Native plants change the endophyte assembly and growth of an invasive plant in response to climatic factors.

Climate change, microbial endophytes, and local plants can affect the establishment and expansion of invasive species, yet no study has been performed to assess these interactions. Using a growth chamber, we integrated the belowground (rhizosphere soils) and aboveground (mixture of mature leaf and leaf litter) microbiota into an experimental framework to evaluate the impacts of four native plants acting as microbial inoculation sources on endophyte assembly and growth of the invasive plant Ageratina adenophora in response to drought stress and temperature change. We found that fungal and bacterial enrichment in the leaves and roots of A. adenophora exhibited distinct patterns in response to climatic factors. Many fungi were enriched in roots in response to high temperature and drought stress; in contrast, many bacteria were enriched in leaves in response to low temperature and drought stress. Inoculation of microbiota from phylogenetically close native plant species (i.e., Asteraceae Artemisia atrovirens) causes the recipient plant A. adenophora (Asteraceae) to enrich dominant microbial species from inoculation sources, which commonly results in a lower dissimilar endophytic microbiota and thus produces more negative growth effects when compared to non-Asteraceae inoculations. Drought, microbial inoculation source, and temperature directly impacted the growth of A. adenophora. Both drought and inoculation also indirectly impacted the growth of A. adenophora by changing the root endophytic fungal assembly. Our data indicate that native plant identity can greatly impact the endophyte assembly and host growth of invasive plants, which is regulated by drought and temperature.IMPORTANCEThere has been increasing interest in the interactions between global changes and plant invasions; however, it remains to quantify the role of microbial endophytes in plant invasion with a consideration of their variation in the root vs leaf of hosts, as well as the linkages between microbial inoculations, such as native plant species, and climatic factors, such as temperature and drought. Our study found that local plants acting as microbial inoculants can impact fungal and bacterial enrichment in the leaves and roots of the invasive plant Ageratina adenophora and thus produce distinct growth effects in response to climatic factors; endophyte-mediated invasion of A. adenophora is expected to operate more effectively under favorable moisture. Our study is important for understanding the interactions between climate change, microbial endophytes, and local plant identity in the establishment and expansion of invasive species.

Ageratina adenophora and Lantana camara in Kailash Sacred Landscape, India: Current distribution and future climatic scenarios through modeling.

The Himalayan region is one of the global biodiversity hotspots. However, its biodiversity and ecosystems are threatened due to abiotic and biotic drivers. One of the major biotic threats to biodiversity in this region is the rapid spread of Invasive Alien Species (IAS). Natural forests and grasslands are increasingly getting infested by IAS affecting regeneration of native species and decline in availability of bio-resources. Assessing the current status of IAS and prediction of their future spread would be vital for evolving specific species management interventions. Keeping this in view, we conducted an in-depth study on two IASs, viz., Ageratina adenophora and Lantana camara in the Indian part of Kailash Sacred Landscape (KSL), Western Himalaya. Intensive field surveys were conducted to collect the presence of A. adenophora (n = 567) and L. camara (n = 120) along an altitudinal gradient between 300 and 3000 m a.s.l. We performed Principal Component Analysis to nullify the multi-colinearity effects of the environmental predictors following MaxEnt species distribution model in the current and future climatic scenarios for both the species. All current and future model precision (i.e., Area Under the Curve; AUC) for both species was higher than 0.81. It is predicted that under the current rate of climate change and higher emission (i.e., RCP 8.5 pathway), A. adenophora will spread 45.3% more than its current distribution and is likely to reach up to 3029 m a.s.l., whereas, L. camara will spread 29.8% more than its current distribution range and likely to reach up to 3018 m a.s.l. Our results will help in future conservation planning and participatory management of forests and grasslands in the Kailash Sacred Landscape-India.

Plant-soil-foliage feedbacks on seed germination and seedling growth of the invasive plant Ageratina adenophora.

