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1.
Acta Pharmaceutica Sinica ; (12): 507-513, 2022.
Article in Chinese | WPRIM | ID: wpr-922927

ABSTRACT

The key factors for producing the best quality Chinese herbal medicines are high-quality germplasm, suitable cultivation area and the proper processing methods for herbal raw materials. Gentiana crassicaulis in Gentiana (Sect. Cruciata), Gentianaceae is one of the original plants of the Chinese herb Qinjiao (Gentianae Macrophyllae Radix), and its type specimen was collected in Lijiang, Yunnan. There is a long planting history of the herb in this area. In this study a sampling plot was designated in these traditional planting areas. G. crassicaulis was planted and herbal raw materials were harvested from the plot. The raw materials were prepared locally and at a pharmaceutical factory in Shanghai using processing methods such as "sweating" or "no sweating", "slicing" or "no slicing" (whole root), and "stoving" or "no stoving" (air drying). The quality of all processed samples was evaluated. In addition, molecular markers were determined for identifying cultivated and wild samples from Lijiang, Yunnan. The results are as follows: ① Samples from the sampling plot and the field are taxonomically identified as Gentiana crassicaulis. ② A total of 270 sequences of trnC-GCA-petN, atpB-rbcL, psbN, ndhB-rps7 and ycf1 were obtained, and three genotypes were determined from the cultivated samples; the type III was shared by both cultivated and wild plants. Based on the molecular markers, a DNA barcoding method to identify cultivated and wild samples of G. crassicaulis from Lijiang, Yunnan was established. ③ Total content of loganic acid and gentiopicroside in all samples was ≥ 2.5%, and above the Chinese Pharmacopoeia (2020) limit. ④ In HPLC fingerprinting, 9 common peaks were assigned and similarity between all samples was > 0.999; and ⑤ In a PCA score plot all slice samples were clustered, while whole root samples were scattered. Therefore, our studies could provide basic data for optimizing the processing method, producing best quality Gentianae Macrophyllae Radix, and evaluating the quality of different ecotype varieties and the multiple origin of herbal medicines.

2.
Braz. j. biol ; 81(4): 1054-1060, Oct.-Dec. 2021. graf
Article in English | LILACS | ID: biblio-1153441

ABSTRACT

Abstract One aquatic coleopteran species from family Dytiscidae and two aquatic coleopteran genera from family Hydrophilidae were recorded in the summer period and represent first records in the Egyptian lakes. Beetles were collected from two northern lakes, Lake Idku and Lake Burullus. They were identified by morphological characteristics as well as the mtDNA barcoding method. A molecular phylogenetic approach was used to determine the genetic identity of the collected samples based on the mitochondrial cytochrome oxidase I (COI). Prodaticus servillianus (Dytiscidae) from Egypt showed no significant difference in the COI region and they are highly similar to P. servillianus from Madagascar. The phylogenetic analysis revealed that the other two coleopteran genera belong to family Hydrophilidae. Based on COI only, there is no clear evidence for their genetic identity at the species level. So, we defined them to the closest taxon and denoted them as Cymbiodyta type A and B. The results indicated that resolving the molecular identity of the aquatic beetles from northern lakes of Egypt need more considerations in the field of biological conservation. We concluded that utilization of COI as a barcoding region for identifying some coleopteran species is not sufficient and additional molecular markers are required to uncover the molecular taxonomy at deep levels.


Resumo Uma espécie de coleópteros aquático da família Dytiscidae e dois gêneros de coleópteros aquáticos da família Hydrophilidae foram registrados no período de verão e representam os primeiros registros nos lagos egípcios. Os besouros foram coletados em dois lagos do norte, o lago Idku e o lago Burullus, e identificados por características morfológicas e pelo método de código de barras mtDNA. Uma abordagem filogenética molecular foi usada para determinar a identidade genética das amostras coletadas com base no citocromo oxidase I mitocondrial (COI). Prodaticus servillianus (Dytiscidae) do Egito não mostrou diferença significativa na região COI e é altamente semelhante a P. servillianus de Madagascar. A análise filogenética revelou que os outros dois gêneros de coleópteros pertencem à família Hydrophilidae. Com base apenas no COI, não há evidências claras de sua identidade genética no nível da espécie. Assim, nós os agrupamos no táxon mais próximo e os denominamos Cymbiodyta tipo A e B. Os resultados indicaram que a identidade molecular dos besouros aquáticos dos lagos do norte do Egito precisa de mais considerações no campo da conservação biológica. Concluímos que a utilização de COI como região de código de barras para identificar algumas espécies de coleópteros não é suficiente, sendo necessários marcadores moleculares adicionais para descobrir a taxonomia molecular em níveis profundos.


