agent: integrating distribution modeling with climate-dependent vital rates.

Species distribution models can predict the suitable climatic range of a potential biological control agent (BCA), but they provide little information on the BCA's potential impact. To predict high population buildup, a prerequisite of biocontrol impact, studies are needed that assess the effect of environmental factors on vital rates of a BCA across the environmental gradient of the BCA's suitable habitats, especially for the region where the BCA is considered for field release. We extended a published species distribution model with climate-dependent vital rates of Ophraella communa, a recently and accidentally introduced potential BCA of common ragweed, Ambrosia artemisiifolia in Europe. In field and laboratory experiments, we collected data on climate-dependent parameters assumed to be the most relevant for the population buildup of O. communa, i.e., temperature driving the number of generations per year and relative humidity (RH) determining egg hatching success. We found that O. communa concluded one generation in 334 cumulative degree days, and that egg hatching success strongly decreased from > 80% to < 20% when RH drops from 55% to 45% during the day. We used these values to spatially explicitly project population densities across the European range suitable for both A. artemisiifolia and the beetle and found that the present distribution of the beetle in Europe is within the range with the highest projected population growth. The highest population density of O. communa was predicted for northern Italy and parts of western Russia and western Georgia. Field observations of high impact on A. artemisiifolia with records of 80% aerial pollen reduction in the Milano area since the establishment of O. communa are in line with these predictions. The relative importance of temperature and RH on the population density of O. communa varies considerably across its suitable range in Europe. We propose that the combined statistical and mechanistic approach outlined in this paper helps to more accurately predict the potential impact of a weed BCA than a species distribution model alone. Identifying the factors limiting the population buildup of a BCA across the suitable range allows implementation of more targeted release and management strategies to optimize biocontrol efficacy.

Predicting abundances of invasive ragweed across Europe using a "top-down" approach.

Common ragweed (Ambrosia artemisiifolia L.) is a widely distributed and harmful invasive plant that is an important source of highly allergenic pollen grains and a prominent crop weed. As a result, ragweed causes huge costs to both human health and agriculture in affected areas. Efficient mitigation requires accurate mapping of ragweed densities that, until now, has not been achieved accurately for the whole of Europe. Here we provide two inventories of common ragweed abundances with grid resolutions of 1 km and 10 km. These "top-down" inventories integrate pollen data from 349 stations in Europe with habitat and landscape management information, derived from land cover data and expert knowledge. This allows us to cover areas where surface observations are missing. Model results were validated using "bottom-up" data of common ragweed in Austria and Serbia. Results show high agreement between the two analytical methods. The inventory shows that areas with the lowest ragweed abundances are found in Northern and Southern European countries and the highest abundances are in parts of Russia, parts of Ukraine and the Pannonian Plain. Smaller hotspots are found in Northern Italy, the Rhône Valley in France and in Turkey. The top-down approach is based on a new approach that allows for cross-continental studies and is applicable to other anemophilous species. Due to its simplicity, it can be used to investigate such species that are difficult and costly to identify at larger scales using traditional vegetation surveys or remote sensing. The final inventory is open source and available as a georeferenced tif file, allowing for multiple usages, reducing costs for health services and agriculture through well-targeted management interventions.

Do invasive alien plants differ from non-invasives in dominance and nitrogen uptake in response to variation of abiotic and biotic environments under global anthropogenic change?

