Abstract
Meiotic homologous recombination plays a central role in creating genetic variability, making it an essential biological process relevant to evolution and crop breeding. In this study, we used pollen-specific fluorescent tagged lines (FTLs) to measure male meiotic recombination frequency during the development of Arabidopsis thaliana. Interestingly, a subset of pollen grains consistently shows loss of fluorescence expression in tested lines. Using nine independent FTL intervals, the spatio-temporal dynamics of male recombination frequency was assessed during plant development, considering both shoot type and plant age as independent parameters. In most genomic intervals assayed, male meiotic recombination frequency is highly consistent during plant development, showing no significant change between different shoot types and during plant aging. However, in some genomic regions, such as I1a and I5a, a small but significant effect of either developmental position or plant age were observed, indicating that the meiotic CO frequency in those intervals varies during plant development. Furthermore, from an overall view of all nine genomic intervals assayed, both primary and tertiary shoots show a similar dynamics of increasing recombination frequency during development, while secondary and lateral shoots remain highly stable. Our results provide new insights in the dynamics of male meiotic recombination frequency during plant development.
Highlights
Meiotic homologous recombination plays a central role in creating genetic variability, making it an essential biological process relevant to evolution and crop breeding
fluorescent tagged lines (FTLs)-based quantification of meiotic recombination frequency in Arabidopsis relies on the segregation analysis of two or three linked hemizygous fluorescent markers (DsRed, eYFP and eCFP) in mature pollen grains
In order to assess putative losses or biases in pollen fluorescence of the widely used FTL marker combinations, we monitored DsRed, eYFP and eCFP expression in tetrad configured pollen grains isolated from quartet1-2−/− plants containing homozygous FTL marker combinations
Summary
Meiotic homologous recombination plays a central role in creating genetic variability, making it an essential biological process relevant to evolution and crop breeding. Francis et al.[10] developed a series of pollen-specific fluorescent markers (FTLs; fluorescent tagged lines) in the model plant Arabidopsis thaliana, that upon combination in a hemizygous state, allow for high-throughput quantification of male meiotic recombination in corresponding genomic intervals[11,12]. FTL-based quantification of meiotic recombination frequency can be performed based on the fluorescent analysis of single pollen grains by flow cytometry (Figure S1) Using this methodology, data acquisition speed can be increased up to 150-fold compared to tetrad analysis, allowing rapid quantification of male meiotic CO rates[12]. We use nine independent FTLs intervals (I1a, I1b, I2a, I2b, I3c, CEN3, I5a, I5c and I5d) to measure the spatio-temporal dynamics of male CO frequency in flowers isolated from primary, secondary, tertiary and lateral shoots, in order to provide an extensive overview of the spatio-temporal dynamics of male meiotic recombination frequency in the genomic intervals delineated by a set of widely used pollen FTL markers
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