Mitchell B. Cruzan

Evidence for multiple sources of invasion and intraspecific hybridization in Brachypodium sylvaticum (Hudson) Beauv. in North America.

We compared the levels and distribution of genetic diversity in Eurasian and North American populations of Brachypodium sylvaticum (Huds.) Beauv. (false brome), a newly invasive perennial bunchgrass in western North America. Our goals were to identify source regions for invasive populations, determine the number of independent invasion events, and assess the possibility that postinvasion bottlenecks and hybridization have affected patterns of genetic diversity in the invaded range. We tested the hypothesis that this Eurasian grass was accidentally introduced into two areas in Oregon and one site in California by examining nuclear microsatellites and chloroplast haplotype variation in 23 introduced and 25 native populations. In the invaded range, there was significantly lower allelic richness (RS), observed heterozygosity (HO) and within‐population gene diversity (HS), although a formal test failed to detect a significant genetic bottleneck. Most of the genetic variation existed among populations in the native range but within populations in the invaded range. All of the allelic variation in the invaded range could be explained based on alleles found in western European populations. The distribution of identified genetic clusters in the North American populations and the unique alleles associated with them is consistent with two historical introductions in Oregon and a separate introduction to California. Further analyses of population structure indicate that intraspecific hybridization among genotypes from geographically distinct regions of western Europe occurred following colonization in Oregon. The California populations, however, are more likely to be derived from one or perhaps several genetically similar regions in the native range. The emergence and spread of novel recombinant genotypes may be facilitating the rapid spread of this invasive species in Oregon.

Contributions of Heterosis and Epistasis to Hybrid Fitness

Early‐generation hybrid fitness is difficult to interpret because heterosis can obscure the effects of hybrid breakdown. We used controlled reciprocal crosses and common garden experiments to distinguish between effects of heterosis and nuclear and cytonuclear epistasis among morphotypes and advanced‐generation hybrid derivative populations in the Piriqueta caroliniana (Turneraceae) plant complex. Seed germination, growth, and sexual reproduction of first‐generation hybrids, inbred parental lines, and outbred parental lines were compared under field conditions. Average vegetative performance was greater for hybrids than for inbred lines, and first‐season growth was similar for hybrids and outbred parental lines. Hybrid survival surpassed that of inbred lines and was equal to or greater than outbred lines’ survival, and more F1 than parental plants reproduced. Reductions in hybrid fitness due to Dobzhansky‐Muller incompatibilities (epistasis among divergent genetic elements) were expressed as differences in vegetative growth, survival, and reproduction between plants from reciprocal crosses for both F1 and backcross hybrid generations. Comparing performance of hybrids against parental genotypes from intra‐ and interpopulation crosses allowed a more robust prediction of F1 hybrids success and more accurate interpretations of the genetic architecture of F1 hybrid vigor.

Postpollination mechanisms influencing mating patterns and fecundity: an example from Eichhornia paniculata

Plant mating systems are influenced by the amount and genetic composition of pollen grains deposited on stigmas and by the ability of recipients to discriminate among pollen from different sources. We describe an experimental procedure that uses limiting and excess pollinations with mixtures of genetically marked pollen to partition the siring success of donors into three components: prefertilization gamete attrition (failure of male gametophytes before fertilization), pollen competitive ability (differences in pollen tube growth rate), and postfertilization gamete attrition (embryo abortion). Regression models for the relationships of pollen load size with each pollens siring success and total recipient fecundity indicate that, for mixtures of self and outcross pollen, differences in gamete attrition, pollen competitive ability, and postfertilization success will have distinct and predictable effects on mating patterns and fecundity. Mating systems that rely on differences in pollen competitive ability result in outcrossing frequencies that increase with pollen load size, with seed production remaining high over a broad range of load sizes. In contrast, for mating systems governed by differences in gamete attrition, the frequency of outcrossed progeny will not vary greatly with pollen load size, but reduced fecundity will be expected over a wider range of pollen load sizes. These predictions were confirmed by analyzing the response of siring success and fecundity in response to pollen load size in the tristylous Eichhornia paniculata (Pontederiaceae). Experimental manipulations of the size and composition of pollen loads allow prediction of the frequency of outcrossed progeny produced under varying pollen environments.