Some exotic plants become invasive because they partially release from soil-borne enemies and thus benefit from positive plant-soil feedbacks (PSFs) in the introduced range. However, reports that have focused only on PSFs may exaggerate the invader's competitiveness. Here, we conducted three experiments to characterize plant-soil-foliage feedbacks, including mature leaves (ML), leaf litter (LL), rhizosphere soil (RS) and leaves plus soil (LS), on the early growth stages of the invasive plant Ageratina adenophora. In general, the feedbacks from aboveground (ML, LL) adversely affected A. adenophora by delaying germination time, inhibiting germination rate and reducing seedling growth. The increased invasion history exacerbated the adverse effects of LL and LS feedbacks on seedling growth. These adverse effects were partially contributed by more abundant fungi (e.g. Didymella) or/and more virulent fungi (e.g. Fusarium) developed in the aboveground part of A. adenophora during the invasion. Interestingly, the aboveground adverse effects can be weakened by microbes from RSs. Our novel findings emphasize the important role of aboveground feedbacks in the evaluation of plant invasiveness, and their commonness and significance remain to be explored in other invasive systems.

Enrichment of soil rare bacteria in root by an invasive plant Ageratina adenophora.

The assembly of the root-associated microbiome provides mutual benefits for the host plant and bacteria in soils. It is interesting how invasive plants interact with the local soil microbial community and establish the soil bacterial community in the endosphere of these plants in the short term. In this study, we compared the bacterial community in the rhizosphere with that in the root endosphere of an invasive plant, Ageratina adenophora, using high-throughput sequencing. The results showed that the roots of A. adenophora selectively accumulated the genera Clostridium and Enterobacter, which are rarely distributed in the rhizosphere. This selective accumulation caused a switch in the bacterial composition at the phylum level from Bacteroidetes predominant in the rhizosphere to Proteobacteria dominant in the root endosphere of A. adenophora. Our data indicated the potential existence of a highly conserved signal recognition in which hosts, either invasive or native, enrich the endosphere bacteria, such as Clostridium, Enterobacter, etc., from the rhizosphere. Moreover, the accumulated bacteria were physiologically and genetically different at the strain level and displayed distinct roles in growth between invasive and native plants. The assembly of the bacterial community in the roots may be an advantageous strategy for A. adenophora in competition with native plants.

Processes affecting altitudinal distribution of invasive Ageratina adenophora in western Himalaya: The role of local adaptation and the importance of different life-cycle stages.

The spread of invasive plants along elevational gradients is considered a threat to fragile mountain ecosystems, but it can also provide the opportunity to better understand some of the basic processes driving the success of invasive species. Ageratina adenophora (Asteraceae) is an invasive plant of global importance and has a broad distribution along elevational gradients in the Western Himalayas. Our study aimed at understanding the role of evolutionary processes (e.g. local adaptation and clinal differentiation) and different life history stages in shaping the distribution pattern of the invasive plant along an elevational gradient in the Western Himalaya. We carried out extensive distributional surveys, established a reciprocal transplant experiment with common gardens at three elevational levels, and measured a suite of traits related to germination, growth, reproduction and phenology. Our results showed a lack of local adaptation, and we did not find any evidence for clinal differentiation in any measured trait except a rather weak signal for plant height. We found that seed germination was the crucial life-cycle transition in determining the lower range limit while winter mortality of plants shaped the upper range limit in our study area, thus explaining the hump shaped distribution pattern. Differences in trait values between gardens for most traits indicated a high degree of phenotypic plasticity. Possible causes such as apomixis, seed dispersal among sites, and pre-adaptation might have confounded evolutionary processes to act upon. Our results suggest that the success and spread of Ageratina adenophora are dependent on different life history stages at different elevations that are controlled by abiotic conditions.

Are invasive plants more competitive than native conspecifics? Patterns vary with competitors.

