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Using molecular data (nrDNA ITS, cpDNA psbA-trnH), the systematic position of the genus Palimbia in theUmbelliferae was studied. It was shown that Palimbia is a monophyletic taxon and belongs to the clade Pyramidoptereae.Taxa closely related to Palimbia, such as Carum buriaticum and Schulzia, are very different carpologically, but they arecharacterized by finely dissected leaves, which is a very rare trait for the Siberian Umbelliferae.

Introducción y objetivos: La presente investigación corresponde a un análisis taxonómico molecular de muestras florales obtenidas de plantas establecidas naturalmente en una zona recóndita del Desierto de Atacama, Región de Arica-Parinacota, Chile. En primera instancia, las muestras morfológicamente se atribuyeron al género Lathyrus, sin embargo, conforme a la literatura actual, no se han registrado individuos de este género en la región, dado esto, se ha generado confusión en la identificación de las muestras colectadas. Por lo cual, la presente investigación tiene por objetivo identificar genéticamente las especies presuntamente de Lathyrus, mediante DNA Barcoding. M&M: En el presente estudio se evaluaron estos individuos mediante un análisis filogenético para determinar su identidad taxonómica a partir de secuencias ITS (nuclear) y secuencias psbA, matK y rpoC1 (plastídicas). Resultados: Los resultados demostraron que los individuos están relacionados a la especie Lathyrus odoratus. Conclusiones: Los cuatro loci de DNA barcoding, permitieron identificar genéticamente las muestras como L. odoratus, el cual se encuentra creciendo de manera natural en Chile.

In Cambodia, medicinal plants are often used to treat various illnesses. However, the identities of many medicinal plants remain unknown. In this study, we collected 50 types of traditional Cambodian medicinal plants that could not be identified by their appearance from a domestic market. We utilized the DNA barcoding technique, combined with the literature survey, to trace their identities. In the end, 33 species were identified at the species level and 7 species were identified at the genus level. The ethnopharmacological information of 33 medicinal plants was documented. The DNA barcoding technique is useful in the identification of medicinal plants with no previous information.

Medicinal plants have been widely used in traditional medicine due to their therapeutic properties. Although they are mostly used as herbal infusion and tincture, employment as ingredients of food supplements is increasing. However, fraud and adulteration are widespread issues. In our study, we aimed at evaluating DNA metabarcoding as a tool to identify product composition. In order to accomplish this, we analyzed fifteen commercial products with DNA metabarcoding, using two barcode regions: psbA-trnH and ITS2. Results showed that on average, 70% (44–100) of the declared ingredients have been identified. The ITS2 marker appears to identify more species (n = 60) than psbA-trnH (n = 35), with an ingredients’ identification rate of 52% versus 45%, respectively. Some species are identified only by one marker rather than the other. Additionally, in order to evaluate the quantitative ability of high-throughput sequencing (HTS) to compare the plant component to the corresponding assigned sequences, in the laboratory, we created six mock mixtures of plants starting both from biomass and gDNA. Our analysis also supports the application of DNA metabarcoding for a relative quantitative analysis. These results move towards the application of HTS analysis for studying the composition of herbal teas for medicinal plants’ traceability and quality control.

The genus Dalbergia contains approximately 250 species with many valuable trees being destroyed by targeted and illegal logging. DNA barcoding is a reliable method for the molecular identification of different species and resources conservation. In the present study, the specimen discrimination ability of internal transcribed spacer (ITS), matK, rbcL and psbA-trnH barcoding were tested on Dalbergia sequences, downloaded from the National Center for Biotechnology Information (NCBI), and the combined barcoding ITS+matK+rbcL was used to identify unknown specimens. It was found that ITS+matK+rbcL have good discrimination rates based on the analysis methods best match (BM) and best close match (BCM). These barcodes also have the best performance concerning barcode gap distribution, and are able to discriminate unknown specimens from South-China. Furthermore, it was demonstrated that D. tamarindifolia and D. rubiginosa are also relatively close to sister-species D. pinnata and D. candenatensis within the phylogenetic Dalbergia tree. Considering the overall performance of these barcodes, we suggest that the ITS+matK+rbcL region is a suitable barcode for identifying Dalbergia species.

