Abstract 【Aim】 Drosophila is one of holometabolous insects. It undergoes the process of dissociation of old tissues and remodeling of adult tissues at the pupal stage. The objective of this study is to investigate whether hindgut enterocytes of Drosophila migrate into the midgut during metamorphosis by G-TRACE (Gal4 technique for real-time and clonal expression) which is a new genetic technique. 【Methods】 engrailed-Gal4 (en-Gal4) line and lineage-tracing line (G-TRACE) of Drosophila melanogaster were hybridized, and tub-gal80ts was introduced to temporally control Gal4 activity. The cell lineage was traced at the larval and pupal stages, respectively. For larval stage tracing, eggs were cultured at 30℃ after egg-laying by the parental generation, and the mid 3rd instar larvae were shifted to 18℃. The adult guts were detected within 1 d after eclosion. For pupal stage tracing, eggs were cultured at 18℃ after egg-laying by the parental generation, pupae were shifted to 30℃ at different stages, and adult guts were detected after eclosion. 【Results】 In larval stage tracing of Drosophila, green intestinal cells appeared in the posterior section of midgut, which is adjacent to the hindgut-midgut boundary and Malpighian tubules. In pupal stage tracing of Drosophila, green intestinal cells appeared at different sections in the midgut and Malpighian tubules, and engrailed gene was expressed in Drosophila intestine at the pupal stage. 【Conclusion】 These results suggest that during pupa formation, part of the hindgut cells migrate into the midgut or Malpighian tubules and are involved in the reformation of adult midgut or Malpighian tubules. This study is of important significance in understanding the mechanisms of insect organ remodeling during metamorphosis.