Comparative Studies of Evolutionary Responses to Light Environments in Arabidopsis

In this article, we compare the reaction norms to foliage shade (changes in light quality, spatially fine‐grained environmental variation) and photoperiod (day length, spatially coarse‐grained environmental variation) in several haplotypes of Arabidopsis thaliana from Scandinavia. We found that both across‐environment means and phenotypic plasticities evolved continuously and very rapidly within this group. Both character means and trait plasticities were highly integrated, in part as predicted by the adaptive plasticity hypothesis for response to foliage shade (the so‐called shade‐avoidance syndrome). We found that a significant but small fraction of the variance in across‐treatment trait means and plasticities in response to one environmental factor is explained by variation of the same traits in response to the other factor. Genetic relatedness based on chloroplast DNA sequence variation among haplotypes was not associated with variation in across‐treatment character means or their plasticities, suggesting that evolution of these characters has occurred on a local geographic scale via reticulation (outcrossing) among maternal lines rather than by the differential survival of selfing lineages.

Shifting dispersal modes at an expanding species' range margin

While it is generally recognized that noncontiguous (long‐distance) dispersal of small numbers of individuals is important for range expansion over large geographic areas, it is often assumed that colonization on more local scales proceeds by population expansion and diffusion dispersal (larger numbers of individuals colonizing adjacent sites). There are few empirical studies of dispersal modes at the front of expanding ranges, and very little information is available on dispersal dynamics at smaller geographic scales where we expect contiguous (diffusion) dispersal to be prevalent. We used highly polymorphic genetic markers to characterize dispersal modes at a local geographic scale for populations at the edge of the range of a newly invasive grass species (Brachypodium sylvaticum) that is undergoing rapid range expansion in the Pacific Northwest of North America. Comparisons of Bayesian clustering of populations, patterns of genetic diversity, and gametic disequilibrium indicate that new populations are colonized ahead of the invasion front by noncontiguous dispersal from source populations, with admixture occurring as populations age. This pattern of noncontiguous colonization was maintained even at a local scale. Absence of evidence for dispersal among adjacent pioneer sites at the edge of the expanding range of this species suggests that pioneer populations undergo an establishment phase during which they do not contribute emigrants for colonization of neighbouring sites. Our data indicate that dispersal modes change as the invasion matures: initial colonization processes appear to be dominated by noncontiguous dispersal from only a few sources, while contiguous dispersal may play a greater role once populations become established.

EVIDENCE OF LOCAL ADAPTATION TO COARSE‐GRAINED ENVIRONMENTAL VARIATION IN ARABIDOPSIS THALIANA

Abstract Plants can achieve an appropriate phenotype in particular conditions either constitutively or plastically, depending in part on the grain size of the environmental conditions being considered. Coarse-grained environmental variation should result in selection for local adaptation and no selection on plasticity to novel levels of the coarse-grained environmental factors. We tested the hypotheses that natural populations of the well-studied model system Arabidopsis thaliana are locally adapted to spatially coarse-grained environmental variation, and that the photoperiodic regime per se is at least partially responsible for that local adaptation, by exposing natural populations to photoperiodic regimes characteristic of their native and foreign (novel) environments. We also tested the hypothesis that plasticity to novel photoperiodic regimes should appear random. We found that populations showed evidence of local adaptation at a spatially coarse grain, although not to photoperiodic regime per se. We also found that the plasticities to novel photoperiodic regimes appeared random and did not generally show evidence of adaptive divergence. Our study highlights the need for caution in extrapolating from the finding of local adaptation to the causes of local adaptation.

Sequencing and De Novo Transcriptome Assembly of Brachypodium sylvaticum (Poaceae)

Premise of the study: We report the de novo assembly and characterization of the transcriptomes of Brachypodium sylvaticum (slender false-brome) accessions from native populations of Spain and Greece, and an invasive population west of Corvallis, Oregon, USA. Methods and Results: More than 350 million sequence reads from the mRNA libraries prepared from three B. sylvaticum genotypes were assembled into 120,091 (Corvallis), 104,950 (Spain), and 177,682 (Greece) transcript contigs. In comparison with the B. distachyon Bd21 reference genome and GenBank protein sequences, we estimate >90% exome coverage for B. sylvaticum. The transcripts were assigned Gene Ontology and InterPro annotations. Brachypodium sylvaticum sequence reads aligned against the Bd21 genome revealed 394,654 single-nucleotide polymorphisms (SNPs) and >20,000 simple sequence repeat (SSR) DNA sites. Conclusions: To our knowledge, this is the first report of transcriptome sequencing of invasive plant species with a closely related sequenced reference genome. The sequences and identified SNP variant and SSR sites will provide tools for developing novel genetic markers for use in genotyping and characterization of invasive behavior of B. sylvaticum.

