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1.
PLoS One ; 15(9): e0239823, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32986773

RESUMO

Houttuynia cordata (Saururaceae), an ancient and relic species, has been used as an important medicinal and edible plant in most parts of Asia. However, because of the lack of genome information and reliable molecular markers, studies on its population structure, or phylogenetic relationships with other related species are still rare. Here, we de novo assembled the complete chloroplast (cp) genome of H. cordata using the integration of the long PacBio and short Illumina reads. The cp genome of H. cordata showed a typical quadripartite cycle of 160,226 bp. This included a pair of inverted repeats (IRa and IRb) of 26,853 bp, separated by a large single-copy (LSC) region of 88,180 bp and a small single-copy (SSC) region of 18,340 bp. A total of 112 unique genes, including 79 protein-coding genes, 29 tRNA genes, and four rRNA genes, were identified in this cp genome. Eighty-one genes were located on the LSC region, 13 genes were located on the SSC region, and 17 two-copy genes were located on the IR region. Additionally, 48 repeat sequences and 86 SSR loci, which can be used as genomic markers for population structure analysis, were also detected. Phylogenetic analysis using 21 cp genomes of the Piperales family demonstrated that H. cordata had a close relationship with the species within the Aristolochia genus. Moreover, the results of mVISTA analysis and comparisons of IR regions demonstrated that the cp genome of H. cordata was conserved with that of the Aristolochia species. Our results provide valuable information for analyzing the genetic diversity and population structure of H. cordata, which can contribute to further its genetic improvement and breeding.


Assuntos
Cloroplastos/genética , Genoma de Cloroplastos , Houttuynia/genética , Plantas Comestíveis/genética , Aristolochia/genética , Evolução Molecular , Genes de Plantas , Genes de RNAr , Loci Gênicos , Variação Genética , Genoma de Planta , Sequências Repetidas Invertidas , Filogenia , Plantas Medicinais/genética , RNA de Transferência/genética
3.
Zhongguo Zhong Yao Za Zhi ; 45(11): 2714-2719, 2020 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-32627508

RESUMO

To breed new varieties of medicinal plants with high resistance is the premise to ensure the production of high-quality medicinal materials. Molecular breeding using modern molecular biology and genetic technology can save time and effort and realize rapid and accurate breeding. Here we are trying to summarize the difference of breeding characteristics between medicinal plants and crops such as genetic background and breeding purpose. The strategy of molecular breeding of medicinal plants was summarized, and the four-phases breeding based on high-throughput sequencing and target gene mining was emphasized. We put forward the current molecular breeding of medicinal plants in the condition of four phases breeding is the optimal technological way of breeding, and gene editing breeding is the direction of medicinal plants breeding.


Assuntos
Plantas Medicinais/genética , Cruzamento , Embaralhamento de DNA , Edição de Genes , Melhoramento Vegetal
4.
Chin J Nat Med ; 18(6): 417-424, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32503733

RESUMO

In recent years, biosynthesis of triterpenoid saponins in medicinal plants has been widely studied because of their active ingredients with diverse pharmacological activities. Various oxidosqualene cyclases, cytochrome P450 monooxygenases, uridine diphosphate glucuronosyltransferases, and transcription factors related to triterpenoid saponins biosynthesis have been explored and identified. In the biosynthesis of triterpenoid saponins, the progress of gene mining by omics-based sequencing, gene screening, gene function verification, catalyzing mechanism of key enzymes and gene regulation are summarized and discussed. By the progress of the biosynthesis pathway of triterpenoid saponins, the large-scale production of some triterpenoid saponins and aglycones has been achieved through plant tissue culture, transgenic plants and engineered yeast cells. However, the complex biosynthetic pathway and structural diversity limit the biosynthesis of triterpenoid saponins in different system. Special focus can further be placed on the systematic botany information of medicinal plants obtained from omics large dataset, and triterpenoid saponins produced by synthetic biology strategies, gene mutations and gene editing technology.


