trnL-F Spacer Research Articles

Molecular phylogenetic reconstruction improves the taxonomic understanding of Indian Dipcadi (Asparagaceae) and reveals a new species from the bank of Hiranyakeshi River, Maharashtra, India.

Dipcadi (Scilloideae: Asparagaceae) is a genus of bulbous monocots with approximately 40 species, of which 13 occur in India. Species delimitation within the genus has been troublesome hindering a comprehensive phylogenetic analysis. The most recent phylogeny of the subfamily Ornithogaloideae included six species of Dipcadi only from Africa. Here, we reconstructed the phylogeny of Ornithogaloideae including 23 accessions comprising 13 recognized taxa (11 species and two varieties) of Indian Dipcadi. The phylogenetic analyses were based on nucleotide sequences of three plastid regions (rbcL, matK and trnL-F spacer) and one nuclear region (ITS). Pseudogaltonia clavata exhibited sister relationship to Dipcadi. Our combined nuclear + plastid dataset analyses revealed a monophyletic Dipcadi with five clades, Clade I-V. Clade I, II and III included mainly Indian species whereas Clade V included mostly African species. Clade IV comprised D. serotinum. Clade I included nine taxa including our newly described species, D. mukaianum. The new species was phylogenetically placed with D. erythraeum, D. saxorum and D. ursulae. Morphologically, the species resembled D. montanum and D. ursulae but differed in characters such as tepal cohesion, number of ovules per locule and foul-smelling flowers. Clade II and III included 11 and six taxa, respectively. D. erythraeum which has a native range from Egypt to western India was found in Clades I and V. The widespread Dipcadi species, viz. D. erythraeum and D. serotinum showed polyphyly however, the monophyly of Dipcadi is established. Our studies suggest that additional molecular markers (plastid as well asnuclear) should be tested for their taxonomy utility. Further work on the historical biogeography of Dipcadi on the subfamily Ornithogaloideae with more genetic data will yield insights how aridification of the landscape would have shaped the evolution of the geographical clades.

Three Newly Described Species of Ziziphus from Maharashtra, India Based on Morphological and Molecular Evidence

Abstract— Three new species of Ziziphus (Rhamnaceae) from Maharashtra, India, Z. bhandarii, Z. naikii, and Z. bhansalii are described here. These species occur in isolated fragments of the Deccan thorn scrub forest ecoregion on the Deccan plateau. Their placement in Ziziphus is confirmed by molecular analyses combining nrITS and trnL-F spacer regions. Given this phylogeny is the most robust to date of Ziziphus, a previously proposed intrageneric classification was tested, and the classification was not supported suggesting a need for additional investigations in this genus. The new species are most closely related to three Indian species, Z. horrida, Z. xylopyrus, and Z. caracutta, respectively, which are morphologically similar and sympatric, and are distinguishable from known species based on several vegetative and reproductive characteristics presented here. A diagnostic key to the Ziziphus species known to occur in Maharashtra is provided.

Phylogenetic, cytogenetic and morphological evidences are critical for recognizing a new genus: Valdesiana, an Iberian intergeneric allopolyploid between Schenkia and Exaculum

The present taxonomic status of Schenkia elegans, endemism recently described in the Iberian Peninsula, and its relationship with the sympatric and the nearest morphological species Schenkia spicata and Exaculum pusillum is revaluated. Different kinds of evidence based on plant morphology, ploidy estimation by flow cytometry, karyotype characterisation, and phylogenetic data have been analysed. Two maternally inherited plastid DNA regions (trnL intron and trnL-F spacer) and biparentally inherited nuclear ribosomal DNA sequence region (nrDNA ITS) have been used. Comparative multivariate analyses show an intermediate morphology of the S. elegans plants between the other two species studied. Flow cytometry and karyotype analyses in S. elegans point to an allopolyploid origin, with the latter constituted by a mixture of those of the diploids S. spicata and E. pusillum. Phylogenetic analyses based on plastid and nuclear DNA regions cluster S. elegans in two different clades, those of S. spicata and E. pusillum, suggesting a possible hybrid origin of S. elegans between both species, acting as maternal or paternal progenitors. In consequence, taking in consideration the taxonomic relationships among genera (Exaculum, Schenkia and the closely related genus Zeltnera found in America), a monotypic genus Valdesiana gen. nov. is proposed to accommodate the allopolyploid species, combined as V. elegans, for which immediate conservation measures must be evaluated.

