Terezine E, bioactive prenylated tryptophan analogue from an endophyte of Centaurea stoebe.

Fungal endophytes are considered promising sources of new bioactive natural products. In this study, a Mucor sp. has been isolated as an endophyte from the medicinal plant Centaurea stoebe. Through bioactivity-guided fractionation, the isolation of the new bioactive terezine E in addition to the previously reported 14-hydroxyterezine D was carried out. The isolated compounds were fully characterised by HRESIMS and 1D and 2D NMR analyses. Both compounds exhibited potent antiproliferative activity against K-562 and HUVEC cell lines and antifungal efficacy against the tested fungal strains.

Native and Invading Yellow Starthistle (Centaurea solstitialis) Microbiomes Differ in Composition and Diversity of Bacteria.

Invasive species could benefit from being introduced to locations with more favorable species interactions, including the loss of enemies, the gain of mutualists, or the simplification of complex interaction networks. Microbiomes are an important source of species interactions with strong fitness effects on multicellular organisms, and these interactions are known to vary across regions. The highly invasive plant yellow starthistle (Centaurea solstitialis) has been shown to experience more favorable microbial interactions in its invasions of the Americas, but the microbiome that must contribute to this variation in interactions is unknown. We sequenced amplicons of 16S rRNA genes to characterize bacterial community compositions in the phyllosphere, ectorhizosphere, and endorhizosphere of yellow starthistle plants from seven invading populations in California, USA, and eight native populations in Europe. We tested for the differentiation of microbiomes by geography, plant compartment, and plant genotype. Bacterial communities differed significantly between native and invading plants within plant compartments, with consistently lower diversity in the microbiome of invading plants. The diversity of bacteria in roots was positively correlated with plant genotype diversity within both ranges, but this relationship did not explain microbiome differences between ranges. Our results reveal that these invading plants are experiencing either a simplified microbial environment or simplified microbial interactions as a result of the dominance of a few taxa within their microbiome. Our findings highlight several alternative hypotheses for the sources of variation that we observe in invader microbiomes and the potential for altered bacterial interactions to facilitate invasion success.IMPORTANCE Previous studies have found that introduced plants commonly experience more favorable microbial interactions in their non-native range, suggesting that changes to the microbiome could be an important contributor to invasion success. Little is known about microbiome variation across native and invading populations, however, and the potential sources of more favorable interactions are undescribed. Here, we report one of the first microbiome comparisons of plants from multiple native and invading populations, in the noxious weed yellow starthistle. We identify clear differences in composition and diversity of microbiome bacteria. Our findings raise new questions about the sources of these differences, and we outline the next generation of research that will be required to connect microbiome variation to its potential role in plant invasions.

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.

Strong indirect herbicide effects on mycorrhizal associations through plant community shifts and secondary invasions.

Million of acres of U.S. wildlands are sprayed with herbicides to control invasive species, but relatively little is known about non-target effects of herbicide use. We combined greenhouse, field, and laboratory experiments involving the invasive forb spotted knapweed (Centaurea stoebe) and native bunchgrasses to assess direct and indirect effects of the forb-specific herbicide picloram on arbuscular mycorrhizal fungi (AMF), which are beneficial soil fungi that colonize most plants. Picloram had no effect on bunchgrass viability and their associated AMF in the greenhouse, but killed spotted knapweed and reduced AMF colonization of a subsequent host grown. Results were similar in the field where AMF abundance in bunchgrass-dominated plots was unaffected by herbicides one year after spraying based on 16:1ω5 phospholipid fatty acid (PLFA) and neutral lipid fatty acid (NLFA) concentrations. In spotted-knapweed-dominated plots, however, picloram application shifted dominance from spotted knapweed, a good AMF host, to bulbous bluegrass (Poa bulbosa), a poor AMF host. This coincided with a 63% reduction in soil 16:1ω5 NLFA concentrations but no reduction of 16:1ω5 PLFA. Because 16:1ω5 NLFA quantifies AMF storage lipids and 16:1ω5 PLFA occurs in AMF membrane lipids, we speculate that the herbicide-mediated reduction in host quality reduced fungal carbon storage, but not necessarily fungal abundance after one year in the field. Overall, in greenhouse and field experiments, AMF were only affected when picloram altered host quantity and quality. This apparent lack of direct effect was supported by our in-vitro trial where picloram applied to AMF mycelia did not reduce fungal biomass and viability. We show that the herbicide picloram can have profound, indirect effects on AMF within one year. Depending on herbicide-mediated shifts in host quality, rapid interventions may be necessary post herbicide applications to prevent loss of AMF abundance. Future research should assess consequences of these potential shifts for the restoration of native plants that differ in mycorrhizal dependency.

