A new species of Haplophyllum A. Juss. (Rutaceae) from the Iberian Peninsula: evidence from morphological, karyological and molecular analyses.
OBJECTIVE
The discovery of a new species, Haplophyllum bastetanum F.B. Navarro, V.N. Suárez-Santiago & Blanca sp. nov., in the south-east of Spain has prompted the comparative study of species of the Iberian Peninsula, and others related, through morphological, cytogenetic, molecular, distributional and ecological characterization.


METHODS
The morphological study involved a quantitative analysis of the species present in the Iberian Peninsula and a comparative analysis of the morphological characteristics between H. bastetanum and other related species. Mitotic analyses were made with root meristems taken from germinating seeds. Phylogenetic analyses of the internal transcribed spacer sequences of nuclear ribosomal DNA were performed using neighbour-joining (NJ) and maximum-parsimony methods.


RESULTS
Haplophyllum bastetanum is a diploid species (2n = 18) distinguished primarily for its non-trifoliate glabrous leaves, lanceolate sepals, dark-green petals with a dorsal band of hairs, and a highly hairy ovary with round-apex locules. The other two Iberian species (H. linifolium and H. rosmarinifolium) are tetraploid (2n = 36) and have yellow petals. Both phylogenetic methods generated a well-supported clade grouping H. linifolium with H. rosmarinifolium. In the NJ tree, the H. linifolium-H. rosmarinifolium clade is a sister group to H. bastetanum, while in the parsimony analysis this occurred only when the gaps were coded as a fifth base and the characters were reweighted according to the rescaled consistency index. This latter group is supported by the sequence divergence among taxa.


CONCLUSIONS
The phylogenies established from DNA sequences together with morphological and cytogenetic analyses support the separation of H. bastetanum as a new species. The results suggest that the change in the number of chromosomes may be the key mechanism of speciation of the genus Haplophyllum in the Iberian Peninsula. An evolutionary scheme for them is propounded.
Departamento de Botánica, Facultad de Ciencias, Universidad de Granada, 18071, Spain. pipo@ugr.es
F B
FB
Navarro
V N
VN
Suárez-Santiago
G
G
Blanca
D001483:Base Sequence; D005075:Biological Evolution; D021901:DNA, Intergenic / Q000737:chemistry; D018744:DNA, Plant / Q000032:analysis; D035264:Flowers / Q000033:anatomy & histology; D005638:Fruit / Q000033:anatomy & histology; D010641:Phenotype; D010802:Phylogeny; D018515:Plant Leaves / Q000033:anatomy & histology; D018547:Plant Stems / Q000033:anatomy & histology; D011003:Ploidies; D027821:Rutaceae / Q000033:anatomy & histology / Q000166:cytology / Q000235:genetics*; D012639:Seeds / Q000648:ultrastructure; D013030:Spain; D013045:Species Specificity
D021901:DNA, Intergenic; D018744:DNA, Plant