Phil S. Allen

Ecological genetics of seed germination regulation in Bromus tectorum L.

Abstract Regulation of seed germination phenology is an important aspect of the life history strategy of invading annual plant species. In the obligately selfing winter annual grass Bromus tectorum, seeds are at least conditionally dormant at dispersal in early summer and lose dormancy through dry-afterripening. Patterns of germination response at dispersal vary among populations and sometimes across years within populations. To assess the relative contribution of genotype and maturation environment to this variation, we grew progeny of ten parental lines from each of six contrasting populations in a common greenhouse environment. We then tested the germination responses of recently harvested seeds of the putative full-sib progeny at five incubation temperatures. Significant germination response differences among populations were observed in greenhouse cultivation, and major differences among full-sib families were evident for some populations and traits. Among-population variation accounted for over 90% of the variance in each trait, while within-family variance accounted for 1% or less. Germination responses of greenhouse-grown progeny were positively correlated with the responses of wild-collected seeds, but there was a tendency for lowered dormancy at higher incubation temperatures. This tendency was more marked in populations from cold desert, foothill, and plains habitats, suggesting a genotype-maturation environment interaction. Differences among populations in the amount of among-family variance were more evident at lower incubation temperatures, while among-family variance was more uniformly low at summer incubation temperatures. Populations from predictable extreme environments (subalpine meadow and warm desert margin) showed significantly less variation among families than populations from less predictable cold desert, foothill, and plains environments. Low among-family variance was not specifically associated with small population size or marginality of habitat, as small marginal populations from unpredictable environments showed variance as high as that of large populations. In populations with high among-family variance for germination traits, germination responses tended to be correlated across incubation temperatures, making it possible to characterize families in terms of their general dormancy status. The results indicate that seed germination regulation in this species is probably under strong genetic control, and that habitats with temporally varying selection are occupied by populations that tend to be more polymorphic in terms of their germination response patterns.

Ecological aspects of seed dormancy loss

Advances in seed biology include progress in understanding the ecological significance of seed dormancy mechanisms. This knowledge is being used to make more accurate predictions of germination timing in the field. For several wild species whose seedlings establish in spring, seed populations show relevant variation that can be correlated with habitat conditions. Populations from severe winter sites, where the major risk to seedlings is frost, tend to have long chilling requirements or to germinate very slowly at low temperatures. Populations from warmer sites, where the major risk is drought, are non-dormant and germinate very rapidly under these same conditions. Seed populations from intermediate sites exhibit variation in dormancy levels, both among and within plants, which spreads germination across a considerable time period. For grasses that undergo dry after-ripening, seed dormancy loss can be successfully modelled using hydrothermal time. Dormancy loss for a seed population is associated with a progressive downward shift in the mean base water potential, i.e., the water potential below which half of the seeds will not germinate. Other parameters (hydrothermal time requirement, base temperature and standard deviation of base water potentials) tend to be constant through time. Simulation models for predicting dormancy loss in the field can be created by combining measurements of seed zone temperatures with equations that describe changes in mean base water potential as a function of temperature. Successful validation of these and other models demonstrates that equations based on laboratory data can be used to predict dormancy loss under widely fluctuating field conditions. Future progress may allow prediction of germination timing based on knowledge of intrinsic dormancy characteristics of a seed population and long-term weather patterns in the field.

Seed germination regulation in Bromus tectorum (Poaceae) and its ecological significance

Bromus tectorum is a winter annual grass that has become extensively naturalized in western North America. Its seeds are usually at least conditionally dormant at dispersal and lose dormancy through dry afterripening. Germination response to temperature for recently harvested seeds and rate of change in germination response during afterripening were examined for collections from 21 western North American populations representing a wide array of habitats. Analysis of variance showed highly significant among-population differences in germination response variables. Principal components analysis of 20 germination variables revealed groups of populations that could be characterized by distinct response syndromes. Degree of dormancy at summer temperatures in recently harvested seeds as well as rate of dormancy loss during dry storage could be related to the risk of premature summer germination in different habitats. Mojave Desert populations showed the most clearly differentiated response. Populations from Intermountain desert and foothill habitats showed intermediate responses and did not form distinct groups. Montane populations showed the widest variation. Fully afterripened seeds from all populations were nondormant and could germinate quickly across a wide temperature range. These results demonstrate the existence of adaptively significant variation in germination response. Such variation probably represents the beginning of genetic differentiation as a result of selection among and within founder populations. Lack of a consistent relationship with habitat reflects the stochastic nature of colonization and the fact that diverse germination strategies may permit persistence, especially in less extreme habitats.

