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REPEAT AND SINGLE COPY SEQUENCES ARE DIFFERENTIALLY CONSERVED IN THE EVOLUTION OF KELP CHLOROPLAST DNA 1
Author(s) -
Fain Steven R.,
Druehl Louis D.,
Baillie David L.
Publication year - 1988
Publication title -
journal of phycology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.85
H-Index - 127
eISSN - 1529-8817
pISSN - 0022-3646
DOI - 10.1111/j.1529-8817.1988.tb04469.x
Subject(s) - biology , chloroplast dna , restriction enzyme , restriction fragment , phylogenetic tree , kelp , restriction site , genetics , botany , dna , gene
A method for the isolation of total blade, chloroplast and nuclear DNAs from kelp is described. Total blade DNA was isolated from various kelp species at yields of about 1.0 μg.g −1 wet weight of tissue extracted. Purified chloroplast and nuclear DNAs were isolated from Macrocystis integrifolia at respective yields of about 0.05 μg.g −1 and 1.0 μg.g −1 wet weight of tissue extracted. Hybridization experiments with a cloned portion of the 23S rRNA gene of M. integrifolia as probe (pMiR23S) showed that the M. integrifolia chloroplast genome contains two copies of the 23S rRNA gene. Three regions of the kelp chloroplast genome were studied by restriction analysis of the cpDNA of five genera. Respective differences were assessed by comparing the restriction fragment patterns produced with five restriction endonucleases for each of three cloned M. integrifolia cpDNA hybridization probes. The kelp cpDNA sequences that hybridized to the repeated sequence probe pMiR23S were less divergent (100 × p = 2.53) than were the sequences that hybridized to the single copy probes pMiR432 and pMiH7 (100 × p = 7.48–7.74). Sequence variation (100 × p) between kelp species ranged from 3.38–6.82. Sequence variation between kelp species and Fucus gardneri ranged from 9.08–12.04. Chloroplast DNA restriction fragment length data were used to derive two most parsimonious phylogenetic trees. Both trees had consistency indexes of 0.78 and indicated that the three lessoniacean genera in the comparison do not comprise a single distinct lineage.