Identification of Murraya koenigii (L.) Spreng Using DNA Barcoding Technique Based on the ITS Sequence

Abstract

OBJECTIVE: To determine pharmacognostical characteristics and DNA marker of Murraya koenigii leaf. METHODS: Leaf tissue of M. koenigii was investigated under light microscope. DNA marker for M. koenigii was established by amplification of the internal transcribed spacer (ITS) of nuclear ribosomal RNA gene using conserved plant sequences as primers. Multiple alignment with ITS sequences of plants in the tribe Clauseneae retrieved from GenBank was then was performed to identify the polymorphic sites. RESULTS: Microscopic results showed the presence of calcium oxalate prism sheath, unicellular trichomes, anisocytic type of stomata and the schizolysigenous oil glands. An alignment result of ITS sequences indicated several polymorphic sites that could be exploited as DNA markers and revealed the presence of a BsrBI restriction site in M. koenigii which could be used to cut the PCR amplified DNA segment to generate fingerprint for identification of M. koenigii. CONCLUSION: Microscopic investigation of M. koenigii leaf exhibited characteristics of Murraya species and the ITS sequence was an appropriate molecular marker for discrimination of M. koenigii from the selected plants.Keywords: Murraya koenigii, microscopic investigation, DNA marker, internal transcribed spacer