Invasive plants are sometimes considered to be more competitive than their native conspecifics, according to the prediction that the invader reallocates resources from defense to growth due to liberation of natural enemies ['Evolution of Increased Competitive Ability' (EICA) hypothesis]. However, the differences in competitive ability may depend on the identity of competitors. In order to test the effects of competitors, Ageratina adenophora plants from both native and invasive ranges competed directly, and competed with native residents from both invasive (China) and native (Mexico) ranges respectively. Invasive A. adenophora plants were more competitive than their conspecifics from native populations when competing with natives from China (interspecific competition), but not when competing with natives from Mexico. Invasive A. adenophora plants also showed higher competitive ability when grown in high-density monoculture communities of plants from the same population (intrapopulation competition). In contrast, invasive A. adenophora plants showed lower competitive ability when competing with plants from native populations (intraspecific competition). Our results indicated that in the invasive range A. adenophora has evolved to effectively cope with co-occurring natives and high density environments, contributing to invasion success. Here, we showed the significant effects of competitors, which should be considered carefully when testing the EICA hypothesis.

ICE1 demethylation drives the range expansion of a plant invader through cold tolerance divergence.

Cold tolerance adaption is a crucial determinant for the establishment and expansion of invasive alien plants into new cold environments; however, its evolutionary mechanism is poorly understood. Crofton weed (Ageratina adenophora), a highly invasive alien plant, is continuously spreading across subtropical areas in China, north-eastward from the first colonized south-western tropical regions, through cold tolerance evolution. Close relations between the cold tolerance levels of 34 populations, represented by 147 accessions, and the latitude, extreme lowest temperature, coldest month average temperature, and invasion period have provided direct insight into its cold tolerance divergence. A comparative study of the CBF pathway, associated with the cold tolerance enhancement of cold-susceptible CBF1-transgenic plant, among four geographically distinct crofton weed populations revealed that the CBF pathway plays a key role in the observed cold tolerance divergence. Four epialleles of the cold response regulator ICE1 ranged from 66 to 50 methylated cytosines, representing a 4.4% to 3.3% methylation rate and significantly corresponding to the lowest to highest cold tolerance levels among these different populations. The significant negative relation between the transcription levels of the primary CBF pathway members, except for CBF2, and the methylation levels among the four populations firstly demonstrates that the demethylation-upregulated transcription level of CBF pathway is responsible for this evolution. These facts, combined with the cold tolerance variation and methylation found among three native and two other introduced populations, indicate that the ICE1-demethylated upregulation of cold tolerance may be the underlying evolutionary mechanism allowing crofton weed to expand northward in China.

Synergistic interactions of CO2 enrichment and nitrogen deposition promote growth and ecophysiological advantages of invading Eupatorium adenophorum in Southwest China.

Global environmental change and ongoing biological invasions are the two prominent ecological issues threatening biodiversity worldwide, and investigations of their interaction will aid to predict plant invasions and inform better management strategies in the future. In this study, invasive Eupatorium adenophorum and native congener E. stoechadosmum were compared at ambient and elevated atmospheric carbon dioxide (CO(2)) concentrations combined with three levels of nitrogen (N; reduced, control and increased) in terms of growth, energy gain, and cost. Compared with E. stoechadosmum, E. adenophorum adopted a quicker-return energy-use strategy, i.e. higher photosynthetic energy-use efficiency and shorter payback time. Lower leaf mass per area may be a pivotal trait for the invader, which contributed to an increased N allocation to Rubisco at the expense of cell walls and therefore to higher photosynthetic energy gain. CO(2) enrichment and N deposition synergistically promoted plant growth and influenced some related ecophysiological traits, and the synergistic effects were greater for the invader than for the native congener. Reducing N availability by applying sugar eliminated the advantages of the invader over its native congener at both CO(2) levels. Our results indicate that CO(2) enrichment and N deposition may exacerbate E. adenophorum's invasion in the future, and manipulating environmental resources such as N availability may be a feasible tool for managing invasion impacts of E. adenophorum.

A congeneric comparison shows that experimental warming enhances the growth of invasive Eupatorium adenophorum.

Rising air temperatures may change the risks of invasive plants; however, little is known about how different warming timings affect the growth and stress-tolerance of invasive plants. We conducted an experiment with an invasive plant Eupatorium adenophorum and a native congener Eupatorium chinense, and contrasted their mortality, plant height, total biomass, and biomass allocation in ambient, day-, night-, and daily-warming treatments. The mortality of plants was significantly higher in E. chinense than E. adenophorum in four temperature regimes. Eupatorium adenophorum grew larger than E. chinense in the ambient climate, and this difference was amplified with warming. On the basis of the net effects of warming, daily-warming exhibited the strongest influence on E. adenophorum, followed by day-warming and night-warming. There was a positive correlation between total biomass and root weight ratio in E. adenophorum, but not in E. chinense. These findings suggest that climate warming may enhance E. adenophorum invasions through increasing its growth and stress-tolerance, and that day-, night- and daily-warming may play different roles in this facilitation.

