Amy C. Blair

Does intraspecific hybridization contribute to the evolution of invasiveness?: an experimental test.

One of the major objectives of research on invasive species is to determine the relative importance of different evolutionary and ecological forces in the invasion process. It was recently suggested that post-introduction intraspecific hybridization between previously isolated genotypes could produce novel and/or heterotic progeny that might express enhanced invasiveness. We tested this hypothesis with Silene latifolia, a European native that has successfully invaded North America and has previously been shown to have undergone genetic change since its introduction. In a common garden experiment we compared the performance of plants derived from within and between population crosses from eight European and 18 North American populations. Results showed that there was no significant effect of crossing distance on progeny phenotype. Furthermore, progeny from within or between population crosses did not differ in size, reproductive output or survival. Collectively, these results suggest that the invasive phenotype of S. latifolia is likely the result of natural selection and/or genetic drift rather than intraspecific hybridization.

The importance of analytical techniques in allelopathy studies with the reported allelochemical catechin as an example

Allelopathy can be challenging to demonstrate. Developing rigorous analytical techniques to detect and quantify compound(s) of interest from soil or liquid media lays the foundation for designing ecologically relevant experiments that incorporate candidate allelochemicals. In this paper, fundamental components of analytical techniques, including method development, validation, and appropriate controls are discussed. Research on the candidate allelochemical from spotted knapweed, catechin, is used as an example to demonstrate the importance of including these components both during data collection and in subsequent publications. This example shows how contrasting results between research groups can be difficult to interpret when information on controls and method validation are not included in publications. Recent research suggests that catechin is not likely driving spotted knapweed’s invasion, and thus future research on this system should focus on alternate candidate toxins from spotted knapweed. By employing appropriate analytical techniques, such as those outlined here, a strong foundation can be laid for ecologically oriented experiments that examine the role of allelochemicals in structuring communities.

Hybridization and invasion: one of North America’s most devastating invasive plants shows evidence for a history of interspecific hybridization

Hybridization has been hypothesized to influence invasion through the generation of novel phenotypes and/or increased levels of genetic variance. Based on morphology, hybrids between diffuse knapweed and spotted knapweed, two invasive plants in North America, are present in the invaded range. Some individuals within most diffuse knapweed sites in North America exhibit intermediate diffuse × spotted floral morphology. We examined hybridization at the molecular level, using amplified fragment length polymorphisms. Approximately a quarter of the assayed North American diffuse knapweed individuals exhibited evidence of introgression from spotted knapweed. However, plants with intermediate morphology did not show evidence of mixed ancestry more often than the plants with typical diffuse knapweed morphology. The high proportion of hybrid individuals in North American diffuse knapweed sites found here, combined with evidence from recent studies, suggests that diffuse knapweed was likely introduced with admixed individuals, and the hybrids are not newly created postintroduction. A century of backcrossing with diffuse knapweed has likely decoupled the relationship between morphology and admixture at the molecular level. In contrast to the scenario encountered in North America, in the native range where diploid diffuse and spotted knapweed overlap, hybrid swarms are common. In such sites, the floral phenotype aligns more closely with the genotype.

Herbivory and novel weapons: no evidence for enhanced competitive ability or allelopathy induction of Centaurea diffusa by biological controls

Biological control of weeds by arthropod herbivores is thought to work by reducing the competitive ability of the weed relative to the surrounding vegetation. However, the assumption that herbivory reduces plant competitive ability has not been tested in most biological control systems, and counter to expectation, recent research on the impact of biological control agents on invasive Centaurea species suggests that this genus may respond to herbivory by increased competitive ability through enhanced plant re-growth and/or by inducing increased production of phytotoxic allelochemicals. We examined the impact of two biological control agents of the invasive plant diffuse knapweed (C. diffusa) to see if feeding by either of these insects would enhance the plant’s competitive ability or allelochemical output. Sub-lethal herbivory by either of the biological control agents significantly reduced knapweed performance when the plant was grown in competition with either of two native species. Competition with knapweed significantly reduced the performance of both native species (Artemisia frigida and Bouteloua gracilis), and herbivory by one of the biocontrol agents resulted in a small but significant increase in both native species’ performance. Diffuse knapweed’s putative allelochemical 8-hydroxyquinoline was not detected in experimental or field collected soils from knapweed-infested sites. In contrast to other studies on the impacts of biological control on other Centaurea species, these data support the premise that biological control agents may reduce invading plant competitive ability. We find no evidence for diffuse knapweed allelopathy mediated by 8-hydroxyquinoline or enhanced allelopathy in response to herbivory by biological control agents.

The case against (–)-catechin involvement in allelopathy of Centaurea stoebe (spotted knapweed)

Proving allelopathic chemical interference is a daunting endeavor, in that production and movement of a phytotoxin from a donor plant to a receiving plant must be demonstrated in the substrate in which the plants grow, which is usually a complex soil matrix. The soil levels or soil flux levels of the compound generated by the donor must be proven to be sufficient to adversely affect the receiving plant. Reports of (–)-catechin to be the novel weapon used by Centaurea stoebe (spotted knapweed) to invade new territories are not supported by the paper featured in this Addendum, nor by papers produced by two other laboratories. These papers find that (–)-catechin levels in soil in which C. stoebe grows are orders of magnitude below levels that cause only minor growth effects on reported sensitive species. Furthermore, the claim that (–)-catechin acts as a phytotoxin through causing oxidative damage is refuted by the fact that the molecule is a strong antioxidant and is quickly degraded by extracellular root enzymes.

