Chloroplast DNA Sequences Research Articles

Investigation of the variability of chloroplast DNA of some Astragalus spp. of sect. Helmia and A. arcanus of sect. Erioceras (Fabaceae)

Analysis of chloroplast DNA sequences revealed a high polymorphism of the length and nucleotide composition of trnF–trnV and the intron trnL for Helmia section species of the genus Astragalus (Fabaceae). Molecular genetic studies of Astragalus species of the Helmia section were previously practically not carried out; sequencing of trnF–trnV and the intron trnL of chloroplast DNA for 7 species of the section, as well as the probable intersection hybrid Astragalus arcanus held for the first time. On the basis of genetic studies, clarified the relationship between species included in the section Helmia. The possible stages of sect. Helmia evolution are discussed. The taxonomic isolation of Astragalus permiensis has been confirmed; it is assumed not to be of Uralic, but of Siberian or Central Asian origin. Astragalus ionae belongs to the section Helmia is confirmed. Astragalus depauperatus s. l. is thought to be a complex of twin species. In this article, one of them is considered in the status of the species A. chakassiensis. 
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Complete chloroplast genome sequences of the ornamental plant Prunus cistena and comparative and phylogenetic analyses with its closely related species

BackgroundPrunus cistena is an excellent color leaf configuration tree for urban landscaping in the world, which has purplish red leaves, light pink flowers, plant shape and high ornamental value. Genomic resources for P. cistena are scarce, and a clear phylogenetic and evolutionary history for this species has yet to be elucidated. Here, we sequenced and analyzed the complete chloroplast genome of P. cistena and compared it with related species of the genus Prunus based on the chloroplast genome.ResultsThe complete chloroplast genome of P. cistena is a 157,935 bp long typical tetrad structure, with an overall GC content of 36.72% and higher GC content in the in the inverted repeats (IR) regions than in the large single-copy (LSC) and small single-copy (SSC) regions. It contains 130 genes, including 85 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The ycf3 and clpP genes have two introns, with the longest intron in the trnK-UUU gene in the LSC region. Moreover, the genome has a total of 253SSRs, with the mononucleotide SSRs being the most abundant. The chloroplast sequences and gene arrangements of P. cistena are highly conserved, with the overall structure and gene order similar to other Prunus species. The atpE, ccsA, petA, rps8, and matK genes have undergone significant positive selection in Prunus species. P. cistena has a close evolutionary relationship with P. jamasakura. The coding and IR regions are more conserved than the noncoding regions, and the chloroplast DNA sequences are highly conserved throughout the genus Prunus.ConclusionsThe current genomic datasets provide valuable information for further species identification, evolution, and phylogenetic research of the genus Prunus.

Conservation Priorities and Demographic History of Saussurea involucrata in the Tianshan Mountains and Altai Mountains.

Rare and vulnerable endemic plants represent different evolutionary units that occur at different times, and protecting these species is a key issue in biological protection. Understanding the impact of the history of endangered plant populations on their genetic diversity helps to reveal evolutionary history and is crucial for guiding conservation efforts. Saussurea involucrata, a perennial alpine species mainly distributed in the Tianshan Mountains, is famous for its medicinal value but has become endangered due to over-exploitation. In the present study, we employed both nuclear and chloroplast DNA sequences to investigate the genetic distribution pattern and evolutionary history of S. involucrata. A total of 270 individuals covering nine S. involucrata populations were sampled for the amplification and sequencing of nrDNA Internal Transcribed Spacer (ITS) and chloroplast trnL-trnF, matK and ndhF-rpl32 sequences. Via calculation, we identified 7 nuclear and 12 plastid haplotypes. Among the nine populations, GL and BA were characterized by high haplotype diversity, whereas BG revealed the lowest haplotype diversity. Molecular dating estimations suggest that divergence among S. involucrata populations occurred around 0.75 Ma, coinciding with the uplift of Tianshan Mountains. Our results reveal that both isolation-by-distance (IBD) and isolation-by-resistance (IBR) have promoted genetic differentiation among populations of S. involucrata. The results from the ecological niche modeling analyses show a more suitable habitat for S. involucrata in the past than at present, indicating a historical distribution contraction of the species. This study provides new insight into understanding the genetic differentiation of S. involucrata, as well as the theoretical basis for conserving this species.

Molecular phylogeography and historical demography of a widespread herbaceous species from eastern North America, Podophyllum peltatum.