Biocontrol attack increases pollen limitation under some circumstances in the invasive plant Centaurea solstitialis.

Herbivore damage often deters pollinator visitation and many invasive plants in North America are pollinator-dependent. This has important implications for the biological control of invasive plants because it means that agents that deter pollinators may have a larger than expected impact on the plant. Yet interactions between pollinators and biocontrol agents are rarely evaluated. Centaurea solstitialis, one of the most problematic invasive species in California, is dependent on pollinators for reproduction. I factorially manipulated infection by a biocontrol pathogen and pollen supplementation to test for (1) pollen limitation in C. solstitialis, (2) whether infection increased pollen limitation, and (3) whether this varied across a soil moisture gradient. Plants growing on north-facing slopes where soil moisture was higher experienced mild pollen limitation in the absence of the pathogen and more pronounced pollen limitation when they were infected. Plants on drier south-facing slopes did not suffer from pollen limitation but instead appeared to suffer from resource limitation. Pathogen infection directly reduced seed set in C. solstitialis by 67-72%. On north-facing slopes, infection had an additional, indirect effect by increasing the degree of pollen limitation plants experienced. The trait that mediates this indirect pathogen-pollinator interaction is the number of inflorescences plants produced: infected plants made fewer inflorescences which led to greater pollen limitation. Although in the present study this outcome is dependent on abiotic factors that vary over small spatial scales, exploiting other invasive plants' dependence on pollinators by selecting agents that deter visitation may enhance agent impact.

Exclusionary interactions among diverse fungi infecting developing seeds of Centaurea stoebe.

Developing seeds are expected to be strongly defended against microbial attack. In keeping with this, only 26% of seeds of Centaurea stoebe from its native and invaded ranges in Eurasia and North America were infected with fungi, and 92.2% of those were infected with a single fungus per seed. Even when developing seeds in flower heads were inoculated under conducive conditions for infection with 14 of these seed-infecting fungi, re-isolation of inoculants was only 16% overall, and again limited to the particular inoculant. Environmental fungi (i.e. those not isolated from seed of C. stoebe) were present in control flower heads under conditions conducive to infection but they were never re-isolated from fully developed seeds in any experiments. When two or three seed isolates were co-inoculated to compete in flower heads, only one inoculant, and always the same one, was re-isolated from all matured seeds, regardless of maternal plant genotype. PCR-based detection methods confirmed that these fungal interactions were exclusionary rather than suppressive. In these strongly defended, developing seeds, we had expected the plant to control not only the overall level of infection but also the outcome of co-inoculations. Consequences for the next plant generation of this exclusionary competition among seed-infecting fungi included effects on seedling emergence, growth and fecundity.

Models of experimental competitive intensities predict home and away differences in invasive impact and the effects of an endophytic mutualist.

Understanding the role of competition in the organization of communities is limited in part by the difficulty of extrapolating the outcomes of small-scale experiments to how such outcomes might affect the distribution and abundance of species. We modeled the community-level outcomes of competition, using experimentally derived competitive effects and responses between an exotic invasive plant, Centaurea stoebe, and species from both its native and nonnative ranges and using changes in these effects and responses elicited by experimentally establishing symbioses between C. stoebe and fungal endophytes. Using relative interaction intensities (RIIs) and holding other life-history factors constant, individual-based and spatially explicit models predicted competitive exclusion of all but one North American species but none of the European species, regardless of the endophyte status of C. stoebe. Concomitantly, C. stoebe was eliminated from the models with European natives but was codominant in models with North American natives. Endophyte symbiosis predicted increased dominance of C. stoebe in North American communities but not in European communities. However, when experimental variation was included, some of the model outcomes changed slightly. Our results are consistent with the idea that the effects of competitive intensity and mutualisms measured at small scales have the potential to play important roles in determining the larger-scale outcomes of invasion and that the stabilizing indirect effects of competition may promote species coexistence.

