DNA Barcoding of Three Species of Bryaceae -Musci

Abstract

DNA barcoding is a relatively novel system designed to provide rapid, accurate and automatable species identifications by using short, standardized gene regions as internal species tags. Bryum (Hedw) is a cosmopolitan moss genus with a reputation for taxonomic difficulty. Fourteen out of 58 collected Egyptian brya have been identified to the specific level while 44 still require precise identification. Three taxa were used, where these taxa represent a part of a greater study which includes different herbarium moss genera in Egypt. Their DNA was extracted, and then subjected to a PCR reaction for two different loci termed chloroplast rbcL and the nuclear ITS regions. They are the most suitable regions for Barcoding in bryophytes. Nucleotide sequences for both regions were determined and analyzed. These sequences were first edited, and then blasted to National Center for Biotechnology Information (NCBI) and Barcoding of Life Database (BOLD) for alignment with published gene’s sequences of same or closest species recorded to verify the identification. If those sequences show similarity less than 95% of two genes we are interested in or don’t meet the located sequence in GenBank, we relied on the morphological approach and submitted all sequences to GenBank with the verified name morphologically and genetically. In this work, DNA Barcoding is used to discriminate and confirm the identification of three Bryum taxa. This is the first time in Egypt, where study of mosses is carried out at the molecular level. Five new genes were recorded from the three taxa, one from Bryum ruderale, two from the probably B. mildeanum, and the last two from the probably B. bornholmense. The existence of B.ruderale in Egypt is confirmed.