Dominant-negative KAI2d paralogs putatively attenuate strigolactone responses in root parasitic plants

Abstract

Many root parasitic plants in the Orobanchaceae use host-derived strigolactones as germination cues. This adaptation facilitates attachment to a host and is particularly important for the success of obligate parasitic weeds that cause substantial crop losses globally. Parasite seeds sense strigolactones through “divergent” KARRIKIN INSENSITIVE2 (KAI2d)/HYPOSENSITIVE TO LIGHT (HTL) α/β-hydrolases that have undergone substantial duplication and diversification in Orobanchaceae genomes. After germination, chemotropic growth of parasite roots toward a strigolactone source also occurs in some species. We investigated which of the seven KAI2d genes found in a facultative hemiparasite, Phtheirospermum japonicum, may enable chemotropic responses to strigolactones. To do so, we developed a triple mutant Nbd14a,b kai2i line of Nicotiana benthamiana in which strigolactone-induced degradation of SMAX1, an immediate downstream target of KAI2 signaling, is disrupted. In combination with a transiently expressed, ratiometric reporter of SMAX1 protein abundance, this mutant forms a system for the functional analysis of parasite KAI2d proteins in a plant cellular context. Using this system, we unexpectedly found three PjKAI2d proteins that do not trigger SMAX1 degradation in the presence of strigolactones. Instead, these PjKAI2d inhibit the perception of low strigolactone concentrations by strigolactone-responsive PjKAI2d in a dominant-negative manner that depends upon an active catalytic triad. Similar dominant-negative KAI2d paralogs were identified in an obligate hemiparasitic weed, Striga hermonthica. These proteins suggest a mechanism for attenuating strigolactone signaling in parasites, which might be used to enhance the perception of shallow strigolactone gradients during root growth toward a host or to restrict germination responses to specific strigolactones.