Climatic Refugia and Geographical Isolation Contribute to the Speciation and Genetic Divergence in Himalayan-Hengduan Tree Peonies (Paeonia delavayi and Paeonia ludlowii)

Himalaya and Hengduan Mountains (HHM) is a biodiversity hotspot, and very rich in endemic species. Previous phylogeographical studies proposed different hypotheses (vicariance and climate-driven speciation) in explaining diversification and the observed pattern of extant biodiversity, but it is likely that taxa are forming in this area in species-specific ways. Here, we reexplored the phylogenetic relationship and tested the corresponding hypotheses within Paeonia subsect. Delavayanae composed of one widespread species (Paeonia delavayi) and the other geographically confined species (Paeonia ludlowii). We gathered genetic variation data at three chloroplast DNA fragments and one nuclear gene from 335 individuals of 34 populations sampled from HHM. We performed a combination of population genetic summary statistics, isolation-with-migration divergence models, isolation by environment, and demographic history analyses. We found evidence for the current taxonomic treatment that P. ludlowii and P. delavayi are two different species with significant genetic differentiation. The significant isolation by environment was revealed within all sampled populations but genetic distances only explained by geographical distances within P. delavayi populations. The results of population divergence models and demographic history analyses indicated a progenitor–derivative relationship and the Late Quaternary divergence without gene flow between them. The coalescence of all sampled cpDNA haplotypes could date to the Late Miocene, and P. delavayi populations probably underwent a severe bottleneck in population size during the last glacial period. Genetic variation in Paeonia subsect. Delavayanae is associated with geographical and environmental distances. These findings point to the importance of geological and climatic changes as causes of the speciation event and lineage diversification within Paeonia subsect. Delavayanae.

Morphological and Biochemical Studies of the Yellow and Purple–red Petal Pigmentation in Paeonia delavayi

Paeonia delavayi is a species endemic to Southwest China and an important genetic resource for flower color breeding of tree peonies. The mechanisms underlying the flower coloration of this plant have not been fully elucidated. In this article, the petals of yellow-colored individual (Pl) and purple–red-colored individual (Pd) of P. delavayi were studied. And anatomical observations revealed that a large amount of yellow protoplasts and a small amount of colorless protoplasts were located in the yellow-colored Pl petals, whereas a mixture of purple, red, and pink protoplasts were observed in the purple–red-colored Pd petals. The Pl cells were subrotund and flat, whereas the Pd cells were irregularly polygon-shaped and bulging. Chemical analyses were performed, and the results indicated that significant differences occurred between the cell sap pH of the Pl and Pd flowers and large differences occurred in the contents of Fe and Al between Pl and Pd. Cyanidin- and peonidin-based anthocyanins with flavones and flavonols as copigments determined the Pd flower color, whereas chalcone 2 ′G with apigenin 7-O-neohesperidoside and chrysoeriol 7-O-glucoside as copigments determined the yellow color of Pl. Correspondingly, the genes dihydroflavonol 4-reductase (DFR) and anthocyanidin synthase (ANS) were significantly highly expressed in Pd, whereas chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), flavone synthase (FNS), flavonol synthase (FLS), flavonoid 7-O-glycosyltransferase (7GT), and 2′4′6′4-tetrahydroxychalcone 2′-glucosyltransferase (THC) had high transcript levels in Pl relative to Pd. The results indicate that the color variation of P. delavayi petals may be related to a delicately controlled balance of the aforementioned factors.