Costs and benefits of self-fertilization in the cleistogamous perennial Ruellia humilis

The degree of self-fertilization in a population determines levels of genetic variation and high selfing rates could thus limit future adaptive potential. Theory predicts that intermediate selfing rates should not persist, yet many plants exhibit mixed mating. Cleistogamy is a floral heteromorphism where individuals produce both showy potentially outcrossing chasmogamous flowers and closed obligately selfing cleistogamous flowers. Reproduction via cleistogamous flowers is thought to be beneficial because of their greater energetic economy compared to chasmogamous flowers but can be costly if selfing leads to inbreeding depression or accelerates the fixation of deleterious mutations within populations. Cleistogamy has evolved independently multiple times and can be used to study the maintenance of adaptive mixed mating. To investigate this, I estimated the costs and benefits of selfing in three populations of Ruellia humilis Nutt (Acanthaceae) in greenhouse common garden experiments. To quantify the costs, I performed hand pollinations and quantified fitness components of progeny resulting from selfing and outcrossing within- and between-populations. To quantify the relative energetic advantage of cleistogamous flowers, I measured dry flower mass, fertility, seed number per fruit, and pollen-ovule ratios of both types of flowers. I found negative cumulative inbreeding depression in all populations, indicative of selfed progeny outperforming outcrossed progeny. While the fitness consequences of between population outcrossing ranged from heterosis to outbreeding depression. When looking at the energetic benefits of selfing, I found that the cost of reproduction via cleistogamous flowers was between 3 and 14-fold less than the cost for outcrossing flowers. Finally, I combined data on inbreeding depression and the energetic costs of reproduction and found that chasmogamous flowers of R. humilis must provide between a 3 to a 45-fold fitness advantage to be maintained, the magnitude of which was dependent upon maternal population. Overall, I conclude that none of the existing hypotheses are sufficient enough to provide the selective advantage needed to explain the persistence of chasmogamous flowers in R. humilis. Without any supported explanations for the maintenance of mixed mating, the exploration of genetic constraints on the loss of chasmogamous flowers could solve this long-standing mystery.