Skip to main content

Advertisement

SpringerLink
Log in

Enhanced thebaine and noscapine production and modulated gene expression of tyrosine/dopa decarboxylase and salutaridinol 7-O-acetyltransferase genes in induced autotetraploid seedlings of Papaver bracteatum Lindl

  • Original Article
  • Published:
Acta Physiologiae Plantarum Aims and scope Submit manuscript

Abstract

Papaver bracteatum, belonging to Papaveraceae, is considered to be a primary ingredient in the production of codeine and anti-drug drugs (e.g., naloxone and Naltrexone) without the involvement of morphine. For polyploidy induction, papaver seedlings were treated with various colchicine concentrations (0, 0.05, 0.1, 0.2 and 0.5% w/v) in three time durations (24, 48 and 72 h). The effects of polyploidy induction on morphological, physiological, alkaloid production and gene expression levels in the pathway of benzylisoquinoline alkaloids were analyzed. High rates of seedling survival were observed at lower colchicine concentrations (0.05 and 0.1% w/v) versus higher concentrations (0.2 and 0.5% w/v). The best polyploidy treatment to induce P. bracteatum shoot tips was observed in plants treated with 0.2% colchicine for 72 h. P. bracteatum-induced tetraploids showed higher stomata and an increase in leaf area (mm2), chlorophyll (a, b and total) and a decrease in stomata density, leaf length (cm) and petiole length. Flow cytometry and chromosome counting analysis revealed that chromosome number was 2n = 2x = 14 in diploid plants and 2n = 4x = 28 in tetraploid plants. Gene expression analysis through semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) showed that the transcription level of sat and tydc genes of the benzylisoquinoline biosynthesis pathway was enhanced in tetraploid plants. The tetraploid plants of P. bracteatum also showed 5.86 and 30.55 times higher thebaine and noscapine contents than the diploid plants, respectively. Regarding the higher commercial value of thebaine and noscapine, tetraploids can efficiently be used to improve the thebaine and noscapine production of P. bracteatum.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

References

  • Abdoli M, Moieni A, Badi HN (2013) Morphological, physiological, cytological and phytochemical studies in diploid and colchicine-induced tetraploid plants of Echinacea purpurea L. Acta Physiol Plant 35(7):1–9

    Google Scholar 

  • Adams KL, Wendel JF (2005) Novel patterns of gene expression in polyploidy plants. Trends Genet 10(21):539–543

    Google Scholar 

  • Adaniya S, Shirai D (2001) In vitro induction of tetraploid ginger (Zingiber officinale Roscoe) and its pollen fertility and germinability. Sci Hort. 88(4):277–287

    Google Scholar 

  • Ajalin I, Kobza F, Dolezel J (2002) Ploidy identification of doubled chromosome number plants in Viola × witrockiana Gams. M1- generation. Hortic Sci. 29(1):35–40

    Google Scholar 

  • Albuzio A, Spettoli P, Cacco G (2006) Changes in gene expression from diploid to autotetraploid status of Lycopersicon esculentum. Physiol Plant 44:77–80

    Google Scholar 

  • Allum JF, Bringloe DH, Roberts AV (2007) Chromosome doubling in a Rosa rugosa Thunb. Hybrid by exposure of in vitro nodes to oryzalin: the effects of node length, oryzalin concentration and exposure time. Plant Cell Rep 26:1977–1984

    CAS  PubMed  Google Scholar 

  • Arnon DI (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant physiol. 24:1–15

    CAS  PubMed  Google Scholar 

  • Berkov S (2001) Size and alkaloid content of seeds in induced autotetraploids of Datura innoxia, Datura stramonium and Hyoscyamus niger. Pharm Biol. 39(5):329–331

    CAS  Google Scholar 

  • Berkov S, Philipov S (2002) Alkaloid production in diploid and autotetraploid plants of Datura stramonium. Pharm Biol. 40(8):617–621

    CAS  Google Scholar 

  • Caruso I, Lepore L, De Tommasi N, Dal Piaz F, Frusciante L, Aversano R, Garramone R, Carputo D (2011) Secondary metabolite profile in induced tetraploids of wild Solanum commersonii Dun. Chem Biodivers 8(12):2226–2237