Glacial/interglacial cycles and topographic complexity are both considered to have shaped today's diverse phylogeographic patterns of taxa from unglaciated eastern North America (ENA). However, few studies have focused on the phylogeography and population dynamics of wide-ranging ENA herbaceous species occurring in forest understory habitat. We examined the phylogeographic pattern and evolutionary history of Podophyllum peltatum L., a widely distributed herb inhabiting deciduous forests of ENA. Using chloroplast DNA (cpDNA) sequences and nuclear microsatellite loci, we investigated the population structure and genetic diversity of the species. Molecular dating, demographic history analyses, and ecological niche modeling were also performed to illustrate the phylogeographic patterns. Our cpDNA results identified three main groups that are largely congruent with boundaries along the Appalachian Mountains and the Mississippi River, two major geographic barriers in ENA. Populations located to the east of the Appalachians and along the central Appalachians exhibited relatively higher levels of genetic diversity. Extant lineages may have diverged during the late Miocene, and range expansions of different groups may have happened during the Pleistocene glacial/interglacial cycles. Our findings indicate that geographic barriers may have started to facilitate the population divergence in P. peltatum before the Pleistocene. Persistence in multiple refugia, including areas around the central Appalachians during the Quaternary glacial period, and subsequent expansions under hospitable climatic condition, especially westward expansion, are likely responsible for the species' contemporary genetic structure and phylogeographic pattern.

Phylogeography and chromosome number variation in Micranthes nelsoniana and related species (Saxifragaceae) in Northeast Asia

Abstract Micranthes nelsoniana possesses multiple different variants and numerous chromosomes. Based on the internal transcribed spacer (ITS) and chloroplast (cp)DNA sequences, the phylogeography of M. nelsoniana and its relatives in Northeast Asia was investigated, with extensive sampling around the Kuril Islands. The Arctic–Asian continent and a clade of marginal islands were the two main groupings that comprised the ITS phylogenetic tree. The island clade was separated into five well-supported clades: Kamchatka and Hokkaido highlands, Kuril–Aleutian Islands, southern Kuril Islands, Japanese archipelago, and Primorye region. Micranthes fusca was found in Japan and in the southern Kuril Islands. It is a separate species that created several types of hybrids between M. nelsoniana in the centre of the Kuril Islands based on a comparison of the ITS and cpDNA networks. Micranthes nelsoniana and M. ohwii appear to have hybridized in the northern Kuril Islands. Cytological investigation on the local species of M. nelsoniana showed that the chromosomal numbers are: 2n = 24, 26, 28, 30, 50, and 80. Among them, two usual numbers to this area, 2n = 24 and 50, appear to encourage interspecific gene exchange. The genomes of Hokkaido plants with high chromosome counts were cloned, revealing that they contained genes of both continental and marginal origins. This study revealed the crucial role of marginal islands along Northeast Asia in the genetic diversity of M. nelsoniana and related species.

The Genetic Diversity of Pleurozium schreberi: A Preliminary Study Based on the atpB-rbcL.

Pleurozium schreberi is a common and widespread species that has been the object of many studies, and its biology and ecology are well known. However, genetic studies on this species are limited or even absent. Because of the lack of any data about the genetic diversity of the moss species P. schreberi in Poland, the present paper describes the results of the studies carrying out for the first time this kind of research based on the atpB-rbcL spacer sequences of chloroplast DNA. A total of 35 specimens of P. schreberi from 19 locations in Poland were sampled. Total genomic DNA was extracted, amplified, and sequenced, and all obtained sequences were analyzed. Our findings suggest the low genetic diversity of P. schreberi in Poland. We detected four different haplotypes, shared between different populations.

No wonder, it is a hybrid. Natural hybridization between Jacobaea vulgaris and J. erucifolia revealed by molecular marker systems and its potential ecological impact.

