Skip to main content

Induction of Polyploidy and Its Effect on Cannabis sativa L.

Abstract

Polyploids are organisms with three or more complete chromosome sets. Polyploidization is widespread in plants, and is an important mechanism of speciation. Polyploids can be formed in various ways. The study of polyploids has both important theoretical significance and valuable applications. The production and application of polyploidy breeding have brought remarkable economic and social benefits. We reported the production of putative tetraploid plants of Cannabis sativa L., with the ultimate aim of improving the medicinal and physiological traits of this widely distributed cultivated plant. The production of tetraploid plant was improved with colchicine at different concentrations and time through dropping method. Flow cytometry analysis was used to confirm the ploidy level. Morphologic, anatomic and biochemical characteristics were compared between tetraploid and diploid control plants. The results showed that 0.2% colchicine for 24 h was the most efficient for production of polyploid plants. The percentage of tetraploid plants and the survival rate were lowered by the increasing the treatment time. In addition, the leaf index and height of tetraploid plants exhibited a significant decrease compared to the diploid plants. The size of stomata on epidermis of leaves were larger in tetraploid plant compared to the diploid ones, in spite of the tetraploid plants have less stomata density. However, the amount of total chlorophyll and carotenoids were almost the same in both tetraploid and diploid plants. In addition, some differences were also observed in the cross section of stem of these plants from a descriptive structural point of view. Overall, the results introduced usage of the stomata parameters as an effective, fast and convenient method for detecting the tetraploid plants. We also investigated polyploidy effects on some primary and secondary metabolites. The results of biochemical analyzes showed that soluble sugars and total protein content increased significantly into mixoploid plants compared to tetraploid and diploid plants. Tetraploid plants had higher amount of total proteins compared with control plants. The results showed that polyploidization could increase the contents of tetrahydrocannabionol only in mixoploid plants but tetraploid plants had lower amounts of this substance in comparison with diploids. Our results suggest that tetraploidization was not useful for production of tetrahydrocannabinol for commercial use but mixoploids were found suitable.

Keywords

  • Ploidy Level
  • Apical Meristem
  • Female Flower
  • Glandular Trichome
  • Soluble Sugar Content

These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

This is a preview of subscription content, access via your institution.

Buying options

Chapter
USD   29.95
Price excludes VAT (USA)
  • DOI: 10.1007/978-3-319-54564-6_17
  • Chapter length: 19 pages
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
eBook
USD   219.00
Price excludes VAT (USA)
  • ISBN: 978-3-319-54564-6
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
Softcover Book
USD   279.99
Price excludes VAT (USA)
Hardcover Book
USD   279.99
Price excludes VAT (USA)
Fig. 17.1
Fig. 17.2
Fig. 17.3
Fig. 17.4
Fig. 17.5
Fig. 17.6
Fig. 17.7
Fig. 17.8
Fig. 17.9
Fig. 17.10
Fig. 17.11

References

  • Allum JF, Bringloe DH, Roberts AV (2007) Chromosome doubling in 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  CrossRef  PubMed  Google Scholar 

  • Bagheri M, Mansouri H (2015) Effect of induced polyploidy on some biochemical parameters in Cannabis sativa L. Appl Biochem Biotechnol. doi: 10.1007/s12010-014-1435-8

  • Bhojwani SS (2004) In vitro production of triploid plants. In: Goodman RM (ed) Encyclopedia of plant and crop science. Marcel Dekker, New York, pp 590–593

    CrossRef  Google Scholar 

  • Blanc G, Wolfe KH (2004) Widespread paleopolyploidy in model plant species inferred from age distributions of duplicate genes. Plant Cell 16:1667–1678

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 7(72):248–254

    CrossRef  Google Scholar 

  • Buchanan BB, Gruissem W, Jones R (2000) Biochemistry and molecular biology of plants. American Society of Plant Physiologists. ISBN 81-88237-11–6

    Google Scholar 

  • Chakraborti SP, Vijayan K, Roy BN, Qadri SM (1998) In vitro induction of tetraploidy in mulberry (Morus alba L.). Plant Cell Rep 17:799–803

    CAS  CrossRef  Google Scholar 

  • Chen C, Hou X, Zhang H, Wang G, Tian L (2011) Induction of Anthurium andraeanum ‘‘Arizona’’ tetraploid by colchicine in vitro. Euphytica 181:137–145

    CrossRef  Google Scholar 

  • Dhawan OP, Lavania UC (1996) Enhancing the productivity of secondary metabolites via induced polyploidy: a review. Euphytica 87(2):81–89

    CAS  CrossRef  Google Scholar 

  • Dickison WC (1974) Trichomes. In: Radford AE, Dickison WC, Massey JR, Bell CR (eds) Vascular plant systematics. Harper and row, New York, pp. 198–202. Oxford, pp 40–53

