Download PDF
Planta Med 2009; 75(15): 1618-1624
DOI: 10.1055/s-0029-1185808
Biochemistry, Molecular Biology and Biotechnology
Original Paper
© Georg Thieme Verlag KG Stuttgart · New York

Changes of Gentiopicroside Synthesis during Somatic Embryogenesis in Gentiana macrophylla

Li-Yu Chen1 [*] , Qian-Liang Chen1 [*] , Dan Xu1 , Jian-Guo Hao1 , Michael Schläppi2 , Zi-Qin Xu1
  • 1Key Laboratory of Resource Biology and Biotechnology in Western China (Ministry of Education), Institute of Life Science, Northwest University, Xi'an, Shaanxi, People's Republic of China
  • 2Department of Biological Sciences, Marquette University, Milwaukee, WI, USA
Further Information

Publication History

received February 19, 2009 revised April 13, 2009

accepted May 17, 2009

Publication Date:
22 June 2009 (online)

    Permissions and Reprints

Abstract

In vitro plant regeneration of Gentiana macrophylla Pall. and determination of gentiopicroside content during somatic embryogenesis are described in the present work. The highest percentage of embryogenic callus formation was observed in Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg/L 6-benzylaminopurine (BA). Calli were subcultured on MS medium containing 1.0 mg/L 2,4-D, 1.0 mg/L BA and 500 mg/L lactalbumin hydrolysate (LH) at intervals of 25 days. A higher frequency of somatic embryo maturation was achieved on MS medium containing B5 vitamins (MB) supplemented with different concentrations of 1-naphthaleneacetic acid (NAA) and BA than with a combination of NAA and kinetin (KT). Addition of AgNO3 improved maturation of somatic embryos while thidiazuron (TDZ) promoted vitrification. The gentiopicroside contents of embryogenic calli and globular-, heart-, torpedo-, and cotyledon-shaped embryoids were determined by high-performance liquid chromatography (HPLC). Gentiopicroside was not detectable in embryogenic calli, but in all types of somatic embryos. The highest gentiopicroside content was observed in cotyledon-shaped embryoids, reaching more than 12 mg/g dry weight.

Key words

Gentianaceae - Gentiana macrophylla - gentiopicroside - HPLC - secondary metabolites - somatic embryo