Hybridization and invasion: an experimental test with diffuse knapweed (Centaurea diffusa Lam.)

A number of studies have suggested a link between hybridization and invasion. In this study, we experimentally test the potential for hybridization to influence invasion through a greenhouse common garden study. Diffuse knapweed (DK) (Centaurea diffusa Lam.) was introduced to North America with admixture from spotted knapweed (SK) (Centaurea stoebe subsp. stoebe L.). Comparisons between North American DK (including hybrid phenotypes) and native (European) DK in a common garden did not reveal enhanced performance or increased phenotypic variance, suggesting that pre‐introduction hybridization or, more generally, post‐introduction evolutionary change has not significantly contributed to the invasion of DK. In contrast, early generation hybrids [artificially created Backcross 1 (BC1) plants] exhibited increased variance for eight of the examined traits, and greater leaf and reproductive shoot production when compared to North American DK. Individual BC1 lines differed for several traits, suggesting the importance of the cross for drawing conclusions from such comparisons. When compared to the parental species (DK and SK), the BC1 plants were not transgressive for any of the measured traits. Overall, these findings suggest that if diploid SK is introduced to North America, interspecific hybridization has the potential to result in even more aggressive invaders.

Geographic Patterns of Interspecific Hybridization between Spotted Knapweed (Centaurea stoebe) and Diffuse Knapweed (C. diffusa)

Abstract Hybridization between species has the potential to change invasion dynamics. Field observations suggest that spotted knapweed and diffuse knapweed, two ecologically and economically destructive invasive plants, hybridize in their introduced range. As a first step towards understanding whether hybridization has affected the dynamics of the invasion of these species, we conducted field surveys in the introduced (North American) and native (European) ranges to discern patterns of hybridization and measured fitness-related traits among field hybrids and parental species. In North America we detected plants with hybrid morphology in 97% of the diffuse knapweed sites (n  =  40); such hybrid plants were taller and more often exhibited polycarpy than plants with typical diffuse knapweed morphology. Hybrids were not detected in North American spotted knapweed sites (n  =  22). In most regions surveyed in Europe, diffuse knapweed and spotted knapweed were isolated from each other and existed as distinct, nonhybridizing species. However, in Ukraine, the two species frequently coexisted within a site, resulting in hybrid swarms. On average, the plants from the North American diffuse knapweed sites (including plants with both diffuse and hybrid morphology), were larger than the apparently pure diffuse knapweed in the native range. The cross-continental patterns of hybridization likely are explained by differences in cytology. It recently has been confirmed that the spotted knapweed in North America is tetraploid whereas the diffuse knapweed is diploid. Genetic incompatibilities associated with these two cytotypes likely prevent ongoing hybridization. We hypothesize that hybrid individuals were introduced to North America along with diffuse knapweed. Because plants with hybrid morphology are found in nearly all North American diffuse knapweed sites, the introduction of hybrids likely occurred early in the invasion of diffuse knapweed. Thus, although the presence of hybrids might facilitate the ongoing invasion of diffuse knapweed into North America, elevated concern regarding their presence might not be warranted. Because such individuals are not likely to represent a new hybridization event, currently effective management strategies used in diffuse knapweed sites should not need alteration. Nomenclature: Diffuse knapweed, Centaurea diffusa Lam.; spotted knapweed, C. stoebe L. ( =  C. maculosa Lam.).

Terrestrial Slugs as a Model Organism for Inquiry-Based Experimentation in a Majors General Biology Laboratory

ABSTRACT Many biology educators at the undergraduate level are revamping their laboratory curricula to incorporate inquiry-based research experiences so that students can directly participate in the process of science and improve their scientific reasoning skills. Slugs are an ideal organism for use in such a studentdirected, hypothesis-driven experience. Slugs are inexpensive, easily collected, and low maintenance. Most students are unfamiliar with slugs, and their curiosity is piqued as they observe and classify these fascinating creatures. The procedures described here offer step-by-step instructions on how slugs are used to guide students through an inquiry-based research project for 7 weeks in a majors general biology course.

Promoting Student Inquiry Using "Zea Mays" (Corn) Cultivars for Hypothesis-Driven Experimentation in a Majors Introductory Biology Course.

Abstract The AAAS Vision and Change report (2011) recommends incorporating student research experiences into the biology curriculum at the undergraduate level. This article describes, in detail, how Zea mays (corn) cultivars were used as a model for a hypothesis-driven short-term research project in an introductory biology course at a small Midwestern university. During the course of this project, student groups generated a research question and hypothesis, designed an experiment, collected data, and reported their findings in a paper modeled after the primary literature. Throughout the project, students experienced first hand the obstacles and accomplishments associated with the process of scientific research and gained a greater understanding of plant biology. By demonstrating biology as a dynamic field centered around hypothesis generation and experimentation, the authors observed an increase in student dedication, interest, and enthusiasm for the course.