Phenotypic and molecular characterization of polyploidy Vicia faba induced by colchicine

Nagat Elhaddad; Kamla  Blash; Hoda Khatab

doi:10.30574/gscbps.2020.11.3.0184

Authors

Nagat Elhaddad Department of Botany, Faculty of Science, University of Omar Al-Mukhtar.
Kamla Blash Department of Botany, Faculty of Science, University of Omar Al-Mukhtar.
Hoda Khatab Department of Botany, Faculty of Science, University of Omar Al-Mukhtar.

DOI:

https://doi.org/10.30574/gscbps.2020.11.3.0184

Keywords:

Colchicine, Spectrophotometer, Vicia faba, Stomatal behavior, Polyploidy, Morphological traits

Abstract

To induce polyploidy in diploid control plant of Vicia faba, seeds were socked in various concentrations of colchicine (0.1, 0.2, 0.3 and 0.4 %) for 9 and18 h. Putative ploidy plants were conﬁrmed by screening morphological characters, measuring DNA content via spectrophotometer and analyzing stomatal behavior. Polyploidy plants showed slower growth rate along with increased potency and thickened large leaves. Three types of polyploidy were achieved; triploid, tetraploid and mixoploid. In induced polyploid plants, the length and width of stomata were increased, while the index of stomata was declined, in comparison to the initial diploid plants. Treatment with 0.3 and 0.4 % colchicine for 9 and 18 h were efficiently induced polyploidy. The highest stomatal size (22.53 µm) and widest stomatal aperture (9.72 µm) and the lowest stomatal index (34.24) where obtained when 0.4 % of colchicine where applied at 18hr tested time with high significant differences at p<0.05 in comparison to dipliod plants. We recommend this method for induction of polyploids in other different crops and possibly will selected these tetraploid plants for further breeding purposes.

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References

Ghani MA, Sun Q, Li J, Cao L, Rao L and Zou X. (2014). Phenotypic and genetic variation occurred during wide hybridisation and allopolyploidisation between Brassica rapa and Brassica nigra‏. Chen, Sci. Hortic, 176, 22–31.

Hannweg K, Sippel A and Bertling IS. (2013) A simple and effective method for the micropropagation and in vitro induction of polyploidy and the effect on floral characteristics of the South African iris, Crocosmia aurea. Afr. J. Bot. 88, 367–372.

Huang H, Gao S, D. Wang P, Huang J and Li J. (2014). Autotetraploidy Induced in Nianmaohuangqin (Radix Scutellariae Viscidulae) With Colchicine in Vitro. Tradit. Chin.Med, 34,199–205.

Sattler MC, Carvalho CR and Clarindo WR. (2016). The polyploidy and its key role in plant breeding. Planta 243(2), 281–296.

Hannweg K, Visser G, de Jager K and Bertling I. (2016). In vitro-induced polyploidy and its effect on horticultural characteristics, essential oil composition and bioactivity of Tetradeniariparia. S Afr J Bot, 106, 186–191.

Urwin NA. (2014). Generation and characterisation of colchicine-induced polyploidy Lavandula× intermedia. Euphytica, 197(3), 331–339.

Lavania U. (2005). Genomic and ploidy manipulation for enhanced production of phyto-pharmaceuticals. Plant Genet. Resour, 3 170–177.

Chen WH, Tang CY, Lin TY, Weng YC and Kao YL. (2011). Changes in the Endopolyploidy Pattern of Different Tissues in Diploid and Tetraploid Phalaenopsis Aphrodite Subsp. Formosana (Orchidaceae) PlantSci, 181, 31–38.

Aina O, Quesenberry K and Gallo M. (2012). In vitro induction of tetraploids in Arachis paraguariensis. Plant Cell Tissue Organ Cult (PCTOC), 111(2), 231–238.

Blakeslee AF and Avery AG. (1937). Methods of inducing doubling of chromosomes by treatment with colchicine. Journal of Heredity, 28, 392–411.

