34Meiotic drive elements like Spore killer-2 (Sk-2) in Neurospora are transmitted through sexual 35 reproduction to the next generation in a biased manner. Sk-2 achieves this biased transmission 36 through spore killing. Here, we identify rfk-1 as a gene required for the spore killing mechanism.
37The rfk-1 gene is associated with a 1,481 bp DNA interval (called AH36) near the right border of 38 the 30 cM Sk-2 element, and its deletion eliminates the ability of Sk-2 to kill spores. The rfk-1 39 gene also appears to be sufficient for spore killing because its insertion into a non-Sk-2 isolate 40 disrupts sexual reproduction after the initiation of meiosis. Although the complete rfk-1 41 transcript has yet to be defined, our data indicate that rfk-1 encodes a protein of at least 39 amino 42 acids and that rfk-1 has evolved from a partial duplication of gene ncu07086. We also present 43 evidence that rfk-1's location near the right border of Sk-2 is critical for the success of spore 44 killing. Increasing the distance of rfk-1 from the right border of Sk-2 causes it to be inactivated 45 by a genome defense process called meiotic silencing by unpaired DNA (MSUD), adding to 46 accumulating evidence that MSUD exists, at least in part, to protect genomes from meiotic drive. 47 48 49 50In eukaryotic organisms, genetic loci are typically transmitted through sexual reproduction to the 51 next generation in a Mendelian manner. However, some loci possess the ability to improve their 52 own transmission rate through meiosis at the expense of a competing locus. These "selfish" loci 53 are often referred to as meiotic drive elements (Zimmering et al. 1970). The genomic conflict 54 caused by meiotic drive elements may impact processes ranging from gametogenesis to 55 speciation (Lindholm et al. 2016). Meiotic drive elements are found across the eukaryote tree of 56 life (Burt and Trivers 2008; Bravo Núñez et al. 2018) and classic examples include SD in fruit 57 flies (Larracuente and Presgraves 2012), the t-complex in mice (Lyon 2003; Sugimoto 2014), 58and Ab10 in Zea mays (Rhoades 1952; Kanizay et al. 2013). In the fungal kingdom, the known 59 meiotic drive elements achieve biased transmission through spore killing (Raju 1994) and a 60 handful of spore killer systems have been studied in detail. While the prion-based spore killing 61 mechanism of het-s in Podospora anserina is the best characterized (Dalstra et al. 2003; Saupe 62 2011), the mechanisms by which other fungal meiotic drive elements kill spores are mostly 63 unknown (e.g., see Grognet et al. 2014; Hu et al. 2017; Nuckolls et al. 2017).
65Two fungal meiotic drive elements have been identified in the fungus Neurospora 66 intermedia (Turner and Perkins, 1979). This species is closely related to the genetic model 67 Neurospora crassa (Davis 2000), and the mating processes in both fungi are essentially identical.
68Mating begins with fertilization of an immature fruiting body called a protoperithecium by a 69 mating partner of the opposite mating type. After ferti...