References

  • 1 Chen L, Liu J C, Zhang X N, Guo Y Y, Xu Z H, Cao W, Sun X L, Sun W J, Zhao M G. Down-regulation of NR2B receptors partially contributes to analgesic effects of gentiopicroside in persistent inflammatory pain.  Neuropharmacology. 2008;  54 1175-1181
    CrossrefPubMedGoogle Scholar
  • 2 Kondo Y, Takano F, Hojo H. Suppression of chemically and immunologically induced hepatic injuries by gentiopicroside in mice.  Planta Med. 1994;  60 414-416
    Article in Thieme ConnectPubMedGoogle Scholar
  • 3 Tan R X, Wolfender J L, Zhang L X, Ma W G, Fuzzati N, Marston A, Hostettmann K. Acyl secoiridoids and antifungal constituents from Gentiana macrophylla.  Phytochemistry. 1996;  42 1305-1313
    CrossrefPubMedGoogle Scholar
  • 4 Piatczak E, Krolicka A, Wysokinska H. Genetic transformation of Centaurium erythraea Rafn by Agrobacterium rhizogenes and the production of secoiridoids.  Plant Cell Rep. 2006;  25 1308-1315
    CrossrefPubMedGoogle Scholar
  • 5 Tiwari R K, Trivedi M, Guang Z C, Guo G-Q, Zheng G-C. Genetic transformation of Gentiana macrophylla with Agrobacterium rhizogenes: growth and production of secoiridoid glucoside gentiopicroside in transformed hairy root cultures.  Plant Cell Rep. 2007;  26 199-210
    CrossrefPubMedGoogle Scholar
  • 6 Mikula A, Rybczynski J J. Somatic embryogenesis of Gentiana genus I. The effect of the preculture treatment and primary explant origin on somatic embryogenesis of Gentiana cruciata (L.), G. pannonica (Scop.), and G. tibetica (King).  Acta Physiol Plant. 2001;  21 15-25
    PubMedGoogle Scholar
  • 7 Mikula A, Skierski J, Rybczynski J J. Somatic embryogenesis of Gentiana genus. III. Characterization of three-year-old embryogenic suspensions of G. pannonica originated from various seedling explants.  Acta Physiol Plant. 2002;  24 311-322
    CrossrefPubMedGoogle Scholar
  • 8 Sharma N, Chandel K PS, Paul A. In vitro propagation of Gentiana kurroo – an indigenous threatened plant of medicinal importance.  Plant Cell Tissue Org. 1993;  34 307-309
    CrossrefPubMedGoogle Scholar
  • 9 Semeniuk P, Griesbach R J. In vitro propagation of prairie gentian.  Plant Cell Tissue Org. 1987;  8 249-253
    PubMedGoogle Scholar
  • 10 Meng Y, Gao Y, Jia J. Plant regeneration from protoplasts isolated from callus of Gentiana crassicaulis.  Plant Cell Rep. 1996;  16 88-91
    CrossrefPubMedGoogle Scholar
  • 11 Chueh F, Chen C, Sagare A P, Tsay H. Quantitative determination of secoiridoid glucosides in in vitro propagated plants of Gentiana davidii var. formosana by high performance liquid chromatography.  Planta Med. 2001;  67 70-73
    Article in Thieme ConnectPubMedGoogle Scholar
  • 12 Sabovljevic A, Rosic N, Jankovic T, Grubisic D. Secoiridoid content of Blackstonia perfoliata in vivo and in vitro.  In Vitro Cell Dev Biol Plant. 2006;  42 427-431
    PubMedGoogle Scholar
  • 13 Chen L Y, Xu Z Q. Somatic embryogenesis pathway for plant regeneration in Qinjiao (Gentiana macrophylla Pall.).  J Mol Cell Biol. 2007;  40 267-271
    PubMedGoogle Scholar
  • 14 Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures.  Physiol Plant. 1962;  15 473-497
    CrossrefPubMedGoogle Scholar
  • 15 Gamborg O L, Millar R A, Ojiama K K. Nutrient requirements of suspension cultures of soybean root cells.  Exp Cell Res. 1968;  50 148-151
    PubMedGoogle Scholar
  • 16 Marchant R, Davey M R, Lucas J A, Power J B. Somatic embryogenesis and plant regeneration in Floribunda rose (Rosa hybrida L.) cvs. Trumpeter and Glad Tidings.  Plant Sci. 1996;  120 95-105
    CrossrefPubMedGoogle Scholar
  • 17 Choi Y E, Katsumi M, Sano H. Triiodobenzoic acid, an auxin polar transport inhibitor, suppresses somatic embryo formation and postembryogenic shoot/root development in Eleutherococcus senticosus.  Plant Sci. 2001;  160 1183-1190
    CrossrefPubMedGoogle Scholar
  • 18 Nhut D T, Hanh N TM, Tuan P Q, Nguyet L TM, Tram N TH, Chinh N C, Nguyen N H, Vinh D N. Liquid culture as a positive condition to induce and enhance quality and quantity of somatic embryogenesis of Lilium longiflorum.  Sci Hortic. 2006;  110 93-97
    CrossrefPubMedGoogle Scholar
  • 19 Yu F R, Yu F H, Li R, Wang R. Inhibitory effects of the Gentiana macrophylla (Gentianaceae) extract on rheumatoid arthritis of rats.  J Ethnopharmacol. 2004;  95 77-81
    CrossrefPubMedGoogle Scholar
  • 20 Coscia C J, Guarnaccia R, Botta L. Monoterpene biosynthesis. I. Occurrence and mevalonoid origin of gentiopicroside and loganic acid in Swertia caroliniensis.  Biochemistry. 1969;  8 5036-5043
    CrossrefPubMedGoogle Scholar
