1.DNA barcoding identification of original plants of a rare medicinal material Resina Draconis and related Dracaena species.
Yue ZHANG ; Mei-Fang SONG ; Hai-Tao LI ; Hui-Fang SUN ; Zhong-Lian ZHANG
China Journal of Chinese Materia Medica 2021;46(9):2173-2181
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.
China
;
DNA Barcoding, Taxonomic
;
DNA, Plant/genetics*
;
Dracaena/genetics*
;
Plants
;
Resins, Plant
;
Sequence Analysis, DNA
2.Identification of Dendrobii Caulis basing on ITS sequence.
Zi YE ; Ye LU ; Zheng-Tao WANG ; Hong XU ; Zhi-Bi HU
China Journal of Chinese Materia Medica 2014;39(20):3928-3935
Isolation of high-quality genomic DNA from dried and processed crude drug is the key for the DNA identification of Dendrobii Caulis. The DNA extract of Dendrobii Caulis was firstly compared using different method to isolate genomic DNA from dried and processed crude drug, including commercial DNA extracted kit and CTAB method. Using modified CTAB method (extracted from large samples), the genomic DNA was successfully isolated from Dendrobii Caulis, including Huangcao and Fengdou. The ITS regions were amplified using the purified DNA as template, and then cloned and sequenced. These ITS sequences were compared with data from Genbank database and our lab, 14 Dendrobium species and five similar species were identified from "Huangcao" and "Huangcao" slice, while six species and three similar species from "Fengdou" according to their sequence similarity. The study demonstrated that the dried Dendrobii Caulis could be identified using DNA molecular method, which could overcome deficiencies and limitations of traditional identification method and has a certain application prospects.
DNA, Intergenic
;
genetics
;
DNA, Plant
;
genetics
;
Dendrobium
;
classification
;
genetics
;
Sequence Analysis, DNA
3.Identification of 23 unknown Li minority medicinal plants based on DNA barcoding.
Xin-Yun CUI ; Wei SUN ; Chao XIONG ; Xiang-Xiao MENG ; Yu-Hua SHI ; Lan WU ; Li-Li CHENG ; Wei-Jie LI ; Xi-Long ZHENG
China Journal of Chinese Materia Medica 2019;44(2):283-292
DNA barcode molecular biological technique is used to identify the species of 23 unknown Li minority medicinal plants.DNA was extracted from 23 unknown medicines using the Plant Genomic DNA Extraction kit. The ITS2 and psbA-trnH regions were amplified and sequenced bi-directionally. The Codon Code Aligner V 7. 0. 1 was used to proofread and assemble the contigs and generated consensus sequences. All the sequences were submitted to Traditional Chinese Medicine DNA Barcode Database and NCBI Gen Bank to get information of the species identifications. If the maximum similarity of the identification result is ≥ 97%,exact species can be known. If it is between 97% and 90%,samples' genus can be confirmed; If it is <90%,then we can only confirm its family. Finally there are 17 samples can be identified to species level,5 can be identified to genus level and 1 can be identified to family level. This shows that DNA barcoding used in medicinal plants molecular identification,can identify unknown species rapidly and accurately.
DNA Barcoding, Taxonomic
;
DNA, Plant
;
genetics
;
Medicine, Chinese Traditional
;
Plants, Medicinal
;
classification
;
Sequence Analysis, DNA
4.Azolla--a model organism for plant genomic studies.
Genomics, Proteomics & Bioinformatics 2003;1(1):15-25
The aquatic ferns of the genus Azolla are nitrogen-fixing plants that have great potentials in agricultural production and environmental conservation. Azolla in many aspects is qualified to serve as a model organism for genomic studies because of its importance in agriculture, its unique position in plant evolution, its symbiotic relationship with the N2-fixing cyanobacterium, Anabaena azollae, and its moderate-sized genome. The goals of this genome project are not only to understand the biology of the Azolla genome to promote its applications in biological research and agriculture practice but also to gain critical insights about evolution of plant genomes. Together with the strategic and technical improvement as well as cost reduction of DNA sequencing, the deciphering of their genetic code is imminent.
Cyanobacteria
;
genetics
;
Ferns
;
Genes, Plant
;
Genome, Plant
;
Genomics
;
methods
;
Nitrogen
;
metabolism
;
Plants
;
genetics
;
Sequence Analysis, DNA
5.De novo sequencing and analysis of root transcriptome to reveal regulation of gene expression by moderate drought stress in Glycyrrhiza uralensis.
