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VARIATION IN THE CHLOROPLAST GENES RPOC1 AND RPOC2 OF THE GENUS ASTRAGALUS (FABACEAE): EVIDENCE FROM RESTRICTION SITE MAPPING OF A PCR‐AMPLIFIED FRAGMENT
Author(s) -
Liston Aaron
Publication year - 1992
Publication title -
american journal of botany
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.218
H-Index - 151
eISSN - 1537-2197
pISSN - 0002-9122
DOI - 10.1002/j.1537-2197.1992.tb13679.x
Subject(s) - biology , restriction site , genetics , restriction enzyme , intergenic region , restriction fragment , phylogenetic tree , gene , restriction map , restriction fragment length polymorphism , intron , genome , polymerase chain reaction , nucleic acid sequence
A 4,100‐base pair (bp) region of the chloroplast genome, amplified via the polymerase chain reaction, was obtained from 14 species of the genus Astragalus and mapped with 23 restriction enzymes. The amplified region encompassed the chloroplast genes RNA polymerase Cl ( rpo Cl; 90.8% of the gene) and RNA polymerase C2 ( rpo C2; 32.7% of the gene) including the intron in rpo C1 and the intergenic spacer between the two genes. Approximately 144 sites (615 bp) were identified; 37 restriction site mutations and one 10‐bp length mutation were detected. Estimated interspecific sequence divergence values ranged from 0.00% to 3.92%. Phylogenetic analysis with Wagner and Dollo parsimony both resulted in a single 41‐step tree with a consistency index of 0.951. The relative positions of 115 restriction sites were mapped. The insertion and ten of the restriction site mutations mapped to the intron in rpo C1, 18 site mutations mapped to the rpo C1 exons, three site mutations mapped to rpo C2, three site changes mapped to the intergenic spacer, and four site changes were not mapped. This study demonstrates the utility of restriction site analysis of PCR‐amplified chloroplast DNA to the study of plant phylogenetic relationships and molecular evolution.