A new species from the Andean region of northwestern Argentina is described and illustrated: Sisyrinchium humahuacense. This species can be distinguish from its closest related species by three main characters: the presence of a conspicuous horizontal rhizome, the colour of the flowers and the presence of rudimentary leaves not splitting in fibers. A phylogenetic analysis based on DNA sequences supported the inclusion of S. humahuacense within the genus and as part of the section Segetia. Also, a key to species to identify S. humahuacense is presented herein. Fil: Zanotti, Christian Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Limnología "Dr. Raúl A. Ringuelet". Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Limnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica Darwinion. Academia Nacional de Ciencias Exactas, Físicas y Naturales. Instituto de Botánica Darwinion; Argentina Fil: Sassone, Agostina Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica Darwinion. Academia Nacional de Ciencias Exactas, Físicas y Naturales. Instituto de Botánica Darwinion; Argentina

Black pepper (Piper nigrum L.) is an important spice. The adulteration of black pepper seeds and powder with papaya seeds, green chili and red chili can be seen and limited studies have been conducted to detect these adulterants. The objectives of the present study were to assess the appropriateness of morphometric methods to discriminate papaya seed and chili adulterations in black pepper and to establish a DNA based strategy to detect these adulterations. A necessary adulteration series of seeds and powders were prepared for the analyses along with commercial samples. The appearance of the seed and powder samples were slightly different but not very distinct among the pure and adulterated samples emphasizing the need of a biochemical approach to detect the adulteration. The adulterated and commercial black pepper samples received lower pungency ranks compared to that of pure samples. QIAGEN DNeasy® Plant Mini Kit was successful in extracting PCR amplifiable DNA from any sample without papaya seeds and the modified CTAB method was able to extract required PCR amenable DNA from any sample with papaya seed material. The universal DNA barcoding primer pair, psbA-trnH, was used to amplify the DNA. Black pepper DNA yielded 200 bp band, chili and papaya DNA yielded 450 bp band and DNA from adulterated samples produced both 200 bp and 450 bp bands. Therefore this strategy can be used to detect papaya / chili adulterations in black pepper.

한국에 분포하는 두릅나무과 식물 대부분은 중요한 약용 식물로 경제적인 가치가 크다. 본 연구는 분자적 방법인 엽록체 DNA 바코드 염기서열 분석을 통해 한국에 자생하고 있는 두릅나무과 식물 14종 전체의 속 및 종간 유연 관계를 파악해 보고 이를 구별할 수 있는 마커를 개발하기 위해 수행되었다. 국제 생물 DNA 바코드 컨소시엄(CBOL, the Consortium for the Barcode of Life)이 DNA barcoding marker로 제안한 엽록체 DNA 7영역의 염기서열을 분석한 결과, psbA-trnH영역에서 가장 많은 삽입, 결실 및 염기치환이 나타났으며 조사된 한국의 두릅나무과 식물 14종 모두 구분 될 수 있었다. 또한 각각의 영역에서 특정속과 종만이 지니는 특이적인 염기서열을 찾을 수 있었다. 인삼의 경우 중국삼과 한국삼의 염기서열에는 차이가 전혀 없었다. 7영역을 모두 유합하여 작성한 계통수에서는 통탈목이 특이성을 나타내며 가장 기부에 분계조를 형성하였다. 두릅나무속과 인삼속은 자매군을 형성하였고, 오갈피속 5 종 역시 서로 높은 유연관계를 나타내었다. 결론적으로 한국에 자생하는 14종의 두릅나무과 식물들이 모두 엽록체 DNA 바코드 마커 개발을 통해 동정이 가능함을 확인하였다.