Chun-rong ZHANG ; Xue-yu SANG ; Meng QU ; Xiao-min TANG ; Xuan-xuan CHENG ; Li-ming PAN ; Quan YANG
China Journal of Chinese Materia Medica 2015;40(24):4817-4823
Moderate drought stress has been found to promote the accumulation of active ingredients in Glycyrrhiza uralensis root and hence improve the medicinal quality. In this study, the transcriptomes of 6-month-old moderate drought stressed and control G. uralensis root (the relative water content in soil was 40%-45% and 70%-75%, respectively) were sequenced using Illumina HiSeq 2000. A total of 80,490 490 and 82 588 278 clean reads, 94,828 and 305,100 unigenes with N50 sequence of 1,007 and 1,125 nt were obtained in drought treated and control transcriptome, respectively. Differentially expressed genes analysis revealed that the genes of some cell wall enzymes such as β-xylosidase, legumain and GDP-L-fucose synthase were down-regulated indicating that moderate drought stress might inhibit the primary cell wall degradation and programmed cell death in root cells. The genes of some key enzymes involved in terpenoid and flavonoid biosynthesis were up-regulated by moderate drought stress might be the reason for the enhancement for the active ingredients accumulation in G. uralensis root. The promotion of the biosynthesis and signal transduction of auxin, ethylene and cytokinins by moderate drought stress might enhance the root formation and cell proliferation. The promotion of the biosynthesis and signal transduction of abscisic acid and jasmonic acid by moderate drought stress might enhance the drought stress tolerance in G. uralensis. The inhibition of the biosynthesis and signal transduction of gibberellin and brassinolide by moderate drought stress might retard the shoot growth in G. uralensis.
Droughts
;
Gene Expression Regulation, Plant
;
Glycyrrhiza uralensis
;
genetics
;
Plant Roots
;
Sequence Analysis, DNA
;
Stress, Physiological
;
Transcriptome
6.Identification of maca (Lepidium meyenii Walp.) and its adulterants by a DNA-barcoding approach based on the ITS sequence.
Jin-Jin CHEN ; Qing-Sheng ZHAO ; Yi-Lan LIU ; Sheng-Hua ZHA ; Bing ZHAO
Chinese Journal of Natural Medicines (English Ed.) 2015;13(9):653-659
Maca (Lepidium meyenii) is an herbaceous plant that grows in high plateaus and has been used as both food and folk medicine for centuries because of its benefits to human health. In the present study, ITS (internal transcribed spacer) sequences of forty-three maca samples, collected from different regions or vendors, were amplified and analyzed. The ITS sequences of nineteen potential adulterants of maca were also collected and analyzed. The results indicated that the ITS sequence of maca was consistent in all samples and unique when compared with its adulterants. Therefore, this DNA-barcoding approach based on the ITS sequence can be used for the molecular identification of maca and its adulterants.
DNA Barcoding, Taxonomic
;
methods
;
DNA, Intergenic
;
analysis
;
DNA, Plant
;
analysis
;
Drug Contamination
;
prevention & control
;
Humans
;
Lepidium
;
genetics
;
Phytotherapy
7.Study on sequence characterized amplified region (SCAR) markers of Polygonum capitatum.
Tao ZHOU ; Yu XIE ; Li-Yan ZHANG ; Sheng-Hua WEI ; Yan-Lei JIN
China Journal of Chinese Materia Medica 2013;38(16):2577-2580
OBJECTIVETo establish sequence characterized amplified region markers of Polygonum capitatum.
METHODThe random primer was screened through RAPD to obtain the specific RAPD marker band, and the band was separated, extracted, cloned and sequenced. The specific primers were designed for conventional PCR reaction on the basis of the specific band, and the SCAR marker was acquired.
RESULTScreening from 50 RAPD primer, only C29 primer had 2 specific bands could distinguish P. capitatum from P. nepalense, then 4 pairs of specific primers were designed based on the 2 sequences of RAPD marker bands, and only 1 pair primer (Z1-2) was successfully converted into SCAR marker after repeated tests.
CONCLUSIONThe Z1-2 primer, could be used as an effective SCAR mark to identify Z300 DNA for P. capitatum. The SCAR mark was established and can be used as a molecular marker to distinguish P. capitatum from P. nepalense
DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; Genetic Markers ; genetics ; Polygonum ; classification ; genetics ; Random Amplified Polymorphic DNA Technique ; Sequence Analysis, DNA
8.Cloning of distinguishing DNA sequences of Gastrodia elata Blume and application of them in identifying gastrodia tuber.