Progressive changes in the environment are related to modifications of the habitat. Introducing exotic species, and interbreeding between species can lead to processes that in the case of rare species or small populations threatens their integrity. Given the declining trends of many populations due to increased hybridization, early recognition of hybrids becomes important in conservation management. Natural hybridization is prevalent in Jacobaea. There are many naturally occurring interspecific hybrids in this genus, including those between Jacobaea vulgaris and its relatives. Although Jacobaea erucifolia and J. vulgaris often co-occur and are considered closely related, apart from the few reports of German botanists on the existence of such hybrids, there is no information on research confirming hybridization between them. Morphologically intermediate individuals, found in the sympatric distributions of J. vulgaris and J. erucifolia, were hypothesized to be their hybrids. Two molecular marker systems (nuclear and chloroplast DNA markers) were employed to test this hypothesis and characterize putative hybrids. Nuclear and chloroplast DNA sequencing results and taxon-specific amplified fragment length polymorphism (AFLP) fragment distribution analysis confirmed the hybrid nature of all 25 putative hybrids. The AFLP patterns of most hybrids demonstrated a closer relationship to J. erucifolia, suggesting frequent backcrossing. Moreover, they showed that several individuals previously described as pure were probably also of hybrid origin, backcrosses to J. erucifolia and J. vulgaris. This study provides the first molecular confirmation that natural hybrids between J. vulgaris and J. erucifolia occur in Poland. Hybridization appeared to be bidirectional but asymmetrical with J. vulgaris as the usual maternal parent.

Tmesipteris alticola (Psilotaceae), a new fern species endemic to New Caledonia

ABSTRACT Tmesipteris alticola is described here as a new fern species endemic to New Caledonia's Grande Terre. It is a fifth species of Tmesipteris for the territory, and the second endemic. The obovate leaves and conic synangia of T. alticola distinguish it morphologically from all other species of Tmesipteris. Phylogenetic analyses of chloroplast DNA sequences support its distinctiveness, and indicate it is most closely related to T. truncata and T. vieillardii among the sampled species. Tmesipteris alticola is known from only four mountainous locations, all with ultramafic substrates in Province Sud, at elevations of 1200 m–1500 m, where it grows terrestrially or as a low epiphyte, seemingly mostly on ridges or around summits within short-statured cloud forest. We propose a provisional IUCN conservation status of Near Threatened for T. alticola because of its limited known occurrences. Tmesipteris alticola adds to a slew of fern species that have been recently newly described, reinstated, or recorded in New Caledonia.

Artemisiaqingheensis (Asteraceae, Anthemideae), a new species from Xinjiang, China.

Artemisiaqingheensis (Asteraceae, Anthemideae), a new species from Qinghe County, Xinjiang, China, is described and illustrated. We investigated its phylogenetic position and relationships with 35 other species of Artemisia using whole chloroplast DNA sequence data. The molecular phylogenetic results and morphological evidence (multi-layered involucral bracts and homogamous capitula with bisexual flowers) showed that the new species belongs to ArtemisiasubgenusSeriphidium. A diagnostic table and discussion of morphological characters are provided to distinguish the new species from A.amoena, A.gracilescens, A.lessingiana and A.terrae-albae.

Are 150 km of open sea enough? Gene flow and population differentiation in a bat-pollinated columnar cactus.

Genetic differentiations and phylogeographical patterns are controlled by the interplay between spatial isolation and gene flow. To assess the extent of gene flow across an oceanic barrier, we explored the effect of the separation of the peninsula of Baja California on the evolution of mainland and peninsular populations of the long-lived columnar cactus Stenocereus thurberi. We analyzed twelve populations throughout the OPC distribution range to assess genetic diversity and structure using chloroplast DNA sequences. Genetic diversity was higher (Hd = 0.81), and genetic structure was lower (GST = 0.143) in mainland populations vs peninsular populations (Hd = 0.71, GST = 0.358 respectively). Genetic diversity was negatively associated with elevation but positively with rainfall. Two mainland and one peninsular ancestral haplotypes were reconstructed. Peninsular populations were as isolated among them as with mainland populations. Peninsular haplotypes formed a group with one mainland coastal population, and populations across the gulf shared common haplotypes giving support to regular gene flow across the Gulf. Gene flow is likely mediated by bats, the main pollinators and seed dispersers. Niche modeling suggests that during the Last Glacial Maximum (c. 130 ka), OPC populations shrank to southern locations. Currently, Stenocereus thurberi populations are expanding, and the species is under population divergence despite ongoing gene flow. Ancestral populations are located on the mainland and although vicariant peninsular populations cannot be ruled out, they are likely the result of gene flow across the seemingly formidable barrier of the Gulf of California. Still, unique haplotypes occur in the peninsula and the mainland, and peninsular populations are more structured than those on the mainland.

