Pollen from common ragweed is an important allergen source worldwide and especially in western and southern Romania. More than 100 million patients suffer from symptoms of respiratory allergy (e.g., rhinitis, asthma) to ragweed pollen. Among the eleven characterized allergens, Amb a 6 is a non-specific lipid transfer protein (nsLTP). nsLTPs are structurally stable proteins in pollen and food from different unrelated plants capable of inducing severe reactions. The goal of this study was to produce Amb a 6 as a recombinant and structurally folded protein (rAmb a 6) and to characterize its physicochemical and immunological features. rAmb a 6 was expressed in Spodoptera frugiperda Sf9 cells as a secreted protein and characterized by mass spectrometry and circular dichroism (CD) spectroscopy regarding molecular mass and fold, respectively. The IgE-binding frequency towards the purified protein was evaluated using sera from 150 clinically well-characterized ragweed-allergic patients. The allergenic activities of rAmb a 6 and the nsLTP from the weed Parietaria judaica (Par j 2) were evaluated in basophil activation assays. rAmb a 6-specific IgE reactivity was associated with clinical features. Pure rAmb a 6 was obtained by insect cell expression. Its deduced molecular weight corresponded to that determined by mass spectrometry (i.e., 10,963 Da). rAmb a 6 formed oligomers as determined by SDS-PAGE under non-reducing conditions. According to multiple sequence comparisons, Amb a 6 was a distinct nsLTP with less than 40% sequence identity to currently known plant nsLTP allergens, except for nsLTP from Helianthus (i.e., 52%). rAmb a 6 is an important ragweed allergen recognized by 30% of ragweed pollen allergic patients. For certain patients, rAmb a 6-specific IgE levels were higher than those specific for the major ragweed allergen Amb a 1 and analysis also showed a higher allergenic activity in the basophil activation test. rAmb a 6-positive patients suffered mainly from respiratory symptoms. The assumption that Amb a 6 is a source-specific ragweed allergen is supported by the finding that none of the patients showing rAmb a 6-induced basophil activation reacted with Par j 2 and only one rAmb a 6-sensitized patient had a history of plant food allergy. Immunization of rabbits with rAmb a 6 induced IgG antibodies which strongly inhibited IgE binding to rAmb a 6. Our results demonstrate that Amb a 6 is an important source-specific ragweed pollen allergen that should be considered for diagnosis and allergen-specific immunotherapy of ragweed pollen allergy.

Rapid adaptive evolution and phenotypic plasticity are two mechanisms that often underlie invasiveness of alien plant species, but whether they can co-occur within invasive plant populations under altered environmental conditions such as nitrogen (N) enrichment has seldom been explored. Latitudinal clines in plant trait responses to variation in environmental factors may provide evidence of local adaptation. Here, we inferred the relative contributions of phenotypic plasticity and local adaptation to the performance of the invasive plant Ambrosia artemisiifolia under different soil N levels, using a common garden approach. We grew A. artemisiifolia individuals raised from seeds that were sampled from six invasive populations along a wide latitudinal cline in China (23°42\' N to 45°43\' N) under three N (0, 5, and 10 g N m-2 ) levels in a common garden. Results show significant interpopulation genetic differentiation in plant height, number of branches, total biomass, and transpiration rate of the invader A. artemisiifolia across the N treatments. The populations also expressed genetic differentiation in basal diameter, growth rate, leaf area, seed width, root biomass, aboveground biomass, stomatal conductance, and intercellular CO2 concentration regardless of N treatments. Moreover, plants from different populations of the invader displayed plastic responses in time to first flower, hundred-grain weight, net photosynthetic rate, and relative biomass allocation to roots and shoots and seed length under different N treatments. Additionally, individuals of A. artemisiifolia from higher latitudes grew shorter and allocated less biomass to the roots regardless of N treatment, while latitudinal cline (or lack thereof) in other traits depended on the level of N in which the plants were grown. Overall, these results suggest that rapid adaptive evolution and phenotypic plasticity in the various traits that we quantified may jointly contribute to invasiveness of A. artemisiifolia under different levels of N availability. More broadly, the results support the idea that phenotypic plasticity and rapid adaptive evolution can jointly enable invasive plants to colonize a wide range of environmental conditions.
快速适应进化和表型可塑性是阐释外来植物入侵的两个机制，但是它们是否可以在变化的环境条件下(比如氮富集)同时作用于入侵种群，这方面的研究还不多。植物性状对环境因子变化响应的纬度趋势可能提供了局域适应的证据。本文采用同质园实验的方法，探究表现可塑性和局域适应对入侵植物豚草(Ambrosia artemisiifolia)在不同土壤氮水平下表现的相对贡献。我们沿中国纬度梯度(23°42′ N-45°43′ N)收集六个豚草入侵种群的种子，分别设置三个氮水平(0，5以及10 g N m−2 )种植在同质园中。结果表明，在不同的氮水平下，入侵豚草的株高、分枝数、总生物量和蒸腾速率存在显著的种群间遗传差异。在三种氮处理水平下，基径、生长速率、叶面积、种子宽度、根生物量、地上生物量、气孔导度和胞间二氧化碳浓度在种群间也存在遗传分化。在不同的氮处理下，来自不同种群的豚草植株在开花时间、百粒重、净光合速率、根冠比以及种子长度上表现出可塑性响应。此外，在所有氮处理水平下，来自更高纬度种群的豚草个体株高较小，分配到根部的生物量较少，而其他性状的纬度差异取决于氮处理水平。总之，研究结果表明，我们测量的各种性状中的快速适应性进化和表型可塑性可能联合作用，共同促进豚草在不同的氮水平下的入侵。广而言之，研究结果支持表型可塑性和快速适应性进化可以共同促进入侵植物在不同环境条件下定殖。.

