Skip to main content

The effect of auxin on plant regeneration of wheat, barley and triticale

Abstract

One of the basic components of a medium influencing somatic embryogenesis of cereals from immature embryos is the type of auxin. According to some researchers, phytohormones can also play an important role during Agrobacterium-mediated transformation. In this first part of research, the influence of three types of auxins used alone or in combination of two on somatic embryogenesis and plant regeneration in three cereal species has been tested. Eight cultivars of barley, five cultivars of wheat and three cultivars of triticale have been used. Efficiency of plant development on two regeneration media, with and without growth regulators has been compared. Efficiency of regeneration characterized by frequency of explants that form embryogenic callus ranged from 25% for wheat cultivar Torka to 100% for two barley cultivars. Mean number of plantlets regenerating per explant differed significantly (from 2 to 58) depending on the type of auxin in inducing media, the type of regenerating media as well as cultivar. The biggest differences in regeneration efficiency were observed between barley cultivars, however regeneration of plants occurred in all combinations tested. The best regeneration coefficients for most barley cultivars were obtained after culture on dicamba or dicamba with 2,4-D. However, in the case of highly regenerating cv Scarlett, the most effective culture media contained picloram or 2,4-D alone. The highest values of regeneration coefficients for two triticale cultivars (Wanad and Kargo) were obtained on picloram (26.1 and 21.4, respectively) and for `Gabo' on picloram with dicamba (12.6). The range of mean number of regenerated plantlets was from 12 to 30. Dicamba alone or lower concentrations of picloram with 2,4-D were the best media influencing embryogenic callus formation in five wheat cultivars. However, the highest values of regeneration coefficients ranging from 10.6 to 26.8 were obtained at lower concentrations of picloram with 2,4-D or picloram with dicamba. R2 regeneration medium containing growth regulators was significantly better for plantlet development in several combinations (cultivar and induction medium) than the one without growth regulators. Generally, regeneration coefficients for all tested cultivars of three cereal species on the best media were high, ranging from 5.5 for barley cultivar Rodion to 51.6 for another barley cultivar Scarlett. Plantlets developed normally, flowering and setting seed.

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

References

  • Ainsley PJ & Aryan AP (1998) Efficient plant regeneration system for immature embryos of Triticale (×Triticosecale Wittmack). Plant Growth Regul. 24: 23-30

    Google Scholar 

  • Arzani A & Mirodjagh S-S (1999) Response of durum wheat cultivars to immature embryo culture, callus induction and in vitro salt stress. Plant Cell Tiss. Org. Cult. 58: 67-72

    Google Scholar 

  • Bahieldin A, Dyer WE & Qu R (2000) Concentration effects of dicamba on shoot regeneration in wheat. Plant Breed. 119: 437-439

    Google Scholar 

  • Barro F, Martin A, Lazzeri PA & Barcel P (1999) Medium optimization for efficient somatic embryogenesis and plant regeneration from immature inflorescences and immature scutella of elite cultivars of wheat, barley and tritordeum. Euphytica 108: 161-167

    Google Scholar 

  • Becker D, Brettschneider R & Lorz H (1994) Fertile transgenic wheat from microprojectile bombardment of scutellar tissue. Plant J. 5: 299-307

    Google Scholar 

  • Bregitzer P (1992) Plant regeneration and callus type in barley: effects of genotype and culture medium. Crop Sci. 32: 1108-1112

    Google Scholar 

  • Bregitzer P, Campbell RD & Wu Y (1995) Plant regeneration from barley callus: Effects of 2,4-dichlorophenoxyacetic acid and phenylacetic acid. Plant Cell Tiss. Org. Cult. 43: 229-235

    Google Scholar 

  • Bregitzer P, Dahleen LS & Campbell RD (1998) Enhancement of plant regeneration from embryogenic callus of commercial barley cultivars. Plant Cell Rep. 17: 941-945

    Google Scholar 

  • Castillo AM, Egana B, Sanz JM & Cistuo L (1998) Somatic embryogenesis and plant regeneration from barley cultivars grow in Spain. Plant Cell Rep. 17: 902-906

    Google Scholar 

  • Chateau S, Sangwan RS & Sangwan-Norreel BS (2000) Competence of Arabidopsis thaliana genotypes and mutants for Agrobacterium tumefaciens-mediated gene transfer: Role of phytohormones. J. Exp. Bot. 51: 1961-1968

    Google Scholar 

  • Cheng M, Fry JE, Pang S, Zhou H, Hironaka CM, Duncan DR, Conner TW & Wan Y (1997) Genetic transformation of wheat mediated by Agrobacterium tumefaciens. Plant Physiol. 115: 971-980

    Google Scholar 

  • Christou P, Ford TL & Kofron M (1991) Production of transgenic rice (Oryza sativa L.) plants from agronomically important Indica and Japonica varieties via electric discharge particle acceleration of exogenous DNA into immature zygotic embryos. Bio/Technology 9: 957-962

    Google Scholar 

  • Dale PJ & Deambrogio E (1979) A comparison of callus induction and plant regeneration from different explants of Hordeum vulgare. Z. Pflanzenphysiol 94: 65-77

    Google Scholar 

  • Elena EB & Ginzo HD (1988) Effect of auxin levels on shoot formation with different embryo tissues from a cultivar and a commercial hybrid of wheat (Triticum aestivum L.). J. Plant Physiol. 132: 600-603