    CAS  PubMed  Google Scholar 

  • Cheng ZM, Korban S (2011) In vitro ploidy manipulation in the genomics era. Plant Cell, Tissue Organ Cult 104:281–282

    Google Scholar 

  • Coffman CB, Bare CE, Gentner WA (1975) Thebaine variations between germplasm sources within one collection of Papaver bracteatum. B narcotics. 27:41–46

    CAS  Google Scholar 

  • Dhooghe E, Van Laere K, Eeckhaut T, Leus V, Van Huylenbroeck J (2011) Mitotic chromosome doubling of plant tissues in vitro. Plant Cell Tiss Organ Cult 104:359–373

    Google Scholar 

  • Facchini PJ, Penzes-Yost C, Samanani N, Kowalchuk N (1998) Expression Patterns Conferred by Tyrosine/Dihydroxyphenylalanine Decarboxylase Promoters from Opium Poppy Are Conserved in Transgenic Tobacco. Plant Physiol 118(1):69–81

    CAS  PubMed  PubMed Central  Google Scholar 

  • Gao S, Zhu D, Cai Z, Xu D (1996) Autotetraploid plants from colchicine-treated bud culture of Salvia miltiorrhiza Bge. Plant Cell Tiss Organ Cult 47(1):73–77

    CAS  Google Scholar 

  • Gao SL, Chen BJ, Zhu DN (2002) In vitro production and identification of autotetraploids of Scutellaria baicalensis. Plant Cell Tiss Organ Cult 70:289–293

    CAS  Google Scholar 

  • Ghotbi Ravandi E, Rezanejad F, Zolala J, Dehghan E (2013) The effects of chromosome-doubling on selected morphological and phytochemical characteristics of Cichorium intybus L. J Hortic Sci Biotechnol. 88(8):701–709

    Google Scholar 

  • Griesbach RJ (1990) Colchicine-induced polyploidy in Eustoma grandiflorum. Sci Hort. 25:1284–1286

    CAS  Google Scholar 

  • Grothe T, Lenz R, Kutchan TM (2001) Molecular characterization of the salutaridinol 7-O-acetyltransferase involved in morphine biosynthesis in opium poppy Papaver somniferum. J Biol Chem 276(33):30717–30723

    CAS  PubMed  Google Scholar 

  • Guo M, Davis D, James A (1996) Birchler dosage effects on gene expression in a maize ploidy series. Genet Soc Am 142(4):1349–1355

    CAS  Google Scholar 

  • Hamill S, Smith M, Dodd W (1992) In vitro induction of banana autotetraploids by colchicine treatment of micropropagated diploids. Aust J Bot 40(6):887–896

    CAS  Google Scholar 

  • He M, Jiang L, Ren Z, Wang G, Wang J (2016) Noscapine targets EGFRp-Tyr1068 to suppress the proliferation and invasion of MG63 cells. Sci. Rep. https://doi.org/10.1038/srep37062

    Article  PubMed  PubMed Central  Google Scholar 

  • Javadian N, Karimzadeh G, Sharifi M, Moieni A, Behmanesh M (2017) In vitro polyploidy induction: changes in morphology, podophyllotoxin biosynthesis, and expression of the related genes in Linum album (Linaceae). Planta. https://doi.org/10.1007/s00425-017-2671-2

    Article  PubMed  Google Scholar 

  • Kaensaksiri T, Soontornchainaksaeng P, Soonthornchareonnon N, Prathanturarug S (2011) In vitro induction of polyploidy in Centella asiatica (L.) Urban. Plant Cell Tiss Organ Cult 107(2):187–194

    Google Scholar 

  • Kamada H, Okamura N, Satake M, Harada H, Shimomura K (1986) Alkaloid production by hairy root cultures in Atropa belladonna. Plant Cell Rep 5(4):239–242

    CAS  PubMed  Google Scholar 

  • Kerr D, Dietze P (2008) Kelly AM Intranasal naloxone for the treatment of suspected heroin overdose. Addiction 103(3):379–386

    PubMed  Google Scholar 

  • Kettenes-van de Bosch JJ, Salemink CA, Khan I (1981) Biological activity of the alkaloids of Papaver bracteatum Lind. J Ethnopharmacol 3(1):21–38