Tulipa edulis (Liliaceae) is the botanical origin of the traditional Chinese medicine (TCM) “Guangcigu”. Due to overexploitation that induced a decline in natural sources, many dried bulbs from other species of Tulipa have been used, adulterating the medicine in recent years. This practice may cause a series of inconsistent therapeutic effects and quality control problems in the herbal medicine industry. Hence, three DNA regions ( mat K, psb A- trn H and rbc L) were evaluated as barcodes for identifying T. edulis and its adulterants. All candidate DNA barcodes were successfully amplified from leaf samples. Based on the sequence divergences, rbc L and psb A- trn H can assign T. edulis and its adulterants to the correct genus, while mat K can accurately differentiate T. edulis and its adulterants. Thus, at the DNA level, the mat K intergenic region is a more suitable, accurate and applicable identification of T. edulis and its adulterants than rbc L and psb A- trn H.

Traded forms of spice and spice powders are often subjected to admixing with inferior substances by design or default, affecting public health and national prestige. Cinnamomum verum (true cinnamon), a high-value spice, is often adulterated with its inferior species such as C. cassia and C. malabatrum. The presence and detection of the spurious species in traded barks (whole or powder) of true cinnamon is posing problems. DNA markers are now used to detect such adulteration. Here we report the application of a DNA barcoding method to detect these adulterants in traded market samples of true cinnamon using the barcoding loci rbcL, matK and psbA-trnH. The PCR success rate, sequencing efficiency, inter and intra specific divergence, and occurrence of single nucleotide polymorphisms (SNPs) were utilized to assess the potential of each barcode loci to authenticate C. verum from its related adulterants. The amplification and sequencing success was 100% for rbcL and psbA-trnH while matK failed to amplify in the ...

This study aims to define the phylogenetic relationship within Korean Pinus L. and to find themolecular markers which resolve the phylogenetic relationship in genus Pinus. cpDNA atpF-H and psbA-trnHregions were used as molecular markers. We performed the molecular phylogenetic study on 17 taxa of Pinus inKorea. The combined analyses of two gene loci showed that Korean Pinus was a monophyletic group supportedby 100% BP. According to the results of separate analyses, psbA-trnH region seems to work better resolvingpower to clarify the phylogenetic ambiguity in Korean Pinus than those of atpF-H region. Also, we tried tochecked the value and resolution of two chloroplast DNA loci on phylogenetic implications. Keywords: Pinus, subgenus Diploxylon & Haploxylon, atpF-H, psbA-trnH 적 DNA 요: 엽록체 atpF-H, psbA-trnH region을 마커로활용하여국내에분포하는 소나무속 식물들 중 17분류군에 대한 분자계통학적 연구를 수행하여 한국산 소나무속의 계통학적 유연관계를 규명하고, 소나무속의유연관계를 잘 나타낼 수 있는 분자마커를 찾아내고자 연구가 수행되었다. atpF-H, psbA-trnH region의 조합분석결과 한국산 소나무속은 100%의 BP로 지지되는 단계통군으로 확인되었으며, 소나무아속과 잣나무아속으로 명확히 구분되어졌다. 본 연구에서 이용된 두 개의 분자마커 중 psbA-trnH region이 atpF-H region보다한국산 소나무속의 계통 및 유연관계를 규명하는데 다소 높은 해상력을 나타내었다.

This study aims to define the phylogenetic relationship within Korean section sabina and findmolecular markers which resolve the phylogenetic relationship in genus Juniperus and section sabina . cpDNAmat K and psb A- trn H were used as molecular markers. The combined analyses of two genes suggested that sec-tion sabina was a clade supported by 100% BP. The relationships of [J. chinensis var. sargentii + J. davurica]clade and [J. chinensis var. chinensis + J. chinensis var. procumbens + J. chinensis var. horizontalis ] clade weresupported by 91% BP and 100% BP, respectively. Thus, the classification of Korean section sabina would beappropriate at follows, (1) J. chinensis var. sargentii + J. davurica, and (2) J. chinensis var. chinensis + J. chin-ensis var. procumbens According to the results of separate analyses, mat K seems to work better resolving powerto clarify the phylogenetic ambiguity in Juniperus and section sabina than psb A- trn H. Keywords: Section Sabina , Molecular phylogeny, mat K, psb A- trn H