Phylogeography of cultivated and wild ophiopogon japonicus based on chloroplast DNA: exploration of the origin and sustainable cultivation

BackgroundOphiopogon japonicus, mainly planted in Sichuan (CMD) and Zhejiang (ZMD) province in China, has a lengthy cultivation history. During the long period of domestication, the genetic diversity of cultivated O. japonicus has substantially declined, which will affect the population continuity and evolutionary potential of this species. Therefore, it is necessary to clarify the phylogeography of cultivated O. japonicus to establish a theoretical basis for the utilization and conservation of the genetic resources of O. japonicus.ResultThe genetic diversity and population structure of 266 O. japonicus individual plants from 23 sampling sites were analyzed based on 4 chloroplast DNA sequences (atpB-rbcL, rpl16, psbA-trnH and rpl20-5’rps12) to identify the effects of domestication on genetic diversity of cultivars and determine their geographic origins. The results showed that cultivated O. japonicus and wild O. japonicus had 4 and 15 haplotypes respectively. The genetic diversity of two cultivars (Hd = 0.35700, π = 0.06667) was much lower than that of the wild populations (Hd = 0.76200, π = 0.20378), and the level of genetic diversity in CMD (Hd = 0.01900, π = 0.00125) was lower than that in ZMD (Hd = 0.06900, π = 0.01096). There was significant difference in genetic differentiation between the cultivated and the wild (FST = 0.82044), especially between the two cultivars (FST = 0.98254). This species showed a pronounced phylogeographical structure (NST > GST, P < 0.05). The phylogenetic tree showed that the genetic difference between CMD and ZMD was not enough to distinguish the cultivars between the two producing areas by using O. amblyphyllus Wang et Dai as an outgroup. In addition, both CMD and ZMD have a closer relationship with wild populations in Sichuan than that in Zhejiang. The results of the TCS network and species distribution model suggested that the wild population TQ located in Sichuan province could serve as the ancestor of cultivated O. japonicus, which was supported by RASP analysis.ConclusionThese results suggest that cultivated O. japonicus has experienced dramatic loss of genetic diversity under anthropogenic influence. The genetic differentiation between CMD and ZMD is likely to be influenced by founder effect and strong artificial selection for plant traits. It appears that wild populations in Sichuan area are involved in the origin of not only CMD but also ZMD. In addition, we also raise some suggestions for planning scientific strategies for resource conservation of O. japonicus based on its genetic diversity and population structure.

Dianthus × belkinii (Caryophyllaceae), a new hybrid taxon from Altai

Dianthus × belkinii is an intersectional hybrid newly described from the Republic of Altai (Talduair massif). This taxon was also noted in Mongolia (Bayan-Ulegei aimag). The article describes the morphological features of the new taxon and discusses its relationship with parental species. The described hybrid has intermediate morphological features between the parental species, most notable are the fringed corollas. The stems are sparsely branched, relatively tall and similar to those of D. superbus. The calyx lobes and bracts are transitional between D. superbus and D. versicolor. The limb of the corolla is cut for ⅓–¼ of its length into a few lobes. Cytometry data showed an identical ploidy level of the new taxon and its putative parent species. The average value of the relative DNA content is almost intermediate, however, somewhat closer to D. versicolor. The ITS sequences of D. × belkinii show characteristic double peaks, which may point to a hybridogeneous origin of the plants. Identical chloroplast DNA sequences with D. versicolor but differences to D. superbus suggests that D. versicolor may be the maternal parent of this hybrid. Based on these polymorphisms, it is possible to state that the ITS sequence and D. versicolor × superbus itself are of a hybrid nature, where D. versicolor and D. superbus are the parent forms.

New Approach to the Systematics of the Section Psammiris (Iris, Iridaceae): What Does Chloroplast DNA Sequence Tell Us?

Iris sect. Psammiris comprises rhizomatous perennials distributed in the north temperate zone of Eurasia. The systematics of the section are currently based on morphology, and the phylogenetic relationships within it still remain unclear. In the framework of Iris systematics, we conducted molecular and morphological analyses of the currently recognized I. sect. Psammiris species to elucidate the taxonomic composition and relationships within the section. The phylogenetic reconstructions based on sequence variation of four noncoding chloroplast DNA regions support the monophyly of I. sect. Psammiris, which includes I. tigridia, while I. potaninii var. ionantha belongs to I. sect. Pseudoregelia. The proposed novel classification of I. sect. Psammiris recognizes three series: an autonymic series with I. humilis, I. bloudowii, and I. vorobievii and two unispecific series (I. ser. Potaninia with I. potaninii and I. ser. Tigridiae with I. tigridia). In addition, the taxonomic statuses of I. arenaria, I. ivanovae, I. kamelinii, I. mandshurica, I. pineticola, I. psammocola, and I. schmakovii are clarified herein. We provide a revised taxonomic treatment for I. sect. Psammiris, including notes on the types; updated information on species synonymy, distributions, habitats, and chromosome numbers; and a new identification key to the species. Three lectotypes are designated here.