Subject(s)
Animals , Lakes , DNA Barcoding, Taxonomic , Phylogeny , Electron Transport Complex IV/genetics , Egypt
3.
Article in Chinese | WPRIM | ID: wpr-906157

ABSTRACT

Objective:DNA barcodes suitable for Lauraceae plants were screened,and 22 Lauraceae plants were identified and classified. Method:The DNA of 22 species of Lauraceae was extracted and amplified by polymerase chain reaction(PCR) with different DNA barcoding primers,followed by electrophoresis and sequencing. Codon Code Aligner was used to proofread,splice, and remove the forward and reverse primer sequences. The sequence was imported into DNAMAN for multiple sequence alignment,and the base mutation sites were analyzed. The Kimura 2-Parameter(K2P) distance of different plants was calculated by MEGA,and the variation degree was analyzed according to the genetic distance. The phylogenetic tree was constructed based on the adjacency method. Result:Three pairs of DNA barcoding primers were used to amplify and sequence the DNA of 22 species of Lauraceae,and 20 species were identified by comparing the specific base sites of gene sequences<bold>.</bold> Conclusion:Four <italic>Litsea</italic> plants could be identified by <italic>matK</italic>, three <italic>Phoebe</italic> plants except for<italic> Cinnamomum validinerve </italic>by<italic> rbcL</italic>, and 20 Lauraceae plants by the combination of<italic> matK</italic>, <italic>rbcL</italic>, and <italic>rpoB</italic>,which provided a theoretical basis for the identification and development of Lauraceae plants. Among them,<italic>matK</italic> was predominant in the identification of Lauraceae plants,and the results also showed that the combination of sequences for plant identification could achieve a better result in DNA barcoding. This study investigated the genetic relationship between Lauraceae plants at the molecular level,aiming at providing a basis for the investigation,cultivation,development,protection, and utilization of medicinal plant resources.

4.
Article in Chinese | WPRIM | ID: wpr-906156

ABSTRACT

Objective:To identify 24 <italic>Rana</italic> species such as <italic>Rana dybowskii</italic> by mitochondrial cytochrome C oxidase subunit I (<italic>CO</italic>Ⅰ) gene-based DNA barcoding and build the neighbour-joining (NJ) tree for hierarchical cluster analysis, so as to provide a basis for the identification and classification of <italic>Rana</italic> species as well as the discovery of new species. Method:<italic>R. dybowskii</italic>, <italic>R. chensinensis</italic>, <italic>R. amurensis</italic>, <italic>R. culaiensi</italic>s, and <italic>R. huanrenesis</italic>, ten for each species, were collected for DNA extraction and polymerase chain reaction (PCR) amplification<italic> </italic>and sequencing. A total of 50 <italic>CO</italic>Ⅰ gene sequences were obtained. Then 163 <italic>CO</italic>Ⅰ gene sequences for 24 species of <italic>Rana</italic> and one <italic>CO</italic>Ⅰ gene sequence for <italic>Pelophylax</italic>,<italic> Odorrana</italic>, <italic>Nidirana</italic>, <italic>Hylarana</italic>, and <italic>Amolops</italic> were harvested from GenBank. After sequence alignment by MEGA X, the parsimony-informative sites of <italic>CO</italic>Ⅰ gene sequences were analyzed and the intraspecific and interspecific genetic distances were calculated, followed by the built of NJ tree and hierarchical cluster analysis. Result:The <italic>CO</italic>Ⅰ gene sequences of 24<italic> Rana</italic> species including <italic>R. dybowskii</italic> were 554 bp in length and there were 210 parsimony-informative sites in total. The intraspecific genetic distance of each species was smaller than 2%. Except that the interspecific genetic distance between <italic>R. sangzhiensis</italic> and <italic>R. zhengi</italic> was 0.004, the genetic distances between the other species ranged from 0.024 to 0.228. <italic>R. sangzhiensis</italic> and <italic>R. zhengi</italic> were clustered into one branch and some <italic>R. dybowskii</italic> and <italic>R. uenoi</italic> into one branch. There were two separate branches for <italic>R. chensinensis</italic> and the other species were all clustered independently. Conclusion:<italic>CO</italic>Ⅰ-based DNA barcoding enabled the identification of 24 species of <italic>Rana</italic> including <italic>R.dybowskii</italic>. The findings supported that <italic>R. sangzhiensis</italic>, <italic>R. zhengi</italic>, <italic>R. coreana</italic>, and <italic>R. kunyuensis</italic> were the same species. One branch of <italic>R. chensinensis </italic>might be one of the four undownloaded species in Ranidae or a new species. The results have demonstrated that <italic>CO</italic>Ⅰ-based DNA barcoding allows not only the identification of 24 species of Rana including <italic>R. dybowskii </italic>but also the classification of ranidae species and the discovery of new species or subspecies.

