Evolutionary breakdown from rigorous outbreeding to self‐fertilization frequently occurs in angiosperms. Since the pollinators are not necessary, self‐compatible populations often reduce investment in floral display characteristics and pollination reward. Primula forbesii is a biennial herb with distribution restricted to southwest China; it was initially a self‐incompatible distylous species, but after 20 years of artificial domestication, homostyly appeared. This change in style provides an ideal material to explore the time required for plant mating systems to adapt to new environmental changes and test whether flower attraction has reduced following transitions to selfing. We did a survey in wild populations of P. forbesii where its seeds were originally collected 20 years ago and recorded the floral morph frequencies and morphologies. The floral morphologies, self‐incompatibility, floral scent, and pollinator visitation between distyly and homostyly were compared in greenhouse. Floral morph frequencies of wild populations did not change, while the cultivated population was inclined to L‐morph and produced homostyly. Evidence from stigma papillae and pollen size supports the hypothesis that the homostyly possibly originated from mutations of large effect genes in distylous linkage region. Transitions to self‐compatible homostyly are accompanied by smaller corolla size, lower amounts of terpenoids, especially linalool and higher amounts of fatty acid derivatives. The main pollinators in the greenhouse were short‐tongued Apis cerana. However, homostyly had reduced visiting frequency. The mating system of P. forbesii changed rapidly in just about 20 years of domestication, and our findings confirm the hypothesis that the transition to selfing is accompanied by decreased flower attraction.

Soil pollution caused by the accumulation of potentially toxic elements (PTEs) has become a serious problem due to industrial and urban pollution as well as pesticide use. Primula forbesii Franch. is a biennial flower native to China with excellent stress resistance and ornamental value. In this study, we examined the phenotypic traits, growth indexes, and physiological properties of P. forbesii in response to five representative PTEs (Cd, Ni, Cr, Cu, and Zn) under hydroponic culture conditions. High concentrations of Zn and Cr had little effect on the growth and physiological properties of P. forbesii, indicating that the species has strong tolerance to Zn and Cr stress. Alternatively, high concentrations of Cd, Ni, and Cu seriously affected plant growth and development, resulting in leaf chlorosis and even death, and therefore may have a serious negative impact on the growth of P. forbesii. However, activity levels of some antioxidant enzymes and osmotic regulatory substances remained high, indicating that P. forbesii resisted heavy metal stress by regulating physiological and biochemical metabolism to a certain extent. Furthermore, principal component analysis and membership function were used to comprehensively evaluate P. forbesii resistance to PTEs. These analyses revealed that P. forbesii exhibits distinct sensitivities and physiological responses to different PTEs and suggested that the resistance to five PTEs in decreasing order is Zn > Cr > Cd > Cu > Ni. These results provide a theoretical basis for the future application of P. forbesii in environments with heavy metal pollution and may expand its practical utilization.

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Primula forbesii Franch. is a unique biennial herb with a strong floral fragrance, making it an excellent material for studying the aroma characteristics of the genus Primula. The floral scent is an important ornamental trait that facilitates fertilization. However, the molecular mechanism regulating the floral scent in Primula is unknown. In order to better understand the biological mechanisms of floral scents in this species, this study used RNA sequencing analysis to discuss the first transcriptome sequence of four flowering stages of P. forbesii, which generated 12 P. forbesii cDNA libraries with 79.64 Gb of clean data that formed 51,849 unigenes. Moreover, 53.26% of the unigenes were annotated using public databases. P. forbesii contained 44 candidate genes covering all known enzymatic steps for the biosynthesis of volatile terpenes, the major contributor to the flower’s scent. Finally, 1-deoxy-d-xylulose 5-phosphate synthase gene of P. forbesii (PfDXS2, MK370094), the first key enzyme gene in the 2-c-methyl-d-erythritol 4-phosphate (MEP) pathway of terpenoids, was cloned and functionally verified using virus-induced gene silencing (VIGs). The results showed that PfDXS2-silencing significantly reduced the relative concentrations of main volatile terpenes. This report is the first to present molecular data related to aroma metabolites biosynthesis pathways and the functional characterization of any P. forbesii gene. The data on RNA sequencing provide comprehensive information for further analysis of other plants of the genus Primula.

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