Assuntos
Plantas Medicinais/química , Plantas Medicinais/genética , Saponinas/biossíntese , Triterpenos/química , Vias Biossintéticas , Sistema Enzimático do Citocromo P-450/fisiologia , Regulação da Expressão Gênica de Plantas , Glucuronosiltransferase/fisiologia , Estrutura Molecular , Plantas Geneticamente Modificadas , Fatores de Transcrição , Difosfato de Uridina/fisiologia
5.
Phytochemistry ; 176: 112400, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32408189

RESUMO

Adulterants in processed food and herbal medicines reduce their safety, quality control, or pharmacological efficacy. Four mistletoe species, including Viscum coloratum, inhabit Korea. Leaves and branches of V. coloratum, defined as edible or medicinal mistletoe species in Korean, are used to prepare Korean herbal medicines as well as leached tea. However, other mistletoe species including Taxillus sutchuenensis var. duclouxii, Korthalsella japonica, and Loranthus tanakae are frequently distributed as authentic V. coloratum in Korean markets because of similarities in the branches morphology and Korean names of these species with V. coloratum. Although herbal medicines and food products prepared from the other mistletoe species are inauthentic, they are sold at high prices because of the rarity of these species. Thus, it is important to distinguish between authentic and inauthentic adulterant mistletoe species. In this study, we developed species-specific primer, based on matK sequences, suitable for both conventional PCR and real time PCR (qPCR) assay. These assays allowed rapid discrimination among all four mistletoe species. Moreover, qPCR assay enabled the detection of trace amounts of adulterant mistletoe species in V. coloratum samples. Furthermore, we used these assays to monitor commercial mistletoe products distributed in Korean markets. Our data suggest that these methods would serve as a reliable genetic tool to prevent adulteration and standardize the quality of commercial V. coloratum products.


Assuntos
Produtos Biológicos , Erva-de-Passarinho , Plantas Medicinais/genética , Reação em Cadeia da Polimerase em Tempo Real , República da Coreia
6.
Artigo em Inglês | MEDLINE | ID: mdl-32460213

RESUMO

Datura innoxia Mill., a traditional Chinese herbal medicine, produces tropane alkaloids such as hyoscyamine and scopolamine. Scopolamine has a larger demand than hyoscyamine due to its stronger pharmacological effects and fewer side reactions. It is extracted from solanaceous plants. However, the content of scopolamine is lower than hyoscyamine in D. innoxia. Hyoscyamine 6ß-hydroxylase (H6H, EC1.14.11.11) is the key enzyme which can catalyze hyoscyamine to form scopolamine. In this study, a cDNA encoding H6H was cloned from D. innoxia roots and named Dih6h. The full-length cDNA is 1413 bp in length with a 1044-bp open reading frame encoding 347 amino acids. The deduced protein sequence of D. innoxia H6H (DiH6H) shared high identity with H6Hs from other plants. The DiH6H was heterologously expressed in Escherichia coli and purified via His-tag affinity technique. The recombinant DiH6H showed activity in transforming hyoscyamine to scopolamine. Despite Dih6h mRNA was detected in various tissues, its levels in roots were higher than that in other tissues. Indeed, scopolamine accumulation was low in roots, but it was very high in aerial parts, especially in flowers and seeds. These observations suggest that scopolamine may be synthesized in the roots and subsequently transported to the aerial parts. To further verify in vivo function of DiH6H, the cDNA of DiH6H was overexpressed in D. innoxia hairy roots. As expected, an increase of scopolamine production was observed in the positive transformants. The results provide a potential strategy for increasing scopolamine yield by metabolic engineering of its biosynthetic pathway in D. innoxia.