5.
Acta Pharmaceutica Sinica ; (12): 1497-1508, 2021.
Article in Chinese | WPRIM | ID: wpr-887087

ABSTRACT

Adulterants and counterfeits were found in some of the commercial traditional Chinese medicine (TCM) decoctions in Hongjin Xiaojie Jiaonang, Hongjin Xiaojie Pian, and Chaihuang Keli during the national drug sampling inspection. However, it was difficult to determine the species of the adulterants and counterfeits by conventional testing methods. Therefore, a total of 184 samples of the TCM decoctions and raw materials belong to the prescriptions of above mentioned traditional Chinese patent medicines, including Bupleuri Radix, Bajiaolian, Heimayi, and Shufuchong, were collected and authenticated by DNA barcoding technology. 111 ITS2 sequences were obtained from 115 commercial TCM decoctions and raw materials of Bupleuri Radix, among which 71 were Bupleurum chinense, three were B. scorzonerifolium, and 31 were closely related species in the same genus. In addition, counterfeits derived from different genera, such as Ailanthus altissima (one sample), Saposhnikovia divaricate (two samples), and Solidago decurrens (three samples), were also detected. 21 ITS2 sequences were obtained from 22 commercial TCM raw materials of Bajiaolian, among which 15 were Diphylleia sinensis and six were Dysosma versipellis and other species in genus Dysosma. For 22 Heimayi samples, PCR amplification of COI sequence was failed due to genomic DNA degradation. Among 38 Shufuchong samples, 24 COI sequences were obtained and only nine of them were the genuine species (Armadillidium vulgare) recorded in the Chinese Pharmacopoeia, 11 were Porcellio laevis, two were Mongoloniscus sinensis, and two samples could not be identified due to the limitation of database. This study demonstrates that DNA barcoding technology is suitable for the species authentication of the decoctions of traditional Chinese patent medicine prescription. It is a conductive way for the establishment of traceability system for the whole TCM industrial chain.

6.
Article in Chinese | WPRIM | ID: wpr-879175

ABSTRACT

Resina Draconis, a rare and precious traditional medicine in China, is known as the "holy medicine for promoting blood circulation". According to the national drug standard, it's derived from the resin extracted from the wood of Dracaena cochinchinensis, a Liliaceae plant. In addition, a variety of Dracaena species all over the world can form red resins, and there is currently no molecular identification method that can efficiently identify the origin of Dracaena medicinal materials. In this study, seven species of Dracaena distributed in China were selected as the research objects. Four commonly used DNA barcodes(ITS2, matK, rbcL and psbA-trnH), and four highly variable regions(trnP-psaJ, psbK-psbI, trnT-trnL, clpP) in chloroplast genome were used to evaluate the identification efficiency of Dracaena species. The results showed that clpP sequence fragment could accurately identify seven species of Dracaena plants. However, due to the long sequence of clpP fragment, there were potential problems in the practical application process. We found that the combined fragment "psbK-psbI+ trnP-psaJ" can also be used for accurate molecular identification of the Resina Draconis origin plants and relative species of Dracaena, which were both relatively short sequences in the combined fragment, showing high success rates of amplification and sequencing. Therefore, the "psbK-psbI+ trnP-psaJ" combined fragment can be used as the DNA barcode fragments for molecular identification of Resina Dracon's origin plants and relative species of Dracaena. Research on the identification of Dracaena species, the results of this study can be used to accurately identify the original material of Resina Draconis, and providing effective means for identification, rational development and application of Resina Draconis base source.


Subject(s)
China , DNA Barcoding, Taxonomic , DNA, Plant/genetics , Dracaena/genetics , Plants , Resins, Plant , Sequence Analysis, DNA
7.
Article in Chinese | WPRIM | ID: wpr-879004

ABSTRACT

Rhei Radix et Rhizoma is a kind of commonly used Chinese medicinal materials. Due to the overharvesting, the wild resource is endangering. Large market demand caused severely adulterant of commercial Rhei Radix et Rhizoma medicinal materials and decoction pieces. This manuscript reviewed the advances of the original species authentication in the industrial chain of Rhei Radix et Rhizoma during the latest decade, including characteristics and microscopic features, phytochemical analysis on anthraquinones, and molecular authentication based on DNA barcoding. Accordingly, an original species authentication route for the industrial chain of Rhei Radix et Rhizoma was summarized:(1)the identification of seeds and seedlings by DNA barcoding;(2) the selection of high variable sites based on the chloroplast genome;(3)biomonitoring of the Rhei Radix et Rhizoma medicinal materials and decoction pieces by two-dimensional DNA barcode;(4)traceability of Chinese patent medicines by third-generation sequencing. In conclusion, the combination of molecular identification and traditional identification methods provides a new idea for the identification of the original species of Rhei Radix et Rhizoma in the industrial chain and a essential guidance for the research of drug safety and efficacy of Rhei Radix et Rhizoma.


