1.Studies on genetic diversity of three Tibetan herbs.
Liang-hong NI ; Zhi-li ZHAO ; Jin-rong WU ; Bo XIONG ; Jia-ni LU ; Dorje GAAWE
China Journal of Chinese Materia Medica 2015;40(19):3883-3888
The genetic diversity of three Tibetan herbs, i. e., Sang-Di, E-Dewa and Ye-Xingba (Tibetan names), was studied based on the field collection, specimen identification and DNA sequence analysis. Swertia hispidicalyx, Gentiana lhassica and Scrophularia dentata, as the original plants of the three Tibetan herbs, were collected and identified. The regions of ITS, matK, rbcL, rpoC1, trnL(UAA), psbA-trnH, atpB-rbcL, trnS (GCU)-trnG(UCC), rpl20-rps12, trnL(UAA)-trnF(GAA) and nadl 2nd intron were amplified and sequenced. The ITS regions of S. hispidicalyx and S. dentata were cloned and sequenced, and the sequences were classified into different genotypes. All the sequences were analyzed and compared with those of closely related species. Our studies may provide reference for the genetic diversity analysis and molecular identification of the three Tibetan herbs.
Genetic Variation
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Gentiana
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classification
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genetics
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Phylogeny
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Plant Proteins
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genetics
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Plants, Medicinal
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classification
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genetics
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Scrophularia
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classification
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genetics
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Swertia
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classification
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genetics
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Tibet
2.DNA-based identification of Gentiana robusta and related species.
Bo XIONG ; Zhi-li ZHAO ; Liang-hong NI ; Dorje GAAWE ; Ma MI
China Journal of Chinese Materia Medica 2015;40(23):4680-4685
The alpine plant Gentiana robusta is an endemic species to the Sino-Himalayan subregion. Also, it is one of the original plants used as traditional Tibetan medicine Jie-Ji. We sequence the nuclear ribosomal internal transcribed spacer (ITS) regions, matK, rbcL, rpoC1, trnL (UAA), psbA-trnH, atpB-rbcL, trnS( GCU)-trnG(UCC), rpl20-rps12, trnL(UAA)-trnF( GAA) fragments of cp DNA in both G. robusta and such relative species as G. straminea, G. crassicaulis and G. waltonii. With Halenia elliptica as the outgroup, molecular systematic analysis reveals that G. robusta is a natural hybrid. G. straminea is the mother of hybrids, but the father is not very clear. In addition, the molecular markers for distinguishing G. robusta from the parental species or closely related species are identified, respectively. Our studies may provide valuable reference for the species identifications of medicinal plants with complex genetic backgrounds.
DNA Barcoding, Taxonomic
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DNA, Plant
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genetics
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Gentiana
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classification
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genetics
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Molecular Sequence Data
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Phylogeny
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Plant Proteins
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genetics
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Plants, Medicinal
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classification
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genetics
3.Molecular Pharmacognosy on Tibetan Traditional Medicine Dong-Na Duan-Chi
Lianghong NI ; Tsering TASHI ; Zhili ZHAO ; Dorje GAAWE ; Qianwan MENG ; Thundrop DAWA
Chinese Journal of Information on Traditional Chinese Medicine 2014;(3):65-67,68
Objective To identify the common Tibetan herb Dong-Na Duan-Chi by molecular pharmacognosy. Methods Ethnopharmacology investigation was carried out and local herbalists were visited to observe which plants were being used as Dong-Na Duan-Chi. Then, nuclear ribosomal DNA ITS sequences were amplified and sequenced for molecular identification. Results The botanic origins of Dong-Na Duan-Chi in Tibet Autonomous Region were authenticated as two species, i.e. Veronica eriogyne H.Winkl. and Veronica ciliata Fisch.. The sequences of ITS for Veronica eriogyne and Veronica ciliata are 551 bp and 552 bp in size. The results showed that nrDNA ITS sequences could be used to discriminate V.eriogyne and V.ciliata from closely related species. Conclusion nrDNA ITS sequences can be used as the molecular markers to identify the Tibetan herb Dong-Na Duan-Chi from other species of the same genus Veronica L.