Deep Insights Into the Plastome Evolution and Phylogenetic Relationships of the Tribe Urticeae (Family Urticaceae).

Urticeae s.l., a tribe of Urticaceae well-known for their stinging trichomes, consists of more than 10 genera and approximately 220 species. Relationships within this tribe remain poorly known due to the limited molecular and taxonomic sampling in previous studies, and chloroplast genome (CP genome/plastome) evolution is still largely unaddressed. To address these concerns, we used genome skimming data—CP genome and nuclear ribosomal DNA (18S-ITS1-5.8S-ITS2-26S); 106 accessions—for the very first time to attempt resolving the recalcitrant relationships and to explore chloroplast structural evolution across the group. Furthermore, we assembled a taxon rich two-locus dataset of trnL-F spacer and ITS sequences across 291 accessions to complement our genome skimming dataset. We found that Urticeae plastomes exhibit the tetrad structure typical of angiosperms, with sizes ranging from 145 to 161 kb and encoding a set of 110–112 unique genes. The studied plastomes have also undergone several structural variations, including inverted repeat (IR) expansions and contractions, inversion of the trnN-GUU gene, losses of the rps19 gene, and the rpl2 intron, and the proliferation of multiple repeat types; 11 hypervariable regions were also identified. Our phylogenomic analyses largely resolved major relationships across tribe Urticeae, supporting the monophyly of the tribe and most of its genera except for Laportea, Urera, and Urtica, which were recovered as polyphyletic with strong support. Our analyses also resolved with strong support several previously contentious branches: (1) Girardinia as a sister to the Dendrocnide-Discocnide-Laportea-Nanocnide-Zhengyia-Urtica-Hesperocnide clade and (2) Poikilospermum as sister to the recently transcribed Urera sensu stricto. Analyses of the taxon-rich, two-locus dataset showed lower support but was largely congruent with results from the CP genome and nuclear ribosomal DNA dataset. Collectively, our study highlights the power of genome skimming data to ameliorate phylogenetic resolution and provides new insights into phylogenetic relationships and chloroplast structural evolution in Urticeae.

Parahellenia, a new genus segregated from Hellenia (Costaceae) based on phylogenetic and morphological evidence

Previous studies recognized three major lineages of the family Costaceae: a South American clade, an Asian clade and a Costus clade. However, the genus Hellenia within the Asian clade has been shown to be non-monophyletic and its morphology has not been studied carefully. Therefore, the complete plastid genomes of Hellenia species were obtained and the monophyly of Hellenia was tested through four different datasets in this study. Plastid phylogenomic analyses of Costaceae revealed that Hellenia is strongly supported as paraphyletic. Two major clades are recovered, namely the Hellenia s.s. subclade and the Parahellenia subclade. Phylogenetic analyses based on an enlarged taxon sampling of the Asian clade using a two chloroplast markers dataset (trnK intron and trnL-F spacer) confirmed the paraphyly of Hellenia. Meanwhile, morphological analyses suggested that members of the Parahellenia subclade differ from the remaining Hellenia species in many characters including inflorescences, bracts, stigma, axillary buds, floral tubes and labellum. According to the present molecular and morphological evidence, the latter subclade is recognized as a new genus, Parahellenia. Two new species are described, four new combinations are made, and identification keys are also provided.

