Cosexuality Reduces Pollen Production and Fitness in Cannabis sativa L.

Cannabis sativa L. is cultivated globally for its cannabinoid-dense inflorescences. Commercial preference for sinsemilla has led to the development of methods for producing feminized seeds through cross-pollination of cosexual (masculinized) female plants. Although the induction of cosexuality in Cannabis plants is common, to date, no work has empirically tested how masculinization of female Cannabis plants impacts male flowering, pollen production, pollen fitness, and related life-history trade-offs. Here, we cultivated a population of Cannabis plants (CFX-2) and explored how the route to cosexuality (drought vs. chemical induction) impacted flowering phenology, pollen production, and pollen fitness, relative to unsexual male plants. Unisexual males flowered earlier and longer than cosexual plants and produced 223% more total pollen (F2,28 = 74.41, p < 0.001), but per-flower pollen production did not differ across reproductive phenotypes (F2,21 = 0.887, p = 0.427). Pollen viability was 200% higher in unisexual males and drought-induced cosexuals (F2,36 = 189.70, p < 0.001). Pollen non-abortion rates only differed in a marginally significant way across reproductive phenotypes (F2,36 = 3.00, p = 0.06). Here, we demonstrate that masculinization of female plants impacts whole-plant pollen production and pollen fitness in Cannabis sativa.

Methods for characterizing pollen fitness in Cannabis sativa L.

Pollen grains are male gametophytes, an ephemeral haploid generation of plants, that commonly engage in competition for a limited supply of ovules. Since variation in reproductive capabilities among male gametophytes may influence the direction and pace of evolution in populations, we must be able to quantify the relative fitness of gametophytes from different sires. To explore this, we estimated the relative fitness of groups of male gametophytes in a dioecious, wind-pollinated model system, Cannabis sativa, by characterizing the non-abortion rate (measured via chemical staining) and viability (measured via in vitro germination) of pollen from multiple sires. Pollen viability quickly declined within two weeks of anther dehiscence, and pollen stored under freezer conditions did not germinate regardless of storage time. In contrast, pollen non-abortion rates declined slowly and persisted longer than the lifetime of a sporophyte plant under both room temperature and freezer conditions. Pollen samples that underwent both viability and non-abortion rate analysis displayed no significant correlation, implying that researchers cannot predict pollen viability from non-abortion rates, nor infer male gametophytic fitness from a single measure. Our work demonstrates two independent, differential approaches to measure proxies of male fitness in C. sativa.

Dioecious hemp (Cannabis sativa L.) plants do not express significant sexually dimorphic morphology in the seedling stage.

Some economically important crop species are dioecious, producing pollen and ovules on distinct, unisexual, individuals. On-the-spot diagnosis of sex is important to breeders and farmers for crop improvement and maximizing yield, yet diagnostic tools at the seedling stage are understudied and lack a scientific basis. Understanding sexual dimorphism in juvenile plants may provide key ecological, evolutionary and economic insights into dioecious plant species in addition to improving the process of crop cultivation. To address this gap in the literature, we asked: can we reliably differentiate males, females, and co-sexual individuals based on seedling morphology in Cannabis sativa, and do the traits used to distinguish sex at this stage vary between genotypes? To answer these questions, we collected data on phenotypic traits of 112 C. sativa plants (50 female, 52 male, 10 co-sexuals) from two hemp cultivars (CFX-1, CFX-2) during the second week of vegetative growth and used ANOVAs to compare morphology among sexes. We found males grew significantly longer hypocotyls than females by week 2, but this difference depended on the cultivar investigated. Preliminary evidence suggests that co-sexual plants may be distinguished from male and female plants using short hypocotyl length and seedling height, although this relationship requires more study since sample sizes of co-sexual plants were small. In one of the cultivars, two-week old male plants tend to produce longer hypocotyls than other plants, which may help to identify these plants prior to anthesis. We call for increased research effort on co-sexual plants, given their heavy economic cost in industrial contexts and rare mention in the literature. Our preliminary data suggests that short hypocotyl length may be an indicator of co-sexuality. These results are the first steps towards developing diagnostic tools for predicting sex using vegetative morphology in dioecious species and understanding how sexual dimorphism influences phenotype preceding sexual maturity.

Growth and fecundity of colonizing hybrid Raphanus populations are environmentally dependent.

