TY - JOUR T1 - The mitotic spindle mediates nuclear migration through an extremely narrow infection structure of the rice blast fungus <em>Magnaporthe oryzae</em> JF - bioRxiv DO - 10.1101/2021.04.07.438902 SP - 2021.04.07.438902 AU - Mariel A. Pfeifer AU - Chang Hyun Khang Y1 - 2021/01/01 UR - http://biorxiv.org/content/early/2021/04/08/2021.04.07.438902.abstract N2 - The blast fungus, Magnaporthe oryzae, causes severe destruction to rice and other crops worldwide. As the fungus infects rice, it develops unique cellular structures, such as an appressorium and a narrow penetration peg, to permit successful invasion of host rice cells. Fundamental knowledge about these cellular structures and how organelles, such as the nucleus, are positioned within them is still emerging. Previous studies show that a single nucleus becomes highly stretched during movement through the narrow penetration peg in an extreme nuclear migration event. Yet, the mechanism permitting this nuclear migration event remains elusive. Here, we investigate the role of the mitotic spindle in mediating nuclear migration through the penetration peg. We find that disruption of spindle function during nuclear migration through the penetration peg prevents development of invasive hyphae and virulence on rice. Furthermore, regulated expression of conserved kinesin motor proteins, MoKin5 and MoKin14, is essential to form and maintain the spindle, as well as, properly nucleate the primary hypha. Overexpression of MoKin5 leads to formation of aberrant microtubule protrusions, which contributes to formation of nuclear fragments within the appressorium and primary hypha. Conversely, overexpression of MoKin14 causes the spindle to collapse leading to the formation of monopolar spindles. These results establish a mechanistic model towards understanding the intricate subcellular dynamics of extreme nuclear migration through the penetration peg, a critical step in the development of rice blast disease.Importance Magnaporthe oryzae, also known as the blast fungus, is a formidable hinderance to global food production, including rice. The destructive fungal pathogen develops highly-specialized cells and structures, such as appressoria and penetration pegs, to permit successful invasion of rice cells. Our understanding of M. oryzae’s fundamental biology during host cell invasion and colonization is still developing. For instance, it is not yet known how organelles, such as the nucleus, migrate through the narrow penetration peg. Moreover, few previous studies examine the role of motor proteins in M. oryzae. In this study, we determined that the mitotic spindle propels a single nucleus through the penetration peg to permit successful development of fungal hyphae inside the first-invaded rice cell. We also identified two conserved kinesin motor proteins, MoKin5 and MoKin14. Our analyses suggested that MoKin5 and MoKin14 exhibit canonical functions in M. oryzae during rice infection. This study addressed long-standing questions in rice blast biology, and our results offer opportunities for future research. ER -