What ecological factors favor the shift from distyly to homostyly? A study from the perspective of reproductive assurance

Aims Distyly is one of the most widespread floral polymorphisms pro-moting cross-fertilization.Evolutionary transition from obligate cross-fertilized distyly to predominantly self-fertilized homostyly is frequently documented in various groups.However,empirical stud-ies concerning the ecological factors connected with this transition are still lacking.Primula chungensis,suggested to be evolving from distyly to homostyly,provides an ideal model for the study of the ecological factors concerned with this transition.We study P.chun-gensis to understand if autonomous self-fertilization would provide reproductive assurance for the self-fertilized homo-styled morph in the field.Methods the incompatibility features of P.chungensis were tested with hand-pollination experiments.We compared the capacity of autonomous self-fertilization between the distylous and homo-styled morph of P.chungensis in the field by excluding the pollinators with bags.In addition,the degrees of herkogamy of some P.chungensis plants were between the short-styled and homo-styled morphs.these plants were studied to understand whether they were able to obtain greater reproductive assurance when the herkogamy in the flowers was reduced.Important Findings All three morphs of P.chungensis were highly self-and intra-morph compatible.the degree of herkogamy positively correlated with the capacity for autonomous self-fertilization.A negative correlation between the degree of herkogamy and the magnitude of pollen limitation was found,but no significant correlation was found between the degree of herkogamy and the contribution of cross-fertilization to overall fertilization.this study suggests that reducing the degree of herkogamy can significantly increase the reproductive assurance for a self-compatible plant.Our results provided evidence that the homo-styled morph of P.chungensis had the highest capacity for autonomous self-fertilization and the highest seed production in the field,because autonomous self-fertilization provided reproduct-ive assurance for the homo-styled morph.this may cause selection towards the transition from distyly to homostyly.

Characterization of 24 microsatellite markers in Primula chungensis(Primulaceae),a distylous-homostylous species,using MiSeq sequencing

Primula chungensis is a species with considerable floral and mating-system variation,including distylous(outcrossing),homostylous(selfing) and mixed populations that contain both outcrossing and selfing forms.We isolated 24 microsatellite markers from P.chungensis using Illumina Mi Seq sequencing.Polymorphism and genetic diversity were then measured based on a sample of 24 individuals from a natural population in southern Tibet.All loci were polymorphic with the number of alleles per locus ranging from 2 to 4.The observed and expected heterozygosity ranged from 0 to 1 and 0.219 to 0.708,respectively.The microsatellite markers we have identified will serve as valuable tools for the investigation of the population genetic structure and phylogeography of P.chungensis and will inform models of the evolutionary history of mating systems in the species.

Fitness-related selection on floral traits in the distylous plant Primula poissonii (Primulaceae)

Aims Reproductive fitness of different floral phenotypes varies within and/or among populations.These variations are important to understand the process of natural selection and the evolution of floral traits.In this study,we focused on a distylous,self-incompat-ible species,Primula poissonii,to investigate fitness-related selec-tion on floral traits.Our aim was to determine how traits vary as targets of natural selection and whether morph-specific selection occurs.Methods This study was conducted at two sites(Yushuizhai at 2700 m and Haligu at 3200 m)in the Lijiang Alpine Botanical Garden,northwest-ern Yunnan,southwestern China.Insects visiting flowers of P.pois-sonii were observed,captured and identified.Randomly selected plants of long-and short-styled morphs were labeled.Five floral/inflorescence traits were measured including floral display,corolla width(CW),floral tube length(FTL),tube opening width(TOW)and floral scape height.Fruit and seed set were recorded.The total num-ber of seeds per individual plant(plant fitness)and seed production per capsule(flower fitness)were calculated.Multiple regression analyses were used to quantify selection gradients.Important Findings The frequencies of the two morphs did not deviate from the expected 1:1 ratio at both sites.Except for FTL,the four other traits did not dif-fer significantly between the long-and short-styled morphs.Floral scape height,floral display and FTL differed between two sites.The selection regimes differed between two morphs and between two sites.At the Yushuizhai site,linear selection for shorter floral tubes was stronger in the short-styled morph.However,nonlinear selec-tion on the floral display was stronger in the long-styled morph than selection on the short-styled morph.At the Haligu site,linear selec-tion for a smaller corolla was stronger in the long-styled morph.A morph-specific nonlinear selection on CW and floral display was also detected.Morph-specific selections were detected through the estimation of flower fitness only in Haligu population.In this site,morph-specific linear selection was also detected for CW and floral display.Morph-specific nonlinear selection on traits was detected only in CW.We found that butterflies and sphingid moths dominated at Yushuizhai,while long-tongued bees dominated at Haligu.The difference in pollinator fauna suggested that selection on floral tubes may be due to differences in pollinator assemblages.Overall,variation of floral and/or inflorescence traits in P.poissonii was probably driven by pollinator selection.Selection regime dif-ferences between two morphs,in part,due the inter-morph diver-gences of sexual functions in distylous plant.