Corrigendum: Geographic population structure in an outcrossing plant invasion after centuries of cultivation and recent founding events.

[This corrects the article DOI: 10.1093/aobpla/ply020.][This corrects the article DOI: 10.1093/aobpla/ply020.].

Geographic population structure in an outcrossing plant invasion after centuries of cultivation and recent founding events.

Population structure and genetic diversity of invasions are the result of evolutionary processes such as natural selection, drift and founding events. Some invasions are also molded by specific human activities such as selection for cultivars and intentional introduction of desired phenotypes, which can lead to low genetic diversity in the resulting invasion. We investigated the population structure, diversity and origins of a species with both accidental and intentional introduction histories, as well as long-term selection as a cultivar. Dyer's woad (Isatis tinctoria; Brassicaceae) has been used as a dye source for at least eight centuries in Eurasia, was introduced to eastern USA in the 1600s, and is now considered invasive in the western USA. Our analyses of amplified fragment length polymorphisms (AFLPs) from 645 plants from the USA and Eurasia did not find significantly lower gene diversity (Hj) in the invaded compared to the native range. This suggests that even though the species was under cultivation for many centuries, human selection of plants may not have had a strong influence on diversity in the invasion. We did find significantly lower genetic differentiation (Fst) in the invasive range but our results still suggested that there are two distinct invasions in the western USA. Our data suggest that these invasions most likely originated from Switzerland, Ukraine and Germany, which correlates with initial biological control agent survey findings. Genetic information on population structure, diversity and origins assists in efforts to control invasive species, and continued combination of ecological and molecular analyses will help bring us closer to sustainable management of plant invasions.

Farnesol-mediated shift in the metabolic origin of prenyl groups used for protein prenylation in plants.

Little is known about how plant cells regulate the exchange of prenyl diphosphates between the two compartmentalized isoprenoid biosynthesis pathways. Prenylation of proteins is a suitable model to study such interactions between the plastidial methylerythritol phosphate (MEP) and the cytosolic mevalonate (MVA) pathways because prenyl moieties used to modify proteins rely on both origins. Tobacco cells expressing a prenylatable GFP were treated with specific MEP and/or MVA pathways inhibitors to block the formation of prenyl diphosphates and therefore the possibility to modify the proteins. Chemical complementation assays using prenyl alcohol precursors restore the prenylation. Indeed, geranylgeraniol (C20 prenyl alcohol) and to a lesser but significant level C15-farnesol restored the prenylation of a protein bearing a geranylgeranylation CaaX motif, which under standard conditions is modified by a MEP-derived prenyl group. However, the restoration takes place in different ways. While geranylgeraniol operates directly as a metabolic precursor, the C15-prenyl alcohol functions indirectly as a signal that leads to shift the metabolic origin of prenyl groups in modified proteins, here from the plastidial MEP pathway in favor of the cytosolic MVA pathway. Furthermore, farnesol interferes negatively with the MEP pathway in an engineered Escherichia coli strain synthesizing isoprenoids either starting from MVA or from MEP. Following the cellular uptake of a fluorescent analog of farnesol, we showed its close interaction with tobacco plastids and modification of plastid homeostasis. As a consequence, in tobacco farnesol supposedly inhibits the plastidial MEP pathway and activates the cytosolic MVA pathway, leading to the shift in the metabolic origin and thereby acts as a potential regulator of crosstalk between the two pathways. Together, those results suggest a new role for farnesol (or a metabolite thereof) as a central molecule for the regulation of isoprenoid biosynthesis in plants.

The effect of MEP pathway and other inhibitors on the intracellular localization of a plasma membrane-targeted, isoprenylable GFP reporter protein in tobacco BY-2 cells.

We have established an in vivo visualization system for the geranylgeranylation of proteins in a stably transformed tobacco BY-2 cell line, based on the expression of a dexamethasone-inducible GFP fused to the carboxy-terminal basic domain of the rice calmodulin CaM61, which naturally bears a CaaL geranylgeranylation motif (GFP-BD-CVIL). By using pathway-specific inhibitors it was demonstrated that inhibition of the methylerythritol phosphate (MEP) pathway with known inhibitors like oxoclomazone and fosmidomycin, as well as inhibition of the protein geranylgeranyltransferase type 1 (PGGT-1), shifted the localization of the GFP-BD-CVIL protein from the membrane to the nucleus. In contrast, the inhibition of the mevalonate (MVA) pathway with mevinolin did not affect the localization. During the present work, this test system has been used to examine the effect of newly designed inhibitors of the MEP pathway and inhibitors of sterol biosynthesis such as squalestatin, terbinafine and Ro48-8071. In addition, we also studied the impact of different post-prenylation inhibitors or those suspected to affect the transport of proteins to the plasma membrane on the localization of the geranylgeranylable fusion protein GFP-BD-CVIL.

