RNA editing is one of the post-transcriptional modification processes in which the bases of a RNA molecule are altered by the addition, deletion and alteration of nucleotides. In most higher plants, RNA editing mainly occurs in mitochondria and plastids and converts from C to U, very rarely from U to C. We investigated RNA editing sites in chloroplasts of Gossypium hirsutum ‘CRRI 10’ by PCR, RT-PCR and sequence alignment. We identified 55 editing sites in 27 protein-coding genes all of which were C-to-U conversion. By comparing editing sites between CRRI 10 and Coker310FR, CRRI 10 had two novel editing sites, accD-468 and rpoC1-163, whereas site psbN-10 was absent. Bioinformatics analysis of the 3 sites revealed that rpoC1-163 and psbN-10 editing might affect the secondary structure of the corresponding protein. Comparison among upstream regions (−30 to −1) of the 55 editing sites of CRRI 10 revealed that 8 pairs share more than 60% sequence similarity suggesting that the sites in each pair may be recognized by the same trans-acting factors.