A revised infrageneric classification of Bellevalia Lapeyr. (Asparagaceae: Scilloideae) based on molecular analysis

Within the genus Bellevalia (Asparagaceae: Scilloideae: Hyacintheae), flower shape and colour, the ratio of leaf to scape length, and the orientation of the pedicels were traditionally used to delimit sections, subsections and species. We conducted a phylogenetic analysis of representative species from sections Bellevalia, Conicae, Nutantes and Oxydontae, representing four out of the six sections that are currently recognised, using the four chloroplast regions rbcL, matK, trnL intron, and trnL-F spacer. Our results indicate that the sections are not monophyletic as currently circumscribed. Our analyses retrieve two major, well-supported clades. The first clade (sect. Conicae) includes only species with ciliate leaves and green-veined perianth lobes, and the second clade (sect. Bellevalia) includes the species lacking these characteristics. Within the second clade, the species with yellow anthers are separated from those with violet anthers. Our molecular analysis does not support the traditional subdivisions of the genus but establishes the value of cilia on the leaf margin, green veins in the perianths lobes, and the colour of the anthers as indicators of relationships among the species. We recommend that the current four sections and six subsections of Bellevalia be reduced to two sections without subsections and we provide an updated and corrected nomenclature for these sections and subsections, designating lectotypes where necessary.

Molecular and Morphological Data Improve the Classification of Plantagineae (Lamiales).

The tribe Plantagineae (Lamiales) is a group of plants with worldwide distribution, notorious for its complicated taxonomy and still unresolved natural history. We describe the result of a broadly sampled phylogenetic study of tribe. The expanded sampling dataset is based on the trnL-F spacer, rbcL, and ITS2 markers across all three included genera (Aragoa, Littorella and Plantago) and makes this the most comprehensive study to date. The other dataset uses five markers and provides remarkably good resolution throughout the tree, including support for all of the major clades. In addition to the molecular phylogeny, a morphology database of 114 binary characters was assembled to provide comparison with the molecular phylogeny and to develop a means to assign species not sampled in the molecular analysis to their most closely related species that were sampled. Based on the molecular phylogeny and the assignment algorithm to place unsampled species, a key to sections is presented, and a revised classification of the tribe is provided. We also include the description of new species from North America.

Molecular phylogenetics of Malesian Diospyros (Ebenaceae) based trnL-F spacer sequences

Abstract. Wanda IF, Djuita NR, Chikmawati T. 2021. Molecular phylogenetics of Malesian Diospyros (Ebenaceae) based trnL-F spacer sequences. Biodiversitas 22: 4106-4114. Diospyros is a genus composed of potential species as an economic commodity with high diversity. However, there is limited information on the phylogenetic relationship of this genus in the Malesian region. This study aimed to provide information on the species diversity through a DNA barcoding approach and revealing the phylogenetic information of Diospyros spp. in the Malesian region. This study used 20 Diospyros accessions from Bogor Botanical Garden collections, 40 Diospyros accessions, and four outgroup accessions obtained from the NCBI database. The DNA barcoding primer utilized comes from plastids, trnL-F intergenic spacer. The phylogenetic trees were constructed using the Maximum-Parsimony method. A total of 20 accessions of Diospyros were validated with sequence data on the genebank. The result showed that all accessions had relationships with 44 other Diospyros species globally. Here, we reported 10 new trnL-F intergenic spacer sequences of Malesian Diospyros species. A phylogenetic tree grouped 64 monophyletic Diospyros species into seven clades. The phylogenetic results supports the biogeographic hypothesis: the Malesian region, the Malesian-Caledonian Region, and Cosmopolite species in almost all bioregions.

A phylogenetic analysis of the Crinum rautanenianum complex (Crininae, Amaryllidaceae) – with a description of C. luangwense sp. nov.

Within genus Crinum L. there has been a lot of confusion, as to species delimitation, particularly in the Crinum rautanenianum complex comprising Crinum carolo-schmidtii, C. euchrophyllum, C. paludosum, C. rautanenianum, and also a specimen referred to as sp. A in Flora of Tropical East Africa. Maximum parsimony and Bayesian inference analyses were conducted on two datasets: nuclear DNA region (ITS) and five plastid regions (trnL-F spacer, trnL intron, rps16 intron, psbA-trnH spacer, and rps12-rpl20 spacer). Additionally, morphological characters based on herbarium specimens, pictures from the field, type specimens, and protologues were studied to establish diagnostic characters within the complex. Results from phylogeny and morphological characters provide evidence for the reinstatement of C. euchrophyllum, recognition of C. rautanenianum, C. carolo-schmidtii, C. paludosum, with a revisited description of C. rautanenianum, and the description of a new taxon: C. luangwense. A taxonomic revision of the C. rautanenianum complex is presented, including diagnostic characters, an identification key and distribution maps.

