The chromatin-associated proteins play important roles in regulating DNA duplication, gene transcription expression etc. in eukaryotic system. Previously, Arabidopsis LFR (leaf and flower related) protein localized in nucleus, and its loss-of-function mutants were reported to have pleiotropic phenotypes in leaf, flower and anther development, but the molecular characteristics of LFR protein should be detected further. Firstly, the fusion protein of GAL4 DNA-binding domain (GBD) with full-length of LFR had transactivation activity in yeast Y190 by yeast one-hybrid assay, and at least 2 ARM-repeat domains in the C terminal and full N teminal of LFR were necessary to its transactivation activity. Next, we further analyzed GBD-LFR transactivation activity in Arabidopsis protoplast system, it showed that the transactivation of full-length LFR was not obvious compared with that of a classical transcription activity domain of VP16. In protoplast transient expression system, fluorescence microscopy was used to observed subcellular localization of YFP fusion proteins of truncated or site-mutation LFR, and the data showed that the N terminal 1～25 amino acids of LFR, particular lysine 22 and arginine 4, 23, 25, was responsible for its nuclear localization. In transient co-expressing onion epidermal cell, LFR obviously co-localized with chromatin stucture protein, histone H4, and chromatin-binding protein, HMGA, in nucleus. All these data suggested that LFR might be as one of chromatin-associated proteins to function in Arabidospis development.