Dhooghe E, Van Laere K, Eeckhaut T, Leus L and Van Huylenbroeck J.(2011). Mitotic chromosome doubling of plant tissues in vitro. Plant Cell Tissue Organ Cult (PCTOC), 104(3), 359–373.

Jaskani M, Kwon S and Kin D. (2005). Flow cytometry of dna contents of colchicine treated watermelon as a ploidy screening method at m1 stage. Pak. J. Bot. 37, 685.

Omidbaigia R, Mirzaeea M, Hassanib M and Moghadamc M. (2010). Induction and identification of polyploidy in basil (Ocimum basilicum L.) medicinal plant by colchicine treatment. Int. J. Plant Prod, 4, 2.

Raza H, Jaskani M, Khan M and Malik T. (2003). In Vitro Induction of Polyploids in Watermelon and Estimation Based on DNA Content. Int. J. Agric. Biol, 5, 298–302.

Liu G, Li Z and Bao M. (2007). Colchicine-induced chromosome doubling in Platanus acerifolia and its effect on plant morphology. Euphytica, 157(1-2), 145-154.

Ye, YM, Tong, J, Shi XP, Yuan W and Li GR. (2010). Morphological and cytological studies of diploid and colchicine-induced tetraploid lines of crape myrtle (Lagerstroemia indica L.). Sci. Hortic., 124(1), 95-101.

Edwards K, Johnstone C and Thompson C. (1991). A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucl. Acid. Res, 19, 1349.

Scarpeci TE, Zanor MI and Valle EM. (2017). Estimation of stomatal aperture in Arabidopsis thaliana using silicone rubber imprints. Bio-protocol, 7(12).

Colijn-Hooymans C, Hakkert J, Jansen J and Custers J. (1994). Competence for regeneration of cucumber cotyledons is restricted to specific developmental stages. Plant Cell, Tissue Organ Cult, 39, 211–217.

Gao S, Zhu D, Cai Z and Xu D. (1997). Autotetraploid plants from colchicine-treated bud culture of Salvia miltiorrhiza. Plant Cell, Tissue Organ Cult, 47, 73–77.

Abreu JC, Davide LC, Pereira AV and Barbosa S (2006). Mixoploidia em híbridos de capim-elefante x milheto tratados com agentes antimitóticos. Pes. Agrop. Bras, 41(11),1629-1635.

Barbosa S. (2004). Micropropagação e duplicação cromossômica de híbridos triplódes de capim-elefante e milheto. Thesis, Universidade Federal de Lavras, Lavras.

Carvalho JF, Carvalho CR and Otoni WC. (2005). In vitro induction of polyploidy in annatto (Bixa orellana). Plant Cell Tissue Org, 80(1),6975.

Poutaraud A and Girardin P. (2005). Influence of chemical characteristics of soil on mineral and alkaloid seed contents of Colchicum autumnale. Environ. Exp. Bot, 54(2),101-108.

Campos JM, Davide LC, Salgado CC, Santos FC, Costa PN, Silva OS, Alves CCS, Viccini LF and Pereira AV (2009). In vitro induction of hexaploid plants from triploid hybrids of Pennisetum purpureum and Pennisetum glaucum. Plant Breed, 128,101-104.

Wannakrairoj S and Wondyifraw T. (2013). In vitro chromosome doubling ink [Aframomum corrorima (Braun) P.C.M. Jansen] using colchicine and oryzalin. Kasetsart J. Nat. Sci, 47,684-694.

Prashant Joshi and Rakesh C. Verma. (2004). High Frequency Production of Colchicine Induced Autotetraploids in Faba Bean (Vicia faba L.). Cytologia, 69(2), 141–147.

Wang L, Zheng SX and Gu ZJ. (2002). In vitro culture of tetraploids of Aloe vera L. Acta Hortic Sinica, 29, 176–178.

Rose JB, Kubba J and Tobutt KR. (2001). Induction of tetraploids for breeding hardy ornamentals. Acta Hortic, 560, 109–112.

