Beginnings of a plant parasite: early development of Rafflesia consueloae inside its Tetrastigma host

Extensive histology of host organs revealed the early events in the vegetative growth of Rafflesia consueloae including initial infection site, endophyte distribution, and other developmental events prior to bud emergence. The early events in the vegetative development of the holoparasite Rafflesia have long remained a mystery. Because its entire vegetative growth occurs within the host body, very little is known about the developmental events prior to emergence of the floral shoot. The goal of this study was to describe the events that occur during the vegetative growth of R. consueloae, particularly in the early stages of infection. We performed extensive microtome sectioning of multiple root and stem segments from different Tetrastigma host individuals to examine the cytology, distribution, and development of the R. consueloae endophyte within the host tissues. We found that R. consueloae infection is restricted to the roots of its host. Infection begins within the vascular cambium where the endophyte appears to initially reside prior to their radial spread to the vascular tissues. The tissues obtained from different host individuals had varying degrees of infection alluding to a possible role of host resistance mechanisms and/or varying levels of parasite infectiousness. Endophyte presence in host vines without external manifestations of infection indicates that the parasite may dwell within the host tissues for prolonged periods as small cell clusters without transitioning to the reproductive stage. Furthermore, we found that floral shoots may develop in scarcely infected host tissues indicating that extensive endophyte growth within the host is not a prerequisite to the onset of reproductive development. Overall, our study describes for the first time the developmental events prior to emergence of R. consueloae buds from its host.