Universal primers were used for PCR amplification of three noncoding regions of chloroplast DNA (cpDNA) in order to study sequence-length variation in the Crassulaceae and in related species. Several length mutations were observed that are of diagnostic value for evolutionary relationships in the Crassulaceae and the Saxifragaceae. Length variation and sequence divergence in the intergenic spacer between the trnL (UAA) 3' exon and the trnF (GAA) gene among 15 species were studied in detail by nucleotide-sequence analysis. A total of 50 insertion/deletion mutations were observed, accounting for a spacer-length variation in the range of 228-360 bp. Eighteen short direct repeat motifs (4-11 bp) and two inverted repeat motifs (7-11 bp) were found to be associated with length variation. Phylogenetic analysis of the sequence data indicated a pattern of relationships that was largely consistent with a previous analysis of cpDNA restriction-site variation. Evaluation of the level of homoplasy in insertion/deletion mutations within a phylogenetic framework revealed that only 1 out of 34 length mutations longer than 2 bp must have had multiple origins. The feasibility of the noncoding chloroplast DNA regions for molecular evolutionary studies is discussed.
A backcross population of the L. peruvianum accession LA 2157, which is resistant to bacterial canker caused by Clavibacter michiganensis ssp. michiganensis, with the susceptible L. peruvianum accession LA 2172 was evaluated for the segregation of C. michiganenis resistance and of RFLP markers in order to map the loci involved in this resistance. The development of symptoms of the disease was scored using an ordinal scale. The mapping of the disease resistance was hampered by distorted segregation ratios of a large number of markers and unexpected quantitative inheritance of the resistance. By means of the Kruskal-Wallis rank-sum test, five regions on chromosomes 1, 6, 7, 8 and 10 were identified that may be involved in C. michiganensis resistance.
In order to map genes determining resistance to bacterial canker in tomato, backcrosses were made between a resistant and a susceptible Lycopersicon peruvianum accession. The linkage study with RFLP markers yielded a genetic map of L. Peruvianum. This map was compared to that derived from a L. esculentum x L. pennellii F2 population, based on 70 shared RFLP markers. The maps showed a good resemblance in both the order of markers and the length of the chromosomes, with the exception of just one relocated marker on chromosome 9. Because backcrosses were made with the F1, either as the pollen parent or as the pistil parent, linkage maps from male and female meioses could be estimated. It was concluded that recombination at male meiosis was reduced, and that gametophytic selection for parental genotypes at more than one locus per chromosome might be partly responsible for the reduction of the estimated male map length.
Phylogenetic relationships between species of Allium section Cepa and A. r q k (section Rhirirideum) have been inferred from nuclear DNA variation (RAPDs; nDNA dataset) and from morphological, pollen epidermis texture, chromosomal and chemical variation (supranuclear dataset). These sets were complemented with data, taken from the literature, on cpDNA variation and crossability. The trees produced with the supranuclear, nDNA and cpDNA datasets were compared by using the topology of the most parsimonious tree of one dataset as the constraint for the construction of a most parsimonious tree of another dataset. The accuracy of the trees were evaluated by calculating several Consistency and Incongruence Indices. The constrained tree of supranuclear-nDNA datasets showed the highest index values. The tree topologies of the supranuclear and cpDNA datasets were the least similar. The cpDNA tree and crossability dendrograms were identical. The most important difference between the nDNAsupranuclear trees and the cpDNA-crossability trees pertains to the position of Allium r q k , which is much closer to the clade A. cepa/A. uauilozii in the cpDNA tree than in the nDNA tree. This difference is considered to be the result of chloroplast capture from one species to another after an introgression event. A shorter distance between species inferred from a cpDNA tree than from a nDNA or comparable tree might be indicative for the level of crossability.
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