Using hydrothermal time concepts to model seed germination response to temperature dormancy loss and priming effects in Elymus elymoides

Hydrothermal time (HTT) describes progress toward seed germination under various combinations of incubation water potential ( ) and temperature ( T ). To examine changes in HTT parameters during dormancy loss, seeds from two populations of the bunchgrass Elymus elymoides were incubated under seven temperature regimes following dry storage at 10, 20 and 30°C for intervals from 0 to 16 weeks. Fully after-ripened seeds were primed for 1 week at a range of s. Data on germination rate during priming were used to obtain a HTT equation for each seed population, while data obtained following transfer to water were used to calculate HTT accumulation during priming. HTT equations accurately predicted germination time course curves if mean base water potential, b ( 50 ), was allowed to vary with temperature. b ( 50 ) values increased linearly with temperature, explaining why germination rate does not increase with temperature in this species. b ( 50 ) showed a linear decrease as a function of thermal time in storage. Slopes for the T × b ( 50 ) relationship did not change during after-ripening. This thermal after-ripening time model was characterized by a single base temperature and a constant slope across temperatures for each collection. Because the difference between initial and final b ( 50 )s was uniform across tempera-tures, the thermal after-ripening requirement was also a constant. When seeds were primed for 1 week at −4 to −20 MPa, accumulation of HTT was a uniform 20% of the total HTT requirement. When primed at 0 to −4 MPa, HTT accumulation decreased linearly with decreasing priming potential, and a hydrothermal priming time model using a constant minimum priming potential adequately described priming effects. Use of these simple HTT relationships will facilitate modelling of germination phenology in the field.

A hydrothermal time model of seed after-ripening in Bromus tectorum L.

Bromus tectorum L. is an invasive winter annual grass with seeds that lose dormancy through the process of dry after-ripening. This paper proposes a model for after-ripening of B. tectorum seeds based on the concept of hydrothermal time. Seed germination time course curves are modelled using five parameters: a hydrothermal time constant, the fraction of viable seeds in the population, base temperature, mean base water potential and the standard deviation of base water potentials in the population. It is considered that only mean base water potential varies as a function of storage duration and incubation temperature following after-ripening. All other parameters are held constant throughout after-ripening and at all incubation temperatures. Data for model development are from seed germination studies carried out at four water potentials (0, −0.5, −1.0 and −1.5 MPa) at each of two constant incubation temperatures (15 and 25°C) following different storage intervals including recently harvested, partially after-ripened (stored for 4, 9 or 16 weeks at 20°C) and fully after-ripened (stored for 14 weeks at 40°C). The model was fitted using a repeated probit regression method, and for the two seed populations studied gave R 2 values of 0.898 and 0.829. Germination time course curves predicted by the model generally had a good fit when compared with observed curves at the incubation temperature/water potential treatment combinations for different after-ripening intervals. Changes in germination time course curves during after-ripening of B. tectorum can largely be explained by decreases in the mean base water potential. The simplicity and good fit of the model give it considerable potential for extension to simulation of after-ripening under field conditions.

A hydrothermal after-ripening time model for seed dormancy loss in Bromus tectorum L.

After-ripening, the loss of dormancy under dry conditions, is associated with a decrease in mean base water potential for germination of Bromus tectorum L. seeds. After-ripening rate is a linear function of temperature above a base temperature, so that dormancy loss can be quantified using a thermal after-ripening time (TAR) model. To incorporate storage water potential into TAR, we created a hydrothermal after-ripening time (HTAR) model. Seeds from two B. tectorum populations were stored under controlled temperatures (20 or 30 °C) and water potentials (−400 to −40 MPa). Subsamples were periodically removed from each storage treatment and incubated at 15 or 25 °C to determine germination time courses. Dormancy status (mean base water potential) was calculated from each time course using hydrothermal time equations developed for each seed collection. Seeds stored at −400 MPa did not after-ripen. At water potentials from −400 to −150 MPa, the rate of after-ripening increased approximately linearly with increasing water potential. Between −150 and −80 MPa, there was no further increase in after-ripening rate, while at −40 MPa seeds did not after-ripen and showed loss of vigour. These results suggest that the concept of critical water potential thresholds, previously shown to be associated with metabolic activity and desiccation damage in partially hydrated seeds, is also relevant to the process of after-ripening. The HTAR model generally improved field predictions of dormancy loss when the soil was very dry. Reduced after-ripening rate under such conditions provides an ecologically relevant explanation of how seeds prolong dormancy at high summer soil temperatures.

Ecology and ecological genetics of seed dormancy in downy brome

Abstract Downy brome, an obligately selfing winter annual, has invaded a variety of habitats in western North America. Seeds are at least conditionally dormant at dispersal in early summer and lose dormancy through dry after-ripening. In the field, patterns of germination response at dispersal vary among populations and across years within populations. Degree of dormancy at summer temperatures in recently harvested seeds, as well as rate of dormancy loss during dry storage, can be related to the risk of premature summer germination in different habitats. Patterns of dormancy loss are predictable and can be modeled using hydrothermal time concepts. To assess the relative contribution of genotype and maturation environment, multiple parental lines from contrasting populations were grown for three generations under manipulated greenhouse conditions. Significant germination response differences among populations were observed, as well as major differences among full-sib families. Among-population variation accounted for over 90% of the variance in germination traits, whereas within-family variance accounted for 1% or less. Populations from predictable extreme environments (subalpine meadow and warm desert margin) showed significantly less variation among families than did populations from less predictable environments (cold desert, foothill, and plains). Environmental conditions that shortened the seed ripening period (water stress and high temperature) resulted in reduced seed dormancy level at maturation, but there were strong inbred line–environment interactions. For fully after-ripened seeds, inbred line and environmental effects were no longer evident, indicating that differences in genotype and maturation environment function mainly to regulate dormancy and dormancy loss rather than to mediate response patterns of nondormant seeds. Nomenclature: Downy brome, Bromus tectorum L. BROTE.