Delimiting the genera of the Ficinia Clade (Cypereae, Cyperaceae) based on molecular phylogenetic data.

Generic delimitations in the Ficinia Clade of tribe Cypereae are revisited. In particular, we aim to establish the placement of annual species currently included in Isolepis of which the phylogenetic position is uncertain. Phylogenetic inference is based on two nuclear markers (ETS, ITS) and five plastid markers (the genes matK, ndhF, rbcL and rps16, the trnL intron and trnL-F spacer) data, analyzed using model based methods. Topologies based on nuclear and plastid data show incongruence at the backbone. Therefore, the results are presented separately. The monophyly of the smaller genera (Afroscirpoides, Dracoscirpoides, Erioscirpus, Hellmuthia, Scirpoides) is confirmed. However, Isolepis is paraphyletic as Ficinia is retrieved as one of its clades. Furthermore, Ficinia is paraphyletic if I. marginata and allies are excluded. We take a pragmatic approach based on the nuclear topology, driven by a desire to minimize taxonomic changes, to recircumscribe Ficinia to include the annual Isolepis species characterized by cartilaginous glumes and formally include all the Isolepis species inferred outside the core Isolepis clade. Consequently, the circumscription of Isolepis is narrowed to encompass only those species retrieved as part of the core Isolepis clade. Five new combinations are made (Ficinia neocapensis, Ficinia hemiuncialis, Ficinia incomtula, Ficinia leucoloma, Ficinia minuta). We present nomenclatural summary at genus level, identification keys and diagnostic features.

Rediscovery of Angiopteris tonkinensis (Marattiaceae) after 100 years, and its revision.

The border area between south-eastern Yunnan, China and northern Vietnam is one of the regions with richest biological diversity including that of the fern genus Angiopteris (Marattiaceae). Based on the analysis of morphology and DNA sequences of multiple chloroplast regions (atpB, rbcL, rps4-trnS spacer and trnL-F spacer), we revised Angiopteris tonkinensis (Hayata) J.M.Camus and proposed a new combination Angiopteris tamdaoensis (Hayata) J.Y.Xiang & T.Wang, comb. nov., which was previously regarded as a synonym of A. tonkinensis. We found support for a monophyletic Angiopteris including Protomarattia. This discovery adds two new distribution sites of A. tonkinensis, one in China (Malipo, Yunnan) and one in Vietnam (Quan Ba, Ha Giang). We suggest A. tonkinensis should be categorised as Critically Endangered (CR) species according to the criteria of IUCN.

Phylogeny and Historical Biogeography of Paphiopedilum Pfitzer (Orchidaceae) Based on Nuclear and Plastid DNA.

The phylogeny and biogeography of the genus Paphiopedilum were evaluated by using phylogenetic trees derived from analysis of nuclear ribosomal internal transcribed spacer (ITS) sequences, the plastid trnL intron, the trnL-F spacer, and the atpB-rbcL spacer. This genus was divided into three subgenera: Parvisepalum, Brachypetalum, and Paphiopedilum. Each of them is monophyletic with high bootstrap supports according to the highly resolved phylogenetic tree reconstructed by combined sequences. There are five sections within the subgenus Paphiopedilum, including Coryopedilum, Pardalopetalum, Cochlopetalum, Paphiopedilum, and Barbata. The subgenus Parvisepalum is phylogenetic basal, which suggesting that Parvisepalum is comprising more ancestral characters than other subgenera. The evolutionary trend of genus Paphiopedilum was deduced based on the maximum likelihood (ML) tree and Bayesian Evolutionary Analysis Sampling Trees (BEAST). Reconstruct Ancestral State in Phylogenies (RASP) analyses based on the combined sequence data. The biogeographic analysis indicates that Paphiopedilum species were firstly derived in Southern China and Southeast Asia, subsequently dispersed into the Southeast Asian archipelagoes. The subgenera Paphiopedilum was likely derived after these historical dispersals and vicariance events. Our research reveals the relevance of the differentiation of Paphiopedilum in Southeast Asia and geological history. Moreover, the biogeographic analysis explains that the significant evolutionary hotspots of these orchids in the Sundaland and Wallacea might be attributed to repeated migration and isolation events between the south-eastern Asia mainland and the Sunda Super Islands.