Two shoot-miners, Ceutorhynchus alliariae and Ceutorhynchus roberti, sharing the same fundamental niche on garlic mustard.

A combination of observational and experimental methods, in both the laboratory and field, were used to assess niche partitioning between Ceutorhynchus alliariae Brisout and C. roberti Gyllenhal (Coleoptera: Curculionidae), two coexisting shoot-boring weevils on garlic mustard, Alliaria petiolata (M. Bieb.) Cavara and Grande (Brassicaceae). We compared their morphology, oviposition behavior, larval development, distribution, abundance, and attack rates in their sympatric range, and of C. alliariae when found alone and in sympatry with C. roberti. Results indicate only very small differences in the fundamental niches of the two species. Comparison of C. alliariae in the range it occurs alone with the range where it co-occurs with C. roberti revealed some evidence for competition between the two species, i.e., attack levels of C. alliariae were reduced in areas where it co-occurred with C. roberti. However, the study showed no character displacement in regard to adult size or shoot choice of C. alliariae and we found no indication for superiority of either of the two species. Clearly, manipulative experiments would be necessary to unambiguously test for competition between the two species. Our results, based on a subset of niche dimensions known to be important in other systems, suggest that C. alliariae and C. roberti may present one of the rare cases, in which niche differentiation is not the main mechanism underlying coexistence.

Herbivore resistance of invasive Fallopia species and their hybrids.

Hybridization has been proposed as a mechanism by which exotic plants can increase their invasiveness. By generating novel recombinants, hybridization may result in phenotypes that are better adapted to the new environment than their parental species. We experimentally assessed the resistance of five exotic Fallopia taxa, F. japonica var. japonica, F. sachalinensis and F. baldschuanica, the two hybrids F. × bohemica and F. × conollyana, and the common European plants Rumex obtusifolius and Taraxacum officinale to four native European herbivores, the slug Arion lusitanicus, the moth Noctua pronuba, the grasshopper Metrioptera roeselii and the beetle Gastrophysa viridula. Leaf area consumed and relative growth rate of the herbivores differed significantly between the Fallopia taxa and the native species, as well as among the Fallopia taxa, and was partly influenced by interspecific variation in leaf morphology and physiology. Fallopia japonica, the most abundant Fallopia taxon in Europe, showed the highest level of resistance against all herbivores tested. The level of resistance of the hybrids compared to that of their parental species varied depending on hybrid taxon and herbivore species. Genotypes of the hybrid F. × bohemica varied significantly in herbivore resistance, but no evidence was found that hybridization has generated novel recombinants that are inherently better defended against resident herbivores than their parental species, thereby increasing the hybrid's invasion success. In general, exotic Fallopia taxa showed higher levels of herbivore resistance than the two native plant species, suggesting that both parental and hybrid Fallopia taxa largely escape from herbivory in Europe.

Invasive knotweed affects native plants through allelopathy.

PREMISE OF STUDY: There is increasing evidence that many plant invaders interfere with native plants through allelopathy. This allelopathic interference may be a key mechanism of plant invasiveness. One of the most aggressive current plant invaders is the clonal knotweed hybrid Fallopia × bohemica, which often forms monocultures in its introduced range. Preliminary results from laboratory studies suggest that allelopathy could play a role in this invasion. METHODS: We grew experimental communities of European plants together with F. × bohemica. We used activated carbon to test for allelopathic effects, and we combined this with single or repeated removal of Fallopia shoots to examine how mechanical control can reduce the species' impact. KEY RESULTS: Addition of activated carbon to the soil significantly reduced the suppressive effect of undamaged F. × bohemica on native forbs. The magnitude of this effect was similar to that of regular cutting of Fallopia shoots. Regular cutting of Fallopia shoots efficiently inhibited the growth of rhizomes, together with their apparent allelopathic effects. CONCLUSIONS: The ecological impact of F. × bohemica on native forbs is not just a result of competition for shared resources, but it also appears to have a large allelopathic component. Still, regular mechnical control successfully eliminated allelopathic effects. Therefore, allelopathy will create an additional challenge to knotweed management and ecological restoration only if the allelochemicals are found to persist in the soil. More research is needed to examine the mechanisms underlying Fallopia allelopathy, and the long-term effects of soil residues.

The plastidial 2-C-methyl-D-erythritol 4-phosphate pathway provides the isoprenyl moiety for protein geranylgeranylation in tobacco BY-2 cells.