Value-added forms of black pepper (Piper nigrum L.) are an important item of trade globally. Adulteration by default or design of the commodity not only leads to economic loss and public health issues but also to self-respect of a nation. DNA barcoding is assuming significance as a quality assurance technique in many agri-food commodities. Three barcoding loci viz., psbA-trnH, rbcL, rpoC1 were used in the study to detect bio adulteration of traded black pepper powder. PCR amplification of P. nigrum and traded black pepper powder was performed for all the three loci. Sequence analysis and BLAST results revealed chilli adulteration in two out of nine market samples, originating probably from exhausted black pepper powder fortified with chilli. Of the three loci, psbA-trnH proved to be the best and ideal for detection of chilli adulteration in black pepper yielding amplicons of size 600 bp and 350 bp, respectively. Cloning and sequencing of the adulterant specific band of both market samples were done to con...

Fabaceae is a huge family that contains a large number of medicinal plants, many of which are commonly used in Chinese traditional medicine. However, traditional taxonomy has not been able to meet the complicated demands of species discrimination within Fabaceae. Thus, we employed a famous DNA barcode, the psbA-trnH region, to discriminate commonly used medicinal species of the family Fabaceae. Here, the psbA-trnH regions derived from 152 samples were amplified. These samples represented 104 Fabaceae medicinal species from 60 genera, including 25 authentic Fabaceae species listed in the Chinese pharmacopoeia and common adulterant species. The results indicate that the psbA-trnH region performed well in terms of its universality and high variability in length and composition. Species discriminative power analysis of the psbA-trnH region showed that 91.3% of species could be identified successfully by the BLAST1 method in conjunction with the nearest distance method. And, the species resolution rate of the TaxonGap method exceeded 93%. The results provide support for the use of the psbA-trnH plastid region as a sensitive marker to the authentication of Fabaceae medicinal plants.

To identify origin of the medicinal materials Dryopteridis Crassirhizomatis Rhizoma by using the psbA-trnH sequence, the polymerase chain reaction (PCR) amplification and product sequencing of the experimental samples were performed. In order to expand the scope of the study, the psbA-trnH sequences of 8 genera and 3 species were downloaded from GenBank for analysis. DNAMAN 8.0 software was used to show splicing and comparison results of the peak diagrams with analysis of them, and MEGA 6.0 software was to calculate K2P genetic distances and establish clustering tree adjacent genus. The results showed that by using the psbA-trnH sequence, Dryopteridis Crassirhizomatis Rhizoma, its original plant and other easy-confused medicinal materials and plants can be distinguished with each other obviously, with the psbA-trnH sequence of Dryopteridis Crassirhizomatis Rhizoma completely consistent with that of its original plant. Consequently, it is revealed that it's feasible to identify Dryopteridis Crassirhizomatis Rhizoma and its original plant, and separate from its adulterants by means of the psbA-trnH sequence, which can provide more scientific bases for the further study of the identification of the ferny medicinal herbs.

Recent research has identified a new species of the genus Scaligeria, the rhizomatous perennial S. alziarii, endemic to cyprus. Prior to the discovery of this species, the biennial S. napiformis was the only identified taxon of the genus in cyprus. Molecular studies corroborate the placement of the new taxon in Scaligeria and its rank as a separate species.

Chase, M. W., Cowan, R. S., Hollingsworth, P. M., van den Berg, C., Madrinan, S., Petersen, G., Seberg, O., Jorgsensen, T., Cameron, K. M., Carine, M., Pedersen, N., Hedderson, T. A. J., Conrad, F., Salazar, G. A., Richardson, J. E., Hollingsworth, M. L., Barraclough, T. G., Kelly, L., Wilkinson, M. (2007). A proposal for a standardised protocol to barcode all land plants. Taxon, 56, (2), 295-299.

