![]() ![]() The effector protein VirE2 has non-specific single-stranded DNA-binding activity and is thought to coat single-stranded T-DNA (T-strands) after entry into the plant cell ( Citovsky et al., 1992), protecting T-strands from nucleolytic degradation ( Yusibov et al., 1994 Rossi et al., 1996). Agrobacterium-mediated plant genetic transformation involves mobilization of transferred-DNA (T-DNA) and five virulence proteins (VirD2, VirE2, VirE3, VirD5, and VirF) from the bacterium into a plant cell ( Gelvin, 2003, 2012). Virulent strains of the soil bacterium Agrobacterium tumefaciens cause the tumorigenic disease crown gall. vip1 mutant and VIP1-SRDX plants show increased tolerance to growth in salt, indicating a role for VIP1 in response to salt stress. ![]() The germination of vip1 mutant and VIP1-SRDX seeds is sensitive to exogenous ABA, suggesting a role for VIP1 in response to ABA. cinerea susceptibility is dependent on abscisic acid (ABA) which is also important for abiotic stress responses. syringae infection, suggesting a role for VIP1 in B. vip1 mutant and VIP1-SRDX plants show increased susceptibility to B. Because VIP1 is involved in innate immune response signaling, we tested the susceptibility of vip1 mutant and VIP1-SRDX plants to Pseudomonas syringae and Botrytis cinerea. All VIP1-SRDX transgenic lines showed wild-type levels of transformation, indicating that neither VIP1 nor its homologs are required for Agrobacterium-mediated transformation. Because of potential functional redundancy among VIP1 homologs, we tested transgenic lines expressing VIP1 fused to a SRDX repression domain. The transformation susceptibility of vip1-2 and wild-type plants is similar. We generated the CRISPR/Cas mutant vip1-2 that lacks this domain. Previously tested vip1-1 mutant plants produce a truncated protein containing the crucial bZIP DNA-binding domain. The bZIP transcription factor VIP1 interacts with the Agrobacterium virulence protein VirE2, but the role of VIP1 in Agrobacterium-mediated transformation remains controversial.
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