PREMISE: Hybrid gene pools harbor more genetic variation than progenitor populations. Thus, we expect hybrid populations to exhibit more dynamic evolutionary responses to environmental variation. We ask how environmental variation experienced by adapted and transplanted populations influence the success of late-generation hybrid populations during invasion. METHODS: For four generations, 20 wild (Raphanus raphanistrum) and 20 hybrid radish (R. sativus × R. raphanistrum) plant populations evolved under experimentally manipulated moisture conditions (dry, wet, control-sheltered, or control-unsheltered plots; i.e., evolutionary environment) in old fields near Toronto, Canada. We planted advanced-generation wild and hybrid radishes in sheltered plots and exposed them to either an evolutionary or a novel watering environment. To determine how soil moisture would influence invasion success, we compared the phenotype and fecundity of plants grown in these various environments. RESULTS: Hybridization produced larger plants. In wet environments, hybrid seedlings emerged more frequently and expressed higher photosynthetic activity. Low-moisture, novel conditions delayed and reduced seedling emergence frequency. Hybrid plants and those that evolved under relatively wet environments exhibited higher aboveground biomass. Hybrid plants from control-sheltered plots colonizing novel moisture environments were more fecund than comparable wild plants. CONCLUSIONS: Dry environments are less likely than other evolutionary environments to contribute colonists. However, relatively wet locations support the evolution of relatively fecund plants, especially crop-wild hybrid populations. Thus, our results provide a strong mechanistic explanation for variation in the relative success of crop-wild hybrids among study locations and a new standard for studies that assess the risk of crop-wild hybridization events.

Optimizing Photoperiod Switch to Maximize Floral Biomass and Cannabinoid Yield in Cannabis sativa L.: A Meta-Analytic Quantile Regression Approach.

Cannabis sativa L. is an annual, short-day plant, such that long-day lighting promotes vegetative growth while short-day lighting induces flowering. To date, there has been no substantial investigation on how the switch between these photoperiods influences yield of C. sativa despite the tight correlation that plant size and floral biomass have with the timing of photoperiod switches in indoor growing facilities worldwide. Moreover, there are only casual predictions around how the timing of the photoperiodic switch may affect the production of secondary metabolites, like cannabinoids. Here we use a meta-analytic approach to determine when growers should switch photoperiods to optimize C. sativa floral biomass and cannabinoid content. To this end, we searched through ISI Web of Science for peer-reviewed publications of C. sativa that reported experimental photoperiod durations and results containing cannabinoid concentrations and/or floral biomass, then from 26 studies, we estimated the relationship between photoperiod and yield using quantile regression. Floral biomass was maximized when the long daylength photoperiod was minimized (i.e., 14 days), while THC and CBD potency was maximized under long day length photoperiod for ~42 and 49-50 days, respectively. Our work reveals a yield trade-off in C. sativa between cannabinoid concentration and floral biomass where more time spent under long-day lighting maximizes cannabinoid content and less time spent under long-day lighting maximizes floral biomass. Growers should carefully consider the length of long-day lighting exposure as it can be used as a tool to maximize desired yield outcomes.

Comparing methods for controlled capture and quantification of pollen in Cannabis sativa.

PREMISE: Precise pollen collection methods are necessary for crop breeding, but anemophilous pollen is notoriously difficult to capture and control. Here we compared a variety of methods for the controlled capture of cannabis pollen, intended to ease the process of cross-fertilization for breeding this wind-pollinated plant, and measured the utility of light spectroscopy for quantifying relative pollen yield. METHODS AND RESULTS: In two independent trials, we compared a control method of pollen collection (hand collection) to either vacuum-, water-, or bag-collection methods. We used visible light spectroscopy to quantify relative pollen yield, and validated this approach using microscopic pollen counts. We determined that pollen yield was highest when using hand collection or vacuum collection, but efficiency did not differ significantly among methods. CONCLUSIONS: To maximize yield, pollen should be collected by hand or vacuum, but all collection methods were equally efficient in a relative sense because yield increased with collection time. We also found that light spectroscopy is an accurate and rapid method of quantifying pollen abundance (R 2 = 0.86) in a liquid suspension.

Cannabinoid Inheritance Relies on Complex Genetic Architecture.