DNA assays for genetic discrimination of three Phragmites australis subspecies in the United States.

To genetically discriminate subspecies of the common reed (Phragmites australis), we developed real-time quantitative (qPCR) assays for identifying P. australis subsp. americanus, P. australis subsp. australis, and P. australis subsp. berlandieri. Utilizing study-generated chloroplast DNA sequences, we developed three novel qPCR assays. Assays were verified on individuals of each subspecies and against two non-target species, Arundo donax and Phalaris arundinacea. One assay amplifies only P. australis subsp. americanus, one amplifies P. australis subsp. australis and/or P. australis subsp. berlandieri, and one amplifies P. australis subsp. americanus and/or P. australis subsp. australis. This protocol enhances currently available rapid identification methods by providing genetic discrimination of all three subspecies. The newly developed assays were validated using P. australis samples from across the United States. Application of these assays outside of this geographic range should be preceded by additional testing.

Comparative chloroplast genomics of three species of Bulbophyllum section Cirrhopetalum (Orchidaceae), with an emphasis on the description of a new species from Eastern Himalaya.

Chloroplast (cp) genomes are useful and informative molecular markers used for species determination and phylogenetic analysis. Bulbophyllum is one of the most taxonomically complex taxa in Orchidaceae. However, the genome characteristics of Bulbophyllum are poorly understood. Based on comparative morphological and genomic analysis, a new species Bulbophyllum pilopetalum from eastern Himalaya belonging to section Cirrhopetalum is described and illustrated. This study used chloroplast genomic sequences and ribosomal DNA (nrDNA) analysis to distinguish the new Bulbophyllum species and determine its phylogenetic position. An additional phylogenetic analysis was conducted using 74 coding sequences from 15 complete chloroplast genomes from the genus Bulbophyllum, as well as nrDNA sequences and two chloroplast DNA sequences from 33 Bulbophyllun species. The new species is morphologically similar to B. pingnanense, B. albociliatum, and B. brevipedunculatum in vegetative and floral morphology, but it can be distinguished by its ovate-triangle dorsal sepal without a marginal ciliate. The chloroplast genome of the new Bulbophyllum species is 151,148 bp in length, and includes a pair of inverted repeats (IRs) of 25,833 bp, a large single-copy region (LSC) of 86,138 bp, and a small single-copy region (SSC) of 13,300 bp. The chloroplast genome includes 108 unique genes encoding 75 proteins, 30 tRNAs, and four rRNAs. Compared with the cp genomes of its two most closely-related species, B. pingnanense and B. albociliatum, this chloroplast genome exhibited great interspecific divergence and contained several Indels that were specific to the new species. The plastid tree showed that B. pilopetalum is most closely-related to B. pingnanense. The phylogenetic tree based on combined nrDNA and chloroplast DNA sequences indicated that section Cirrhopetalum was monophyletic and B. pilopetalum was a member of this section. The taxonomic status of the new species is strongly supported by cp genome data. Our study highlights the importance of using the complete cp genome to identify species, elucidate the taxonomy, and reconstruct the phylogeny of plant groups with complicated taxonomic problems.