    Google Scholar 

  • Dijkstra H, Speckmann GJ (1980) Autotetraploidy in caraway (Carum carvi L.) for the increase of the aetheric oil content of the seed. Euphytica 29(1):89–96

    CrossRef  Google Scholar 

  • ElSohly MA, Slade D (2005) Chemical constituents of marijuana: the complex mixture of natural cannabinoids. Life Sci 78:539–548

    CAS  CrossRef  PubMed  Google Scholar 

  • Gao SL, Zhu DN, Cai ZH, Xu DR (1996) Autotetraploid plants from colchicine treated bud culture of Salvia miltiorrhiza Bge. Plant Cell, Tissue Organ Cult 47:73–77

    CAS  CrossRef  Google Scholar 

  • Grange S, Leskovar DI, Pike LM, Cobb BG (2003) J Am Soc Hort Sci 128:253–259

    Google Scholar 

  • Grant V (1981) Plant speciation, 2nd edn. Columbia University Press, New York

    Google Scholar 

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

    CAS  Google Scholar 

  • Gu XF, Yang AF, Meng H, Zhang JR (2005) In vitro induction of tetraploid plants from diploid Zizyphus jujuba Mill. cv. Zhanhua. Plant Cell Rep 24:671–676

    CAS  CrossRef  PubMed  Google Scholar 

  • Hamill SD, Smith MK, Dodd WA (1992) In vitro Induction of banana autotetraploids by colchicine treatment of micropropagated diploids. Aust J Botany 40:887–896

    CAS  CrossRef  Google Scholar 

  • Haslam E (1986) Secondary metabolism-Fact or fiction. Nat Prod Rep 3:217–249

    CAS  CrossRef  Google Scholar 

  • Hull-Sanders HM, Johnson RH, Owen HA, Meyer GA (2009) Effects of polyploidy on secondary chemistry, physiology, and performance of native and invasive genotypes of Solidago gigantea (Asteraceae). Am J Bot 96:762–770

    CrossRef  PubMed  Google Scholar 

  • Leitch AR, Leitch IJ (2008) Genomic plasticity and the diversity of polyploid plants. Science 320:481–483

    CAS  CrossRef  PubMed  Google Scholar 

  • Li HL (1973) An archaeological and historical account of Cannabis in China. Econ Bot 28:437–444

    CrossRef  Google Scholar 

  • Liu G, Li Z, Bao M (2007) Colchicine-nduced chromosome doubling in Platanus acerifolia and its effect on plant morphology. Euphytica 157:145–154

    CrossRef  Google Scholar 

  • Loureiro J, Pinto G, Lopes T, Dolezel C, Santos C (2005) Assessment of ploidy stability of somatic embryogenesis process in Quercus suber L. using flow cytometry. Planta 221:815–822

    CAS  CrossRef  PubMed  Google Scholar 

  • Madon M, Clyde MM, Hashim H, Mohd Yusuf Y, Mat H, Saratha S (2005) Polyploidy induction of oil palm through colchicine and oryzalin treatments. J Oil Palm Res 17:110–123

    CAS  Google Scholar 

  • Majdi M, Karimzadeh G, Omidbaigi R (2010) Induction of tetraploidy to Feverfew (Tanacetum parthenium Schulz-Bip.): morphological, physiological, cytological, and phytochemical Changes. J Hort Sci 45:16–21

    Google Scholar 

  • Mansouri H, Rohani M (2014) Response of Cannabis sativa L. To foliar application of 2-chloroethyltrimethylammoniumchoride. Iran. J Plant Physiol 6(1):1225–1234

    Google Scholar 

  • Mansouri H, Asrar Z, Mehrabani M (2009) Effect of gibberellic acid on primary terpenoids and Δ9-tetrahydrocannabinol in Cannabis sativa at flowering stage. J Int Plant Biol 51(6):553–561

    CAS  CrossRef  Google Scholar 

  • Mehmedic Z, Chandra S, Slade D, Denham H, Foster S, Patel AS, Ross SA, Khan IA, ElSohly MA (2010) Potency trends of Δ9-THC and other cannabinoids in confiscated Cannabis preparations from 1993 to 2008. J Forensic Sci 55:1209–1710

    CAS  CrossRef  PubMed  Google Scholar 

  • Ming R, Bendahmane A, Renner SS (2011) Sex chromosomes in land plants. Annu Rev Plant Biol 62:485–514

    CAS  CrossRef  PubMed  Google Scholar 

  • Nakano MT, Nomizu K, Mizunashi M, Suzuki S, Mori S, Kuwayama M, Hayashi H, Umehara E, Kobayashi H, Asano M, Sugawara S, Takagi H, Saito H, Nakata M, Godo T, Hara Y, Amano J (2006) Somaclonal variation in Tricyrtis hirta plants regenerated from 1-year-old embryogenic callus cultures. J Hort Sci 110:366–371

