For most of their existence, stars are fuelled by the fusion of hydrogen into helium. Fusion proceeds via two processes that are well understood theoretically: the proton-proton (pp) chain and the carbon-nitrogen-oxygen (CNO) cycle1,2. Neutrinos that are emitted along such fusion processes in the solar core are the only direct probe of the deep interior of the Sun. A complete spectroscopic study of neutrinos from the pp chain, which produces about 99 per cent of the solar energy, has been performed previously3; however, there has been no reported experimental evidence of the CNO cycle. Here we report the direct observation, with a high statistical significance, of neutrinos produced in the CNO cycle in the Sun. This experimental evidence was obtained using the highly radiopure, large-volume, liquid-scintillator detector of Borexino, an experiment located at the underground Laboratori Nazionali del Gran Sasso in Italy. The main experimental challenge was to identify the excess signal-only a few counts per day above the background per 100 tonnes of target-that is attributed to interactions of the CNO neutrinos. Advances in the thermal stabilization of the detector over the last five years enabled us to develop a method to constrain the rate of bismuth-210 contaminating the scintillator. In the CNO cycle, the fusion of hydrogen is catalysed by carbon, nitrogen and oxygen, and so its rate-as well as the flux of emitted CNO neutrinos-depends directly on the abundance of these elements in the solar core. This result therefore paves the way towards a direct measurement of the solar metallicity using CNO neutrinos. Our findings quantify the relative contribution of CNO fusion in the Sun to be of the order of 1 per cent; however, in massive stars, this is the dominant process of energy production. This work provides experimental evidence of the primary mechanism for the stellar conversion of hydrogen into helium in the Universe.