Leaf morphological responses to variation in water availability for plants in the Piriqueta caroliniana complex

Distribution of plants and the expression of traits associated with environmental variation can be affected by both average conditions and the variance in conditions including extreme climatic events. We expect that these same factors should affect the distribution of plants in hybrid zones between ecologically distinct species where the hybrids should occupy ecotones or intermediate habitats. We evaluated water availability and leaf morphological differences among parental and hybrid populations of herbaceous perennial plants in the Piriqueta caroliniana complex along environmental gradients in Southeastern North America. We focus on two taxa in this group; the viridis morphotype, which occurs in southern Florida, and the caroliniana morphotype, which is distributed from northern Florida to southern Georgia. Advanced-generation hybrid derivatives of these morphotypes occupy a broad geographic region that extends across much of central Florida. Overall, we found that hybrid populations occurred in significantly drier locations, indicating that their habitat requirements are transgressive (i.e., exceeding parental values) rather than intermediate to the parental morphotypes. Water availability differed between the two sampling years, and plants displayed morphological changes in response to these changes in moisture. During the drier year, leaves were narrower and more hirsute, corroborating experimental results that these leaf traits are plastic, and confirming that plasticity occurs in natural habitats. Hybrids exhibited intermediate leaf traits (shape and size) across both years, and displayed transgressive (hair density) leaf traits during the drier year. The apparent canalization of the hybrids’ leaf morphological traits may contribute to their tolerance of variable environmental conditions and may partially explain why they have displaced the caroliniana morphotype in central Florida.

The influence of fertilizer level and spore density on arbuscular mycorrhizal colonization of transgenic Bt 11 maize (Zea mays) in experimental microcosms

Crop plants genetically modified for the expression of Bacillus thuringiensis (Bt) insecticidal toxins have broad appeal for reducing insect damage in agricultural systems, yet questions remain about the impact of Bt plants on symbiotic soil organisms. Here, arbuscular mycorrhizal fungal (AMF) colonization of transgenic maize isoline Bt 11 (expressing Cry1Ab) and its non-Bt parental line (Providence) was evaluated under different fertilizer level and spore density scenarios. In a three-way factorial design, Bt 11 and non-Bt maize were inoculated with 0, 40, or 80 spores of Glomus mosseae and treated weekly with No (0 g L(-1) ), Low (0.23 g L(-1) ), or High (1.87 g L(-1) ) levels of a complete fertilizer and grown for 60 days in a greenhouse. While no difference in AMF colonization was detected between the Bt 11 and Providence maize cultivars in the lower spore/higher fertilizer treatments, microcosm experiments demonstrated a significant reduction in AMF colonization in Bt 11 maize roots in the 80 spore treatments when fertilizer was limited. These results confirm previous work indicating an altered relationship between this Bt 11 maize isoline and AMF and demonstrate that the magnitude of this response is strongly dependent on both nutrient supply and AMF spore inoculation level.

Field Evaluation of Arbuscular Mycorrhizal Fungal Colonization in Bacillus thuringiensis Toxin-Expressing (Bt) and Non-Bt Maize

ABSTRACT The cultivation of genetically engineered Bacillus thuringiensis toxin-expressing (Bt) maize continues to increase worldwide, yet the effects of Bt crops on arbuscular mycorrhizal fungi (AMF) in soil are poorly understood. In this field experiment, we investigated the impact of seven different genotypes of Bt maize and five corresponding non-Bt parental cultivars on AMF and evaluated plant growth responses at three different physiological time points. Plants were harvested 60 days (active growth), 90 days (tasseling and starting to produce ears), and 130 days (maturity) after sowing, and data on plant growth responses and percent AMF colonization of roots at each harvest were collected. Spore abundance and diversity were also evaluated at the beginning and end of the field season to determine whether the cultivation of Bt maize had a negative effect on AMF propagules in the soil. Plant growth and AMF colonization did not differ between Bt and non-Bt maize at any harvest period, but AMF colonization was positively correlated with leaf chlorophyll content at the 130-day harvest. Cultivation of Bt maize had no effect on spore abundance and diversity in Bt versus non-Bt plots over one field season. Plot had the most significant effect on total spore counts, indicating spatial heterogeneity in the field. Although previous greenhouse studies demonstrated that AMF colonization was lower in some Bt maize lines, our field study did not yield the same results, suggesting that the cultivation of Bt maize may not have an impact on AMF in the soil ecosystem under field conditions.