With applications of Tree of Life data becoming ever more prevalent in everyday contexts, tree thinking has emerged as a vital component of scientific literacy. This article reports a study testing the hypothesis that instruction in natural selection, which is the primary focus of US evolution education at the high school and introductory college levels, does little to promote tree-thinking skills and that explicit instruction in understanding evolutionary tree diagrams is required. Testing this contention required the creation of a novel tree-thinking assessment and an instructional intervention, both guided by deep knowledge of evolutionary biology and of science education research. College students (N ¼ 124) with weaker versus stronger backgrounds in biology were randomly assigned to control versus instructional conditions and were also assessed for their knowledge of natural selection. Although knowledge of natural selection and ability to engage in tree thinking were correlated, a short instructional booklet that provided a basic introduction to evolutionary trees predicted tree-thinking success more strongly than did either knowledge of natural selection or previous college coursework in biology. Clearly, tree thinking and natural selection are dissociable constructs that must both be taught for students to grasp the full gamut of evolutionary patterns and processes. # Just as beginning students in geography need to be taught how to read maps, so beginning students in biology should be taught how to read trees and to understand what trees communicate (O'Hara, 1998, p. 327). Humans are most closely related, evolutionarily, to chimpanzees. Humans, chimps, and the other primates, in turn, share an ancestor with all other mammals. Mammals share a most recent common ancestor (MRCA) with reptiles. Mammals and reptiles share an ancestor with arthropods. As a group, animals share an ancestor with fungi. Animals and fungi, together, share an ancestor with plants. And so on. If we go back far enough in time, we find that all living things have a single ancestor in common. This history of "descent with modification" (Darwin, 1859) can be represented in the form of a very large, branching Tree of Life (ToL). The set of skills required to understand and reason with such information depicted in diagrammatic representations of the ToL is referred to as tree thinking. Tree thinking is an important aspect of 21st century science literacy.