    CrossRef  Google Scholar 

  • Ortiz R, Vuylsteke D (1995) Factors in Sequencing seed set in triploid Musa spp. L. and production of euploid hybrids. Ann Bot 75:151–155

    CrossRef  Google Scholar 

  • Otto F (1990) DAPI staining of fixed cells for high-resolution flow cytometry of nuclear DNA. In: Darzynkiewicz Z, Crissman H (eds) Methods in cell biology, vol 33., Academic PressNew York, USA, pp 105–110

    Google Scholar 

  • Otto SP, Whitton J (2000) Polyploid incidence and evolution. Annu Rev Genet 34:401–437

    CAS  CrossRef  PubMed  Google Scholar 

  • Ramsey J, Schemske DW (1998) Pathways, mechanisms, and rates of polyploid formation in flowering plants. Annu Rev Ecol Syst 29:467–501

    CrossRef  Google Scholar 

  • Ranney TG (2006) Polyploidy: From evolution to new plant development. Combined Proc Int Plant Propagators’ Soc 56:137–142

    Google Scholar 

  • Roe JH (1955) The determination of sugar in blood and spinal fluid with anthrone reagent. J Biol Chem 212(1):335–343

    CAS  PubMed  Google Scholar 

  • Russo EB, Jiang HE, Li X, Sutton A, Carboni A, Bianco F, Mandolino G, Potter DJ, Zhao YX, Bera S, Zhang YB, Lü E-G, Ferguson DK, Hueber F, Zhao LC, Liu CJ, Wang YF, Schultes RE, Klein WM, Plowman T, Lockwood TE (1974) Cannabis: an example of taxonomic neglect. Bot Mus Leaf Harvard Univ 23:337–367

    Google Scholar 

  • Rustichelli C, Ferioli V, Vezzalini F, Rossi MC, Gamberini G (1996) Simultaneous separation and identification of hashish constituents by coupled liquid chromatography-mass spectrometry (HPLC-MS). Chromatographia 43(3–4):129–134

    CAS  CrossRef  Google Scholar 

  • Schultes RE, Klein WM, Plowman T, Lockwood TE (1974) Cannabis: an example of taxonomic neglect. Harvard Univ. Bot. Mus. Leafl. 23:337–367

    Google Scholar 

  • Seigler DS (1998) Plant secondary metabolism. Originally published by Kluwer Academic Publishers. ISBN 978-1-4615-4913-0 (eBook)

    Google Scholar 

  • Soltis DE, Soltis PS, Tate JA (2003) Advances in the study of polyploidy since plant speciation. New Phytol 161:173–191

    CrossRef  Google Scholar 

  • Song C, Liu S, Xiao J, He W, Zhou Y, Qin QB, Zhang C, Liu Y (2012) Polyploid organisms. Sci China. 55(4):301–311

    CrossRef  Google Scholar 

  • Thao NTP, Ureshino K, Miyajima I, Ozaki Y, Okubo H (2003) Induction of tetraploids in ornamental Alocasia through colchicine and oryzalin treatments. Plant Cell, Tissue Organ Cult 72:19–25

    CAS  CrossRef  Google Scholar 

  • Treier UA, Broennimann O, Normand S, Guisan A, Schaffner U, Steinger T, Müller-Schärer H (2009) Shift in cytotype frequency and niche space in the invasive plant Centaurea maculosa. Ecology 90(5):1366–1377

    CrossRef  PubMed  Google Scholar 

  • Verpoort R (1999) Bioassay methods in natural product research and drug development © Kluwer Academic Publishers. pp 11–23

    Google Scholar 

  • Xu L, Najeeb U, Ms Naeem, Mk Daud, Cao JS, Gong HJ, Shen WQ, Zhou WJ (2010) Induction of tetraploidy in Juncus effusus by colchicine. Biol Planta 54:659–663

    CAS  CrossRef  Google Scholar 

  • Zias J, Stark H, Sellgman J, Levy R, Werker E, Breuer A, Mechoulam R (1993) Early medical use of Cannabis. Nature 363:215

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Hakimeh Mansouri .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and Permissions

Copyright information

© 2017 Springer International Publishing AG

About this chapter

Cite this chapter

Mansouri, H., Bagheri, M. (2017). Induction of Polyploidy and Its Effect on Cannabis sativa L.. In: Chandra, S., Lata, H., ElSohly, M. (eds) Cannabis sativa L. - Botany and Biotechnology. Springer, Cham. https://doi.org/10.1007/978-3-319-54564-6_17

Download citation