Predicting seed dormancy loss and germination timing for Bromus tectorum in a semi-arid environment using hydrothermal time models

A principal goal of seed germination modelling for wild species is to predict germination timing under fluctuating field conditions. We coupled our previously developed hydrothermal time, thermal and hydrothermal afterripening time, and hydration‐dehydration models for dormancy loss and germination with field seed zone temperature and water potential measurements from early summer through autumn to develop predictions of germination timing for Bromus tectorum at a semi-arid site in north-central Utah, USA. Model predictions were tested with a validation dataset based on concomitant seed retrieval experiments in 2 years. Predictions were generally in agreement with observed field germination time courses, even though integration across multiple precipitation events was necessary. Success of the modelling effort hinged on two factors. First, we used a soil capacitance sensor that measured seed zone (5mm soil depth) water content accurately over a wide range. Second, simulations were built using physiologically based threshold models that can incorporate differences in germination timing for multiple germination fractions and for multiple stages of dormancy loss. Our results suggest that simulation models using hydrothermal time concepts can predict field germination phenology accurately. Seeds in this study integrated their experiences in a widely fluctuating environment in a manner consistent with the assumptions of hydrothermal time. Such threshold-based models also have the advantage of generality, as these concepts can be applied to many different species, environments and weather scenarios.

Environmental factors influencing Pyrenophora semeniperda-caused seed mortality in Bromus tectorum

Temperature and water potential strongly influence seed dormancy status and germination of Bromus tectorum. As seeds of this plant can be killed by the ascomycete fungus Pyrenophora semeniperda, this study was conducted to learn how water potential and temperature influence mortality levels in this pathosystem. Separate experiments were conducted to determine: (1) if P. semeniperda can kill dormant or non-dormant seeds across a range of water potentials (0 to 22 MPa) at constant temperature (208C); and (2) how temperature (5–208C) and duration at reduced water potentials (0–28 days) affect the outcome. When inoculated with the fungus at 208C, all dormant seeds were killed, but fungal stromata appeared more quickly at higher water potentials. For non-dormant seeds, decreasing water potentials led to reduced germination and greater seed mortality. Results were similar at 10 and 158C. Incubation at 58C prevented stromatal development on both non-dormant and dormant seeds regardless of water potential, but when seeds were transferred to 208C, dormant seeds evidenced high mortality. For non-dormant seeds, exposure to low water potential at 58C resulted in secondary dormancy and increased seed mortality. Increasing incubation temperature, decreasing water potential and increasing duration at negative water potentials all led to increased mortality for non-dormant seeds. The results are consistent with field observations that pathogen-caused mortality is greatest when dormant seeds imbibe, or when non-dormant seeds experience prolonged or repeated exposure to low water potentials. We propose a conceptual model to explain the annual cycle of interaction in the Bromus tectorum–Pyrenophora semeniperda pathosystem.

Fire Effects on the Cheatgrass Seed Bank Pathogen Pyrenophora semeniperda

Abstract The generalist fungal pathogen Pyrenophora semeniperda occurs primarily in cheatgrass (Bromus tectorum) seed banks, where it causes high mortality. We investigated the relationship between this pathogen and its cheatgrass host in the context of fire, asking whether burning would facilitate host escape from the pathogen or increase host vulnerability. We used a series of laboratory and field experiments to address the ability of host seeds and pathogen life stages to survive fire. First, we determined the thermal death point (TDP50; temperature causing 50% mortality) of seeds and pathogen propagules at two time intervals using a muffle furnace. We then measured peak fire temperatures in prescribed burns at sites in Utah and Washington and quantified seed and fungal propagule survival using pre- and postburn seed bank sampling and inoculum bioassays. Finally, we investigated the survival of both seeds and pathogen after wildfires. We found that radiant heat generated by both prescribed and wild cheatgrass monoculture fires was generally not sufficient to kill either host seeds or pathogen propagules; most mortality was apparently due to direct consumption by flames. The 5-min mean TDP50 was 164°C for pathogen propagules and 148°C for host seeds, indicating that the pathogen is more likely to survive fire than the seeds. Peak fire temperature at the surface in the prescribed burns averaged 130°C. Fire directly consumed 85–98% of the viable seed bank, but prescribed burns and wildfires generally did not lead to dramatic reductions in pathogen inoculum loads. We conclude that the net effect of fire on this pathosystem is not large. Rapid postburn recovery of both host and associated pathogen populations is the predicted outcome. Postfire management of residual cheatgrass seed banks should be facilitated by the persistent presence of this seed bank pathogen.