Assuntos
Datura/enzimologia , Oxigenases de Função Mista/genética , Proteínas de Plantas/genética , Clonagem Molecular , Datura/genética , Plantas Medicinais/enzimologia , Plantas Medicinais/genética
7.
Zhongguo Zhong Yao Za Zhi ; 45(2): 341-346, 2020 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-32237316

RESUMO

Pinellia ternata belongs to the Araceae family and is a medicinal herb. The tuber is the medicinal organ with antitussive, antiemetic and anti-tumor activities. It is easy to encounter high temperature environment during the growth periods, leading to decrease of tuber production. At present, the mechanism of response to high temperature stress in P. ternata is still unknown. DNA methylation plays a vital role in plant protection against adversity stress as a way of epigenetic regulation. In this study, P. ternata was used as material for treatment of high temperature stress at 0 h, 6 h and 80 h, and methylation sensitive amplification polymorphism(MSAP) analysis was conducted on the changes of DNA methylation in its genome. The results showed that 20 pairs of MSAP primers were selected from 100 MSAP primers with multiple clear and uniform bands, and 353, 355 and 342 loci were amplified from materials of P. ternata treated in the high temperature stress 0 h, 6 h and 80 h, respectively. Cytosine methylation levels of CCGG context in the above materials were characterized as 60.91%, 44.79% and 44.74%, respectively. And the full methylation ratios were 16.71%, 22.25% and 29.24, respectively. It demonstrated that high temperature stress significantly induced the down-regulation of DNA methylation level and up-regulation of the full methylation rate in P. ternata genome. This study provides a preliminary theoretical reference for analyzing the mechanism of P. ternata responding to high temperature stress from the epigenetic perspective.


Assuntos
Metilação de DNA , Epigênese Genética , Temperatura Alta , Pinellia/genética , Plantas Medicinais/genética
8.
Zhongguo Zhong Yao Za Zhi ; 45(6): 1311-1315, 2020 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-32281341

RESUMO

Pinellia ternata is a medicinal herb of Araceae, and its tubers are used as medicines. It is a common Chinese herbal medicine in China and has a large market demand. When exposing to strong light intensity and high temperature during the growth process, P. ternata withers in a phenomenon known as "sprout tumble", which largely limits tuber production. Shade can effectively delay sprout tumble formation and increase its yield, however the relevant regulation mechanism is unclear. DNA methylation, as a self-modifying response to environmental changes, is often involved in the regulation of plant growth and development. In this study, P. ternata grown under natural light and 90% shading were selected as the control group and the experimental group for genomic DNA methylation analysis by using methylate sensitive amplification polymorphism(MSAP). The results showed that a total of 617 loci were detected with 20 pairs of primers, of which 311 were in the natural light group and 306 in the shading group. The methylation sites in the light and shading groups accounted for 58.2% and 71.57%, respectively, and the methylation ratios in the methylation sites were 27.65% and 29.41%, respectively, indicating that shading significantly induced the genome DNA methylation of P. ternata. Compared to the natural light group, shading promoted 32.51% of the genes methylation, while inducing 16.25% gene demethylation. This study reveals the DNA methylation variation of P. ternata under shading conditions, which lays a preliminary theoretical foundation for further analysis of the mechanism of shading regulation of P. ternata growth from epigenetic level.


Assuntos
Metilação de DNA , Escuridão , Pinellia/genética , Pinellia/efeitos da radiação , Luz Solar , China , Epigênese Genética , Plantas Medicinais/genética , Plantas Medicinais/efeitos da radiação
9.
Sci Rep ; 10(1): 4614, 2020 03 12.
Artigo em Inglês | MEDLINE | ID: mdl-32165650

RESUMO

Pinellia (Pinellia ternata (Thunb.) Breit.), as important medicinal plant, has been used to treat various ailments for a long time. The sixteen ploid plant (2n = 16 * 13 = 208) Pinellia T2Plus line was obtained from an octoploid (2n = 8 * 13 = 104) T2 line by chromosome-doubling technique. Compared with T2 line, the content of various medicinal components (polysaccharide, guanosine, adenosine and ephedrine) was increased in T2Plus line. In this study, the transcriptome of T2 line and T2Plus line were characterized by RNA sequencing (RNA-seq) technology. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways enrichment analysis on differential expressed unigenes (DEGs) revealed that multiple metabolic pathway were enriched significantly, such as 'Starch and sucrose metabolism', 'Purine metabolism', 'Photosynthesis' and six transcription factors (MYB, WRKY, bHLH, lateral organ boundaries domain (LBD), homeodomain-zipper (HD-ZIP) and Ethylene-responsive factor (ERF)) play a key role in difference of transcriptome between T2 line and T2Plus line. These metabolic pathways and transcription factors may play an important role in the difference of medicinal components and epigenetic features between these two Pinellia cultivars. This conclusion provides a robust theoretical basis for the mechanism of the formation of medicinal ingredients in Pinellia cultivars.