Soil biota reduce allelopathic effects of the invasive Eupatorium adenophorum.

Allelopathy has been hypothesized to play a role in exotic plant invasions, and study of this process can improve our understanding of how direct and indirect plant interactions influence plant community organization and ecosystem functioning. However, allelopathic effects can be highly conditional. For example allelopathic effects demonstrated in vivo can be difficult to demonstrate in field soils. Here we tested phytotoxicity of Eupatorium adenophorum (croftonweed), one of the most destructive exotic species in China, to a native plant species Brassica rapa both in sand and in native soil. Our results suggested that natural soils from different invaded habitats alleviated or eliminated the efficacy of potential allelochemicals relative to sand cultures. When that soil is sterilized, the allelopathic effects returned; suggesting that soil biota were responsible for the reduced phytotoxicity in natural soils. Neither of the two allelopathic compounds (9-Oxo-10,11-dehydroageraphorone and 9b-Hydroxyageraphorone) of E. adenophorum could be found in natural soils infested by the invader, and when those compounds were added to the soils as leachates, they showed substantial degradation after 24 hours in natural soils but not in sand. Our findings emphasize that soil biota can reduce the allelopathic effects of invaders on other plants, and therefore can reduce community invasibility. These results also suggest that soil biota may have stronger or weaker effects on allelopathic interactions depending on how allelochemicals are delivered.

Multiple mechanisms underlie rapid expansion of an invasive alien plant.

• With growing concerns over serious ecological problems, a particular challenge is to reveal the complex mechanisms underlying rapid expansion of invasive species. Ageratina adenophora is of particular interest in addressing this question. • We used geographic information systems and logistic regression to identify the geographic and environmental factors contributing to the presence of A. adenophora. Join-count spatial statistics with reproduction mode examination were employed to elucidate the spatiotemporal dispersal mechanisms. • Multiple factors have significantly contributed to the rapid expansion of A. adenophora. Its biological traits, favoring dispersal by water and wind coupled with local spatiotemporally heterogeneous geography and ecology, promote invasion downstream and upstream along river valleys, while other factors associated with human activities facilitate its invasion over high mountains and across river valleys, providing new scope for progressive invasions. We further identified an unusual invasion event of A. adenophora subsequent to a great flood that amplified its dispersal ability from vegetative propagules and seeds. • These findings suggest that dynamic interactions of multiple factors in heterogeneous ecogeographical environments - a 'combinatorial' invasion mechanism - would generate an unexpected invasion rate of an alien species or a seemingly stochastic invasion event.

Molecular cloning of allelopathy related genes and their relation to HHO in Eupatorium adenophorum.

In this study, conserved sequence regions of HMGR, DXR, and CHS (encoding 3-hydroxy-3-methylglutaryl-CoA reductase, 1-deoxyxylulose-5-phosphate reductoisomerase and chalcone synthase, respectively) were amplified by reverse transcriptase (RT)-PCR from Eupatorium adenophorum. Quantitative real-time PCR showed that the expression of CHS was related to the level of HHO, an allelochemical isolated from E. adenophorum. Semi-quantitative RT-PCR showed that there was no significant difference in expression of genes among three different tissues, except for CHS. Southern blotting indicated that at least three CHS genes are present in the E. adenophorum genome. A full-length cDNA from CHS genes (named EaCHS1, GenBank ID: FJ913888) was cloned. The 1,455 bp cDNA contained an open reading frame (1,206 bp) encoding a protein of 401 amino acids. Preliminary bioinformatics analysis of EaCHS1 revealed that EaCHS1 was a member of CHS family, the subcellular localization predicted that EaCHS1 was a cytoplasmic protein. To the best of our knowledge, this is the first report of conserved sequences of these genes and of a full-length EaCHS1 gene in E. adenophorum. The results indicated that CHS gene is related to allelopathy of E. adenophorum.

