Planta Med 2020; 86(16): 1176-1184
DOI: 10.1055/a-1194-0471
Natural Product Chemistry and Analytical Studies
Original Papers

Molecular Identification Based on Chloroplast Sequences and Anti-complementary Activity Comparison of Juniperus Samples from the Qinghai-Tibet Plateau

Yi-Ting Yang
1   School of Pharmacy, Institutes of Integrative Medicine, Fudan University, Shanghai, Peopleʼs Republic of China
,
Ji De
2   College of Science, Tibet University, Lhasa, Peopleʼs Republic of China
,
Ze-Long Fu
1   School of Pharmacy, Institutes of Integrative Medicine, Fudan University, Shanghai, Peopleʼs Republic of China
,
Xie-Li Wang
1   School of Pharmacy, Institutes of Integrative Medicine, Fudan University, Shanghai, Peopleʼs Republic of China
,
Dao-Feng Chen
1   School of Pharmacy, Institutes of Integrative Medicine, Fudan University, Shanghai, Peopleʼs Republic of China
,
Hui Xie
3   Human Phenome Institute, Fudan University, Shanghai, Peopleʼs Republic of China
,
Yan Lu
1   School of Pharmacy, Institutes of Integrative Medicine, Fudan University, Shanghai, Peopleʼs Republic of China
› Author Affiliations

Abstract

Juniperus (Cupressaceae, Pinales) plants are widely distributed in the Qinghai-Tibet Plateau of China. The leaves and twigs of at least 8 Juniperus species (J. pingii, J. pingii var. wilsonii, J. squamata, J. recurva var. coxii, J. saltuaria, J. indica, J. tibetica and J. convallium var. microsperma) have been used as the Tibetan medicine Xuba. At present, it is difficult to distinguish among the original species of Xuba based only on their similar morphological characteristics. However, in our previous studies, 4 Xuba samples from different Juniperus species exhibited significant differences in both anticomplementary activity in vitro and anti-inflammatory effects on acute lung injury in vivo. To identify the effective original species of Xuba reliably, in this study, we developed a sequencing-based DNA molecular technology to distinguish 14 populations of 8 Juniperus species collected from Tibet region, using trnS-G, trnD – T, and petN-psbM genomic regions to build phylogenetic trees. In addition, their anticomplementary activities were evaluated. The results showed that combined sequence of these 3 genomic regions could identify 8 Juniperus species clearly and clustered individuals of one species but from different locations, whichever phylogenetic tree was constructed. Moreover, the anticomplementary activities of the 8 species were clustered into 2 groups. Among them, J. saltuaria and J. recurva var. coxii, which formed an independent branch apart from the other 6 species in phylogenetic trees, were the most potent (CH50: 0.029 – 0.032 mg/mL). Consequently, DNA identification of Juniperus using the combined sequence could provide beneficial guidance for further efficacy evaluation and quality control of Xuba.