Objective To evaluate the ability of several hotspot candidate sequence of DNA barcodes for identifying medicinal plant species in Verbenaceae. Method Using universal primers, three chloroplast sequences, psbA-trnH, rbcL, matK, two nuclear ribosomal DNA ITS2 and ITS were amplified and sequenced. PCR amplification and sequencing efficiency, intra- and inter-specific variation, barcoding gap and identification efficiency (with BLAST 1 and Nearest Distance methods) were used to evaluate these loci. Result The rate of successful amplification using matK was too low to further analyze, and the rate of successful amplification and sequencing using psbA-trnH, ITS, and ITS2 and rbcL sequence was 83.6%, 83.6%, 96.4%, 98.2%, respectively. The rate of successful identification using psbA-trnH, ITS, and ITS2 was 100% at the species level except that rbcL was 77.8%, 75.9% for 55 samples belonging to 32 species, but ITS2 did better in intra- and inter-specific variation, barcoding gap than the other loci. The rate of successful identification of ITS2 was 89.5%, 87.6% even when joining the date of 165 samples from GenBank. Conclusion It proposes that the combination of ITS2 and psbA-trnH senquence is promising for the identification of the species in Verbenaceae.

Phylogenetic relationships within the genus Rhododendron were studied using psbA-trnH intergenic spacer sequences. In the analysis, forty samples of thirty-six Rhododendron species, which belong to seven subgenus according to taxonomy system of Sleumer in "Flora of China" were analyzed. The results indicated that: all species are grouped into seven clades and all samples come from every same subgenus formed into a monophyletic clade with higher bootstrap value which supported the taxonomy system of Sleumer in "Flora of China". Pseudorhodorastrum as an independent subgenus was supported. The opinion of Chamberlian proposed the subg. Pseudazalea should be placed in subg. Rhododendron as a subsection Trichoclada was not supported by the reconstructed phylogenetic tree.

Objective To determine the candidate sequences which can be used as DNA barcode to identify species in Caprifoliaceae family by screening out from four different DNA fragments sequences. Method PCR amplification, sequencing efficiency, differential intra- and interspecific divergences, the DNA barcoding gap and identification efficiency were used to evaluate these loci. Result The ITS2 was used as a candidate sequence of DNA barcode to identify the species in Caprifoliaceae family, whose rate of success in identification in genera level was 100% and in species 96.6%, and psbA-trnH as a complementary barcode to ITS2 for Caprifoliaceae.

Aconite is one of the most toxic plants, and is known world-wide. To confirm cases of poisoning by aconite, it would be helpful to be able to identify aconite DNA from samples such as stomach contents. In this study, we designed a tetraplex PCR targeted to the ITS region of the aconite nuclear DNA and three loci (matK, trnL-F and psbA-trnH) in chloroplast DNA. Four amplification products were specifically detected from all Japanese aconite samples we tested, substances with suspected aconite contamination, and a medicine, as well as in highly degraded materials. Furthermore, the sequencing analysis of psbA-trnH revealed that seven types of aconite were observed in Japanese samples. This method is a simple, sensitive and reliable method that can be used to identify Aconitum species.

A 1887-nucleotide chloroplast-DNA region from Arabidopsis thaliana was analyzed. It contains the conserved genes psbA for the precursor of the D1 reaction-centre protein of photosystem II, trnH for tRNAHis, and rps19' for the 6.8-kDa protein of the small ribosomal subunit. Northern hybridization and RNase protection experiments suggest co-transcription of a minor RNA fraction over the full lengths of psbA and the preceding trnK-UUU gene, but not including downstream trnH sequences. In front of the mapped 5' end of the major 1.2-kb psbA transcript is a DNA region that shows the typical architecture of a psbA promoter, consisting of the prokaryotic-type '-35' and '-10' elements as well as the eukaryotic-type 'TATA' motif. The common 3' end of psbA transcripts seems to be located immediately after a stem-loop structure downstream from the coding region.