Flow cytometric seed screen data are consistent with models of chromosome inheritance in asymmetrically compensating allopolyploids

Angiosperms have evolved a mechanism of double fertilization, which results in the production of a separate embryo (new individual) and endosperm (nutritive tissue). The flow cytometric seed screen (FCSS) was developed to infer plant reproduction modes based on endosperm-to-embryo DNA content ratio (P-ind). A ratio of 1.5 indicates sexual reproduction, whereas higher values of 2.0 are consistent with apomixis. Although FCSS has been successfully applied to the study of sexual and asexual plants, the limits of FCSS and particularly its potential for determination of reproduction modes in hemisexual plants have not been explored. Here, we evaluated the application of FCSS to the study of reproduction modes in two asymmetrically compensating allopolyploids (ACAs), Onosma arenaria and Rosa canina. These two species are characterized by the presence of asexually inherited univalent-forming and sexually inherited bivalent-forming chromosome sets. They both use asymmetric meiosis, which eliminates univalent-forming chromosome sets from the male gamete and retains them in the female gamete. Different chromosomal behavior in male and female meiosis in these plants is reflected in different theoretically derived P-ind values, which deviate from a sexual 1.5 value. Here, we determined P-ind FCSS-based values in seeds of ACAs, and compared the results to sexual species. As expected, we determined that the mean P-ind is 1.51, 1.52, and 1.52 in the sexual plants, that is, Capsella bursa-pastoris, Crataegus monogyna, and O. pseudoarenaria, respectively. In the ACAs, different mean P-ind values were determined for O. arenaria (1.61) and R. canina (1.82). These values are consistent with the theoretical P-ind values determined based on models of chromosome inheritance. This study highlights the precision of flow cytometry in determining DNA content and it's utility in screening reproduction modes. Additionally, it advocates for more in-depth investigations into rapid screening of accessions where the P-ind ratio has deviated from the 1.5 value typical of sexual species, which may indicate meiotic irregularities.