Hartung, Frank

Sprink, Thorben

English

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Nachwuchswissenschaftlerforum / Young Scientists Meeting 2014 Sprink & Hartung     Initiation of meiotic double strand breaks in Arabidopsis depends on two different SPO11 proteins   Thorben Sprink1 and Frank Hartung1 1 Julius Kühn-Institut, Institute for Biosafety in Plant Biotechnology Email of corresponding author: thorben.sprink@jki.bund.de  Pairing and balanced distribution of allelic chromosomes during meiosis depends in most organisms on the initi-ation of double strand breaks (DSBs) by the protein SPO11. SPO11 is an evolu-tionary conserved meiotic transesterase, which introduces DSBs to the DNA during early prophase I of meiosis. Whereas in animals and fungi only one single SPO11 is present, plants have at least two meiotic active SPO11 proteins (SPO11-1 and SPO11-2) which are essential for proper chromosome distribution and recombination pro-cesses. Single knock out mutants of SPO11-1 as well as SPO11-2 are nearly sterile due to random chromosome segregation during meiosis.   In all so far sequenced land plants orthologous genes to Arabidopsis SPO11-1 and SPO11-2 can be found. Our aim is to investigate whether the function of SPO11-1 and -2 is species specific or interchangeable between different near or far related species.  We were able to show that it is, in some cases, possible to interchange SPO11-1 as well as SPO11-2 between Arabidopsis and a different species.   Furthermore we were able to identify a species specific splicing pattern of SPO11-1 and SPO11-2. We will present results on heterologous complementa-tion approach as well as splicing  patterns of different plants SPO11-1 and -2 genes, as well as patterns from Arabidopsis plants carrying SPO11 from a different species. Additionally we in-vestigate if the function of SPO11-1 and -2 is sequence specific and if they work together. For this purpose we inter-changed regions between SPO11-1 and -2. We will present results on these swapped gene approach.   Additionally we designed and produced antibodies against Arabidopsis SPO11-1 and -2 for use in immunofluorescence-microscopy to get a closer look on the behavior and the distribution of SPO11-1 and -2 during meiosis. We will show images using these and other antibod-ies on different Arabidopsis mutant lines from the complementation ap-proaches.   By analyzing the results of these exper-iments we should be able to answer the question if there is an sequence and/or species specific function of   SPO11-1 and -2 and if the specific splicing pat-tern has influence on the function if SPO11. With the new antibodies we should also be able to get a closer look on the behavior of SPO11-1 and -2 dur-ing meiosis. Berichte aus dem Julius Kühn-Institut 177 15

Initiation of meiotic double strand breaks in Arabidopsis depends on two different SPO11 proteins

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Initiation of meiotic double strand breaks in Arabidopsis depends on two different SPO11 proteins

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