Subject(s)
Animals , Anthraquinones , Drugs, Chinese Herbal , Plant Roots , Rheum , Rhizome
8.
Acta Pharmaceutica Sinica ; (12): 879-889, 2021.
Article in Chinese | WPRIM | ID: wpr-876534

ABSTRACT

Although the guiding principles for molecular identification of traditional Chinese medicines (TCM) using DNA barcoding have been recorded in the Chinese Pharmacopoeia, there is still a lack of systematic research on its application to commercial TCM decoctions. In this study, a total of 212 commercial TCM decoctions derived from different medicinal parts such as root and rhizome, fruit and seed, herb, flower, leaf, cortex, and caulis were collected to verify applicability and accuracy of the method. DNA barcodes were successfully obtained from 75.9% (161/212) of the samples, while other samples failed to be amplified due to genomic DNA degradation. Among the 161 samples, 85.7% of them were identified as recorded species in the Chinese Pharmacopoeia (2020 edition). In addition, 14 samples could be identified as species recorded in the Chinese Pharmacopoeia and their closely related species in the same genus. Morphological identification for the unconfirmed samples showed that eight were genuine species and three were adulterants, while the other three were unidentifiable due to lack of morphological characteristics. Furthermore, the DNA barcodes of seven samples accurately mapped to the sequences of adulterants. Remarkably, counterfeit products were detected in two samples. These results demonstrate that DNA barcoding is suitable for the identification of commercial TCM decoctions. The method can effectively detect adulterants and is appropriate for use throughout the industrial chain of TCM production and distribution, and by the supervisory agencies as well.

9.
Neotrop. ichthyol ; 19(1): e200102, 2021. tab, graf, ilus, mapas
Article in English | LILACS, VETINDEX | ID: biblio-1154969

ABSTRACT

A new species of Hyphessobrycon belonging to the Hyphessobrycon heterorhabdus species-group from the lower rio Tapajós, state of Pará, Brazil, is described. The new species is allocated into the Hyphessobrycon heterorhabdus species-group due to its color pattern, composed by an anteriorly well-defined, horizontally elongated humeral blotch that becomes diffuse and blurred posteriorly, where it overlaps with a conspicuous midlateral dark stripe that becomes blurred towards the caudal peduncle and the presence, in living specimens, of a tricolored longitudinal pattern composed by a dorsal red or reddish longitudinal stripe, a middle iridescent, golden or silvery longitudinal stripe, and a more ventrally-lying longitudinal dark pattern composed by the humeral blotch and dark midlateral stripe. It can be distinguished from all other species of the group by possessing humeral blotch with a straight or slightly rounded ventral profile, lacking a ventral expansion present in all other species of the group. The new species is also distinguished from Hyphessobrycon heterorhabdus by a 9.6% genetic distance in the cytochrome c oxidase I gene. The little morphological distinction of the new species when compared with its most similar congener, H. heterorhabdus, indicates that the new species is one of the first truly cryptic fish species described from the Amazon basin.(AU)


Uma nova espécie de Hyphessobrycon pertencente ao grupo Hyphessobrycon heterorhabdus é descrita da região do baixo rio Tapajós, estado do Pará, Brasil. A nova espécie é incluída no grupo Hyphessobrycon heterorhabdus devido ao seu padrão de coloração, composto por uma mancha umeral alongada, anteriormente bem definida, que se torna difusa e borrada posteriormente, onde se sobrepõe a uma conspícua faixa escura médio-lateral que se torna borrada próxima ao pedúnculo caudal, e pela presença, em exemplares vivos, de um padrão longitudinal tricolor, composto por uma faixa longitudinal vermelha ou avermelhada dorsal, uma faixa média iridescente dourada ou prateada e, mais ventralmente, o padrão longitudinal escuro composto pela faixa escura médio-lateral e mancha umeral. A espécie pode ser distinguida das outras espécies pertencentes ao grupo por possuir uma mancha umeral com região ventral retilínea ou levemente arredondada, sem uma expansão ventral presente nas demais espécies do grupo. A espécie também se diferencia de Hyphessobrycon heterorhabdus por uma distância genética de 9,6% no gene citocromo c oxidase I. A sutil diferença morfológica da nova espécie quando comparada ao seu congênere mais similar, H. heterorhabdus, indica que a nova espécie é uma das primeiras espécies de peixes verdadeiramente crípticas descritas da Bacia Amazônica.(AU)


Subject(s)
Animals , DNA , Cytochromes c , Biodiversity , Characidae , Amazonian Ecosystem
10.
Neotrop. ichthyol ; 19(2): e200109, 2021. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1279484

ABSTRACT

The fishes of the Haemulidae family are currently allocated to 19 genera with a worldwide distribution in the tropical and subtropical waters of the world's oceans. Brachygenys and Haemulon are important genera of reef fish in Brazil, as they occur in large shoals, which are both ecologically and commercially valuable. This study identified the Brazilian species of the genera Brachygenys and Haemulon using DNA barcodes. While we found only a single lineage in Brachygenys chrysargyrea, Haemulon melanurum, H. parra, and H. squamipinna, more than one molecular operational taxonomic unit (MOTU) was identified in H. atlanticus, H. aurolineatum, and H. plumieri, indicating the possible existence of discrete populations or cryptic species.(AU)


