The Dunhuang tectonic belt (DTB) is of great importance for understanding the tectonic evolution of the southernmost Central Asian Orogenic Belt (CAOB). In this study, the temporal-spatial distribution, petrogenesis and tectonic setting of the Paleozoic representative intrusive rocks from the DTB were systematically investigated to discuss crustal evolution history and tectonic regime of the DTB during Paleozoic.Our results reveal that the Paleozoic magmatism within the DTB can be broadly divided into two distinct episodes of early Paleozoic and late Paleozoic. The early Paleozoic intrusive rocks, represented by a suite metaluminous-slight peraluminous and medium- to high-K calc-alkaline I-type granitoids crystallized at Silurian (ca. 430–410Ma), are predominantly distributed along the northern part of the DTB. They were probably produced with mineral assemblage of eclogite or garnet+amphibole+rutile in the residue, and were derived from magma mixing source of depleted mantle materials with various proportions of Archean-Mesoproterozoic continental crust. The late Paleozoic intrusive rocks can be further subdivided into two stages of late Devonian stage (ca. 370–360Ma) and middle Carboniferous stage (ca. 335–315Ma). The former stage is predominated by metaluminous to slight peraluminous and low-K tholeiite to high-K calc-alkaline I-type granitic rocks distributed in the central part of the DTB. They were also generated with mineral assemblage of amphibolite- to eclogite-facies in the residue, and originated from magma source of depleted mantle materials mixed with different degrees of old continental crust. The later stage is represented by adakite and alkali-rich granite exposed in the southern part of the DTB. The alkali-rich granites studied in this paper were possibly produced with mineral assemblage of granulite-facies in the residue and were generated by partial melting of thickened lower continental crust. Zircon Hf isotopes and field distribution of those Paleozoic intrusive rocks reveal that both the Silurian and the late Devonian magmatic activities predominantly represent crustal growth processes in the DTB, accompanied by different degrees of reworking of pre-existing continental crust. However, the middle Carboniferous (ca. 335–315Ma) magmatic activity reflects a crustal reworking process. The Silurian and late Devonian intrusive rocks were most likely formed in the arc-related subduction zones, whereas, the middle Carboniferous intrusive rocks were possibly formed in a transitional tectonic setting from compression to extension, representing the final stage of Paleozoic orogeny in the DTB.These Paleozoic magmatic rocks further suggest that the DTB has reactivated from a stable block to an orogen and undergone two episodes (the early Paleozoic and the late Paleozoic) of orogeny during Paleozoic. It represents a Paleozoic accretionary orogen of the southernmost margin of the CAOB between the Tarim Craton and North China Craton, and tectonically extends northward to the Beishan orogen and westward to the eastern South Tianshan Belt.