Assuntos
Perfilação da Expressão Gênica , Fenótipo , Pinellia/genética , Transcriptoma , Biologia Computacional/métodos , Metabolismo Energético , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Redes e Vias Metabólicas , Fotossíntese , Pinellia/metabolismo , Plantas Medicinais/genética
11.
PLoS One ; 15(2): e0229490, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32107496

RESUMO

Application of plant growth regulators has become one of the most important means of improving yield and quality of medicinal plants. To understand the molecular basis of phytohormone-regulated oleanolic acid metabolism, RNA-seq was used to analyze global gene expression in Achyranthes bidentata treated with 2.0 mg/L 1-naphthaleneacetic acid (NAA) and 1.0 mg/L 6-benzyladenine (6-BA). Compared with untreated controls, the expression levels of 20,896 genes were significantly altered with phytohormone treatment. We found that 13071 (62.5%) unigenes were up-regulated, and a lot of differentially expressed genes involved in hormone or terpenoid biosynthesis, or transcription factors were significantly up-regulated. These results suggest that oleanolic acid biosynthesis induced by NAA and 6-BA occurs due to the expression of key genes involved in jasmonic acid signal transduction. This study is the first to analyze the production and hormonal regulation of medicinal A. bidentata metabolites at the molecular level. The results herein contribute to a better understanding of the regulation of oleanane-type triterpenoid saponins accumulation and define strategies to improve the yield of these useful metabolites.


Assuntos
Achyranthes/efeitos dos fármacos , Achyranthes/metabolismo , Compostos de Benzil/farmacologia , Ciclopentanos/metabolismo , Ácidos Naftalenoacéticos/farmacologia , Ácido Oleanólico/biossíntese , Oxilipinas/metabolismo , Purinas/farmacologia , Achyranthes/crescimento & desenvolvimento , Vias Biossintéticas/efeitos dos fármacos , Vias Biossintéticas/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas , Medicina Tradicional Chinesa , Filogenia , Reguladores de Crescimento de Planta/metabolismo , Plantas Medicinais/efeitos dos fármacos , Plantas Medicinais/genética , Plantas Medicinais/metabolismo , RNA-Seq , Saponinas/metabolismo
12.
J Genet ; 992020.
Artigo em Inglês | MEDLINE | ID: mdl-32089530

RESUMO

The endangered medicinal plant Glehnia littoralis is one of the important natural source of furanocoumarin, which has been used as mucolytic, antitussive, antitumour and antibacterial. However, the genetic information of furanocoumarin biosynthesis in G. littoralis is scarce at present. The objective of this study was to mine the putative candidate genes involved in the biosynthesis pathwayof furanocoumarin and provide references for gene identification, and functional genomics of G. littoralis. We carried out the transcriptome analysis of leaves and roots in G. littoralis, which provided a dataset for gene mining. Psoralen, imperatorin and isoimperatorin were detected in G. littoralis by high performance liquid chromatography analysis. Candidate key genes were mined based on the annotations and local BLAST with homologous sequences using BioEdit software. The relative expression of genes was analysed using quantitative real-time polymerase chain reaction. Further, the CYP450 genes were mined using phylogenetic analyses using MEGA 6.0 software. Atotal of 156,949 unigenes were generated, of which 9021 were differentially-expressed between leaves and roots. A total of 82 unigenes encoding eight enzymes in furanocoumarin biosynthetic pathway were first obtained. Seven genes that encoded key enzymes in the downstream furanocoumarin biosynthetic pathway and expressed more in roots than leaves were screened. Twenty-six candidate CYP450 unigenes expressed abundantly in roots and were chiefly concentrated in CYP71, CYP85 and CYP72 clans. Finally, we filtered 102 differentially expressed transcription factors (TFs) unigenes. The transcriptome of G. littoralis was characterized which would help to elucidate the furanocoumarin biosynthetic pathway in G. littoralis and provide an invaluable resource for further study of furanocoumarin.


