Allopolyploids are believed to inherit the genetic characteristics of its progenitors and exhibit stronger adaptability and vigor. The allotetraploid Isoetes sinensis was formed by the natural hybridization and polyploidization of two diploid progenitors, Isoetes taiwanensis and Isoetes yunguiensis, and was believed to have the potential to adapt to plateau environments. To explore the expression pattern of homoeologous genes and their contributions to altitude adaptation, we transplanted natural allotetraploid I. sinensis (TnTnYnYn) along the altitude gradient for a long‐term, and harvested them in summer and winter, respectively. One year after transplanting, it still lived well, even in the extreme environment of the Qinghai‐Tibet Plateau. Then, we performed high‐throughput RNA sequencing to measure their gene expression level. A total of 7801 homoeologous genes were expressed, among which 5786 were identified as shared expression in different altitudes and seasons. We further found that altitude variations could change the subgenome bias trend of I. sinensis, but season could not. Moreover, the functions of uniquely expressed genes indicated that temperature might be an important restrictive factor during the adaptation process. Through the analysis of DEGs and uniquely expressed genes, we found that Y subgenome provided more contributions to high altitude adaptation than T subgenome. These adaptive traits to high altitude may be inherited from its plateau progenitor I. yunguiensis. Through weighted gene co‐expression network analysis, pentatricopeptide repeats gene family and glycerophospholipid metabolism pathway were considered to play important roles in high‐altitude adaptation. Totally, this study will enrich our understanding of allopolyploid in environmental adaptation. Long‐term transplanting experiment was performed to simulate real local habitats. The genomes of Isoetes taiwanensis and Isoetes yunguiensis (diploid progenitors) were used as references to separately obtain the two subgenomes transcriptome expression data of Isoetes sinensis (subgenome T and Y).Through transcriptomic analysis, we demonstrated that the potential adaptation of I. sinensis to high altitude inherited from its plateau progenitor, I. yunguiensis.