Os peixes da família Haemulidae estão atualmente distribuídos em 19 gêneros, com distribuição mundial em águas oceânicas tropicais e subtropicais. Brachygenys e Haemulon são importantes gêneros de peixes recifais do Brasil, visto que ocorrem em grandes cardumes, de valores ecológicos e comerciais. Este estudo identificou as espécies brasileiras dos gêneros Brachygenys e Haemulon usando o código de barras de DNA. Embora apenas uma única linhagem de Brachygenys chrysargyrea, Haemulon melanurum, H. parra e H. squamipinna tenha sido encontrada em nosso conjunto de dados, mais de uma unidade taxonômica operacional molecular (MOTU) foi identificada em H. atlanticus, H. aurolineatum e H. plumieri, indicando a possível existência de populações discretas ou espécies crípticas.(AU)


Subject(s)
Animals , Perciformes , Products Distribution , Molecular Biology , Sequence Analysis, DNA , Fishes
11.
Braz. j. biol ; 81(2): 258-267, 2021. tab, graf
Article in English | MEDLINE, LILACS, VETINDEX | ID: biblio-1153355

ABSTRACT

The ichthyofauna diversity of the Jebba Hydroelectric Power (HEP) Dam, Jebba, North-central Nigeria was studied. Fishes were sampled for 24 months using gill net, hook and line, and cast net. Individuals were identified using morphological and molecular (mitochondrial Cytochrome c Oxidase subunit I) data. A total of 9605 freshwater fishes were recorded during the sampling period. The use of an integrative taxonomic approach enabled the identification of 83 species belonging to 42 genera. Additionally, the study recorded three unidentified species ­ Ctenopoma sp, Malapterurus sp., and Protopterus sp. Analyses showed that individuals belonging to families Cichlidae and Mochokidae dominated the dam. The diversity analyses revealed relatively high fish diversity during the rainy season at the downstream section of Jebba HEP dam compared to the upstream section. The study, therefore, showed the presence of a diverse fish community comprising high species richness and diversity across the Jebba HEP dam. Finally, we recommend proper biodiversity monitoring and assessment of freshwater fish diversity across Nigeria. In addition, the use of an integrated taxonomic approach is recommended for appropriate species' identification and studies of freshwater fishes from Nigeria.


A diversidade da ictiofauna da hidrelétrica de Jebba (HEP), Jebba, centro-norte da Nigéria foi estudada. Os peixes foram amostrados por 24 meses, utilizando rede de emalhar, anzol e linha, e rede de arrasto. Os indivíduos foram identificados usando a abordagem combinada morfológica e molecular (citocromo c Oxidase mitocondrial subunidade I). Um total de 9605 peixes de água doce foram registrados durante o período de amostragem. A identificação das espécies utilizando a abordagem taxonômica integrada possibilitou a identificação de 83 espécies pertencentes a 42 gêneros. Além disso, o estudo registrou três espécies não identificadas - Ctenopoma sp, Malapterurus sp e Protopterus sp. Análises mostraram que indivíduos pertencentes às famílias Cichlidae e Mochokidae dominaram a barragem. As análises dos índices de diversidade revelaram uma diversidade de peixes relativamente alta durante a estação chuvosa na seção a jusante da barragem Jebba HEP em comparação com a seção a montante. O estudo mostrou, portanto, a presença de diversas comunidades de peixes, que incluem alta riqueza e diversidade de espécies através da barragem Jebba HEP. Finalmente, recomendamos o monitoramento adequado da biodiversidade e a avaliação da diversidade de peixes de água doce em toda a Nigéria. Além disso, recomenda-se o uso de abordagem taxonômica integrada para a identificação adequada das espécies e estudos de peixes de água doce da Nigéria.


Subject(s)
Humans , Animals , Catfishes/genetics , DNA Barcoding, Taxonomic/veterinary , Fishes , Seasons , Hydroelectric Power Plants (Environmental Health) , Biodiversity , Fresh Water , Nigeria
12.
Rev. bras. entomol ; 65(2): e20210040, 2021. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1280016

ABSTRACT

Abstract The integration of field work with DNA barcoding is useful to disentangle lepidopteran interactions in nature. Adults of the little-known geometrid moth Cosmophyga cortesi Vargas, 2008 (Lepidoptera: Geometridae) were reared from folivorous larvae collected on the native tree Schinus areira L. (Anacardiaceae) at about 260 and 1500 m elevation in the transverse valleys of the Atacama Desert of northern Chile. Similarity of two DNA barcodes of C. cortesi was 99.7 and 100% with one sequence of an unidentified geometrid larva previously collected in a cell of the potter wasp Hypodynerus andeus (Packard, 1869) (Hymenoptera: Vespidae). This finding represents the first host plant record for C. cortesi, expands its previously documented altitudinal range by more than 1200 m and confirms that its larvae are captured by females of a potter wasp.

