Plasticity in floral longevity and sex-phase duration of Lobelia siphilitica in response to simulated pollinator declines.

PREMISE: Pollinator declines can reduce the quantity and quality of pollination services, resulting in less pollen deposited on flowers and lower seed production by plants. In response to these reductions, plant species that cannot autonomously self-pollinate and thus are dependent on pollinators to set seed could plastically adjust their floral traits. Such plasticity could increase the opportunity for outcross pollination directly, as well as indirectly by affecting inflorescence traits. METHODS: To test whether plants can respond to pollinator declines by plastically adjusting their floral traits, we simulated declines by experimentally reducing pollinator access to Lobelia siphilitica plants and measuring two traits of early- and late-season flowers: (1) floral longevity; and (2) sex-phase duration. To test whether plasticity in these floral traits affected inflorescence traits, we measured daily display size and phenotypic gender. RESULTS: We found that experimentally reducing pollination did not affect female-phase duration, but did extend the male-phase duration of early-season flowers by 13% and the longevity of late-season flowers by 12.8%. However, plants with an extended male phase did not have a more male-biased phenotypic gender, and plants with an extended floral longevity did not have a larger daily display. CONCLUSIONS: Our results suggest that plants can respond to pollinator declines by plastically adjusting both the longevity and sex-phase duration of their flowers. If this plasticity increases the opportunity for outcross pollination, then it could be one mechanism by which pollinator-dependent plant species maintain seed production as pollinators decline.

Physiological effects of temperature do not explain prevalence of females in populations of gynodioecious Lobelia siphilitica growing in warmer climates.

PREMISE OF THE STUDY: Gynodioecy is a sexual polymorphism whereby female and hermaphroditic plants co-occur within populations. In many gynodioecious species, stressful abiotic environments are associated with higher frequencies of females. This association suggests that abiotic stress affects the relative fitness of females and hermaphrodites and, thus, the maintenance of gynodioecy. METHODS: To test whether abiotic stress affects the fitness of females and hermaphrodites, we grew open-pollinated Lobelia siphilitica families in temperature regimes characteristic of the southern portion of the species' range (where females are common) and the northern portion of the range (where females are rare). We measured physiological and phenological traits that are indicative of heat stress, and fitness components of females and hermaphrodites that could affect the maintenance of gynodioecy. KEY RESULTS: Contrary to expectations if growth at high temperatures is stressful, we found that the hot treatment increased leaf chlorophyll content, decreased the percentage of plants that delayed flowering initiation, and did not affect the quantum efficiency of photosystem II. Growth at high temperatures did not affect the magnitude of the difference in rosette size (a correlate of flower number) between females and hermaphrodites, or the variance in pollen viability among hermaphrodites. CONCLUSIONS: We found that growing-season temperatures typical of high female L. siphilitica populations were not stressful and did not affect either the fitness of females compared to hermaphrodites or variation in fitness among hermaphrodites. Consequently, further research is necessary to explain correlations between abiotic environmental factors and the frequency of females in this and other gynodioecious species.

Frequency-dependent fitness in gynodioecious Lobelia siphilitica.

Selection is frequency dependent when an individual's fitness depends on the frequency of its phenotype. Frequency-dependent selection should be common in gynodioecious plants, where individuals are female or hermaphroditic; if the fitness of females is limited by the availability of pollen to fertilize their ovules, then they should have higher fitness when rare than when common. To test whether the fitness of females is frequency dependent, we manipulated the sex ratio in arrays of gynodioecious Lobelia siphilitica. To test whether fitness was frequency dependent because of variation in pollen availability, we compared open-pollinated and supplemental hand-pollinated plants. Open-pollinated females produced more seeds when they were rare than when they were common, as expected if fitness is negatively frequency dependent. However, hand-pollinated females also produced more seeds when they were rare, indicating that variation in pollen availability was not the cause of frequency-dependent fitness. Instead, fitness was frequency dependent because both hand- and open-pollinated females opened more flowers when they were rare than when they were common. This plasticity in the rate of anthesis could cause fitness to be frequency dependent even when reproduction is not pollen limited, and thus expand the conditions under which frequency-dependent selection operates in gynodioecious species.

Testing models of sex ratio evolution in a gynodioecious plant: female frequency covaries with the cost of male fertility restoration.

In many gynodioecious species, cytoplasmic male sterility genes (CMS) and nuclear male fertility restorers (Rf) jointly determine whether a plant is female or hermaphrodite. Equilibrium models of cytonuclear gynodioecy, which describe the effect of natural selection within populations on the sex ratio, predict that the frequency of females in a population will primarily depend on the cost of male fertility restoration, a negative pleiotropic effect of Rf alleles on hermaphrodite fitness. Specifically, when the cost of restoration is higher, the frequency of females at equilibrium is predicted to be higher. To test this prediction, we estimated variation in the cost of restoration across 26 populations of Lobelia siphilitica, a species in which Rf alleles can have negative pleiotropic effects on pollen viability. We found that L. siphilitica populations with many females were more likely to contain hermaphrodites with low pollen viability. This is consistent with the prediction that the cost of restoration is a key determinant of variation in female frequency. Our results suggest that equilibrium models can explain variation in sex ratio among natural populations of gynodioecious species.

A novel approach to estimating the cost of male fertility restoration in gynodioecious plants.

*In many gynodioecious plants, sex is determined by cytoplasmic male sterility genes (CMS) and nuclear male fertility restorers (Rf). Models predict that the costs of restoration are important determinants of population sex ratios. However, current approaches to the estimation of these costs require prior identification of CMS genotypes, information that is available for few species. *We tested a novel approach to estimating the cost of restoration in natural populations without determining CMS or Rf genotypes. We used estimates of pollen viability and offspring sex ratios from open- and hand-pollinated families of Lobelia siphilitica to test whether the cost of restoration, expressed as low pollen viability, is higher in populations with more females. *Among populations with CMS, we found that variation in pollen viability was higher in small populations with more females, as expected if the proportion of females within populations increases with the maximum cost of restoration. In controlled crosses, families with low pollen viability also produced fewer females, suggesting that variation in viability is primarily determined by the number and frequency of Rf alleles carried. *This approach to estimating the cost of restoration can be applied to other cytonuclear gynodioecious species, offering new opportunities for testing gynodioecy models in the wild.