CcLBD25functions as a key regulator of haustorium development inCuscuta campestris

Parasitic plants reduce yield of crops worldwide. Cuscuta campestris is a stem parasite that attaches to its host, using haustoria to extract nutrients and water. We analyzed the transcriptome of six C. campestris tissues and identified a key gene, CcLBD25 , as highly expressed in prehaustoria and haustoria. Our gene co-expression networks (GCN) from different tissue types and laser-capture microdissection (LCM) RNA-Seq data indicate that CcLBD25 could be essential for regulating cell wall loosening and organogenesis. We employed host-induced gene silencing (HIGS) by generating transgenic tomato hosts that express hairpin RNAs to target and down-regulate CcLBD25 in the parasite. Our results showed that C. campestris growing on CcLBD25 RNAi transgenic tomatoes transited to the flowering stage earlier and had less biomass compared with C. campestris growing on wild type host. This suggests that the parasites growing on the transgenic plants were stressed due to insufficient nutrient acquisition. Anatomy of C. campestris haustoria growing on CcLBD25 RNAi tomatoes showed reduced pectin digestion and lack of searching hyphae, which interfered with haustorium penetration and the formation of vascular connections. We developed an in vitro haustorium (IVH) system to assay the number of prehaustoria produced on strands from C. campestris . When C. campestris was grown on CcLBD25 RNAi tomatoes or wild type tomatoes, the former produce fewer prehaustoria than the latter, indicating that down-regulating CcLBD25 may affect haustorium initiation. The results of this study shed light on the role of CcLBD25 in haustorium development and might help to develop a parasite-resistant system in crops. One-sentence summary CcLBD25 plays a pivotal role in haustorium initiation, regulating pectin digestion, and searching hyphae development during the haustorium penetration process.