    Google Scholar 

  • Kim CS, Park JB, Kim KJ, Chang SJ, Ryoo SW, Jeon BH (2002) Effect of Korea red ginseng (Panax ginseng) on cerebral blood flow and superoxide production. Acta Pharmacol. 23:1152–1156

    CAS  Google Scholar 

  • Kondorosi E, Roudier F, Gendrau E (2000) Plant cell-size: growing by ploidy. Curr Opin Plant Biol 3:488–492

    CAS  PubMed  Google Scholar 

  • Lin X, Zhou Y, Zhang J, Lu X, Zhang F, Shen Q, Wu S, Chen Y, Wang T, Tang K (2011) Enhancement of artemisinin content in tetraploid Artemisia annua plants by modulating the expression of genes in artemisinin biosynthetic pathway. Biotechnol Appl Biochem 58(1):50–57

    CAS  PubMed  Google Scholar 

  • Liu S, Chen S, Chen Y, Guan Z, Yin D, Chen F (2011) In vitro induced tetraploid of Dendranthema nankingense (Nakai) Tzvel. Shows an improved level of abiotic stress tolerance. Sci Hort. 127(3):411–419

    CAS  Google Scholar 

  • Machara A, Werner L, Ann Endoma-Arias M, Cox DF, Hudlicky T (2012) Improved synthesis of buprenorphine from thebaine and/or oripavine via palladium-catalyzed N-demethylation/acylation and/or concomitant O-demethylation. Adv Synth Catal 354:613–626

    CAS  Google Scholar 

  • Madani H, Hosseini B, Dehghan E, Rezaei- chiyaneh E (2015) Enhanced production of scopolamine in induced autotetraploid plants of Hyoscyamus reticulatus L. Acta Physiol Plant 37:55

    Google Scholar 

  • Mathura S, Fossey A, Beck SL (2006) Comparative study of chlorophyll content in diploid and tetraploid black wattle (Acacia mearnsii). Forestry 79:381–388

    Google Scholar 

  • Milo J, Levy A, Palevitch D, Ladizinsky G (1987) Thebaine content and yield in induced tetraploid and triploid plants of Papaver bracteatum Lindl. Euphytica 36:361–367

    CAS  Google Scholar 

  • Milo J, Levy A, Ladizinsky G, Palevitch D (1988) Phylogenetic and genetic studies in Papaver section Oxytona: cytogenetics, isozyme analysis and chloroplast DNA variation. Theor Appl Genet 75:795–802

    CAS  Google Scholar 

  • Mishra BK, Pathak S, Sharma A, Trivedi PK, Shukla S (2010) Modulated gene expression in newly synthesized auto tetraploid of Papaver somniferum L. S Afr J Bot 76:447–452

    CAS  Google Scholar 

  • Neubauer D, Mothes K (1963) Uber Papaver bracteatum I. Mitteilung, ein neuer weg zur gewinnung von morphinanen auf pflanztichef rohstoffbasis. Planta Med 11:387–391

    CAS  Google Scholar 

  • Omidbaigi R, Mirzaee M, Hassani ME, Sedghi Moghadam M (2010a) Induction and identification of polyploidy in basil (Ocimum basilicum L.) medicinal plant by colchicine treatment. Int J Plant Prod. 4:1735–8043

    Google Scholar 

  • Omidbaigi R, Yavari S, Hassani ME, Yavari S (2010b) Induction of autotetraploidy in dragonhead (Dracocephalum moldavica L.) by colchicine treatment. J. Fruit Ornam. Plant Res. 18:23–35

    CAS  Google Scholar 

  • Palevitch D, Levy A (1983) Cultural and genetic factors affecting thebaine yield in Papaver bracteatum Lindl. Acta Hort 132:189–195

    Google Scholar 

  • Rey H, Sansberro B, Collavino M, Davina J, Gonzalez A, Mroginski L (2002) Colchicine, trifluralin and oryzalin promoted development of somatic embryos in Ilex paraguariensis (Aquifoliaceae). Euphytica 123:49–56

    CAS  Google Scholar 

  • Rinner U, Hudlicky T (2012) Synthesis of morphine alkaloids and derivatives. Top Curr Chem 309:33–66

    CAS  PubMed  Google Scholar 

  • Rowson JM (1949) Increased alkaloid contents of induced polyploid of Datura. Nature Rev Genet. 154:81–82

