OsMAPKKK69 Negatively Regulates Resistance to Blast and Bacterial Blight Diseases in Rice (Oryza sativa L.)

Rice blast is one of the main diseases of rice, causing severe economic losses to agricultural production; thus, the search for blast resistance is a top priority for rice breeding. When challenged by the blast causal fungus Magnaporthe oryzae the expression level of OsMAPKKK69 gene in rice cultivar Nipponbar was found to increase significantly. Such an induction was also found in a different genetic material, cultivar Shufanggaonuo, indicating that OsMAPKKK69 plays an important role in blast disease response. However, the function of OsMAPKKK69 remains unclear. In this study, wild type ZH11 was selected as the background material to investigate the expression and functions of OsMAPKKK69 in rice disease resistance by constructing knockout mutants. The results showed that OsMAPKKK69 is mainly expressed in four-week-old shoots and localized in cell membrane, cytoplasm, and nucleus. The two allelic knockout mutants, osmapkkk69-1 and osmapkkk69-2, were more resistant to M. oryzae and bacterial blight Xanthomonas oryzae pv. Oryzae (Xoo). Further agronomic trait analysis revealed that the osmapkkk69-1 and osmapkkk69-2 mutants had reduced plant height, smaller grain size, a significant increase in tillering number, but also a significant increase in yield per plant. Our results show that OsMAPKKK69 is involved in the immune response of rice by negatively regulating the resistance to rice blast and blight diseases, and in regulating important agronomic traits. This study lays a foundation for revealing the molecular mechanism of OsMAPKKK69 in the immune response to rice diseases and provides novel genetic resources for rice breeding.