Jun TAO ; Tie-Xiang FU ; Zhi-Yong LUO ; Li WEN ; Zhi-Cheng WANG ; Xiao-Shun SHU ; Shui-Ping LIU ; Yao TAO ; Wei-Xin HU
Chinese Journal of Biotechnology 2006;22(4):587-591
Gastrodia elata Bl. is a famous and costful traditional Chinese medicine. Their genomic DNA fingerprints were investigated using a modified Randomly Amplified Polymorphic DNA method. DNA fragments common to all or to fine populations were identified and recovered. Five DNA fragments were proven not to be reported through DNA cloning, PCR identifying, nucleotide sequencing and bioinformatics analyses and were received in and recorded by NCBI GenBank. Gastrodine contents of the Gastrodia tuber samples were determined using high performance liquid chromatography technique. The distribution of the five DNA fragments in 9 Gastrodia elata Blue populations and the correlation with gastromedicine content were studied. The results show the distribution of these DNA sequences varied greatly among the populations whereby DNA Sequence 1 was the common and distinguishing molecular marker for all the populations studied and DNA Sequence 2 may relate to higher gastrodine content. In conclusion, these DNA marker sequences can be employed to identify genuine gastrodia tubers, better varieties and optimize their selection and cultivating.
Base Sequence
;
Benzyl Alcohols
;
analysis
;
Cloning, Molecular
;
Computational Biology
;
DNA, Plant
;
chemistry
;
Gastrodia
;
genetics
;
Glucosides
;
analysis
;
Plant Tubers
;
genetics
9.Between Hengshanhuangqi and Chuanhuangqi based on metabolomics and ITS2 sequences.
Pan HE ; Zhen-Yu LI ; Sheng-Ci FAN ; Fu-Sheng ZHANG ; Xue-Mei QIN ; Guo-Jun DU
Acta Pharmaceutica Sinica 2013;48(10):1595-1601
To compare the differences between Hengshanhuangqi (HH) and Chuanhuangqi (CH) at molecular level, 1H NMR based plant metabolomics approach was used to reveal the chemical difference between HH and CH. Then, the contents of astragaloside IV and calycosin-7-O-beta-D-glucoside, the marker compounds specified in China Pharmacopoeia, were determined. In addition, the ITS2 fragments of HH and CH were sequenced. Twenty-three metabolites were identified in the 1H NMR spectrum, and the principal component analysis showed CH and HH could be separated clearly. HH contained more aspartic acid, GABA, citric acid, astragaloside IV and calycosin-7-O-beta-D-glucoside, while CH contained more threonine, alanine, acetic acid, choline, arginine, fructose and sucrose. And the astragaloside IV is almost undetectable in CH. In addition, the ITS2 fragment sequences of HH and CH were different at eight bases. Thus, the HH and CH showed significant differences chemically and genetically.
Astragalus membranaceus
;
chemistry
;
classification
;
genetics
;
Base Sequence
;
DNA, Plant
;
genetics
;
DNA, Ribosomal Spacer
;
genetics
;
Glucosides
;
analysis
;
Isoflavones
;
analysis
;
Metabolomics
;
Plant Roots
;
chemistry
;
Plants, Medicinal
;
chemistry
;
classification
;
genetics
;
Principal Component Analysis
;
Saponins
;
analysis
;
Sequence Analysis, DNA
;
Species Specificity
;
Triterpenes
;
analysis
10.Analysis of intraspecific variation of Chinese Carthamus tinctorius L. using AFLP markers.
Lei ZHANG ; Bei-Bei HUANG ; Guo-Yin KAI ; Mei-Li GUO
Acta Pharmaceutica Sinica 2006;41(1):91-96
AIMTo investigate the intraspecific variation of Carthamus tinctorius L. (safflower) and establish foundation for further breeding of safflower germplasm resource and screening the quality correlation genes.
METHODSAmplified fragment length polymorphism (AFLP) was carried out to analyze genetic variation of 28 safflower populations collected in China. Unweighed pair-group method of with arithmetical averages (UPGMA) cluster analysis was used to construct a dendrogram and to estimate the genetic distances among the populations.
RESULTSAll populations could be uniquely distinguished using 12 selected primer combinations. Similarity coefficients ranged from 0.48 to 0.96 among the populations. Dendrogram revealed distinct segregation of all the cultivars into three main groups and one midst group.
CONCLUSIONLimited genetic diversity exists within the tested 28 collections at intra specific level and AFLP-based phyiogeny was not absolutely consistent with that based on morphological characters may be due to the interaction effect between genotype and environment.
Carthamus tinctorius ; genetics ; Cluster Analysis ; DNA, Plant ; analysis ; Genetic Variation ; Phylogeny ; Plants, Medicinal ; genetics ; Polymorphism, Genetic