Introduction: Understanding the inheritance of cannabinoid compounds in Cannabis sativa will facilitate effective crop breeding and careful regulation of controlled substances. The production of two key cannabinoids, Δ-9-tetrahydrocannabinol (THC) and cannabidiol (CBD), is partially controlled by two additive loci. Here, we present the first study to search for evidence of alternate genetic models describing the inheritance and expression of cannabinoids. Materials and Methods: Using an information-theoretic approach, we estimated composite genetic effects (CGEs) of four cultivars with pure CBD or pure THC chemotypes, their F1 and F2 hybrid progeny, to identify genetic models that explain cannabinoid inheritance patterns. We also estimated the effective number of genetic factors that control differences in cannabinoid concentration (THC, CBD, and cannabichromene [CBC]). Results: Unlike previous research, we note nonadditive components of cannabinoid inheritance. Concentration of THC is a polygenic trait (three to four genetic factors). Both additive and dominance CGEs best explained THC expression patterns. In contrast, cytoplasmic genomes and additive genes may influence CBD concentration. Maternal additive effects and additive genetic effects apparently influence CBC expression. Conclusions: Cannabinoid inheritance is more complex than previously appreciated; among other genetic effects, cytogenetic and maternal contributions may be undervalued influences on cannabinoid ratios and concentrations. Further research on the environmental sensitivity of cannabinoid production is advised.

Correlates of hybridization in plants.

Hybridization is a biological phenomenon increasingly recognized as an important evolutionary process in both plants and animals, as it is linked to speciation, radiation, extinction, range expansion and invasion, and allows for increased trait diversity in agricultural and horticultural systems. Estimates of hybridization frequency vary across taxonomic groups, but causes of this variation are unknown. Here, we ask on a global scale whether hybridization is linked to any of 11 traits related to plant life history, reproduction, genetic predisposition, and environment or opportunity. Given that hybridization is not evenly distributed across the plant tree of life, we use phylogenetic generalized least squares regression models and phylogenetic path analysis to detect statistical associations between hybridization and plant traits at both the family and genus levels. We find that perenniality and woodiness are each weakly associated with an increased frequency of hybridization in univariate analyses, but path analysis suggests that the direct linkage is between perenniality and increased hybridization (with woodiness having only an indirect relationship with hybridization via perenniality). Weak associations between higher rates of hybridization and higher outcrossing rates, abiotic pollination syndromes, vegetative reproductive modes, larger genomes, and less variable genome sizes are detectable in some cases but not others. We argue that correlational evidence at the global scale, such as that presented here, provides a robust framework for forming hypotheses to examine and test drivers of hybridization at a more mechanistic level.

Phenotypic plasticity influences the success of clonal propagation in industrial pharmaceutical Cannabis sativa.

The burgeoning cannabis market requires evidence-based science such that farmers can quickly and efficiently generate new plants. In part, horticultural operations are limited by the success of cloning procedures. Here, we measured the role of environmental conditions and cultivar identity on the success of generating long branch material with many meristems in planting stock (mothers) and in rooting success of stem-derived clones. To evaluate the influence of lighting treatments on the optimal production of branching mothers, four lighting conditions (Fluorescent High Output T5s [T5], Metal halide lamps [MH], Plasma lamps [PL], or Metal halide lamps augmented with far red LED lights [MH+FR]) were applied to two cultivars of container grown plants (Cannabis sativa L. 'Bubba Kush', 'Ghost Train Haze') grown in peat-based organic substrates in mylar grow tents. To evaluate the influence of lighting, cutting tool (secateurs or scalpels), and stem wounding (present/absent) on optimal rooting of stems, three lighting conditions (Fluorescent T8s, T5, PL) were applied to three cultivars of peat pellet grown plants (C. sativa L. 'Bubba Kush', 'Ghost Train Haze', 'Headband'). Mothers grown under T5 and MH (vs MH+FR) produced ~30% more meristems. However, growing mothers under MH+FR were 19% taller than mothers under T5, with ~25% longer internodes on dominant stems than plants under any other lighting condition. Canopies were denser under T5 because petiole length was ~30% shorter under T5 and fan leaves were longer and narrower under MH+FR and MH+FR and PL, respectively, than under other lighting conditions. Cultivar Ghost Train Haze stems rooted most frequently and most quickly. Wounded stems were 162% more likely to root than unwounded stems and rooted 1.5 days earlier. Our results will guide producers attempting to maximize the rate of clone production in licensed facilities; although results may differ among cultivars, where cultivars differed in their average phenotype as mother plants, and their propensity to root from cuttings, and the speed with which they produced those roots.

Pollen sleuthing for terrestrial plant surveys: Locating plant populations by exploiting pollen movement.