13.
Article | IMSEAR | ID: sea-209914

ABSTRACT

Groupers (Perciformes: Serranidae: Epinephelinae) are a significant group of marine and estuarine fishes havingecological and commercial importance in tropical and subtropical waters. The genus Epinephelus is identifiedbased on the color configuration and morphological characteristics. The homogeneity in the morphologicalcharacteristics has created confusion in the species identification of Epinephelus. Epinephelus areolatus withits delicious taste has high economic value; however, its population showed a remarkable decline in recenttimes due to overfishing. Due to morphological resemblance, it has often been confused with Epinepheluschlorostigma and Epinephelus bleekeri, which leads to misidentification. Hence, correct identification ofspecies is needed for the proper management of fish resources. This study aims to identify and validate E.areolatus by using morpho-meristic characteristics, including the number and pattern of pyloric caeca, vertebraecount through radiograph imaging, scale and otolith morphology, and DNA barcoding. The morphologicaland meristic characteristics of Epinephelus species observed in Nizampatnam coast are strikingly comparableto those reported earlier as E. areolatus. Comparative molecular analysis was also carried along with othermorphologically similar species, viz. E. chlorostigma and E. bleekeri, using mitochondrial cytochrome oxidaseI gene sequences. The mean interspecific genetic distance revealed high similarity with E. bleekeri (0.109 ±0.012) and low similarity with E. chlorostigma (0.079 ± 0.009). The neighbor-joining tree has shown distinctclades for three species with high bootstrap values. The partial DNA sequence of the mitochondrial cytochromeoxidase I was established for the first time for E. areolatus from the Nizampatnam coast in this study

14.
Article | IMSEAR | ID: sea-215774

ABSTRACT

Aims: To review the phytomedicinal researches on endemic plants of Northern Cyprus and to assess the plants of their DNA barcoding status.Study Design:A review.Methodology:This work reviewed available and accessible original articles in EBSCO, Ovid MEDLINE®, PubMed®, ScienceDirectTM, Scopus®and Web of ScienceTMdatabases on phytomedicinal investigations and BOLD System, MMDBD version 1.5 and GenBank®on DNA barcodes of the endemic plants of Northern Cyprusuntil May, 2020. Using keywords searches related to phytochemistry, biological activity and DNA barcoding, DNA Sequences and the data obtain evaluated and the information that does not meet the inclusion criteria were excluded. We believe that this information would tentatively help researchers to ethically explore these plants for their Medicinal and Aromatic potentials.Results:Only 6 of the 20 endemic plants of Northern Cyprus were phytopharmaceutically investigated,while DNA sequences of 5 were foundto be deposited in the publicly accessible databases accounting for30% and 25% of the total plants respectively Conclusion:Endemism is related to uniqueness in features including the phytomedicinal features, thus Northern Cyprus endemic plants holdample of such. However the results of this review showed that only few were harnessed for their medicinal properties and hence the need for their pharmacological properties and comprehensive barcoding for proper authentication, detection of adulteration, and quality control.

15.
Bol. latinoam. Caribe plantas med. aromát ; 19(3): 300-313, mayo 2020. ilus, tab
Article in English | LILACS | ID: biblio-1116300

ABSTRACT

Every 3 to 7 year angiosperms species of the flowering desert appear in the Atacama Region of Chile, as a result of the climatic phenomenon "El Niño". Our objective was to evaluate the universality of matK and rbcL barcode markers of these species, and validate their taxon through phylogenetic relationships. Argemone hunnemannii, Oenothera coquimbensis, Malesherbia humilis, Leucocoryne appendiculata, Loasa elongata, Nicotiana solanifolia, Stachys grandidentata, Aristolochia chilensis, Alstroemeria kingii and Adesmia eremophila, almost all classified as endemic to Chile, were collected in Pan de Azúcar and Llanos de Challe National Park (Atacama Region, Chile) at the end of October 2017. The phylogeny of these ten angiosperm species from the flowering desert was analyzed using rbcL and matK markers with the maximum likelihood and Bayesian inference methods. The results showed that 70% of the species can be distinguished with the matK or rbcL locus, however, 100% were distinguished using both loci. The phylogenetic results showed that the species formed clades with high reliability and high support with both the matK and rbcL genes, when comparing our results with sequences obtained from GenBank. The matK and rbcL genes are efficient markers for analyzing phylogenetic relationships and validating the taxonomy of flowering species.


Las especies de angiospermas del Desierto Florido de la Región de Atacama de Chile aparecen cada 3 a 7 años, influenciado por el fenómeno climático "El Niño". Nuestro objetivo fue evaluar la universalidad de los marcadores de código de barra matK y rbcL de estas especies, y validar su taxón por medio de relaciones filogenéticas. Las especies Argemone hunnemannii, Oenothera coquimbensis, Malesherbia humilis, Leucocoryne appendiculata, Loasa elongata, Nicotiana solanifolia, Stachys grandidentata, Aristolochia chilensis, Alstroemeria kingii y Adesmia eremophila son clasificadas la mayoría endémicas de Chile. Estas especies fueron colectadas en el Parque Nacional Pan de Azúcar y Llanos de Challe, Región de Atacama, Chile. La colecta se realizó a fines de octubre de 2017. Con los marcadores rbcL y matK se analizó la filogenia con los métodos máxima verosimilitud e inferencia bayesiana en diez especies de angiosperma del Desierto Florido. Los resultados mostraron que el 70% de las especies pueden ser distinguidas con un locus matK o rbcL, sin embargo, el 100% se distinguió usando ambos locus. Los resultados filogenéticos mostraron que las especies formaron clados con alta fiabilidad y alto soporte tanto con los genes matK y rbcL, al comparar con accesos de secuencias obtenidas de GenBank. Lo genes matK y rbcL son marcadores eficientes para analizar relaciones filogenéticas y validar el taxón de las especies de flor.