Fungal endophytes directly increase the competitive effects of an invasive forb.

Competitive outcomes among plants can vary in different abiotic and biotic conditions. Here we tested the effects of two phylotypes of Alternaria endophytes on the growth, competitive effects, and competitive responses of the exotic invasive forb Centaurea stoebe. Centaurea stoebe was a better competitor against North American grass species than grasses from its European home range in the absence of endophytes. However, one endophyte both increased the biomass of C. stoebe and reduced the competitive effect of North American grasses on C. stoebe. The competitive effects of C. stoebe on grass species native to North America were enhanced by both fungal endophytes, but not for native European grasses. We do not know the mechanism by which endophytes increased C. stoebe's competitive ability, and particularly against biogeographically new neighbors, but one endophyte increased the competitive ability of C. stoebe without increasing its size, suggesting mechanisms unrelated to increased growth. We tested only a fraction of the different endophytic fungi that have been found in C. stoebe, only scratching the surface of understanding their indirect effects. However, our results are the first to demonstrate such effects of a fungal endophyte infecting an invasive forb, and one of the few to show that endophyte effects on competition do not have to be mediated through herbivory.

[Interactions between invasive plants and arbuscular mycorrhizal fungi: a review].

The invasion of invasive plants changes the biological community structure in their invaded lands, leading to the biodiversity loss. As an important component of soil microorganisms in terrestrial ecosystem, arbuscular mycorrhizal (AM) fungi can affect the growth performance of invasive plants. This kind of specific relations between AM fungi and invasive plants also implies that AM fungi can affect plant invasion. On the other hand, the invasion of invasive plants can affect the community structure and function of AM fungi. This paper summarized the species and harms of invasive plants in China, and discussed the relationships between AM fungi and invasive plants invasion, including the roles of AM fungi in the processes of invasive plants invasion, the effects of the invasion on AM fungi, and the interactive mechanisms between the invasion and AM fungi.

Synchytrium solstitiale: reclassification based on the function and role of resting spores.

Studies were made about resting spores of Synchytrium solstitiale, a chytrid that causes false rust disease of yellow starthistle (YST). During evaluation of this fungus for biological control of YST, a protocol for resting spore germination was developed. Details of resting spore germination and study of long-term survival of the fungus were documented. Resting spores from dried leaves germinated after incubating them on water agar at least 7 d at 10-15 C. Resting spores were viable after storage in air-dried leaves more than 2 y at room temperature, suggesting they have a role in off-season and long-term survival of the fungus. Each resting spore produced a single sorus that contained a single sporangium, which on germination released zoospores through a pore. YST inoculated with germinated resting spores developed symptoms typical of false rust disease. All spore forms of S. solstitiale have been found to be functional, and the life cycle of S. solstitiale has been completed under controlled laboratory and greenhouse conditions. Resting spore galls differ from sporangial galls both morphologically and biologically, and in comparison, each sporangial gall cleaves into several sori and each sorus produces 5-25 sporangia that rupture during release of zoospores. For this reason S. solstitiale should be reclassified as diheterogallic sensu Karling (Am J Bot 42:540-545). Because resting spores function as prosori and produce an external sorus, S. solstitiale is best placed in into the subgenus Exosynchytrium.

Species-specific effects of polyploidisation and plant traits of Centaurea maculosa and Senecio inaequidens on rhizosphere microorganisms.

