In collaboration with Payame Noor University and Iranian Biotechnology Society

Document Type : Research Paper

Authors

1 M.Sc. Agricultural Biotechnology, University of Mohaghegh Ardabili, Ardabil, Iran.

2 Assistant Professor, Department of Agronomy & Plant Breeding, Faculty of Agricultural Sciences, University of Mohaghegh Ardabili, Ardabil, Iran

3 Plant biotechnology researcher, Plant breeding department, Wageningen UR.

4 Associate Professor, Department of Plant Protection, Faculty of Agricultural Sciences, University of Mohaghegh Ardabili, Ardabil, Iran

Abstract

DNA barcoding is a simple way to identify species using a very short genetic sequence from a standard part of the genome. This technique used to identify eight medicinal plants collected from the Ardabil province. DNA extraction was performed by modified CTAB method and PCR was performed with primers designed based on rbcL, trnH-psbA, matK Chloroplast barcodes and ITS nuclear barcode. Then, PCR products purified and sequenced. The percentage of amplification and sequencing success were assumed in samples respectively, 87 and 62, 75 and 37, 62 and 12, 75 and 37. The sequences were blasted with samples existed in NCBI database and Bioinformatics analyses were performed. In phylogenetic tree, the species belonging to the same genus were separated from other genus based on rbcL and trnH-psbA barcode sequences. Also, in ITS barcode only G. glabra organized with plants from same genus. In this study, barcoding of L. ledebourii with rbcL was done for the first time. SNPs were counted for barcodes of rbcL (less than 30), trnH-psbA (less than100), ITS (more than 200) and matK (less than 20). Thus, rbcL barcode due to high separation ability, low number of SNPs and universality in most species, was introduced as the best barcode. However, trnH-psbA and ITS barcodes due to related problem with direct sequencing of PCR products and lack of access to high quality sequences were identified as complementary barcodes. MatK barcode is not recommended for these samples because of the low ability of amplification and sequencing.

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Main Subjects

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