Assessment of the taxonomic status of the members of genus Artocarpus (Moraceae) in Sri Lanka

The genus Artocarpus J.R. Forster & G. Forster shows a higher diversity along the tropics of Indo Malayan region to Australasia. The Artocarpus phylogeny is resolved in previous studies. According to morpho-genetic assessments, Artocarpus spp. have been subdivided into subgenera and sections. However, the published phylogenies did not comprise the Artocarpus spp. found in Sri Lanka, leaving an ambiguity of their phylogenetic positions. The morphology was observed in many trees in natural range, and the line diagrams were drawn to showcase the morphological features that can be used to classify the Artocarpus spp. into subgenera and sections. We collected leaf samples from the Artocarpus spp. in Sri Lanka. The genomic DNA was extracted, and PCR amplified for five markers (trnH-psbA, rbcL, trnL-F spacer, trnSGCU-trnGUUC, and ITS1-4) followed by sequencing. The Artocarpus phylogeny was constructed in Maximum Likelihood and Bayesian framework using the generated sequences and the related sequence-popsets. Moreover, a divergence dating analysis was also carried out to check the evolutionary history of Sri Lankan Artocarpus spp. According to the morphology and phylogeny, the subgeneric classification of A. heterophyllus Lam, A. altilis (Parkinson) Fosberg, and, A. camansi Blanco was in line with previous studies. The dating analysis revealed a recent introduction of A. heterophyllus, A. altilis, and, A. camansi into Sri Lanka because of the associated food values. We positioned A. nobilis Thwaites, an endemic species to Sri Lanka, in the Artocarpus phylogeny under section Duricarpus. The nomenclatural status of A. gomezianus Wall. Ex Trecul coming under subspecies zeylanicus was verified as it does not clade closely with true A. gomezianus. However, we kept the species name as A. gomezianus as both true A. gomezianus, and Sri Lankan species got 10–13 pairs of lateral veins and the whitish hair underneath the leaf. We also recognized three ancient lineages of subgenera Artocarpus and Pseudojaca. These species naturally occur in India and Sri Lanka where they might be close descendants of the common ancestors of the subgenera Artocarpus and Pseudojaca.

Molecular Phylogeography of <i>Ficus benghalensis</i> Linnaeus using nrDNA ITS 1, cpDNA trnL and cpDNA rps16 from the Indian Subcontinent

Ficus benghalensis Linnaeus (Moraceae) is the national tree of India and is well known for its pharmacolo-gical properties. The present study was aimed to determine the genetic diversity of F. benghalensis from the Indian subcontinent using sequence-based multi-locus phylogeography. A total of 20 geograph-ical isolates were collected from different regions, cov-ering major parts of its species range within the country. Sequence data from nuclear-encoded internal transcribed spacer region (ITS1), plastid-encoded trnL-F spacer region (trnL) and ribosomal intron re-gion (rps16) were generated. The trnL-based maxi-mum likelihood phylogram revealed the existence of two haplotypes, whereas ITS1 and rps16-based maxi-mum likelihood analysis did not reveal much variation for this species distributed in the Indian subcontinent. These results depict long-distance random gene flow across the subcontinent, and support the post-glacial population contraction events. To validate the impact of palaeo-historic climatic events on current geo-graphic and genetic distribution, species distribution modelling-coupled phylogeography is suggested.