Subject(s)
Phylogeny , Plants/genetics , Desert , DNA Barcoding, Taxonomic/methods , Ribulose-Bisphosphate Carboxylase , Chile , Sequence Analysis, DNA
16.
Rev. peru. biol. (Impr.) ; 27(2): 139-148, abr.-jun 2020. tab, graf
Article in Spanish | LILACS-Express | LILACS | ID: biblio-1144944

ABSTRACT

Resumen En la Amazonia Peruana los caracoles dulceacuícolas de la familia Ampullariidae son conocidos como churos y originalmente han sido descritas para Perú alrededor de 20 especies. Aunque son muy usadas para alimentación, medicina tradicional y objeto de muchos estudios para su cultivo e industrialización, solamente es mencionada en la literatura la especie Pomacea maculata. Se llevó a cabo la identificación molecular sobre la base del marcador mitocondrial COI, de individuos de churos negros (Pomacea) comercializados en los mercados de Iquitos, así como los usados en platos a la carta en la ciudad de Lima, contrastados con otros individuos de procedencia de su hábitat natural. Se encontró que estos especímenes expendidos corresponden a la especie Pomacea nobilis (Reeve, 1856). El análisis filogenético molecular mostró que P. nobilis es especie hermana de P. guyanensis, en el grupo de P. glauca, distantemente relacionada de P. maculata. Las distancias no corregidas encontradas entre ellas, para el marcador mitocondrial COI, fueron de 11.33% a 13.17%, mientras que con P. maculata fueron de 13.67% a 15.33%. Estos resultados demostraron la eficacia del código de barras de ADN para la identificación y autenticación de la especie, lo que le da un valor agregado para su eventual comercio de exportación.


Abstract In the Peruvian Amazon, freshwater snails of the Ampullariidae family are known as churos, and around 20 species have originally been described for Peru. Although they are widely used for food, traditional medicine and the object of many studies for their cultivation and industrialization, only the species Pomacea maculata is mentioned in the literature. Molecular identification was carried out based on the mitochondrial marker COI of individuals of "churo negro" apple snails (Pomacea) commercialized in the markets of Iquitos, as well as those used in restaurant dishes in the city of Lima, and contrasted with specimens from their natural habitat. It was found that these specimens, correspond to the species Pomacea nobilis (Reeve, 1856). The molecular phylogenetic analysis showed P. nobilis as the sister species of P. guyanensis, in the P. glauca group, distantly related to P. maculata. The uncorrected distances found between them, for the mitochondrial marker COI, were from 11.33% to 13.17%, while with P. maculate were from 13.67% to 15.33%. These results demonstrated the effectiveness of the DNA barcode for the identification and authentication of the species, which gives it added value for its eventual export trade.

17.
Article in Chinese | WPRIM | ID: wpr-872963

ABSTRACT

Objective::To select volatile oils from 16 species of plants (Cymbopogon citratus, Pelargonium graveolens, Pinus tabulieformis, Litsea cubeba, Mentha haplocalyx, Zingiber officinale, Syzygium aromaticum, Curcuma longa, Zanthoxylum bungeanum, Cinnamomum cassia, Ocimum basilicum, Rosmarinus officinalis, Zanthoxylum schinifolium, Zanthoxylum armatum, Illicium verum, Myristica fragrans) that have good inhibitory effect on the growth of Aspergillus flavus. Method::Aspergillus flavus was isolated from the surface of Platycladi Semen medicinal materials by plate culture method. The volatile oils of 16 plants were extracted by steam distillation. The colony diameter of Aspergillus flavus was determined by fumigation of filter paper, and the effect of volatile oils on the growth of Aspergillus flavus was systematically studied. Result::Aspergillus flavus was successfully isolated from Platycladi Semen by means of morphological, microscopic and DNA barcoding identification methods, the bacteriostatic rates of the above 16 kinds of volatile oils against Aspergillus flavus were 2.93%, 0.05%, 0.37%, 76.07%, 0.34%, 0.15%, 50.05%, 8.51%, 1.43%, 58.20%, 0.07%, 2.60%, 8.73%, 100.00%, 52.62%, 0.07%, respectively. Conclusion::The volatile oils of 16 plants all have different degrees of antibacterial activities for Aspergillus flavus, and volatile oils of Zanthoxylum armatum, Litsea cubeba and Cinnamomum cassia have good inhibitory effect. This study can provide a theoretical basis for the prevention and control of Aspergillus flavus in the growth and storage of Platycladi Semen, and provide a basis for further research on plant volatile oil as bacteriostatic agents in the storage process of traditional Chinese medicine.