Assuntos
Apiaceae/genética , Apiaceae/metabolismo , Vias Biossintéticas/genética , Furocumarinas/biossíntese , Perfilação da Expressão Gênica , Plantas Medicinais/genética , Plantas Medicinais/metabolismo , Apiaceae/classificação , Biologia Computacional/métodos , Regulação da Expressão Gênica de Plantas , Anotação de Sequência Molecular , Filogenia , Plantas Medicinais/classificação , Transcriptoma
13.
Nat Commun ; 11(1): 971, 2020 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-32080175

RESUMO

Triptolide is a trace natural product of Tripterygium wilfordii. It has antitumor activities, particularly against pancreatic cancer cells. Identification of genes and elucidation of the biosynthetic pathway leading to triptolide are the prerequisite for heterologous bioproduction. Here, we report a reference-grade genome of T. wilfordii with a contig N50 of 4.36 Mb. We show that copy numbers of triptolide biosynthetic pathway genes are impacted by a recent whole-genome triplication event. We further integrate genomic, transcriptomic, and metabolomic data to map a gene-to-metabolite network. This leads to the identification of a cytochrome P450 (CYP728B70) that can catalyze oxidation of a methyl to the acid moiety of dehydroabietic acid in triptolide biosynthesis. We think the genomic resource and the candidate genes reported here set the foundation to fully reveal triptolide biosynthetic pathway and consequently the heterologous bioproduction.


Assuntos
Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Diterpenos/metabolismo , Fenantrenos/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Tripterygium/genética , Tripterygium/metabolismo , Abietanos/metabolismo , Antineoplásicos Fitogênicos/biossíntese , Vias Biossintéticas/genética , Medicamentos de Ervas Chinesas/metabolismo , Compostos de Epóxi/metabolismo , Perfilação da Expressão Gênica , Genoma de Planta , Humanos , Engenharia Metabólica , Metaboloma , Oxirredução , Filogenia , Plantas Medicinais/genética , Plantas Medicinais/metabolismo
14.
PLoS One ; 15(1): e0227610, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31990944

RESUMO

Atractylodes lancea, A. chinensis, and A. macrocephala are the three most widely used medicinal species of the Atractylodes genus. Their similar morphological features cause disagreement as whether they are three unique species, leading to their frequent misuses in medical products. Our study aimed to understand their relationships through both the complete plastome sequences and nuclear sequences, to identify molecular markers for their differentiation and explore the evolutionary relationships among three species. We sequenced, annotated, and analyzed the plastomes of these three species. The plastomes are 153,201, 153,258, and 153,265 bps in length for A. lancea, A. chinensis, and A. macrocephaly, respectively. Similar to other Asteraceae species, their plastomes exhibit typical quadripartite structures. Each plastome consists of 119 distinct genes, including 78 protein-coding, 37 tRNA, and 4 rRNA genes. Analyses of indels, single-nucleotide polymorphisms and simple sequence repeats, and comparison of plastomes showed high degree of conservation, leading to difficulty in the discovery of differentiating markers. We identified eleven potential molecular markers using an algorithm based on interspecific and intraspecific nucleotide diversity gaps. Validation experiments with fifty-five individuals from the three species collected from the botanical garden and fields confirmed that the marker cz11 could effectively distinguish samples from the three different species. Analysis of the several nuclear sequences suggests that the species of A. macrocephala may be a hybrid of A. lancea and A. chinensis. In summary, the results from this study highlight the complex relationships among of these three medicinal plants.