Plant invasion is the outcome of complicated interactions of both biotic and abiotic environments (i.e. eutrophication and human-induced propagules) under global anthropogenic change. Here, we want to know why some alien clonal plant species become invasive and others do not in the introduced range with variations of both abiotic and biotic environments under global anthropogenic change. We selected three invasive alien and three co-occurring, non-invasive alien clonal plant species in China, and grew them under the constant or variable soil nutrient environments in a native community with low or high vegetative propagule pressure (i.e. simulating pressure of anthropogenic alien propagules). Invasive alien species produced more biomass than non-invasives. Interestingly, invasive species benefited significantly greater from high propagule pressure than non-invasives. Biomass and evenness of native communities were greater with non-invasive than with invasive target species. Invasive plants showed a greater increase of leaf N and decrease of leaf C: N ratio when subject to variable nutrients in comparison to constant nutrients than non-invasives. The negative effects of variable nutrients on evenness of native communities were significantly greater under invasive than non-invasive target species. Moreover, biomass of native communities was significantly negatively related to biomass of invasive species. Variable nutrients significantly promoted the negative biomass relationship between non-invasive species and native communities under high propagule pressure and the negative biomass-evenness relationship between invasive species and native communities. Our study suggests that soil nutrient variability and vegetative propagule pressure influence the growth and leaf C and N uptake of alien clonal plant species in native experimental communities, especially under the high propagule pressure and nutrient variability. Invasive alien clonal species have higher performance and advantages over non-invasives. Future studies should also test the mechanisms that invasive and non-invasive or native plants differ in native communities of native or introduced ranges in the field.

Sympatric diploid and tetraploid cytotypes of Centaurea stoebe s.l. do not differ in arbuscular mycorrhizal communities and mycorrhizal growth response.

PREMISE OF THE STUDY: Genome duplication is associated with multiple changes at different levels, including interactions with pollinators and herbivores. Yet little is known whether polyploidy may also shape belowground interactions. METHODS: To elucidate potential ploidy-specific interactions with arbuscular mycorrhizal fungi (AMF), we compared mycorrhizal colonization and assembly of AMF communities in roots of diploid and tetraploid Centaurea stoebe s.l. (Asteraceae) co-occurring in a Central European population. In a follow-up greenhouse experiment, we tested inter-cytotype differences in mycorrhizal growth response by combining ploidy, substrate, and inoculation with native AMF in a full-factorial design. KEY RESULTS: All sampled plants were highly colonized by AMF, with the Glomeraceae predominating. AMF-community composition revealed by 454-pyrosequencing reflected the spatial distribution of the hosts, but not their ploidy level or soil characteristics. In the greenhouse experiment, the tetraploids produced more shoot biomass than the diploids did when grown in a more fertile substrate, while no inter-cytotype differences were found in a less fertile substrate. AMF inoculation significantly reduced plant growth and improved P uptake, but its effects did not differ between the cytotypes. CONCLUSIONS: The results do not support our hypotheses that the cytotype structure in a mixed-ploidy population of C. stoebe is mirrored in AMF-community composition and that ploidy-specific fungal communities contribute to cytotype co-existence. Causes and implications of the observed negative growth response to AMF are discussed.

Direct effects of insecticides on common ragweed-implications for natural enemy exclusion trials.

Experimentally applying pesticides is an important method to assess the efficacy of weed biocontrol agents, but potential direct effects of the chemicals on plant performance are controversial or unknown. We assessed how three broad-spectrum insecticides applied in combination affect the performance of the widely invasive, crop-yield reducing, allergenic common ragweed (Ambrosia artemisiifolia L.) in an insect-free environment. Spraying insecticides had no significant effects on aboveground dry weight, seed and pollen output or pollen allergenicity, and only explained 1-8% of variation in these parameters. Our insecticide treatment can hence be applied to assess biocontrol impact on biomass and reproductive output of common ragweed. As our insecticide treatment delayed senescence, however, other methods of insect exclusion should be preferred when studying common ragweed phenology.

Deciphering the biology of Cryptophyllachora eurasiatica gen. et sp. nov., an often cryptic pathogen of an allergenic weed, Ambrosia artemisiifolia.