Polyphyletic origin in Pimpinella (Apiaceae): evidence in Western Europe.

The genus Pimpinella L. comprises about 150 species, being one of the largest genera within the family Apiaceae (subfamily Apioideae). Previous molecular phylogenetic studies have shown that Pimpinella is a taxonomically complex group. In this study, evolutionary relationships among representatives from Western Europe have been inferred from phylogenetic analyses of nuclear ribosomal DNA internal transcribed spacer (ITS 1 and ITS 2) and plastid sequences (trnL intron and the trnL-F spacer), with a representative sampling included (168 accessions in the ITS analysis, representing 158 species; and 42 accessions in the cpDNA analysis representing 35 taxa of Pimpinella and closely related species). All analyses resolved that Pimpinella is a non-monophyletic group, and Pimpinella's taxa that grow in Western Europe are part of phylogenetically independent groups that correspond to three different tribes of the subfamily Apioideae: Pimpinelleae (core group), Pyramidoptereae and Smyrnieae.

Coumarin Content, Morphological Variation, and Molecular Phylogenetics of Melilotus.

Melilotus albus and Melilotus officinalis are widely used in forage production and herbal medicine due to the biological activity of their coumarins, which have many biological and pharmacological activities, including anti-HIV and anti-tumor effects. To comprehensively evaluate M. albus and M. officinalis coumarin content (Cou), morphological variation, and molecular phylogeny, we examined the Cou, five morphological traits and the molecular characterization based on the trnL-F spacer and internal transcribed spacer (ITS) regions of 93 accessions. Significant (p < 0.05) variation was observed in the Cou and all five morphological traits in both species. Analysis of population differentiation (Pst) of the phenotypic traits showed that powdery mildew resistance (PMR) had the greatest Pst, meaning that this trait demonstrated the largest genetic differentiation among the accessions. The Pst values of dry matter yield (DMY) and Cou were relatively high. Biplot analysis identified accessions with higher DMY and higher and lower Cou. Analysis of molecular sequence variation identified seven haplotypes of the trnL-F spacer and 13 haplotypes of the ITS region. Based on haplotype and sequence analyses, the genetic variation of M. officinalis was higher than that of M. albus. Additionally, ITS sequence analysis showed that the variation among accessions was larger than that among species across three geographical areas: Asia, Europe, and North America. Similarly, variation among accessions for both the trnL-F and ITS sequences were larger than the differences between the geographical areas. Our results indicate that there has been considerable gene flow between the two Melilotus species. Our characterization of Cou and the morphological and genetic variations of these two Melilotus species may provide useful insights into germplasm improvement to enhance DMY and Cou.

Unidentifiable by morphology: DNA barcoding of plant material in local markets in Iran.

Local markets provide a rapid insight into the medicinal plants growing in a region as well as local traditional health concerns. Identification of market plant material can be challenging as plants are often sold in dried or processed forms. In this study, three approaches of DNA barcoding-based molecular identification of market samples are evaluated, two objective sequence matching approaches and an integrative approach that coalesces sequence matching with a priori and a posteriori data from other markers, morphology, ethnoclassification and species distribution. Plant samples from markets and herbal shops were identified using morphology, descriptions of local use, and vernacular names with relevant floras and pharmacopoeias. DNA barcoding was used for identification of samples that could not be identified to species level using morphology. Two methods based on BLAST similarity-based identification, were compared with an integrative identification approach. Integrative identification combining the optimized similarity-based approach with a priori and a posteriori information resulted in a 1.67, 1.95 and 2.00 fold increase for ITS, trnL-F spacer, and both combined, respectively. DNA barcoding of traded plant material requires objective strategies to include data from multiple markers, morphology, and traditional knowledge to optimize species level identification success.

