Abstract :In order to verify the function of IPT gene in Psathyrostachys juncea, the tillering nodes of DT type and ST type were used as materials. The relative expression of IPT was verified by RNA-seq analysis and qRT-PCR test, and GO enrichment analysis was carried out. The IPT gene was cloned by PCR method, and the 1300-cYFP-IPT overexpression vector was constructed and analyzed by bioinformatics. The subcellular localization and expression of IPT protein were identified by tobacco transient transfection and Western blot. The results showed that IPT was related to the activity of tRNA dimethylallyl transferase and up-regulated in the tillering of Psathyrostachys juncea. The full-length IPT gene of Psathyrostachys juncea was cloned, and the 1300-cYFP-IPT overexpression vector was constructed with an open reading frame (ORF) of 1362 bp. Multiple sequence alignment and conserved domain analysis showed that the IPT protein of Psathyrostachys juncea had a conserved domain of PLN02840 protein, which was closely related to the IPT protein of Triticum dicoccoides, Triticum aestivum and Triticum turgidum subsp. Durum. Protein secondary structure prediction showed that the secondary structure of IPT protein was composed of alpha helix, extended strand, beta turn and random coil. Analysis showed that serine phosphorylation sites were most likely to be potential phosphorylation sites for protein function. Transient transfection of tobacco showed that IPT protein was normally expressed and localized in chloroplasts. Western blot results showed that the target protein IPT of the connected vector was normally expressed, with a size of 76.8 kDa. The results showed that IPT gene up-regulated the number of tillers during the tillering process of Psathyrostachys juncea, and the 1300-cYFP-IPT vector could be normally expressed in the chloroplast of plants. This study provides experimental materials and theoretical basis for further functional verification of IPT gene in Psathyrostachys juncea.