We analysed 87 species of Onosma (Boraginaceae) from throughout its distribution range to investigate its evolutionary history. Using nrDNA ITS and two plastid (rpl32‐trnL(UAG) and trnH–psbA) markers, we reconstructed phylogenetic relationships within Onosma by conducting maximum parsimony, maximum likelihood, Bayesian, and BEAST analyses. The analyses revealed that Onosma as currently circumscribed is not monophyletic. However, the vast majority of Onosma species appear to belong to a single clade, the so‐called Onosma s.s. Outside of this core clade is a clade containing O. rostellata, a subclade of Sino‐Indian species and Maharanga emodii. Podonosma orientalis (as O. orientalis) appear only distantly related to Onosma but is more closely related to Alkanna, as also suggested in previous molecular studies. The Onosma s.s. clade includes all representatives of O. sect. Onosma, and encompasses three subsections, i.e. Onosma, Haplotricha and Heterotricha, corresponding to asterotrichous, haplotrichous and heterotrichous groups, respectively, but none of these subsections was retrieved as monophyletic. We observed significant incongruence between nuclear and chloroplast phylogenies regarding the phylogenetic status of the heterotrichous group. A dozen of the Iranian haplotrichous species formed a lineage which may not hybridize with asterotrichous species. Divergence time estimates suggested that the early radiation of Onosma s.l. took place at the Oligocene‐Miocene boundary and the diversification within Onosma s.s. occurred during middle to late Miocene and Pliocene.

Hedera, with 12 extant species, is a genus of evergreen climbers native to Europe, north Africa, and south Asia. In this study, the micromorphological, anatomical structure and molecular evidences of 11 populations from two species of Hedera (H. helix and H. pastuchovii) have been considered to evaluate the relationships in Hedera. In total, seven quantitative and qualitative characters of pollen were selected and measured. Based on this study, the anticlinal wall and surface sculpturing of seed support for separation of two species of Hedera. Micromorphology of epidermis illustrated two types of epidermal cells: puzzle-shaped and polygonal cells. Using nuclear (nrDNA ITS) marker, we reconstructed phylogenetic relationships within two species of Hedera. This data set was analyzed by phylogenetic methods including Bayesian inference, maximum likelihood, and maximum parsimony. In phylogenetic analyses, all members of two species formed a well-supported clade (PP = 1; ML/BS = 100/100) and divided into two major clades (A and B). Neighbor Net diagram demonstrated separation of the studied populations. The results showed that these taxa differ in taxonomically important micromorphological, anatomical and molecular characteristics and these data provide reliable evidence for separation of these two species.

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Sambucus is a medicinal plant with approximately 25 accepted taxa in the world from which two are present in Iran, S. nigra and S. ebulus. In order to evaluate the affinities and relationships in Sambucus and resurrect subspecies or verities in these two species, the micro-morphological, anatomical and molecular properties of Sambucus species of Iran have been studied. Pollen and seed surfaces of two species were examined with scanning electron microscope. In micro-morphological studies, the pollen shape is prolate-spheroidal in S. nigra but subprolate in S. ebulus. The exine sculpture is microperforate in S. nigra and is reticulate-microechinate in S. ebulus. The surface of the seed in S. nigra is smooth, not elongated and no clear organizational pattern is discernable and the anticlinal walls are shallowly undulate. In S. ebulus, the seeds have a microperforate surface and the anticlinal walls are observed deeply undulate. Petiole anatomical studies show that vascular bundles and existence of druse crystals are diagnostic characters in studied species. Using nuclear (nrDNA ITS) marker, we reconstructed phylogenetic relationships within two species of Sambucus. The ITS dataset was analyzed by phylogenetic methods including Bayesian inference, maximum likelihood and maximum parsimony. In phylogenetic analyses, all members of this genus form a well-supported clade (PP = 1, ML/BS = 100/100). The Sambucus clade is composed of two clades. Clade I includes the populations of S. nigra and the clade II comprises the rest of the species of Sambucus (S. ebulus). NeighborNet diagram demonstrated separation of the studied populations. In general, molecular studies supported micro-morphological findings. Pollen, seed-coat characters and molecular phylogeny support the separation of S. nigra and S. ebulus .