Abstract: 22 kinds of fresh leaves of Camellia nitidissima were collected, and DNA was extracted by centrifugal column plant genome kit. ITS2, matK, rbcL, trnL-trnF, psbA-trnH sequence universal primers were used for PCR amplification, electrophoresis detection and biaxial sequencing. The sequences of ribosomal DNA second internal transcriptional spacer（ITS2）, RNA transcriptor type Ⅱ intron cut enzyme gene（matK）, chloroplast ribulose-1, 5-diphosphate carboxylase large subunit（rbcL）, and chloroplast non-coding region（trnL-trnF, psbA-trnH） were used to study 22 kinds of Camphor tea DNA barcoding molecular identification, and to determine the relationship. The results showed that the DNA of 22 species of Camellia nitidissima samples could be extracted successfully, the amplification of ITS2 primer failed, and the amplification of other primers was successful and the sequencing success rate reached 100%. matK, rbcL, trnL-trnF, psbA-trnH and their combination sequences can distinguish different kinds of golden tea successfully. matK, rbcL, trnL-trnF, psbA-trnH and their combination sequences had good application value in molecular identification and genetic relationship determination of 22 Camellia nitidissima species.