Sexually different morphological, physiological and molecular responses of Fraxinus mandshurica flowers to floral development and chilling stress.

Fraxinus mandshurica is considered a dioecious hardwood, and the temporal separation of the maturation of the male and female flowers is one reason that F. mandshurica has become an endangered species in China. Rainfall and low temperature influence pollen formation and dispersal and the blooming of female flowers. Therefore, low fertilization efficiency strongly influences the population of F. mandshurica. Nevertheless, few studies have investigated the sex-specific morphological, physiological and molecular differentiation of F. mandshurica during flowering and its responses to low temperature. In this study, we investigated the sexual differences in the morphological, physiological, and biochemical parameters of F. mandshurica during flowering and determined the physiological and biochemical parameters and expression levels of related genes in response to low-temperature stress induced by exposure to 4 °C (chilling stress) during pollen dispersal and fertilization. Our study supports the hypothesis that male flowers suffer more severe injuries while female flowers are more adaptable to environmental stress during flower development in F. mandshurica. The results showed higher physiological and biochemical levels of malondialdehyde, proline, and soluble sugar, as well as the expression of genes involved in calcium signaling, cold shock and DNA methylation in male flowers compared with female flowers, which suggested that male flowers suffer from more serious peroxidation than female flowers. In contrast, higher antioxidant capacity and FmaCAT expression were detected in female flowers, providing preliminary evidence that male flowers rapidly fade after pollination and further demonstrating that female flowers need a much stronger antioxidant enzyme system to maintain embryonic growth. Most peaks related to physiological and molecular responses were observed at 2-4 h and 8-10 h of exposure to chilling stress in the female and male flowers, respectively. This trend implies that female flowers have higher adaptability to low temperature during fertilization.