Assuntos
Atractylodes/genética , Filogenia , Plastídeos/genética , Alelos , Núcleo Celular/genética , Genes de Plantas , Marcadores Genéticos , Genoma de Planta , Mutação INDEL , Plantas Medicinais/genética , Reprodutibilidade dos Testes
15.
PLoS One ; 15(1): e0227921, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31971962

RESUMO

Low temperature induces changes in plants at physiological and molecular levels, thus affecting growth and development. The Lanzhou lily (Lilium davidii, var. unicolor) is an important medicinal plant with high economic value. However, the molecular mechanisms underlying its photosynthetic and antioxidation responses to low temperature still remain poorly understood. This study subjected the Lanzhou lily to the two temperatures of 20°C (control) and 4°C (low temperature) for 24 h. Physiological parameters related to membrane integrity, photosynthesis, antioxidant system, and differentially expressed genes were investigated. Compared with control, low temperature increased the relative electrical conductivity by 43.2%, while it decreased net photosynthesis rate, ratio of variable to maximal fluorescence, and catalase activity by 47.3%, 10.1%, and 11.1%, respectively. In addition, low temperature significantly increased the content of soluble protein, soluble sugar, and proline, as well as the activity of superoxide dismutase and peroxidase. Comparative transcriptome profiling showed that a total of 238,109 differentially expressed genes were detected. Among these, 3,566 were significantly upregulated while 2,982 were significantly downregulated in response to low temperature. Gene Ontology enrichment analysis indicated that in response to low temperature, the mostly significantly enriched differentially expressed genes were mainly involved in phosphorylation, membrane and protein kinase activity, as well as photosynthesis, light harvesting, light reaction, and alpha,alpha-trehalose-phosphate synthase activity. Kyoto Encyclopedia of Genes and Genomes enrichment analysis also indicated that the most significantly enriched pathways involved ribosome biogenesis in eukaryotes, phenylalanine metabolism, circadian rhythm, porphyrin and chlorophyll metabolism, photosynthesis of antenna proteins, photosynthesis, and carbon fixation in photosynthetic organisms. Moreover, the expression patterns of 10 randomly selected differentially expressed genes confirmed the RNA-Seq results. These results expand the understanding of the physiological and molecular mechanisms underlying the response of the Lanzhou lily to low temperature stress.


Assuntos
Lilium/genética , Raízes de Plantas/genética , Plantas Medicinais/genética , Transcriptoma/genética , Temperatura Baixa/efeitos adversos , Regulação da Expressão Gênica de Plantas/genética , Ontologia Genética , Lilium/fisiologia , Fotossíntese/genética , Fotossíntese/fisiologia , Plantas Medicinais/fisiologia , RNA-Seq , Estresse Fisiológico/genética
16.
Genome ; 63(1): 53-60, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31580739

RESUMO

Arctium lappa, commonly called burdock, has a long medicinal and edible history. It has recently gained increasing attention because of its economic value. In this study, we obtained the complete chloroplast genome of A. lappa by Illumina Hiseq. The complete chloroplast genome of A. lappa is a typical circular structure with 152 708 bp in length. The GC content in the whole chloroplast genome of A. lappa is 37.7%. A total of 37 tRNA genes, 8 rRNA genes, and 87 protein-coding genes were successfully annotated. And the chloroplast genome contains 113 unique genes, 19 of which are duplicated in the inverted repeat. The distribution of 39 simple sequence repeats was analysed, and most of them are in the large single-copy (LSC) sequence. An inversion comprising 16 genes was found in the LSC region, which is 26 283 bp long. We performed multiple sequence alignments using 72 common protein-coding genes of 29 species and constructed a Maximum Parsimony (MP) tree. The MP phylogenetic result shows that A. lappa grouped together with Carthamus tinctorius, Centaurea diffusa, and Saussurea involucrata. The chloroplast genome of A. lappa is a valuable resource for further studies in Asteraceae.


