This book constitutes the proceedings of the 25th International Conference on Developments in Language Theory, DLT 2021, which was held in Porto, Portugal, during August 16-20, 2021. The conference took place in an hybrid format with both in-person and online participation.The 27 full papers included in these proceedings were carefully reviewed and selected from 48 submissions. The DLT conference series provides a forum for presenting current developments in formal languages and automata. Its scope is very general and includes, among others, the following topics and areas: grammars, acceptors and transducers for words, trees and graphs; algebraic theories of automata; algorithmic, combinatorial, and algebraic properties of words and languages; variable length codes; symbolic dynamics; cellular automata; polyominoes and multidimensional patterns; decidability questions; image manipulation and compression; efficient text algorithms; relationships to cryptography, concurrency, complexity theory, and logic; bio-inspired computing; quantum computing. The book also includes 3 invited talks in full paper length.