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Background: Plant invasions are causing habitat degradation in Galapagos. Problems are concentrated on the four inhabited islands. Plants introduced to rural areas in the humid highlands and urban areas on the arid coast act as foci for invasion of the surrounding Galapagos National Park.

Methodology/Principal Findings: Here we present results of the most comprehensive inventory to date of alien vascular plants in the inhabited areas of Galapagos. The survey was conducted between 2002 and 2007, in 6031 properties (97% of the total) on Floreana, Isabela, San Cristobal and Santa Cruz Islands. In total 754 alien vascular plant taxa were recorded, representing 468 genera in 123 families. Dicotyledons represented 554 taxa, monocotyledons 183, there were 7 gymnosperms and 10 pteridophytes. Almost half (363) of the taxa were herbaceous. The most represented families were Fabaceae (sensu lato), Asteraceae and Poaceae. The three most recorded species in the humid rural areas were Psidium guajava, Passiflora edulis and Bryophyllum pinnatum, and in the dry urban areas, Aloe vera, Portulaca oleracea and Carica papaya. In total, 264 (35%) taxa were recorded as naturalized. The most common use for taxa was ornamental (52%).

Conclusions/Significance: This extensive survey has increased the known alien vascular flora of Galapagos by 257 species, giving a ratio of alien to native taxa of 1.57:1. It provides a crucial baseline for plant invasion management in the archipelago and contributes data for meta analyses of invasion processes worldwide. A repeat of the survey in the future would act as an effective early detection tool to help avoid further invasion of the Galapagos National Park.