The development and cell death domain-containing protein HaB2 enhances sunflower resistance to Orobanche cumana via the ABA signaling pathway

Abstract Broomrape (Orobanche cumana) is a major threat to sunflower (Helianthus annuus) production. Although B2 proteins, a class of development and cell death (DCD) domain-containing proteins, are reported to play a role in responses to a variety of abiotic stresses in plants, their function in biotic stress remains unclear. This study explored the role of HaB2 in sunflower resistance to O. cumana, focusing on its regulation through the ABA signaling pathway. Our results show that O. cumana infection and exogenous ABA treatment strongly induce HaB2 expression in sunflower root. Silencing HaB2 increases parasite attachment, while over-expression of HaB2 reduces the formation of haustoria and xylem bridges. Further, the DCD domain is both essential and sufficient for HaB2 resistance function. In addition, we found a physiological interaction between HaB2 and the phosphatase HaFyPP3, while HaFyPP3 interacts with the ABA-responsive transcription factor HaABI5. Exogenous application of ABA leads to a resistance phenotype consistent with that of HaB2 over-expression plants. HaABI5 promotes the expression of HaB2 by binding to the ABRE element on its promoter and therefore enhances sunflower resistance, while HaFyPP3 negatively regulates HaB2-mediated resistance by possibly dephosphorylating both ABI5 and HaB2. These results suggest that HaB2 acts as a positive regulator to enhance sunflower resistance to broomrape through involvement in the ABA signaling pathway, providing new insights into ABA-mediated biotic stress resistance and potential targets for the exploitation of superior O. cumana-resistant sunflower varieties by molecular breeding.