Cytological Characterization of Arabidopsis arenosa Polyploids by SIM
Arabidopsis arenosa has recently become established as a model organism for investigating how meiosis has evolved to overcome the meiotic challenges faced by newly formed autotetraploids. Here, we describe a protocol for the preparation of spread, immunolabeled prophase I chromosomes from established A. arenosa autotetraploids for imaging with three-dimensional structured illumination microscopy (3D-SIM). This technique allows us to dissect the unique synaptic behavior in A. arenosa and identify synaptic partner switch sites that are unresolvable with conventional widefield microscopy.
Key wordsArabidopsis arenosa Meiosis Synaptonemal complex Synaptic partner switch Structured illumination microscopy
We thank Dr. Kirsten Bomblies (John Innes Centre) for supporting the development of this protocol and Prof. Chris Franklin (University of Birmingham) for kindly supplying antibodies. This work was supported by a European Research Council Consolidator grant to K.B. (CoG EVO-MEIO 681946) and a UK Biological and Biotechnology Research Council (BBSRC) studentship to C.M. (DTP BB/MO1116 x/1 M1BTP). This work has also been supported by the BBSRC via grant BB/P013511/1 to the John Innes Centre.
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