Four sugarcane ScDIR genes contribute to lignin biosynthesis and disease resistance to Sporisorium scitamineum

Abstract Sugarcane ( Saccharum spp.) is a major sucrose and bioenergy crop in the world. The fungal pathogen Sporisorium scitamineum causes sugarcane smut, a devastating disease that destroys stalks and reduces sugar content in sugarcane. This disease can be controlled most effectively by applying smut-resistant sugarcane varieties. Previous studies have shown that Dirigent (DIR ) genes are involved in the synthesis of the lignin precursor pinoresinol, which plays a crucial role in plant resistance to biotic stresses. However, the immune response of the DIR homologs in sugarcane ( ScDIR ) has not been reported yet. In this study, we found that the lignin content of smut-resistant sugarcane varieties (ZZ1, ZZ6, and ZZ9) was significantly higher than that of smut-susceptible varieties (GT42, ROC22, and FN41), and the lignin content of sugarcane increased after smut infection. The smut-resistant and smut-susceptible clones derived from the same genetic population (ROC25 × YZ89-7) showed similar patterns. Quantitative real-time PCR assays revealed that among the 64 DIR genes in sugarcane, ScDIR5 , ScDIR7 , ScDIR11 , and ScDIR40 showed elevated expression after S. scitamineum infection. In vitro coupling reactions showed that the four corresponding ScDIR proteins could mediate the coupling of coniferyl alcohol and its conversion into the lignin precursor pinoresinol. Overexpression of the four ScDIR genes in Nicotiana benthamiana enhanced disease resistance to the fungal pathogens Sclerotium rolfsii , Rhizoctonia solani , and Botrytis cinerea . Moreover, transgenic sugarcane overexpressing these ScDIR genes showed enhanced resistance to smut disease. Taken together, our findings provide evidence that sugarcane ScDIR genes can improve the resistance of plants to fungal pathogens and highlight their potentials in sugarcane breeding for disease resistance.