PREMISE OF THE STUDY: We present an innovative technique for sampling, identifying, and locating plant populations that release pollen, without extensive ground surveys. This method (1) samples pollen at random locations within the target species' habitat, (2) detects species' presence using morphological pollen analysis, and (3) uses kriging to predict likely locations of populations to focus future search efforts. METHODS: To demonstrate, we applied the pollen sleuthing system to search for artificially constructed populations of Brassica rapa in an old field. Population size varied from 0-100 flowers labeled with artificial pollen (paint pellets). After characterizing the landscape, we pan-trapped 2762 potential insect vectors from random locations across the field and washed particulate matter from their bodies to assess artificial pollen abundance with a microscope. RESULTS: Population size greatly influenced artificial pollen detection success; following random pollen trap sampling and interpolation, ground surveys would be best focused on identified areas with high pollen density and low variation in pollen density. Sampling sites most successfully detected artificial pollen when they were located at higher elevations, near showy flowering plants that were not grasses. DISCUSSION: Detection of nascent populations using the proposed system is possible but accuracy will depend on local environmental factors (e.g., wind, elevation). Conservation and invasive species control programs may be improved by using this approach.

The Effect of Altered Soil Moisture on Hybridization Rate in a Crop-Wild System (Raphanus spp.).

Since plant mating choices are flexible and responsive to the environment, rates of spontaneous hybridization may vary across ecological clines. Developing a robust and predictive framework for rates of plant gene flow requires assessing the role of environmental sensitivity on plant reproductive traits, relative abundance, and pollen vectors. Therefore, across a soil moisture gradient, we quantified pollinator movement, life-history trait variation, and unidirectional hybridization rates from crop (Raphanus sativus) to wild (Raphanus raphanistrum) radish populations. Both radish species were grown together in relatively dry (no rain), relatively wet (double rain), or control soil moisture conditions in Ohio, USA. We measured wild and crop radish life-history, phenology and pollinator visitation patterns. To quantify hybridization rates from crop-to-wild species, we used a simply inherited morphological marker to detect F1 hybrid progeny. Although crop-to-wild hybridization did not respond to watering treatments, the abundance of hybrid offspring was higher in fruits produced late in the period of phenological overlap, when both species had roughly equal numbers of open flowers. Therefore, the timing of fruit production and its relationship to flowering overlap may be more important to hybrid zone formation in Raphanus spp. than soil moisture or pollen vector movements.

Assessing the effects of hybridization and precipitation on invasive weed demography using strength of selection on vital rates.

BACKGROUND: As global climate change transforms average temperature and rainfall, species distributions may meet, increasing the potential for hybridization and altering individual fitness and population growth. Altered rainfall specifically may shift the strength and direction of selection, also manipulating population trajectories. Here, we investigated the role of interspecific hybridization and selection imposed by rainfall on the evolution of weedy life-history in non-hybrid (Raphanus raphanistrum) and hybrid (R. raphanistrum x R. sativus) populations using a life table response experiment. RESULTS: In documenting long-term population dynamics, we determined intrinsic (r) and asymptotic (λ) population growth rates and sensitivities, a measure of selection imposed on demographic rates. Hybrid populations experienced 8.7-10.3 times stronger selection than wild populations for increased seedling survival. Whereas crop populations generally exhibit little dormancy and wild populations often exhibit dormancy, non-hybrid populations experienced 10% stronger selection than hybrid populations for exhibiting seed dormancy. Selection on survival-to-flowering in wild, not hybrid, populations declined marginally with increasing soil moisture. Hybrid populations exhibited greater r, but not λ, than wild populations regardless of moisture environment. In general, fecundity contributed most to differences in λ but fecundity only contributed positively to hybrid λ relative to wild λ when precipitation was altered (either higher or lower than control) and not under control watering conditions. CONCLUSIONS: Selection on key demographic traits may not change dramatically in response to rainfall, and hybridization may more strongly influence the demography of these weedy species than rainfall. If hybrid populations can respond to selection for increased dormancy, this may make it more difficult to deplete weed seed banks and increase the persistence of crop genes in weed populations.

Contemporary evolution and the dynamics of invasion in crop-wild hybrids with heritable variation for two weedy life-histories.