ANALYSIS OF THE TAXONOMIC STATUS OF LIMONIUM CASPIUM (WILLD.) P. FOURN. USING THE ITS1-5.8S-ITS2 REGION

There are 15 representatives of the genus Limonium Mill in Ukraine. The taxonomic status of some of them requires clarification. In particular, some taxonomists consider L. caspium (Willd.) P. Fourn. to be a separate species, while others tend to consider this name as a synonym for L. bellidifolium Dumort. Until now, only one sample of L. caspium from eastern Iran has been analyzed using molecular genetic approaches: for this accession, the ITS1-5.8S-ITS2 of 35S rDNA region was sequenced. For both species, L. caspium and L. bellidifolium, samples from the territory of Ukraine have not yet been examined using molecular taxonomy methods. In this work, we amplified and sequenced ITS1-5.8S-ITS2 region for two L. caspium and two L. bellidifolium samples from southern Ukraine. All four samples appeared to be identical in this region. They also do not differ from previously sequenced samples of L. bellidifolium from France, Turkey and Greece. On the other hand, the sequence of L. caspium from Iran has five nucleotide substitu-tions and four indels compared to the mentioned samples. The phylogenetic analysis revealed that representatives of the so-called "L. bellidifolium complex", which, in addition to the two species mentioned, also includes L. iconium and L. tamaricoides analyzed in this work, are grouped together with representatives of the Nephrophyllum section. A similar result was obtained in the studies of other authors using the ITS1-5.8S-ITS2 sequence and regions of chloro-plast DNA. However, according to the IGS 5S rDNA sequence, representatives of the "L. bellidifolium complex" in our previous work were grouped together with the species of the Limonium section, to which they were previously as-signed based on morphological characters. Such a discrepancy between phylogenies using different genomic regions indicates the reticular nature of the evolution of the genomes of representatives of this group and indicates probable hybridization events in their past. Within the clade of the "L. bellidifolium complex" that we obtained in our analysis, the sample of L. caspium from Iran forms a sister branch to all its other representatives. Thus, the comparison of the ITS1-5.8S-ITS2 region shows that the Ukrainian samples identified as L. caspium do not differ from L. bellidifolium. However, to finally clarify the taxonomic status of L. caspium, it is necessary to analyze more samples from different geographic origins.

Chloroplast DNA Markers on the Phylogeography Study of Roburoid Oaks (&lt;i&gt;Quercus&lt;/i&gt; L. sect. &lt;i&gt;Quercus&lt;/i&gt;, Fagaceae) in the Crimean-Caucasian Region

The variability of five fragments of chloroplast DNA (ASq, CDq, TFq, trnH-psbA and trnK-matK) with a total length of more than 10,000 bp in three species of roburoid oaks of the Crimean-Caucasian region (Q. robur, Q. petraea, Q. pubescens) was studied. A total of 11 haplotypes were found in 290 individuals from 20 populations, 10 of which differed from the haplotypes of the main range. The obtained results show a pronounced structure of the geographical distribution of haplotypes of roburoid oaks in the Crimea and the Caucasus, and the chloroplast haplotypes of the region belong to several divergent phylogenetic lines. A comparison with data from other studies was carried out, with an assessment of the degree of effectiveness of the use of various chloroplast fragments and markers. The isolated phylogenetic position of the haplotypes of the Crimean-Caucasian populations and deep differences from the haplotypes of the main area allows us to conclude that robouroid oaks have historically been isolated for a long time in the Caucasus and Crimea. At the same time, the presence of common haplotypes indicates closer historical ties between the Crimean-Caucasian populations with Asia Minor and the Balkans. All haplotypes are identified by three fragments (ASq, CDq and TFq), with a sufficient level of variability and suitable for solving problems of oak phylogeography in the Crimean-Caucasian region. The trnH-psbA and trnK-matK markers had a low level of variability and did not reveal additional haplotypes. Optimized genotyping options have been proposed, including both step by step sequencing, and a combination of analysis of chloroplast DNA microsatellite loci (cpSSR), restriction analysis (PCR-RFLP), and sequencing. To assess the variability of markers in a larger taxonomic range, representatives of other sections of the genus Quercus were included in the analysis. It was shown that the oak-specific primers proposed for ASq, CDq, and TFq fragments are suitable for species of other sections.

Molecular barcoding of Melissa officinalis L. (badranjboye) in Iran and identification of adulteration in its medicinal services

Medicinal plants are an important source for treatment of diseases in many countries. Today, controlling the quality of the products of medicinal plants is an important task. Customer health may be in danger due to the fraud and misconduct by the sales associates in the sales centers. Melissa officinalis L. (Lamiaceae) is an important medicinal plant used for treatment of several diseases. In Iran, the species of Dracocephalum, Hymencrater, Nepeta and Stachys are mistakenly sold under the name of Badranjboye that have different pharmaceutical properties. For avoiding this mistake, this investigation was performed with the following aims: 1) Checking for the cheating and identifying the Badranjboye in the Iran’s market of medicinal plants, 2) Providing the molecular barcode for the medicinal species of Melissa. For this purpose, Market-sold plant samples (leaves) and original reference plant species were compared by morphology, odor as well as Internal transcribed spacer (ITS) and chloroplast DNA ((psbA–trnH) and (trnL–trnF)) sequences. Various molecular analyses, such as sequencing, determination of genetic distance, and construction of phylogenetic tree were performed. These reports have shown that internal transcribed spacer (ITS) and chloroplast DNA (psbA–trnH) sequences are an efficient molecular marker to produce barcode gap and differentiating Melissa officinalis from other species.