18.
Article in Chinese | WPRIM | ID: wpr-872962

ABSTRACT

Objective::The complete chloroplast genome of Pyrrosia assimilis was sequenced, its sequence characteristics was analyzed and herbgenomics of P. assimilis was discussed. Method::Its complete chloroplast genome sequence was determined through high-throughput sequencing technology, and its structural characteristics and phylogenetic relationships were analyzed by bioinformatics. Result::The chloroplast genome of P. assimilis was a circular double-chain structure with a total length of 154 964 bp, and the total content of guanine and cytosine (GC) was 41.2%. A total of 131 genes were annotated, including 88 protein-coding genes, 35 transfer RNA (tRNA) genes and 8 ribosomal RNA (rRNA) genes. A total of 43 dispersed repetitive sequences and 56 simple sequence repeats (SSRs) were detected. The frequency of codon encoding leucine was the highest, while the number of codon encoding tryptophan was the lowest. Five highly divergent regions (psbA, rrn16, petA-psbJ, ndhC-trnM, and psbM-petN) were screened, phylogenetic analysis showed that P. assimilis was closely related to P. bonii. Conclusion::Comparative analysis of the complete chloroplast genome of P. assimilis reveals that non-coding regions exhibited a higher divergence than the coding regions, the large single copy region (LSC) and small single copy region (SSC) are more divergent than the reverse repeat region (IR), the selected five highly variable regions can be used as specific DNA barcodes for identification of Pyrrosia species. Study on the chloroplast genome of P. assimilis can provide a reference for the molecular identification, genetic transformation, expression of resistance protein and secondary metabolism pathway analysis of other Pyrrosia medicinal plants.

19.
Article in Chinese | WPRIM | ID: wpr-872932

ABSTRACT

Objective:Five popular DNA barcoding sequences,namely ITS,ITS2,rbcL,matK and psbA-trnH,were employed to evaluate the identification efficiency of multi-source ethnodrug Rodgersiae Rhizoma,and the most suitable sequence was then screened out. Method:Efficiency of polymerase chain reaction(PCR) amplification,success rate of sequencing,intra- and inter-specific distances calculated by rank sum test,phylogenetic tree constructed with neighbor-joining(NJ) method and identification efficiency assessed by Blast 1 and NJ method were adopted in this study. Result:Efficiency of PCR amplification and success rate of sequencing for ITS,ITS2,psbA-trnH,rbcL and matK were 100%,96.61% ,100%,98.31%,100%,100%,100%,100%,98.31% and 98.31%,respectively. Intra- and inter-specific genetic distances and identification achievement rate for psbA-trnH were the highest among the five candidate sequences. Besides,the average coalescent depth was less than the smallest interspecific distance for psbA-trnH. Phylogenetic tree also illustrated that Rodgersiae Rhizoma could be distinguished based on psbA-trnH. Conclusion:According to the findings,psbA-trnH was superior to other DNA barcodes. Therefore, psbA-trnH was recommended as the ideal DNA barcode for the identification of multi-source ethnodrug Rodgersiae Rhizoma.

20.
Article in Chinese | WPRIM | ID: wpr-872836

ABSTRACT

Objective:To construct a systematic identification system of Anemonis Flaccidae Rhizoma, and to evaluate the comprehensive quality of Anemonis Flaccidae Rhizoma from 16 regions in China, so as to lay a foundation for its origin selection and clinical medication safety. Method:The authenticity of Anemonis Flaccidae Rhizoma was quickly identified by traditional identification method and DNA barcode molecular identification technology, and HPLC-UV was used to determine the contents of 5 active ingredients in Anemonis Flaccidae Rhizoma. All high pressure chromatographic separations were performed with a Welch Ultimate XB-C18 column (4.6 mm×250 mm, 5 μm), the mobile phase consisted of acetonitrile-0.01% trifluoroacetic acid aqueous solution (30∶70) at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 210 nm and the column temperature was maintained at 30 ℃. Result:The authenticity of Anemonis Flaccidae Rhizoma could be precisely and rapidly identified by ribosomal DNA internal transcribed spacer 2 (ITS2) sequence and traditional identification methods. BLAST comparative analysis found that medicinal materials from 16 areas were all Anemone flaccida. Based on the contents of multi-index components, it was shown that the total content of 5 triterpenoid saponins in Anemonis Flaccidae Rhizoma from Banqiao, Enshi, Hubei was the highest (10.59%), followed by Hezhang, Bijie, Guizhou (6.28%) and Duzhenwan, Changyang, Hubei (5.64%). Conclusion:DNA barcoding can be used as an effective supplement to the traditional identification technology, it can ensure the authenticity of Anemonis Flaccidae Rhizoma and the safety of clinical use. The comprehensive evaluation of multi-index components of HPLC and cluster analysis show that the quality of medicinal materials in Enshi, Changyang, Wufeng of Hubei, Bijie of Guizhou and Jinfoshan of Chongqing is superior, which can be considered as important origin of Anemonis Flaccidae Rhizoma.

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