Invasive plant species represent a threat to terrestrial ecosystems, but their effects on the soil biota and the mechanisms involved are not yet well understood. Many invasive species have undergone polyploidisation, leading to the coexistence of various cytotypes in the native range, whereas, in most cases, only one cytotype is present in the introduced range. Since genetic variation within a species can modify soil rhizosphere communities, we studied the effects of different cytotypes and ranges (native diploid, native tetraploid and introduced tetraploid) of Centaurea maculosa and Senecio inaequidens on microbial biomass carbon, rhizosphere total DNA content and bacterial communities of a standard soil in relation to plant functional traits. There was no overall significant difference in microbial biomass between cytotypes. The variation of rhizosphere total DNA content and bacterial community structure according to cytotype was species specific. The rhizosphere DNA content of S. inaequidens decreased with polyploidisation in the native range but did not vary for C. maculosa. In contrast, the bacterial community structure of C. maculosa was affected by polyploidisation and its diversity increased, whereas there was no significant change for S. inaequidens. Traits of S. inaequidens were correlated to the rhizosphere biota. Bacterial diversity and total DNA content were positively correlated with resource allocation to belowground growth and late flowering, whereas microbial biomass carbon was negatively correlated to investment in reproduction. There were no correlations between traits of the cytotypes of C. maculosa and corresponding rhizosphere soil biota. This study shows that polyploidisation may affect rhizosphere bacterial community composition, but that effects vary among plant species. Such changes may contribute to the success of invasive polyploid genotypes in the introduced range.

Puccinia jaceae var. solstitialis teliospore priming on yellow starthistle.

Following the introduction of Puccinia jaceae var. solstitialis to California for biological control of yellow starthistle (Centaurea solstitialis, Asteraceae), teliospores, pycnia, and multiple urediniospore generations have been observed in the field. Because urediniospores have a relatively short life span in the field, functioning teliospores are expected to be necessary for the permanent establishment of P. jaceae var. solstitialis in California. To determine if conditions in California were conducive to this, teliospore emergence and priming were evaluated in the field. A factorial experiment in the laboratory with five incubation times and three incubation temperatures was used to determine teliospore priming requirements. Teliospore production coincided with plant senescence in August and September at two sites in 2 years; fewer teliospores were produced in 2006, suggesting inconsistent teliospore production may limit population growth and contribute to local extinctions in some areas. When teliospores were primed in the field, germination was low through the fall and abruptly peaked in January during both years. In the laboratory, teliospore germination increased as incubation time increased from 2 to 6 weeks and temperatures decreased from 12 to 4 degrees C. A degree-hour model derived from laboratory data accurately predicts when teliospores are primed for germination in the field. Based on the results obtained in this study, it is apparent that teliospore germination can occur over a range of priming conditions. However, lower temperatures and longer incubation periods are superior in breaking teliospore dormancy.

Soil fungal abundance and diversity: another victim of the invasive plant Centaurea maculosa.

Interactions between plants and soil microbes are important determinants of both above- and belowground community composition, and ultimately ecosystem function. As exotic plants continue to invade and modify native plant communities, there has been increasing interest in determining the influence of exotic invasives on native soil microbial communities. Here, using highly sensitive molecular techniques, we examine fungal abundance and diversity in the soil surrounding a particularly aggressive invasive plant species in North America, Centaurea maculosa Lam. In mixed stands, we show that this invasive weed can alter the native fungal community composition within its own rhizosphere and that of neighboring native plants. At higher densities, the effect of C. maculosa on native soil fungal communities was even greater. Our results demonstrate that this invasive weed can have significant effects not only on visible aboveground biodiversity but also on the native soil microbial community that extends beyond its rhizosphere.

Finally, proof of weapons of mass destruction.

Allelopathy (one species' use of chemicals to harm other species) may be a key ingredient in successful invasions of alien plants into established communities. Bais et al. show that in response to elicitation by common soil fungi, spotted knapweed (Centaurea maculosa) launches an ineffective defense against the fungi that results in extensive collateral damage to neighboring plants. Specifically, the flavonoid (-)-catechin, released from the roots of knapweed, produces a massive reactive oxygen species (ROS)-mediated oxidative burst, Ca2+ signaling, and rapid cell death in those unadapted native species whose habitats the plant regularly invades. The roots release both (+) and (-) enantiomers, but only the (-) enantiomer functions as a weapon of mass destruction; the (+) enantiomer inhibits the growth of numerous common soil-borne bacterial pathogens. Eliciting apoptotic response for a competitive advantage is an example of signal cross-talk between the genomes of interacting organisms and highlights how the internal signaling of one organism can be used by others to adjust their phenotypes in an adaptive manner. The study provides strong circumstantial evidence for an allelopathic interaction, but the genetic manipulation of (-)-catechin release would allow researchers to determine if these responses occur in nature. Precise genetic control over the release of secondary metabolites from plants would benefit ecological research.