4.The identification of Sect. Cruciata (Gentiana ) species using mtDNA nad 1/b-c and nad 5/d-e fragments
Jia-ni LU ; Zhi-li ZHAO ; Liang-hong NI ; Dorje GAAWE ; Ma MI
Acta Pharmaceutica Sinica 2019;54(1):166-172
italic>Gentiana section Cruciata (Gentianaceae) is a medicinally important section of herbs, including Chinese traditional medicine Gentianae Macrophyllae Radix and Tibetan herb Jieji. Here, we assess the taxonomic significance using mtDNA
5.A strategy for identifying six species of Sect. Cruciata (Gentiana) in Gansu using DNA barcode sequences
Liang-hong NI ; Zhi-li ZHAO ; Bo XIONG ; Dorje GAAWE ; Ma MI
Acta Pharmaceutica Sinica 2016;51(5):821-
Located in the transition zone between the Qinghai-Tibet Plateau and the Loess Plateau, Gansu province is one of the distribution centers of Sect. Cruciata, Gentiana (Gentianaceae) in China. Six species in the section, G. crassicaulis, G. straminea, G. siphonantha, G. officinalis, G. dahurica and G. macrophylla, are native to Gansu. In this paper, samples of 6 species and Halenia elliptica (outgroup) were collected. Nuclear DNA ITS, chloroplast DNA matK, rbcL, rpoC1, trnL (UAA) intron, psbA-trnH, atpB-rbcL, trnS (GCU)-trnG (UCC), rpl20-rps12 and trnL (UAA)-trnF (GAA) were sequenced from these samples. Based on the sequence analyses, high intragenomic polymorphisms were detected in ITS regions of G. crassicaulis, G. straminea, G. siphonantha, G. officinalis and G. dahurica, and they showed incomplete concerted evolution. A methodological study to identifying such close-related species as G. macrophylla, G. officinalis and G. dahurica was carried out based on the special genotypes. The results showed that 7 cp DNA sequence fragments could be used to identify G. crassicaulis, G. straminea and G. siphonantha. With nr ITS genotype II, III and IV of G. dahurica, the species can be distinguished from the close-related G. officinalis using 12 cloned sequences in a sample (with statistical significance). The cp DNA sequences of G. macrophylla were classified into two genotypes, and with genotype II, the species can be distinguished from the close-related G. officinalis and G. dahurica using 6 test samples each (with statistical significance). Furthermore, DNA barcode sequences were determined for all 6 species in Gansu. Also, the studies provide some basic data for analyses of genetic diversity and identification of Gentiana species.
6.Analysis of transcriptomes to explore genes contributing to iridoid biosynthesis in Gentiana waltonii and Gentiana robusta (Gentianaceae)
Liang-hong NI ; Zhi-li ZHAO ; Jin-rong WU ; Dorje GAAWE ; Ma MI
Acta Pharmaceutica Sinica 2019;54(5):944-953
The roots and flowers of
7.Molecular markers based upon whole chloroplast genomes and identifying alpine Gentiana waltonii and G. lhassica (Gentianaceae)
Bo-ran DONG ; Zhi-li ZHAO ; Liang-hong NI ; Dorje GAAWE ; Tong-hua LIU
Acta Pharmaceutica Sinica 2021;56(9):2584-2591
As two original plants of Tibetan herb Jieji,
8.DNA fingerprinting and genetic diversity in a threatened alpine medicinal plant, Gentiana crassicaulis (Gentianaceae) using AFLP
Fen-fen ZONG ; Zhi-li ZHAO ; Liang-hong NI ; Dorje GAAWE ; Tong-hua LIU
Acta Pharmaceutica Sinica 2020;55(8):1941-1950
Jieji Nabao is a common Tibetan herb. According to our ethnobotanical studies, one of its original plants is identified as
9.SNP double peak methods through chloroplast genome profiling for identifying Gentiana crassicaulis and two closely related species
Na-na GAO ; Liang-hong NI ; Zhi-li ZHAO ; Dorje GAAWE
Acta Pharmaceutica Sinica 2022;57(8):2520-2527
italic>Gentiana crassicaulis Duthie ex Burk. is one of the plant sources of Gentianae Macrophyllae Radix (QinJiao).
10.Identification of medicinal plants used as Tibetan traditional medicine Chuan-Bu based on nrDNA markers.
Yao-Hua YANG ; Zhi-Li ZHAO ; Dorje GAAWE ; Ma MI ; Qian-Wan MENG ; Liang-Hong NI
China Journal of Chinese Materia Medica 2013;38(21):3773-3775
OBJECTIVETo identify the common Tibetan herb Chuan-Bu.
METHODLocal herbalists were visited to observe which plants were being used as Chuan-Bu. Samples of the indigenous plants were collected at the same time. Leaf materials were collected from field surveys. Total genomic DNA was extracted from silica gel-dried leaf samples. The PCR products were purified and directly sequenced.
RESULTAs the origin of Chuan-Bu in Tibet autonomous region was authenticated, two species were determined, i. e. Euphorbia stracheyiand E. wallichii. Also, based on our earlier research, the origin of Chuan-Bu in Gansu province, is from E. kansuensis. The sequences of ITS1 for E. stracheyi and E. wallichii were 261 bp in size, and 221 bp in ITS2, respectively. The size of the 5.8S coding region was 164 bp for all species examined in the genus. Especially, there was a heterozygous locus in ITS1 (C/G; position 72) for E. stracheyi. The nucleotide divergence between sequences of the 6 species in pairwise comparisons was calculated and the result showed that the variable site could be detected in each pairwise comparison of sequences. Also, there were 8 point mutations in the 5.8S coding region.
CONCLUSIONnrDNA ITS sequences can be used as the molecular markers to identify the Tibetan herb Chuan-Bu and such Traditional Chinese Medicines from the same genus Euphorbia as E. lathyris, E. humifusa and E. pekinensis.
DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Euphorbia ; classification ; genetics ; Genetic Markers ; Molecular Sequence Data ; Phylogeny ; Tibet