Revised molecular phylogeny, global biogeography, and diversification of palms subfamily Coryphoideae (Arecaceae) based on low copy nuclear and plastid regions.

Coryphoideae are palmate-leaved palms from the family Arecaceae consisting of 46 genera representing 421 species. Although several phylogenetic analyses based on different genomic regions have been carried out on Coryphoideae, a fully resolved molecular phylogenetic tree has not been reported yet. To achieve this, we applied two phylogenetic reconstruction methods: Maximum Likelihood and Bayesian Inference, using amplified sampling by retrieving chloroplast and nuclear DNA sequences from NCBI and adding newly produced sequences from Indian accession into the dataset. The same dataset (chloroplast + nuclear DNA sequences) was used to estimate divergence times and the evolutionary history of Coryphoideae with a Bayesian uncorrelated, lognormal relaxed-clock approach and a Statistical Divergence-Vicariance Analysis method,respectively. The phylogenetic analyses based on a combined chloroplast and nuclear DNA sequence dataset showed well-resolved relationships within the subfamily. Both phylogenetic trees divide Coryphoideae into two main groups: CSPT (Crysophileae, Sabaleae, Phoeniceae, and Trachycarpeae) and the Syncarpous group. These main groups are segregated into eight tribes (Trachycarpeae, Phoeniceae, Sabaleae, Crysophileae, Borasseae, Corypheae, Caryoteae, and Chuniophoeniceae) and four subtribes (Rhapidine, Livistoninae, Hyphaeninae, and Lataniinae) with strong support-values. Most previously unresolved and doubtful relationships within tribes Trachycarpeae and Crysophilieae are now resolved and well-supported. The reconstructed phylogenetic trees support all previous systematic revisions of the subfamily. All Indian sampled species of Arenga, Bentinckia, Hyphaene, and Trachycarpus show close relation with their respective congeneric species. Molecular dating results and integration of biogeography suggest that Coryphoideae originated in Laurasia at ~95.12Ma and then diverged into the tropical and subtropical regions of the whole world. This study offers the correct combination of nuclear and plastid regions to test the current and future systematic revisions.

Allopatric Lineage Divergence of the East Asian Endemic Herb Conandron ramondioides Inferred from Low-Copy Nuclear and Plastid Markers

The evolutionary histories of ornamental plants have been receiving only limited attention. We examined the origin and divergence processes of an East Asian endemic ornamental plant, Conandron ramondioides. C. ramondioides is an understory herb occurring in primary forests, which has been grouped into two varieties. We reconstructed the evolutionary and population demography history of C. ramondioides to infer its divergence process. Nuclear and chloroplast DNA sequences were obtained from 21 Conandron populations on both sides of the East China Sea (ECS) to explore its genetic diversity, structure, and population differentiation. Interestingly, the reconstructed phylogeny indicated that the populations should be classified into three clades corresponding to geographical regions: the Japan (Honshu+Shikoku) clade, the Taiwan-Iriomote clade, and the Southeast China clade. Lineage divergence between the Japan clade and the Taiwan-Iriomote and Southeast China clades occured 1.14 MYA (95% HPD: 0.82-3.86), followed by divergence between the Taiwan-Iriomote and Southeast China clades approximately 0.75 MYA (95% HPD: 0.45-1.3). Furthermore, corolla traits (floral lobe length to tube length ratios) correlated with geographical distributions. Moreover, restricted gene flow was detected among clades. Lastly, the lack of potential dispersal routes across an exposed ECS seafloor during the last glacial maximum suggests that migration among the Conandron clades was unlikely. In summary, the extant Conandron exhibits a disjunct distribution pattern as a result of vicariance rather than long-distance dispersal. We propose that allopatric divergence has occurred in C. ramondioides since the Pleistocene. Our findings highlight the critical influence of species' biological characteristics on shaping lineage diversification of East Asian relic herb species during climate oscillations since the Quaternary.