Supporting Information



Publication History

Received: 28 December 2019

Accepted after revision: 31 May 2020

Article published online:
09 July 2020

© 2020. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 
  • References

  • 1 Al-Shehbaz IA, Bartholomew B, Boufford DE, Brach AR. Flora of China 4. Beijing: Science Press (Beijing), St. Louis: Missouri Botanical Garden Press (St. Louis); 1999: 69-77
  • 2 Luo DS. Xin Xiu Jingzhubencao. Chengdu: Sichuan Science and Technology Publishing House; 2004: 173-178
  • 3 Emami SA, Asgary S, Naderi GA, Ardekani MRS, Aslani S, Airin A, Kasher T, Sahebkar A. Investigation of antioxidant and anti-glycation properties of essential oils from fruits and branchlets of Juniperus oblonga . Rev Bras Farmacogn 2012; 22: 985-993
  • 4 Ennajar M, Bouajila J, Lebrihi A, Mathieu F, Abderraba M, Raies A, Romdhane M. Chemical composition and antimicrobial and antioxidant activities of essential oils and various extracts of Juniperus phoenicea L. (Cupressacees). J Food Sci 2009; 74: M364-M371
  • 5 Cabral C, Francisco V, Cavaleiro C, Goncalves MJ, Cruz MT, Sales F, Batista MT, Salgueiro L. Essential oil of Juniperus communis subsp. alpina (Suter) Celak needles: chemical composition, antifungal activity and cytotoxicity. Phytother Res 2012; 26: 1352-1357
  • 6 Li BB, Dong MX, De J, Ye L, Chen DF, Lu Y. Structural characterization and anti-proliferation activities against tumor cells of an Arabinogalactan from Juniperus convallium . Molecules 2019; 24: 1850
  • 7 Morgan BP, Harris CL. Complement therapeutics; history and current progress. Mol Immunol 2003; 40: 159-170
  • 8 Whaley K, Loos M, Weiler JM. Complement in Health and Disease. Dordrecht: Springer; 1987: 965-975
  • 9 Xia L, Deji. Zhu MX, Chen DF, Lu Y. Juniperus pingii var. wilsonii acidic polysaccharide: Extraction, characterization and anticomplement activity. Carbohydr Polym 2020; 231: 115728
  • 10 Fu ZL, Xia L, De J, Zhu MX, Li H, Lu Y, Chen DF. Beneficial effects on H1N1-induced acute lung injury and structure characterization of anti-complementary acidic polysaccharides from Juniperus pingii var. wilsonii . Int J Biol Macromol 2019; 129: 246-253
  • 11 Carrero-Sarmiento D, Hoyos-Lopez R. Molecular identification and genetic diversity of Lutzomyia gomezi (Diptera: Psychodidae) using DNA-barcodes in Cordoba, Colombia. Trop Biomed 2018; 35: 100-110
  • 12 Donga TK, Meadow R. Determination of genetic diversity in Chilo partellus, Busseola fusca, and Spodoptera frugiperda infesting sugarcane in southern Malawi using DNA barcodes. Insects 2018; 9: 74
  • 13 Mao K, Hao G, Liu J, Adams RP, Milne RI. Diversification and biogeography of Juniperus (Cupressaceae): variable diversification rates and multiple intercontinental dispersals. New Phytol 2010; 188: 254-272
  • 14 Adams RP, Schwarzbach AE. Taxonomy of the turbinate shaped seed cone taxa of Juniperus, section Sabina: sequence analysis of nrdna and four cpdna regions. Phytologia 2012; 94: 388-403
  • 15 Li ZH, Fang MF, Yue M, Liu WZ, Liu ZL, Zhao P, Zhao GF. Method for identification of Juniperus convallium var. microsperma, Juniperus saltuaria and Juniperus tibetica. CN 104263846 A 20150107; 2015.
  • 16 Chen SL. DNA Barcode Molecular Identification of TCM. Beijing: Peopleʼs Medical Publishing House; 2012: 1-5
  • 17 Fazekas AJ, Burgess KS, Kesanakurti PR, Graham SW, Newmaster SG, Husband BC, Percy DM, Hajibabaei M, Barrett SCH. Multiple multilocus DNA barcodes from the plastid genome discriminate plant species equally well. PLoS 2008; 3: e2802
  • 18 Doyle JJ, Doyle DJ. Isolation of plant DNA from fresh tissue. Focus 1990; 12: 13-15
  • 19 Xu DH, Abe J, Sakai M, Kanazawa A, Shimamoto Y. Sequence variation of non-coding regions of chloroplast DNA of soybean and related wild species and its implications for the evolution of different chloroplast haplotypes. Theor Appl Genet 2000; 101: 724-732
  • 20 Adams RP, Bartel JA, Price RA. A new genus, Hesperocyparis, for the cypresses of the Western Hemisphere (Cupressaceae). Phytologia 2009; 91: 160-185
  • 21 Kress WJ, Wurdack KJ, Zimmer EA, Weigt LA, Janzen DH. Use of DNA barcodes to identify flowering plants. Proc Natl Acad Sci U S A 2005; 102: 8369-8374
  • 22 Taberlet P, Gielly L, Pautou G, Bouvet J. Universal primers for amplification of 3 noncoding regions of chloroplast DNA. Plant Mol Biol 1991; 17: 1105-1109
  • 23 Little DP, Schwarzbach AE, Adams RP, Hsieh CF. The circumscription and phylogenetic relationships of Callitropsis and the newly described genus Xanthocyparis (Cupressaceae). Am J Bot 2004; 91: 1872-1881
  • 24 Di HY, Zhang YY, Chen DF. An anti-complementary polysaccharide from the roots of Bupleurum chinense . Int J Biol Macromol 2013; 58: 179-185
  • 25 Mayer MM. Complement and complement fixation. In: Experimental Immunochemistry. Second edition. Kalamazoo: Springfield Publications; 1961: 133-240
  • 26 Lu Y, Zhang JJ, Huo JY, Chen DF. Structural characterization and anti-complementary activities of two polysaccharides from Houttuynia cordata . Planta Med 2019; 85: 1098-1106
  • 27 Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 1997; 25: 4876-4882
  • 28 Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol 2018; 35: 1547-1549