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Species identification of red and brown seaweeds using ITS ribosomal DNA amplification and RFLP patterns
Author(s) -
Antoine Elisabeth,
Fleurence Joël
Publication year - 2003
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.1362
Subject(s) - biology , porphyra , restriction fragment length polymorphism , algae , brown algae , restriction enzyme , ribosomal dna , laminaria , red algae , botany , laminaria digitata , internal transcribed spacer , ribosomal rna , polymerase chain reaction , dna , gene , biochemistry , phylogenetics
The identity of five macroalgae used as sea vegetables or food ingredients has been determined by amplification of the ribosomal DNA ITS region and RFLP analysis. This allows the detection of specificity patterns for each species and provides an alternative method, when morphological or biochemical methods fail, for control of their use as food ingredients. Alga‐specific PCR primers have been used to determine the ITS rDNA sequences of DNAs extracted from dried and ground algae up to 5 years old. The seaweeds studied were the red algae Palmaria palmata (dulse), Porphyra umbilicalis (nori), and Chondrus crispus (pioca) and the brown algae Himanthalia elongata (sea spaghetti) and Laminaria sp (konbu). Total genomic DNA suitable for amplification was extracted from the alga powder following the CTAB method. This methodology allowed the sequencing of the amplified product and the drawing of theoretical restriction maps useful in the choice of restriction enzymes for RFLP analysis. Enzymes that appeared useful included Mbo I and Alu I . Cutting with a single enzyme was sufficient to obtain characteristic patterns for the red algae P palmata, P umbilicalis and C crispus . For the two brown algae H elongata and Laminaria sp the ITS rDNA sequence showed a lack of suitable restriction site, contrary to other species for which characteristic patterns were obtained. © 2003 Society of Chemical Industry