Assuntos
Arctium/genética , Genoma de Cloroplastos , Arctium/classificação , Uso do Códon , DNA de Plantas/química , Genes de Plantas , Sequências Repetidas Invertidas , Repetições de Microssatélites , Filogenia , Plantas Medicinais/genética , Sequências Repetitivas de Ácido Nucleico
17.
J Nat Med ; 74(1): 282-293, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31587135

RESUMO

The dried fruits of Terminalia plant (Combretaceae) called "Samo" have been used as herbal medicine in Thai traditional medicine. Four "Samo" crude drugs, namely, Samo thai, Samo thed, Samo dee-ngu, and Samo phiphek, are used as the main ingredients in Triphala and Trisamo recipes. Their commercial products are available in processed and powdered form, but are difficult to authenticate by conventional methods. In this study, we aimed to discriminate species of genus Terminalia for the identification of their crude drugs by a DNA barcoding technique. A total of 208 closely related nucleotide sequences were obtained from nine Terminalia species collected from Thailand and the DDBJ/EMBL/GenBank database. An effective DNA barcode marker was selected from six DNA loci (matK, rbcL, psbA-trnH, ITS, ITS1, and ITS2) and their two-locus combination. All sequences were analyzed by three major methods: (1) BLAST search; (2) the genetic divergence method using Kimura 2-parameter (K2P) distance matrices; and (3) tree topology analysis based on the neighbor-joining method. Comparison of the six candidate DNA loci indicated that ITS identified Terminalia with 100% accuracy at the species and genus levels in the BLAST1 method. ITS2 showed the highest K2P variability. The data from the single markers and the two-locus combinations revealed that only the two-locus combinations, namely, the combinations of rbcL, ITS, ITS1, and ITS2 with psbA-trnH, clearly discriminated all the species. From the results of DNA sequence analysis and the three methods, ITS2 is recommended for the identification of Terminalia species to supplement psbA-trnH.


Assuntos
Código de Barras de DNA Taxonômico/métodos , DNA Intergênico/genética , Complexo de Proteína do Fotossistema II/genética , Terminalia/classificação , Terminalia/genética , Sequência de Bases , DNA de Plantas/genética , Marcadores Genéticos/genética , Fitoterapia , Extratos Vegetais/química , Plantas Medicinais/genética , Análise de Sequência de DNA , Tailândia
18.
J Ethnopharmacol ; 250: 112495, 2020 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-31877364

RESUMO

ETHNOPHARMALOGICAL RELEVANCE: In Africa, traditional medicine is important for local healthcare and plants used for these purposes are commonly traded. Identifying medicinal plants sold on markets is challenging, as leaves, barks and roots are often fragmented or powdered. Vernacular names are often homonymic, and identification of material lacking sufficient morphological characters is time-consuming, season-dependent and might lead to incorrect assessments of commercialised species diversity. AIM OF THE STUDY: In this study, we identified cases of vernacular heterogeneity of medicinal plants using a tiered approach of literature research, morphology and DNA barcoding. MATERIAL AND METHODS: A total of 870 single ingredient medicinal plant samples corresponding to 452 local names were purchased from herbal markets in Dar-es-Salaam and Tanga, Tanzania, and identified using conventional methods as well as DNA barcoding using rbcL, matK and nrITS. RESULTS: Using conventional methods, we could identify 70% of samples to at least family level, while 62% yielded a DNA barcode for at least one of the three markers. Combining conventional methods and DNA barcoding, 76% of the samples could be identified to species level, revealing a diversity of at least 175 species in 65 plant families. Analysis of the market samples revealed 80 cases of multilingualism and over- and under-differentiation. Afzelia quanzensis Welw., Zanthoxylum spp., Allophylus spp. and Albizia anthelmintica Brongn. were the most evident cases of multilingualism and over-differentiation, as they were traded under 8-12 vernacular names in up to five local languages. The most obvious case of under-differentiation was mwingajini (Swahili), which matched to eight scientific species in five different plant families. CONCLUSIONS: Use of a tiered approach increases the identification success of medicinal plants sold in local market and corroborates findings that DNA barcoding can elucidate the identity of material that is unidentifiable based on morphology and literature as well as verify or disqualify these identifications. Results of this study can be used as a basis for quantitative market surveys of fragmented herbal medicine and to investigate conservation issues associated with this trade.