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

Forni-Martins ER and Cruz ND. (1985). Dados preliminaries sobre induc¸a ˜o de poliploidia em Jatropha curcas (Euphorbiaceae). Congresso da Sociedade Bota `nica de Sa ˜o Paulo. V. Botucatu. Resumos, 167.

Shao JZ, Chen CL and Deng XX. (2003). In vitro induction of tetraploid in pomegranate (Punica granatum). Plant Cell Tiss Organ Cult, 75, 241–246.

Wright JW. (1976). Introduction of forest genetics. Academic Press, New York.

Stebbins GL. (1984). Polyploidy and the distributed of arctic-alpine ﬂora: a new evidence and new approaches. Bot Helv, 94, 1–13.

Avery GS and Pottorf L. (1945). Polyploidy, auxin and nitrogen in green plants. American Journal of Botany, 32, 669–671.

Larsen P and Mintung S. (1950). Growth promoting and growth retarding substances in pollen from 2n and 3n apple varieties. Botanical Gazette, 3, 436–447.

Yadav AK, Singh S, Dhyani D and Ahuja PS. (2011). A review on the improvement of stevia [Stevia rebaudiana (Bertoni)]. Can J Plant Sci, 91, 1–27.

Yadav AK, Singh S, Yadav SC, Dhyani D, Bhardwaj G, Sharma A and Singh B. (2013). Induction and morpho-chemical characterization of Stevia rebaudiana colchiploids. Indian J Agric Sci, 83, 159–169.

Gomes SS, Saldanha CW, Neves CS, Trevizani M, Raposo NR, Notini MM, de Oliveira Santos M, Campos JMS, Otoni WC and Viccini LF. (2014). Karyotype, genome size, and in vitro chromosome doubling of Pfaffia glomerata (Spreng.) Pedersen. Plant Cell, Tissue and Organ Cult (PCTOC), 118(1), 45–56.

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

Miguel TP and Leonhardt KW. (2011). In vitro polyploid induction of orchids using oryzalin. Sci. Hortic, 130, 314–319.

Chakraborti S, Vijayan K, Roy B and Qadri S. (1998). In vitro induction of tetraploidy in mulberry (Morus alba L.) Plant Cell Rep,17, 799–803.

Gantait S, Mandal N, Bhattacharyya S and Das PK. (2011). Induction and identification of tetraploids using in vitro colchicine treatment of Gerbera jamesonii Bolus cv. Sciella. Plant Cell Tissue Organ Cult (PCTOC), 106(3), 485.

Majdi M, Karimzadeh G, Malboobi MA, Omidbaigi R and Mirzaghaderi G. (2010). Induction of tetraploidy to feverfew (Tanacetum parthenium Schulz-Bip.): Morphological, physiological, cytological, and phytochemical changes. HortScience, 45(1), 16–21.

Xu C, Huang Z, Liao T, Li Y and Kang X. (2016). In vitro tetraploid plants regeneration from leaf explants of multiple genotypes in Populus. Plant Cell Tissue Organ Cult (PCTOC), 125(1), 1–9.

Boaventura YM, Medina DM, Vieira MJ and Arruda HV. (1981). Nu ´mero de cloroplastos e nıˇvel de ploidia em espe ´cies de Coffea L. Rev Bras Bot, 4,15–21.

Carvalho R, Guerra M and Carvalho PC. (1999). Occurrence of spontaneous triploids in Manihot esculenta Cruntz. Cytologia, 64, 137–140.

Evans AM. (1955). The production and identiﬁcation of polyploids in red clover, white clover and Lucerne. New Phytol, 514, 149–162.

De Oliveira VM, Forni-Martins ER, Magalha ˜es PM and Alves MN. (2004). Chromosomal and morphological studies of diploid and polyploid cytotypes of Stevia rebaudiana (Bertoni) (Eupatorieae, Asteraceae). Genet Mol Biol, 27, 215–222.