A little known, unculturable ascomycete, referred to as Phyllachora ambrosiae, can destroy the inflorescences of Ambrosia artemisiifolia, an invasive agricultural weed and producer of highly allergenic pollen. The fungus often remains undetectable in ragweed populations. This work was conducted to understand its origin and pathogenesis, a prerequisite to consider its potential as a biocontrol agent. The methods used included light and transmission electron microscopy, nrDNA sequencing, phylogenetic analyses, artificial inoculations, and the examination of old herbarium and recent field specimens from Hungary, Korea, Ukraine and USA. The Eurasian and the North American specimens of this fungus were to represent two distinct, although closely related lineages that were only distantly related to other lineages within the Ascomycota. Consequently, we describe a new genus that includes Cryptophyllachora eurasiatica gen. et sp. nov. and C. ambrosiae comb. nov., respectively. The pathogenesis of C. eurasiatica was shown in A. artemisiifolia. No evidence was found for either seed-borne transmission or systemic infection. Two hypotheses were developed to explain the interaction between C. eurasiatica and A. artemisiifolia: (i) as yet undetected seed-borne transmissions and latent, systemic infections; or (ii) alternative hosts.

Areas of high conservation value at risk by plant invaders in Georgia under climate change.

Invasive alien plants (IAP) are a threat to biodiversity worldwide. Understanding and anticipating invasions allow for more efficient management. In this regard, predicting potential invasion risks by IAPs is essential to support conservation planning into areas of high conservation value (AHCV) such as sites exhibiting exceptional botanical richness, assemblage of rare, and threatened and/or endemic plant species. Here, we identified AHCV in Georgia, a country showing high plant richness, and assessed the susceptibility of these areas to colonization by IAPs under present and future climatic conditions. We used actual protected areas and areas of high plant endemism (identified using occurrences of 114 Georgian endemic plant species) as proxies for AHCV. Then, we assessed present and future potential distribution of 27 IAPs using species distribution models under four climate change scenarios and stacked single-species potential distribution into a consensus map representing IAPs richness. We evaluated present and future invasion risks in AHCV using IAPs richness as a metric of susceptibility. We show that the actual protected areas cover only 9.4% of the areas of high plant endemism in Georgia. IAPs are presently located at lower elevations around the large urban centers and in western Georgia. We predict a shift of IAPs toward eastern Georgia and higher altitudes and an increased susceptibility of AHCV to IAPs under future climate change. Our study provides a good baseline for decision makers and stakeholders on where and how resources should be invested in the most efficient way to protect Georgia's high plant richness from IAPs.

Biological invasion of oxeye daisy (Leucanthemum vulgare) in North America: Pre-adaptation, post-introduction evolution, or both?

Species may become invasive after introduction to a new range because phenotypic traits pre-adapt them to spread and become dominant. In addition, adaptation to novel selection pressures in the introduced range may further increase their potential to become invasive. The diploid Leucanthemum vulgare and the tetraploid L. ircutianum are native to Eurasia and have been introduced to North America, but only L. vulgare has become invasive. To investigate whether phenotypic differences between the two species in Eurasia could explain the higher abundance of L. vulgare in North America and whether rapid evolution in the introduced range may have contributed to its invasion success, we grew 20 L. vulgare and 21 L. ircutianum populations from Eurasia and 21 L. vulgare populations from North America under standardized conditions and recorded performance and functional traits. In addition, we recorded morphological traits to investigate whether the two closely related species can be clearly distinguished by morphological means and to what extent morphological traits have changed in L. vulgare post-introduction. We found pronounced phenotypic differences between L. vulgare and L. ircutianum from the native range as well as between L. vulgare from the native and introduced ranges. The two species differed significantly in morphology but only moderately in functional or performance traits that could have explained the higher invasion success of L. vulgare in North America. In contrast, leaf morphology was similar between L. vulgare from the native and introduced range, but plants from North America flowered later, were larger and had more and larger flower heads than those from Eurasia. In summary, we found litte evidence that specific traits of L. vulgare may have pre-adapted this species to become more invasive than L. ircutianum, but our results indicate that rapid evolution in the introduced range likely contributed to the invasion success of L. vulgare.