Following the novel weapon hypothesis, the invasiveness of non-native species, such as common ragweed (Ambrosia artemisiifolia L.) can result from a loss of natural competitors due to the production of chemical compounds, which negatively affect native communities. Particularly the genus Ambrosia produces several types of organic compounds, which have the potential to inhibit germination and growth of other plants. Subsequent to an assessment of the chemical content of three different ragweed extracts (aqueous shoot and root extracts, as well as essential oil), two different trials on the effects of different concentrations of these extracts, as well as ragweed residues, were conducted on two different mediums (Petri dish vs. soil). In addition, we investigated the impact on the infection potential of Bradyrhizobium japonicum on soybean roots in three different soil types (arable soil, potting soil, and sand). The results showed that the exposure to common ragweed extracts and residues induced changes in the biomass and root production of crops and ragweed itself. Even though crops and ragweed differed in their response behavior, the strongest negative impact on all crops and ragweed was observed with ragweed residues, leading to reductions in biomass and root growth of up to 90%. Furthermore, we found a decrease in the number of rhizobial nodules of up to 48% when soybean was exposed to ragweed root extract.

As multiple invaders often co-occur, understanding the interactions between different invasive species is important. Previous studies have reported on invasional meltdown and neutral and interference relationships between invasive species. However, interspecific interactions may vary with environmental change owing to the different responses of interacting invaders. To better understand the interaction of notorious invasive alien plants under CO2 enrichment and N deposition, the growth characteristics of common ragweed (Ambrosia artemisiifolia) and redroot pigweed (Amaranthus retroflexus) were studied when they were planted in monoculture (4Rag and 4Pig) or mixture (1Rag:3Pig, 2Rag:2Pig, 3Rag:1Pig) under four environmental treatments: elevated CO2, increased N, elevated CO2 + increased N and a control. Increased N positively affected almost all the traits (basal stem diameter, height, shoot biomass, root biomass and total biomass) of common ragweed, except for branch number and root-shoot ratio. But increased N only promoted redroot pigweed\'s height and basal stem diameter. interspecific competition promoted basal stem diameter and number of branches but decreased root biomass of common ragweed, and the basal stem diameter was significantly higher in 1Rag:3Pig and 2Rag:2Pig compared to the other two treatments. interspecific competition inhibited almost all the characteristics of redroot pigweed. The interaction between elevated CO2 and increased N also increased the biomass characteristics (shoot biomass, root biomass and total biomass) of common ragweed. However, elevated CO2 inhibited the root biomass of redroot pigweed. The results indicated that common ragweed was a superior competitor under conditions of elevated CO2 and increased N. Moreover, environmental change might strengthen the super-invasive plant common ragweed\'s competitive ability.

The beetle Ophraella communa is an effective biological control agent against the invasive common ragweed spread across various ecosystems with variable temperature ranges. The trend in climate change attributed to fluctuating temperatures and abrupt rainfalls is expected to continue. This study aimed to better understand the effects of thermal fluctuation on O. communa by exposing all their life stages to heat stress under different treatments. Repeated exposure to high temperatures, relative to constant milder temperatures, increased the duration of immature development, mean generation time, and the adult longevity, decreased the intrinsic rate of increase, finite rate of population increase, net reproductive rate, survival rate, overall longevity, body length, and mass of adults and positively affected overall fecundity by prolonging the oviposition period, biasing sex ratio towards females. After exposure to heat stress, the mating success and production of viable offspring were higher in O. communa. Our findings demonstrate that exposure to heat stress negatively affects ragweed beetles, but they were able to survive and reproduce.

