reliable-method-for-assessing-seed-germination-dormancy-mortality

Reliable Method for Assessing Seed Germination, Dormancy, and Mortality under Field Conditions

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester ...

Ohio State University

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.

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

• Premise of the study: The fitness of an offspring may depend on its nuclear genetic composition (via both parental genotypes) as well as on genetic maternal effects (via only the maternal parent). Understanding the relative importance of these two genetic factors is particularly important for research on crop-wild hybridization, since traits with important genetic maternal effects (e.g., seed size) often differ among crops and their relatives. We hypothesized that the effects of these genetic factors on fitness components would change across the life cycle of hybrids.• Methods: We followed seed, plant size, and reproductive traits in field experiments with wild and four crop-wild hybrids of sunflower (Helianthus annuus), which differed in nuclear genetic composition and maternal parent (wild or F1 hybrid).• Key results: We identified strong genetic maternal effects for early life cycle characteristics, with seeds produced on an F1 mother having premature germination, negligible seed dormancy, and greater seedling size. Increased percentages of crop alleles also increased premature germination and reduced dormancy in seeds produced on a wild mother. For mature plants, nuclear genetic composition dominated: greater percentages of crop alleles reduced height, branching, and fecundity.• Conclusions: Particular backcrosses between hybrids and wilds may differentially facilitate movement of crop alleles into wild populations due to their specific features. For example, backcross seeds produced on wild mothers can persist in the seed bank, illustrating the importance of genetic maternal effects, whereas backcross individuals with either wild or F1 mothers have high fecundity, resulting from their wild-like nuclear genetic composition.

Roles of maternal effects and nuclear genetic composition change across the life cycle of crop-wild hybrids.

Understanding the likelihood and extent of introgression of novel alleles in hybrid zones requires comparison of lifetime fitness of parents and hybrid progeny. However, fitness differences among cross types can vary depending on biotic conditions, thereby influencing introgression patterns. Based on past work, we predicted that increased competition would enhance introgression between cultivated and wild sunflower (Helianthus annuus) by reducing fitness advantages of wild plants. To test this prediction, we established a factorial field experiment in Kansas, USA where we monitored the fitness of four cross types (Wild, F1, F2, and BCw hybrids) under different levels of interspecific and intraspecific competition. Intraspecific manipulations consisted both of density of competitors and of frequency of crop-wild hybrids. We recorded emergence of overwintered seeds, survival to reproduction, and numbers of seeds produced per reproductive plant. We also calculated two compound fitness measures: seeds produced per emerged seedling and seeds produced per planted seed. Cross type and intraspecific competition affected emergence and survival to reproduction, respectively. Further, cross type interacted with competitive treatments to influence all other fitness traits. More intense competition treatments, especially related to density of intraspecific competitors, repeatedly reduced the fitness advantage of wild plants when considering seeds produced per reproductive plant and per emerged seedling, and F2 plants often became indistinguishable from the wilds. Wild fitness remained superior when seedling emergence was also considered as part of fitness, but the fitness of F2 hybrids relative to wild plants more than quadrupled with the addition of interspecific competitors and high densities of intraspecific competitors. Meanwhile, contrary to prediction, lower hybrid frequency reduced wild fitness advantage. These results emphasize the importance of taking a full life cycle perspective. Additionally, due to effects of exogenous selection, a given hybrid generation may be especially well-suited to hastening introgression under particular environmental conditions.

Fitness of crop-wild hybrid sunflower under competitive conditions: implications for crop-to-wild introgression.

Domestication has resulted in selection upon seed traits found in wild populations, yet crop-wild hybrids retain some aspects of both parental phenotypes. Seed fates of germination, dormancy, and mortality can influence the success of crop allele introgression in crop-wild hybrid zones, especially if crop alleles or crop-imparted seed coverings result in out-of-season germination. We performed a seed burial experiment using crop, wild, and diverse hybrid sunflower (Helianthus annuus) cross types to test how a cross type's maternal parent and nuclear genetic composition might affect its fate under field conditions. We observed higher maladaptive fall germination in the crop- and F1- produced seeds than wild-produced seeds and, due to an interaction with percent crop alleles, fall germination was higher for cross types with more crop-like nuclear genetics. By spring, crop-produced cross types had the highest overwintering mortality, primarily due to higher fall germination. Early spring germination was identical across maternal types, but germination continued for F1-produced seeds. In conclusion, the more wild-like the maternal parent or the less proportion of the cross type's genome contributed by the crop, the greater likelihood a seed will remain ungerminated than die. Wild-like dormancy may facilitate introgression through future recruitment from the soil seed bank.

Article	Year
The influence of fertilizer level and spore density on arbuscular mycorrhizal colonization of transgenic Bt 11 maize (Zea mays) in experimental microcosms. FEMS microbiology ecology\| PubMed ID: 21198682	2011
Roles of maternal effects and nuclear genetic composition change across the life cycle of crop-wild hybrids. American journal of botany\| PubMed ID: 25016007	2014
Fitness of crop-wild hybrid sunflower under competitive conditions: implications for crop-to-wild introgression. PloS one\| PubMed ID: 25295859	2014
Seed fates in crop-wild hybrid sunflower: crop allele and maternal effects. Evolutionary applications\| PubMed ID: 25685189	2015

Brian A. Pace

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