A new species of Ilex (Aquifoliaceae) from Jiangxi Province, China, based on morphological and molecular data

A new species, Ilex venusta, is described and illustrated from the Luoxiao Mountains, Jiangxi Province, China. It is included in a phylogenetic analysis using nuclear ribosomal nrITS and two plastid DNA markers (the atpB-rbcL and trnL-F spacers). This species is a member of Ilex sect. Paltoria., which can be distinguished from other sections in Ilex by their solitary pistillate inflorescences. The new species is similar to I. viridis and I. triflora, but it differs in its narrow leaves and slender trunk and branches.

Nuclear and Plastid DNA Sequence-based Molecular Phylogeography of Salvadora oleoides (Salvadoraceae) in Punjab, India Reveals Allopatric Speciation in Anthropogenic Islands Due to Agricultural Expansion

Salvadora oleiodes is a tropical tree species belonging to the little-known family Salvadoraceae and distributed in the arid regions of Africa and Asia. Aims of our study were to trace the microevolutionary legacy of this tree species with the help of sequence-based multi-local phylogeography and to find the comparative placement of family Salvadoraceae within angiosperm clade malvids. A total 20 geographical isolates were collected from different regions of North India, covering a major part of its species range within the Indian Subcontinent. Sequence data from nuclear-encoded Internal Transcribed Spacer region (ITS1-5.8S-ITS2) and plastid-encoded trnL-F spacer region, were generated for this species for the first time in the world. ITS-based Bayesian phylogeographic analysis revealed the existence of four clades while trnL-F spacer based Bayesian analysis revealed one clade for this species distributed in the Indian subcontinent. Between these two loci, ITS revealed more distinct phylogeographic clades, indicating the phylogeographic utility of this locus for the systematics of Salvadoraceae. Interestingly, the ITS phylogeny indicated the existence of allopatric speciation in this tree species. Factors such as habitat destruction through agricultural expansion of plains might have forced the remaining population of this threatened tree species to isolated patches of thorn forest biotas, the ‘anthropogenic islands’, analogous to an archipelago. Phylogenetic analyzes based on trnL-F spacer suggested a synonymy of this species with Salvadora angustifolia. Maximum Likelihood gene tree based on ITS sequence data revealed that Salvadoraceae belongs to Sapindales rather than Brassicales. However, in the gene tree based on trnL-F spacer sequence, this family clustered within Brassicales. An evolutionary congruence of S. oleoides isolates across its range in North India is revealed in this study. Given the conflicting results on the relative placement of Salvadoraceae in Brassicales and Sapindales, the need for further phylogenetic analyses of malvids using supermatrix approach is highlighted.

EL trnL-trnF DE cpADN CONTRIBUYE A LA SEPARACIÓN DE LOS SUBGÉNEROS Persea y Eriodaphne (Lauraceae; Persea) COMO GÉNEROS INDEPENDIENTES

The genus Persea has two subgenera, Persea and Eriodaphne. There is controversy about the taxonomy between species of the subgenus Persea, and its relationships with the subgenus Eriodaphne. Because of its complex evolutionary patterns, the subgenus Persea has considerable variability with uncertain taxonomic affinities. In this study, we analyzed the sequences of the trnLtrnF region of chloroplast DNA from leaves of 38 accessions: 7 of the subgenus Eriodaphne and 31 from the subgenus Persea; one accession of the subgenus Beilschmiedia was also included. The first alignment with the trnL-trnF intergenic spacer included 12 individuals of the genus Persea and one of Beilschmiedia, which were aligned with 26 sequences from the GenBank of 22 genera of the Lauraceae family and one of the Hernandiaceae family. In a second alignment with a fragment close to 900 pb that comprises the end 3’ of the trnLuaa gene, which includes an intron, the trnL-F spacer and the end 5’ of the trnFgaa gene, four sequences from the GenBank were included: one of Perseaamericana (AY841669), and three species of the genus Sassafras. The aim of this study was to infer the possible phylogenetic relationships, particularly those of the two subgenera. The phylogenetic analysis was not consistent with the current classification of the genus as it clearly separated the two subgenera Persea and Eriodaphne, while in the clade of subgenus Persea did not show species grouping or affinity. The findings of this study support the hypothesis that the subgenera Persea and Eriodaphne should be regarded as independent genera.