[Ecological adaptability of different provenance Eupatorium adenophorum Spreng].

By using reciprocal transplant method, Eupatorium adenophorum Spreng. seedlings of five different provenances were reciprocally transplanted into six different sites in Yunnan Province, with the survival rate, plant height, branch number, biomass, flower number per plant, and seed production measured during the period from April 2007 to May 2008. The seedlings growth and reproductive traits of different provenance E. adenophorum all showed strong plasticity to environmental conditions. With increasing latitude and altitude of transplant site, plant height, branch number, biomass, flower number per plant, and seed production of different provenance E. adenophorum decreased, and the differences in these growth and reproductive traits were significant among the transplant sites. However, there were no significant differences in these traits among different provenance E. adenophorum. Provenance and transplant site had no significant interactive effects on the above-mentioned traits except seed production. At each site, the survival rate, growth potential, and reproductive capability of local provenance E. adenophorum didn't have any superiority, illustrating that the success in the invasion of E. adenophorum in Yunnan Province was mainly due to the phenotypic plasticity of the plant, while local adaptability only played lesser important role.

Inhibition of Ageratina adenophora on spore germination and gametophyte development of Macrothelypteris torresiana.

Allelopathy of Ageratina adenophora plays an important role in its invasion. However, we have little knowledge of its allelpathic effects on ferns. In Petri dish bioassays, the inhibitory potential of aqueous leachates from roots, stems and leaves of A. adenophora was studied on the spore germination and gametophyte development of Macrothelypteris torresiana. All leachates inhibited the spore germination and growth of the first rhizoid of M. torresiana and inhibitory effects increased with increasing leachate concentrations. Root leachates proved most inhibitory. Gametophyte rhizoids of M. torresiana treated with stem and leaf leachates of A. adenophora were erect, which was similar to those of the control. However, gametophyte rhizoids of M. torresiana treated with root leachates of A. adenophora were erect, but also curving or swollen. Moreover, curving and swollen rhizoids increased with increasing concentrations. As time went by, rhizoids treated with root leachates were not so curved and the swelling almost disappeared. Possible causes are discussed in the present study. The increasing concentrations of leaf leachates also delayed the stages of gametophyte development. With the treatment of root leachates, the delay was more obvious. Thus A. adenophora inhibited the spore germination and gametophyte development of M. torresiana and the root leachates were most inhibitory.

[Effects of soil phosphorus level on morphological and photosynthetic characteristics of Ageratina adenophora and chromolaena odorata].

In this paper, a comparative study was made on the growth, morphology, biomass allocation, and photosynthesis of two invasive plant species Ageratina adenophora and Chromolaena odorata under five soil phosphorus levels, aimed to know how the test plant species acclimate to the changes of soil phosphorus level, evaluate which plant traits were associated with the invasiveness of the two species, and know whether the increased level of soil phosphorus could facilitate their invasion. The results showed that the two species had considerable phenotypic plasticity and ? phosphorus acclimation ability. At low phosphorus levels, their root mass ratio increased, which could enhance the nutrient capture ability, while at high phosphorus levels, their specific leaf area, maximum net photosynthetic rate, light saturation point, and chlorophyll and carotenoid contents per unit area were high, and the assimilative capacity and area increased, which could facilitate their carbon gain. A. adenophora had higher phosphorus acclimation ability than C. odorata. With the increase of phosphorous level, the relative growth rate, total biomass, branch number, leaf area index, and maximum net photosynthetic rate of the two species increased significantly, and most of the parameters were not decreased significantly under over-optimal phosphorus level. The two species could grow better under high phosphorus levels which were usually excessive and/or harmful for most native species, and enhanced soil phosphorus level might promote their invasion. At high phosphorus levels, the two invasive plant species might shade out native species through increasing their plant height, branch number, and leaf area index. The two species could maintain relatively high growth rate under high phosphorus levels in dry season when native plant species almost stopped growing. The ability that the invasive plant species could temporally use natural resources which native plant species could not use was also associated with their invasiveness.