Assuntos
Código de Barras de DNA Taxonômico , Medicina Tradicional Africana , Plantas Medicinais/classificação , Plantas Medicinais/genética , Tanzânia
19.
Zhongguo Zhong Yao Za Zhi ; 44(18): 3960-3966, 2019 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-31872731

RESUMO

Buplewrum falcatum is a traditional Chinese medicine,which is mainly used for the treatment of cold and liver protection. B. falcatum is dominantly cultivated in Japan as well as planted in China,Korea and other countries and regions. In order to determine the appropriate sequencing strategy,the genome survey before large-scale genome sequencing is needed. This survey can provide information about the size and complexity of the whole genome of the target species. In the present study,the next generation sequencing technology( Illumina Hiseq 2000) was used to analyze the genome size and complexity of B. falcatum. In addition,SSR loci were analyzed from the sequenced data. Primer 3 was used to design specific primers and 33 pairs of primers were randomly selected for PCR with template DNA of B. falcatum,and the PCR system and optimal annealing temperature were screened. A total of 288. 64 G genome sequence data was obtained,and the estimated genome size of B. falcatum was 2 119. 58 Mb. The measured genome data depth was138×; the rate of heterozygosity was 1. 84%; and the ratio of repeat sequence was 83. 89%. It is speculated that the genome of B. falcatum is complex. The preliminary assembly was performed with K-mer = 41,and the contig N50 was 224 bp,the total length 896. 97 Mb,the scaffold N50 313 bp,and the total length was 922. 67 Mb. A total of 91 377 SSR sequences were detected in the sequenced genome data which were distributed in 70 809 unigenes.The main type is dinucleotide repeats,with 49 680 sequences,accounting for70. 16%. Among the 33 pairs of primers randomly synthesized according to the obtained SSR sequences,21 pairs were successfully amplifying the target sequences. The results will be helpful for later large scale genome sequencing and SSR molecular markers development for germplasm identification and trait mapping.


Assuntos
Bupleurum/genética , Genoma de Planta , Repetições de Microssatélites , Plantas Medicinais/genética , Polimorfismo Genético
20.
Sci Rep ; 9(1): 20171, 2019 12 27.
Artigo em Inglês | MEDLINE | ID: mdl-31882930

RESUMO

Anoectochilus roxburghii and Anoectochilus formasanus are the major species of genus Anoectochilus used in traditional Chinese medicine for their abundant content of flavonoids and some other medicinal constituents. In recent years, their wild resources are gradually exhausted due to over-collection and ecological deterioration. Artificial cultivation and tissue culture are employed to increase production. In this study, the open reading frame, promoter and genomic sequences of the chalcone synthase (CHS) gene were cloned from these two species according to their transcriptome information, and used for expression analysis in response to the induction of phenylalanine, ultraviolet light and NaCl, and its effect investigation on accumulation of flavonoids. The results showed that the expression of the CHS genes was upregulated in response to these inductions and resulted in increasing accumulation of total flavonoids. However, the increased flavonoids induced by phenylalanine and ultraviolet light were mainly allocated into the anthocyanidin branch of flavonoids biosynthesis. Not only did it improved the medicinal value, but might have inhibitory effect on plant growth because of the increased malondialdehyde accumulation. Under the induction of appropriate concentration of NaCl, the medicinal constituents of flavonoids were increased without inhibition to plant growth.


Assuntos
Aciltransferases/genética , Aciltransferases/metabolismo , Flavonoides/metabolismo , Regulação da Expressão Gênica de Plantas , Plantas Medicinais/genética , Plantas Medicinais/metabolismo , Aciltransferases/química , Sequência de Aminoácidos , Clonagem Molecular , Filogenia
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