    Google Scholar 

  • Rubuluza T, Nikolova RV, Smith MT, Hannweg K (2007) In vitro induction oftetraploids in Colophospermum mopane by colchicine. South Afri J Bot. 73:259–261

    Google Scholar 

  • Sadat Noori SA, Norouzi M, Karimzadeh G, Shirkool K, Niazian M (2017) Effect of colchicine-induced polyploidy on morphological characteristics and essential oil composition of ajowan (Trachyspermum ammi L.). Plant Cell Tiss Organ Cult 130(3):543–551

    CAS  Google Scholar 

  • Saminathan T, Nimmakayala P, Manohar S, Malkaram S, Almeida A, Cantrell R, Tomason Y, Abburi L, Rahman MA, Vajja VG, Khachane A, Kumar B, Rajasimha HK, Levi A, Wehner T, Reddy UK (2015) Differential gene expression and alternative splicing between diploid and tetraploid watermelon. J Exp Bot 66(5):1369–1385

    CAS  PubMed  Google Scholar 

  • Seddigh M, Jolliff GD, Calhoun W, Crane JM (1982) Papaver bracteatum, potential commercial source of codeine. Econ Bot 36:433–441

    CAS  Google Scholar 

  • Shahriari-Ahmadi F, Dehghan E, Farsi M, Azizi M (2009) Tetraploid Induction of Hyoscyamus muticus L. using Colchicine Treatment. Pak J Biol Sci 11(24):2653–2659

    Google Scholar 

  • Swanson CP (1957) Cytol cytogenet. Carl P, Swanson, p 596

    Google Scholar 

  • Talebi SF, Saharkhiz MJ, Jafarkhani Kermani M, Sharafi Y, Raouf Fard F (2016) Effect of different antimitotic agents on polyploid induction of anise hyssop (Agastache foeniculum L.). Inte J Cytol Cytosyst Cytogen. 70(2):184–193

    Google Scholar 

  • Tang ZQ, Chen DL, Song ZJ, He YC, Cai DT (2010) In vitro induction and identification of tetraploid plants of Paulownia tomentosa. Plant Cell Tiss Organ Cult 102(2):213–220

    Google Scholar 

  • Tarkesh Esfahani S, Karimzadeh G, Naghavi MR (2016) 2C DNA value of persian poppy (Papaver bracteatum Lindl.) medicinal plant as revealed by flow cytometry analysis; a quick effective criteria for distinguishing unidentified papaver species. Int J Adv Biotechnol Res. 7(2):573–578

    Google Scholar 

  • Wallaart TE, Pras N, Quax WJ (1999) Seasonal variations of artemisinin and its biosynthetic precursors in tetraploid Artemisia annua plants compared with the diploid wild-type. Planta Med 65:723–728

    CAS  Google Scholar 

  • Wendel JF, Doyle JJ (2005) Polyploidy and evolution in plants. In: Henry RJ (ed) Plant diversity and evolution. CABI Publishing, Wallingford, UK, pp 97–117

    Google Scholar 

  • Zahedi AA, Hosseini B, Fattahi M, Dehghan E, Parastar H, Madani H (2014) Overproduction of valuable methoxylated flavones in induced tetraploid plants of Dracocephalum kotschyi Boiss. Bot Stud. 55:22

    PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Horticulture, Faculty of Agriculture, Urmia University, Po.Box 165, Urmia, Iran

    Hadi Madani & Bahman Hosseini

  2. Department of Plant Genetics and Breeding, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

    Ghasem Karimzadeh

  3. Department of Agronomy, Faculty of Agriculture, Urmia University, Urmia, Iran

    Amir Rahimi

Authors
  1. Hadi Madani
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bahman Hosseini
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ghasem Karimzadeh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Amir Rahimi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bahman Hosseini.

Additional information

Communicated by Y. Wang.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Madani, H., Hosseini, B., Karimzadeh, G. et al. Enhanced thebaine and noscapine production and modulated gene expression of tyrosine/dopa decarboxylase and salutaridinol 7-O-acetyltransferase genes in induced autotetraploid seedlings of Papaver bracteatum Lindl. Acta Physiol Plant 41, 194 (2019). https://doi.org/10.1007/s11738-019-2984-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11738-019-2984-9

Keywords

Search

Navigation