Gene flow in crop-wild complexes between phenotypically differentiated ancestors may transfer adaptive genetic variation that alters the fecundity and, potentially, the population growth (λ) of weeds. We created biotypes with potentially invasive traits, early flowering or long leaves, in wild radish (Raphanus raphanistrum) and F5 crop-wild hybrid (R. sativus × R. raphanistrum) backgrounds and compared them to randomly mated populations, to provide the first experimental estimate of long-term fitness consequences of weedy life-history variation. Using a life table response experiment design, we modeled λ of experimental, field populations in Pellston, MI, and assessed the relative success of alternative weed strategies and the contributions of individual vital rates (germination, survival, seed production) to differences in λ among experimental populations. Growth rates (λ) were most influenced by seed production, a trait altered by hybridization and selection, compared to other vital rates. More seeds were produced by wild than hybrid populations and by long-leafed than early-flowering lineages. Although we did not detect a biotype by selection treatment effect on lambda, lineages also exhibited contrasting germination and survival strategies. Identifying life-history traits affecting population growth contributes to our understanding of which portions of the crop genome are most likely to introgress into weed populations.

Long-Chain Omega-3 Polyunsaturated Fatty Acids Have Developmental Effects on the Crop Pest, the Cabbage White Butterfly Pieris rapae.

Nutritional enhancement of crops using genetic engineering can potentially affect herbivorous pests. Recently, oilseed crops have been genetically engineered to produce the long-chain omega-3 polyunsaturated fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) at levels similar to that found in fish oil; to provide a more sustainable source of these compounds than is currently available from wild fish capture. We examined some of the growth and development impacts of adding EPA and DHA to an artificial diet of Pieris rapae, a common pest of Brassicaceae plants. We replaced 1% canola oil with EPA: DHA (11:7 ratio) in larval diets, and examined morphological traits and growth of larvae and ensuing adults across 5 dietary treatments. Diets containing increasing amounts of EPA and DHA did not affect developmental phenology, larval or pupal weight, food consumption, nor larval mortality. However, the addition of EPA and DHA in larval diets resulted in progressively heavier adults (F 4, 108 = 6.78; p = 0.011), with smaller wings (p < 0.05) and a higher frequency of wing deformities (R = 0.988; p = 0.001). We conclude that the presence of EPA and DHA in diets of larval P. rapae may alter adult mass and wing morphology; therefore, further research on the environmental impacts of EPA and DHA production on terrestrial biota is advisable.

An ecological approach to measuring the evolutionary consequences of gene flow from crops to wild or weedy relatives.

PREMISE OF THE STUDY: Agricultural practices routinely create opportunities for crops to hybridize with wild relatives, leading to crop gene introgression into wild genomes. Conservationists typically worry this introgression could lead to genetic homogenization of wild populations, over and above the central concern of transgene escape. Alternatively, viewing introgression as analogous to species invasion, we suggest that increased genetic diversity may likewise be an undesirable outcome. METHODS: Here, we compare the sensitivity of conventional population genetic metrics with species diversity indices as indicators of the impact of gene flow on genetic diversity. We illustrate this novel approach using multilocus genotype data (12 allozyme loci) from 10 wild (Beta vulgaris subsp. maritima) and eight putative crop-wild hybrid beet populations (B. vulgaris subsp. vulgaris × B. vulgaris subsp. maritima) scattered throughout Europe. RESULTS: Conventional population genetic metrics mostly failed to detect shifts in genetic composition of putative hybrid populations. By contrast, species diversity indices unambiguously revealed increased genetic diversity in putative hybrid populations. DISCUSSION: We encourage other workers to explore the utility of our more sensitive approach for risk assessment prior to the release of transgenic crops, with a view toward widespread adoption of our method in studies aimed at detecting allelic invasion.

Gender-heterogeneous working groups produce higher quality science.

Here we present the first empirical evidence to support the hypothesis that a gender-heterogeneous problem-solving team generally produced journal articles perceived to be higher quality by peers than a team comprised of highly-performing individuals of the same gender. Although women were historically underrepresented as principal investigators of working groups, their frequency as PIs at the National Center for Ecological Analysis and Synthesis is now comparable to the national frequencies in biology and they are now equally qualified, in terms of their impact on the accumulation of ecological knowledge (as measured by the h-index). While women continue to be underrepresented as working group participants, peer-reviewed publications with gender-heterogeneous authorship teams received 34% more citations than publications produced by gender-uniform authorship teams. This suggests that peers citing these publications perceive publications that also happen to have gender-heterogeneous authorship teams as higher quality than publications with gender uniform authorship teams. Promoting diversity not only promotes representation and fairness but may lead to higher quality science.

Hybridization alters early life-history traits and increases plant colonization success in a novel region.