Protein farnesylation and geranylgeranylation are important posttranslational modifications in eukaryotic cells. We visualized in transformed Nicotiana tabacum Bright Yellow-2 (BY-2) cells the geranylgeranylation and plasma membrane localization of GFP-BD-CVIL, which consists of green fluorescent protein (GFP) fused to the C-terminal polybasic domain (BD) and CVIL isoprenylation motif from the Oryza sativa calmodulin, CaM61. Treatment with fosmidomycin (Fos) or oxoclomazone (OC), inhibitors of the plastidial 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway, caused mislocalization of the protein to the nucleus, whereas treatment with mevinolin, an inhibitor of the cytosolic mevalonate pathway, did not. The nuclear localization of GFP-BD-CVIL in the presence of MEP pathway inhibitors was completely reversed by all-trans-geranylgeraniol (GGol). Furthermore, 1-deoxy-d-xylulose (DX) reversed the effects of OC, but not Fos, consistent with the hypothesis that OC blocks 1-deoxy-d-xylulose 5-phosphate synthesis, whereas Fos inhibits its conversion to 2-C-methyl-d-erythritol 4-phosphate. By contrast, GGol and DX did not rescue the nuclear mislocalization of GFP-BD-CVIL in the presence of a protein geranylgeranyltransferase type 1 inhibitor. Thus, the MEP pathway has an essential role in geranylgeranyl diphosphate (GGPP) biosynthesis and protein geranylgeranylation in BY-2 cells. GFP-BD-CVIL is a versatile tool for identifying pharmaceuticals and herbicides that interfere either with GGPP biosynthesis or with protein geranylgeranylation.

Demographic models inform selection of biocontrol agents for garlic mustard (Alliaria petiolata).

Nonindigenous invasive plants pose a major threat to natural communities worldwide. Biological control of weeds via selected introduction of their natural enemies can affect control over large spatial areas but also risk nontarget effects. To maximize effectiveness while minimizing risk, weed biocontrol programs should introduce the minimum number of host-specific natural enemies necessary to control an invasive nonindigenous plant. We used elasticity analysis of a matrix model to help inform biocontrol agent selection for garlic mustard (Alliaria petiolata (M. Bieb.) Cavara and Grande). The Eurasian biennial A. petiolata is considered one of the most problematic invaders of temperate forests in North America. Four weevil species in the genus Ceutorhynchus (Coleoptera: Curculionidae) are currently considered potential biocontrol agents. These species attack rosettes (C. scrobicollis), stems (C. roberti, C. alliariae), and seeds (C. constrictus) of A. petiolata. Elasticity analyses using A. petiolata demographic parameters from North America indicated that changes in the rosette-to-flowering-plant transition and changes in fecundity consistently had the greatest impact on population growth rate. These results suggest that attack by the rosette-feeder C. scrobicollis, which reduces overwintering survival, and seed or stem feeders that reduce seed output should be particularly effective. Model outcomes differed greatly as A. petiolata demographic parameters were varied within ranges observed in North America, indicating that successful control of A. petiolata populations may occur under some, but not all, conditions. Using these a priori analyses we predict: (1) rosette mortality and reduction of seed output will be the most important factors determining A. petiolata demography; (2) the root-crown feeder C. scrobicollis will have the most significant impact on A. petiolata demography; (3) releases of single control agents are unlikely to control A. petiolata across its full range of demographic variability; (4) combinations of agents that simultaneously reduce rosette survival and seed production will be required to suppress the most vigorous A. petiolata populations. These predictions can be tested using established long-term monitoring sites coupled with a designed release program. If demographic models can successfully predict biocontrol agent impact on invasive plant populations, a continued dialogue and collaboration between empirical and theoretical approaches may be the key to the development of successful biocontrol tactics for plant invaders in the future.

A review of tobacco BY-2 cells as an excellent system to study the synthesis and function of sterols and other isoprenoids.

In plants, two pathways are utilized for the synthesis of isopentenyl diphosphate (IPP), the universal precursor for isoprenoid biosynthesis. In this paper we review findings and observations made primarily with tobacco BY-2 cells (TBY-2), which have proven to be an excellent system in which to study the two biosynthetic pathways. A major advantage of these cells as an experimental system is their ability to readily take up specific inhibitors and stably- and/or radiolabeled precursors. This permits the functional elucidation of the role of isoprenoid end products and intermediates. Because TBY-2 cells undergo rapid cell division and can be synchronized within the cell cycle, they constitute a highly suitable test system for determination of those isoprenoids and intermediates that act as cell cycle inhibitors, thus giving an indication of which branches of the isoprenoid pathway are essential. Through chemical complementation; and use of precursors, intracellular compartmentation can be elucidated, as well as the extent to which the plastidial and cytosolic pathways contribute to the syntheses of specific groups of isoprenoids (e.g., sterols) via exchange of intermediates across membranes. These topics are discussed in the context of the pertinent literature.