  • 21 Croteau R, Kutchan T M, Lewis N G. Natural products (secondary metabolites). Buchanan B, Gruissem W, Jones R Biochemistry and molecular biology of plants. Maryland; American Society of Plant Physiologists 2000: 1250-1318
    Google Scholar
  • 22 Lin P C, Ye Z H, Lu Y C, Zhao S Q, Hu F Z, Ji L J. Content determination of loganic acid and gentiopicroside in Tibetan herbal medicines Gentiana macrophylla and G. straminea by HPLC.  J Chin Med Mater. 2004;  27 819-821
    PubMedGoogle Scholar
  • 23 Kumarasamy Y, Nahar L, Sarker S D. Bioactivity of gentiopicroside from the aerial parts of Centaurium erythraea. .  Fitoterapia. 2003;  74 151-154
    CrossrefPubMedGoogle Scholar
  • 24 Wang D, Xu M, Zhu H T, Chen K K, Zhang Y J, Yang C R. Biotransformation of gentiopicroside by asexual mycelia of Cordyceps sinensis.  Bioorg Med Chem Lett. 2007;  17 3195-3197
    CrossrefPubMedGoogle Scholar
  • 25 Hikage T, Saitoh Y, Tanaka-Saito C, Hagami H, Satou F, Shimota Y, Nakano Y, Takahashi M, Takahata Y, Tsutsumi K. Structure and allele-specifc expression variation of novel α/β hydrolase fold proteins in gentian plants.  Mol Genet Genomics. 2007;  278 95-104
    CrossrefPubMedGoogle Scholar
  • 26 Shimazu T, Kurata K. Dynamic dissolved oxygen concentration control for enhancing the formation rate of torpedo-stage embryos in carrot somatic embryo culture.  J Biosci Bioeng. 2003;  95 384-390
    PubMedGoogle Scholar
  • 27 Linossier L, Veisseire P, Cailloux F, Coudret A. Effects of abscisic acid and high concentrations of PEG on Hevea brasiliensis somatic embryos development.  Plant Sci. 1997;  124 183-191
    CrossrefPubMedGoogle Scholar
  • 28 Shigeta J, Satoa K, Mii M. Effects of initial cell density, pH and dissolved oxygen on bioreactor production of carrot somatic embryos.  Plant Sci. 1996;  115 109-114
    CrossrefPubMedGoogle Scholar
  • 29 Faure O, Aarrouf J, Nougarède A. Ontogenesis, differentiation and precocious germination in anther-derived somatic embryos of grapevine (Vitis vinifera L.): embryonic organogenesis.  Ann Bot. 1996;  78 29-37
    CrossrefPubMedGoogle Scholar
  • 30 Liu H, Wang Y. Embryological studies in Gentiana macrophylla.  Acta Bot Boreal Occident Sin. 1994;  14 243-248
    PubMedGoogle Scholar
  • 31 West M AL, Harada J J. Embryogenesis in higher plants: an overview.  Plant Cell. 1993;  5 1361-1369
    PubMedGoogle Scholar
  • 32 Zimmerman J L. Somatic embryogenesis: a model for early development in higher plants.  Plant Cell. 1993;  5 1411-1423
    CrossrefPubMedGoogle Scholar
  • 33 Sharma P, Pandey S, Bhattacharya A, Nagar P K, Ahuja P S. ABA associated biochemical changes during somatic embryo development in Camellia sinensis (L.) O. Kuntze.  J Plant Physiol. 2004;  161 1269-1276
    CrossrefPubMedGoogle Scholar
  • 34 McGarvey D J, Croteau R. Terpenoid metabolism.  Plant Cell. 1995;  7 1015-1026
    CrossrefPubMedGoogle Scholar
  • 35 Caliş I, Rüegger H, Chun Z, Sticher O. Secoiridoid glucosides isolated from Gentiana gelida.  Planta Med. 1990;  56 406-409
    Article in Thieme ConnectPubMedGoogle Scholar
  • 36 Wang C, Xing X F, Ma L X, Guan Y, Dai S J, Liu M Y. Determination of gentiopicroside in different morphological types of cultivated Gentiana manshurica population.  Zhongguo Zhong Yao Za Zhi. 2004;  29 841-844
    PubMedGoogle Scholar
  • 37 Zhao Y, Zhang R, Sun W J. Enrichment process of gentiopicroside from the leaves of Gentiana macropylla with macroporous resin.  Zhong Yao Cai. 2007;  30 1583-1586
    PubMedGoogle Scholar
  • 38 Chen Y, Qiu D Y, Guo F X, Wang E J, Liu F Z. Investigation on explovitage of Gentiana straminea.  Zhong Yao Cai. 2007;  30 1214-1216
    PubMedGoogle Scholar
  • 39 Bianco A, Ramunno A, Melchioni C. Iridoids from seeds of Gentiana lutea.  Nat Prod Res. 2003;  17 221-224
    CrossrefPubMedGoogle Scholar
  • 40 Chueh F S, Chen C C, Sagare A P, Tsay H S. Quantitative determination of secoiridoid glucosides in in vitro propagated plants of Gentiana davidii var. formosana by high performance liquid chromatography.  Planta Med. 2001;  67 70-73
    Article in Thieme ConnectPubMedGoogle Scholar
  • 41 Menković N, Savikin-Fodulović K, Momcilović I, Grubisić D. Quantitative determination of secoiridoid and gamma-pyrone compounds in Gentiana lutea cultured in vitro.  Planta Med. 2000;  66 96-98
    PubMedGoogle Scholar

1 These authors made equal contributions to this work.

Zi-Qin Xu

Institute of Life Science
Northwest University

Taibaibeilu 229

Xi'an

710069 Shaanxi

People's Republic of China

Phone: + 86 29 88 30 34 84

Email: ziqinxu@nwu.edu.cn

      >