DOYLE, JEFF J.1*, JANE L. DOYLE1, AND A. H. D. BROWN2. 1L. H. Bailey Hortorium, Cornell University, Ithaca, NY 14853, and 2CSIRO Division of Plant Industry, Canberra, ACT, Australia. - Incongruence in the diploid B-genome species complex of Glycine (Leguminosae) revisited: Histone H3-D alleles vs. chloroplast haplotypes.
The B-genome of Glycine subg. Glycine (Leguminosae:
Papilionoideae) comprises three named Australian diploid species and a
number of distinctive but as yet not formally recognized taxa.
Chloroplast DNA restriction site studies of this group revealed a
monophyletic set of haplotypes that were polymorphic within taxa and
transgressed apparent taxonomic boundaries. We have surveyed over 40
accessions from that study for sequence variation at the single-copy,
intron-containing nuclear locus histone H3-D. As expected for these
largely inbreeding taxa, there was little evidence of heterozygosity,
facilitating direct sequencing of PCR products. A total of 21 alleles
were found, differing by as many as 17 nucleotide substitutions in the
approximately 500 bp region analyzed. Phylogenetic analysis
identified two shortest trees, and revealed very little homoplasy (ci
> 0.95). Four major groups of alleles were well-supported in the H3-D
trees, those from: 1) G. microphylla; 2) an unnamed but
morphologically distinctive taxon supported by cpDNA as basal in the
diploid B-genome group (G. sp. aff. G. tabacina); 3) two
accessions of a second unnamed taxon from an ecologically distinctive
region of high Glycine diversity; and 4) the remaining
accessions, including G. latifolia, several unnamed taxa, and
the "diploid G. tabacina complex." Several subclades
appeared in the fourth group. The distribution of H3-D alleles agreed
much better with species boundaries than did the distribution of cpDNA
haplotypes. The agreement of H3-D alleles with morphology implicates
cpDNA as the incongruent marker, and leaves us with the usual
processes--lineage sorting or introgression--as possible explanations,
as usual without distinguishing decisively between them. The H3-D
data themselves pose some interesting problems, however, such as how
there came to be only a single identical allele throughout our
geographically broad sample of G. microphylla.
Key words: cpDNA, Glycine, histone gene sequences, incongruence, Leguminosae, phylogeny