Larose, Hailey Lee Ann2019-10-102019-10-102018-04-17vt_gsexam:14424http://hdl.handle.net/10919/94423Seeds of the root parasitic plants of the genus Orobanche germinate specifically in response to host-derived germination signals, which enables parasites to detect and attack preferred hosts. The best characterized class of germination stimulants is the strigolactones (SLs), although some species respond to non-SL compounds, such as dehydrocostus lactone (DCL). Recent work indicates that SLs are perceived by members of the KARRIKIN-INSENSITIVE2 (KAI2) gene family, and suggests that within parasitic Orobanchaceae the KAI2 genes have undergone duplication and specialization. The "diverged" clade of these genes, termed KAI2d, has been shown to bind SL germination stimulants in model system assays, but the precise role for KAI2d in regulating germination specificity in a parasitic plant has not been demonstrated. To address this issue, we used genetic and genomic approaches involving two closely related species, Orobanche cernua and O. cumana, which differ primarily in host range and stimulant preference. Orobanche cernua parasitizes tomato (and other Solanaceous crops) and responds to orobanchol, the major SL from tomato roots, whereas O. cumana specifically parasitizes sunflower and responds to DCL. Crosses between O. cernua and O. cumana produced hybrid populations that segregate for stimulant specificity, creating a tractable genetic system. Orobanche cernua contains four KAI2d genes (numbered OrceKAI2d1-4), while O. cumana contains six genes (OrcuKAI2d1-6). The DNA from 94 F2 hybrids was genotyped to identify the KAI2d gene composition and these were correlated with germination phenotype. The pattern of segregation indicated that the KAI2d genes are linked, but pointed to OrceKAI2d2 as a likely orobanchol receptor. Response to DCL was associated with inheritance of all O. cumana KAI2d genes together. Each KAI2d gene was expressed in the Arabidopsis thaliana kai2 mutant background and tested for ability to recover the mutant phenotype when exposed to SLs (including orobanchol, 5-deoxystrigol and GR24) or DCL. One O. cernua gene, OrceKAI2d2, responded to all SLs, but not DCL in this system. No DCL-specific KAI2 genes were identified. In summary, we have identified the likely SL receptor in O. cernua, and show evidence that the DCL receptor is either not a KAI2d protein, or uses KAI2d in combination with other signaling pathway components.ETDIn CopyrightParasitic plantsgermination stimulantgerminationOrobanche cumanaOrobanche cernuaOrobanchaceaestrigolactonedehydrocostus lactoneKAI2D14Dissertation