To predict and mitigate the effects of climate change on communities and ecosystems, the joint effects of extreme climatic events on species interactions need to be understood. Using the common ragweed (Ambrosia artemisiifolia L.)-leaf beetle (Ophraella communa) system, we investigated the effects of heat wave and elevated CO2 on common ragweed growth, secondary metabolism, and the consequent impacts on the beetle. The results showed that elevated CO2 and heat wave facilitated A. artemisiifolia growth; further, A. artemisiifolia accumulated large amounts of defensive secondary metabolites. Being fed on A. artemisiifolia grown under elevated CO2 and heat wave conditions resulted in the poor performance of O. communa (high mortality, long development period, and low reproduction). Overall, under elevated CO2, heat wave improved the defensive ability of A. artemisiifolia against herbivores. On the other hand, enhanced adaptability to climatic changes may aggravate invasive plant distribution, posing a challenge to the control of invasive plants in the future.

Herbicides are the most commonly used means of controlling the growth of Ambrosia artemisiifolia L. Their constant use has led to the development of resistant populations. They can be evaluated by studying seed germination and the corresponding grown plants, but A. artemisiifolia exhibits seed dormancy, preventing germination and delaying research. Here, we developed a simple and rapid method to stimulate germination of freshly collected or stored A. artemisiifolia seeds. The germination of A. artemisiifolia freshly collected/stored seeds was evaluated after storage, stratification, and chemical treatments (ethephon, gibberellic acid (GA3), thiourea, KNO3). Ethephon or ethephon + GA3 improved freshly collected seed germination by 88 and 95%, respectively, and germination of stored seeds by 78 and 80%, respectively. In addition, placing the seeds of A. artemisiifolia in ethephon, GA3, ethephon + GA3, or thiourea solutions caused the freshly collected seeds to germinate faster than stored seeds or nontreated seeds. In contrast, the conditioning of seeds in these solutions favored germination of stored seeds, especially when ethephon + GA3 or GA3 was used. Imbibition of the freshly collected A. artemisiifolia seeds in a mixture of ethephon and GA3 can effectively overcome primary dormancy when rapid experimental results are needed. For seeds requiring prolonged storage, conditioning in ethephon, GA3, or thiourea solutions may be applied to promote germination.

This study aimed to investigate whether different doses of specific phenolic acids (chlorogenic acid, caffeic acid, ferulic acid, gallic acid, protocatechuic acid, p-hydroxybenzoic acid, syringic acid, vanillic acid, and p-coumaric acid), alone or in combination, can inhibit the early growth of the common ragweed (Ambrosia artemisiifolia L., Asterales: Asteraceae). A seed bioassay was performed in Petri dishes and placed in a climate chamber to assess the effects of five dose levels of phenolic acids to radicle and shoot length, as well seedling biomass of A. artemisiifolia. The lowest dose of phenolic acid corresponded to the natural phenolic acid concentration previously reported in dry plant tissue samples from Brassicaceae cover crop plants. Results show that the inhibition of the early growth of A. artemisiifolia depends strongly on phenolic acid. Across different treatments, high doses of phenolic acids significantly shortened shoots and radicles, as well as reduced seedling biomass. Treating seeds with ferulic acid alone, vanillic acid alone, p-hydroxybenzoic acid alone, or a mixture of all phenolic acids significantly reduced all early growth parameters. The estimated effective dose for the 50% inhibition (ED50) of radicle growth in A. artemisiifolia seedlings was 368.39 ± 59.85 × 10-8 mol with ferulic acid, 135.41 ± 17.65 × 10-8 mol with p-coumaric acid, 810.36 ± 134.15 × 10-8 mol with p-hydroxybenzoic acid, and 160.11 ± 12.30 × 10-8 mol with the combination of all phenolic acids.

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.

Several cover crops (CCs) exert allelopathic effects that suppress weed growth. The aim of the present study was to evaluate the effects of aqueous extracts containing different concentrations [0, 0.5, 1, 2.5, 5, 7.5 and 10% (w/v)] of Brassicaceae CCs (Sinapis alba, Raphanus sativus, Camellina sativa) and of the CCs Fagopyrum esculentum and Guizotia abyssinica on germination and early growth of Ambrosia artemisiifolia L. The allelopathic effects were species and concentration-dependent. C. sativa, for example, caused the greatest potential to inhibit germination, shoot, radicle length and fresh seedling weight, whereas S. alba and R. sativus inhibited germination and early growth of A. artemisiifolia only at concentrations ≥7.5%. In contrast, no inhibition was observed when aqueous extracts of F. escultneum and G. abyssinica were added at any of tested concentration. Liquid chromatography-tandem mass spectrometry detected 15 phenolic compounds in Brassicaceae CCs with the highest content (µg/g) of vanillin (48.8), chlorogenic acid (1057), vanilic acid (79), caffeic acid (102.5) and syringic acid (27.3) in C. sativa. Our results suggest that C. sativa is the most allelopathic CCs and that the fruits of C. sativa are the plant organs richest in allelochemicals.