Hybridization is hypothesized to promote invasiveness, but empirical tests comparing the performance of hybrid taxa versus parental taxa in novel regions are lacking. We experimentally compared colonization ability of populations of wild radish (Raphanus raphanistrum) with populations of advanced-generation hybrids between wild radish and cultivated radish (Raphanus sativus) in a southeast Texas pasture, well beyond the known invasive range of hybrid radish. We also manipulated the strength of interspecific competition to better generalize across variable environments. In both competitive environments, hybrid populations produced at least three times more seeds than did wild radish populations, a distinction that was driven by greater hybrid seedling emergence, earlier hybrid emergence, and more hybrid seedlings surviving to flower, rather than by greater individual fecundity. Flowering duration in hybrids was less negatively affected by competition than it was in wild radish, while early emergence was associated with subsequent high seed output in both biotypes. Our data show that hybridization can enhance colonization success in a novel region and, by comparison with previous studies, that the life-history traits enhancing hybrid success can differ across regions, even for lineages originating from the same hybridization event. These results imply a much larger arena for hybrid success than previously appreciated.

When divergent life histories hybridize: insights into adaptive life-history traits in an annual weed.

*Colonizing weed populations face novel selective environments, which may drive rapid shifts in life history. These shifts may be amplified when colonists are hybrids of species with divergent life histories. Selection on such phenotypically diverse hybrids may create highly fecund weeds. We measured the phenotypic variation, strength of natural selection and evolutionary response of hybrid and nonhybrid weeds. *We created F(1) hybrids of wild radish, an early flowering, small-stemmed weed, and its late-flowering, large-stemmed, crop relative (Raphanus spp.). Replicate wild and hybrid populations were established in an agricultural landscape in Michigan, USA. The consequences of three generations of natural selection were measured in a common garden experiment. *Hybrid populations experienced strong selection for larger, earlier flowering plants whereas selection was relatively weak on wild populations. Large plant size evolved two to three times faster in the hybrid populations than in wild populations, yet hybrid populations did not evolve earlier flowering. Strong selection on size and phenotypic correlations between age at reproduction and size may have limited the response of flowering phenology. *Our findings demonstrate hybridization between species with divergent life histories may catalyse the rapid evolution of certain adaptive, weedy traits while tradeoffs limit the evolution of others.

Rapid evolution in crop-weed hybrids under artificial selection for divergent life histories.

When species hybridize, offspring typically exhibit reduced fitness and maladapted phenotypes. This situation has biosafety implications regarding the unintended spread of novel transgenes, and risk assessments of crop-wild hybrids often assume that poorly adapted hybrid progeny will not evolve adaptive phenotypes. We explored the evolutionary potential of early generation hybrids using nontransgenic wild and cultivated radish (Raphanus raphanistrum, Raphanus sativus) as a model system. We imposed four generations of selection for two weedy traits - early flowering or large size - and measured responses in a common garden in Michigan, USA. Under selection for early flowering, hybrids evolved to flower as early as wild lineages, which changed little. These early-flowering hybrids also recovered wild-type pollen fertility, suggesting a genetic correlation that could accelerate the loss of crop traits when a short life cycle is advantageous. Under selection for large size at reproduction, hybrids evolved longer leaves faster than wild lineages, a potentially advantageous phenotype under longer growing seasons. Although early generation hybrid offspring have reduced fitness, our findings provide novel support for rapid adaptation in crop-wild hybrid populations. Biosafety risk assessment programs should consider the possibility of rapid evolution of weedy traits from early generations of seemingly unfit crop-wild hybrids.

Can feral weeds evolve from cultivated radish (Raphanus sativus, Brassicaceae)?

Cultivated plants that cannot survive on their own often have maladaptive domestication traits. Unharvested crop seeds may generate feral populations, at times causing serious weed problems, but little is known about the evolution of ferality. We explored the potential for cultivated radish, Raphanus sativus, to become feral, given that closely related taxa (e.g., R. raphanistrum and crop-wild hybrids) are well-documented weeds. First, we measured the population growth of five experimental, cultivated, self-seeding radish populations in Michigan, USA, for three generations. Three late-flowering populations went extinct, and two others apparently hybridized with local R. raphanistrum. A common garden experiment showed that the two surviving populations had earlier flowering, smaller root diameters, and greater individual fecundity than did nonhybridized populations. We also used artificial selection to measure the evolutionary potential for earlier flowering. After two generations of strong selection, two of three lineages flowered earlier and produced more seeds than control lineages, but insufficient genetic variation prevented dramatic evolution of crop phenotypes. In summary, it seems unlikely that radishes could spontaneously become feral in our study area without gene flow from R. raphanistrum. Applying these approaches to other cultivated species may provide a better understanding of mechanisms promoting the evolution of feral weeds.