Plant-parasitic nematodes are severe pests in crop production worldwide. Chemical control of nematodes has been continuously reduced in recent decades owing to environmental and health concerns. Therefore, breeding nematode-resistant crops is an important aim if we are to secure harvests. The beet cyst nematode impairs root development and causes severe losses in sugar beet production. The only sources for resistance are distantly related wild species of the genus Patellifolia. Nematode resistance had been introduced into the beet genome via translocations from P. procumbens. We sequenced three translocations and identified the translocation breakpoints. By comparative sequence analysis of three translocations, we localized the resistance gene Hs4 within a region c. 230 kb in size. A candidate gene was characterized by CRISPR-Cas-mediated knockout and overexpression in susceptible roots. The gene encodes a rhomboid-like protease, which is predicted to be bound to the endoplasmic reticulum. Gene knockout resulted in complete loss of resistance, while overexpression caused resistance. The data confirm that the Hs4 gene alone protects against the pest. Thus, it constitutes a previously unknown mechanism of plants to combat parasitic nematodes. Its function in a nonrelated species suggests that the gene can confer resistance in crop species from different plant families.
Keywords: Beta vulgaris; Heterodera schachtii; CRISPR-Cas; chromosome translocation; pest resistance; sugar beet; translocation breakpoint.
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