    Google Scholar 

  • Fennell S, Bohorova N, Ginkel M van, Crossa J & Hoisington DA (1996) Plant regeneration from immature embryos of 48 elite CIMMYT bread wheats. Theor. Appl. Genet. 92: 163-169

    Google Scholar 

  • Gamborg OL, Miller RA & Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell. Res. 50: 151-158

    Google Scholar 

  • Goldstein CS & Kronstad WE (1986) Tissue culture and plant regeneration from immature embryo explants of barley, Hordeum vulgare L. Theor. Appl. Genet. 71: 631-636

    Google Scholar 

  • He DG, Yang YM & Scott KJ (1989) The effect of macroelements in the induction of embryogenic callus from immature embryos of wheat (Triticum aestivum L.). Plant Sci. 64:251-258

    Google Scholar 

  • Hiei Y, Ohta S, Komari T & Kumashiro T (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J. 6: 271-282

    Google Scholar 

  • Immonen AST (1996) Influence of media and growth regulators on somatic embryogenesis and plant regeneration for production of primary Triticales. Plant Cell Tiss. Org. Cult. 44: 45-52

    Google Scholar 

  • Ishida Y, Saito H, Ohta S, Hiei Y, Komari T & Kumashiro T (1996) High efficiency transformation of maize (Zea mays L.) mediated by Agrobacterium tumefaciens. Nature Biotech. 14: 745-750

    Google Scholar 

  • Kachhwaha S & Kothari SL (1996) Plant regeneration from immature embryo explants of Hordeum spontaneum and Hordeum vulgare. Cereal Res. Comm. 24: 27-32

    Google Scholar 

  • Kachhwaha S, Varshney A & Kothari SL (1997) Somatic embryogenesis and long term high plant regeneration from barley (Hordeum vulgare L.) using picloram. Cereal Res. Comm. 25: 117-126

    Google Scholar 

  • Koziel MG, Beland GL, Bowman C, Carozzi NB, Crenshaw R, Crossland L, Dawson J, Desai N, Hill M, Kadwell S, Launis K, Lewis K, Maddox D, McPerson K, Meghji MR, Merlin E, Rhodes R, Warren GW, Wright M & Evola SV (1993) Field performance of elite transgenic maize plants expressing an insecticidal protein derived by Bacillus thuringiensis. Bio/Technology 11: 194-200

    Google Scholar 

  • Lazar MD, Collins GB & Vian WE (1983) Genetic and environmental effects on the growth and differentiation of wheat somatic cell cultures. J. Hered. 74: 353-357

    Google Scholar 

  • Linacero R & Vasquez AM (1990) Somatic embryogenesis from immature inflorescences of rye. Plant Sci. 72: 253-258

    Google Scholar 

  • Lührs R & Lörz H (1987) Plant regeneration in vitro from embryogenic cultures of spring-and winter-type barley (Hordeum vulgare L.) varieties. Theor. Appl. Genet. 75: 16-25

    Google Scholar 

  • Machii H, Mizuno H, Hirabayashi T, Li H & Hagio T (1998) Screening wheat genotypes for high callus induction and regeneration capability from anther and immature embryo cultures. Plant Cell Tiss. Org. Cult. 53: 67-74

    Google Scholar 

  • Maddock SE, Lancaster VA, Risiott R & Franklin J (1983) Plant regeneration from cultured immature embryos and inflorescences of 25 cultivars of wheat (Triticum aestivum L.) J. Exp. Bot. 34: 915-926

    Google Scholar 

  • Mathias RJ & Simpson ES (1986) The interaction of genotype and culture medium on the tissue culture responses of wheat (Triticum aestivum L.) callus. Plant Cell Tiss. Org. Cult. 7: 31-37

    Google Scholar 

  • Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473-497

    Google Scholar 

  • Nadolska-Orczyk A, Orczyk W & Przetakiewicz A (2000) Agrobacterium-mediated transformation of cereals - from technique development to its application. Acta Physiol. Plant. 22: 77-88

    Google Scholar 

  • Nonohay JS, Mariath JEA & Winge H (1999) Histological analysis of somatic embryogenesis in Brazilian cultivars of barley, Hordeum vulgare vulgare Poaceae. Plant Cell Rep. 18: 929-934

    Google Scholar 

  • Özgen M, Taret M, Altinok S & Sancak C (1998) Efficient callus induction and plant regeneration from mature embryo culture of winter wheat (Triticum aestivum L.) genotypes. Plant Cell Rep. 18: 331-335

    Google Scholar 

  • Ozias-Akins P & Vasil IK (1982) Plant regeneration from cultured immature embryos and inflorescences of Triticum aestivum L. (wheat): Evidence for somatic embryogenesis. Protoplasma 110: 95-105

    Google Scholar 

  • Papenfuss JM & Carman JG (1987) Enhanced regeneration from wheat callus cultures using dicamba and kinetin. Crop Sci. 27: 588-593

    Google Scholar 

  • Rakoczy-Trojanowska M & Malepszy S (1995) Genetic factors influencing the regeneration ability of rye (Secale cereale L.). II. Immature embryos. Euphytica 83: 233-239

    Google Scholar 

  • Rashid A. & Vikrant (2001) Comparative study of somatic embryogenesis from immature and mature embryos and organogenesis from leaf-base of Triticale. Plant Cell Tiss. Org. Cult. 64: 33-38

    Google Scholar 

  • Redway FA, Vasil V, Lu D & Vasil IK (1990) Identification of callus types for long-term maintenance and regeneration from commercial cultivars of wheat (Triticum aestivum L.). Theor. Appl. Genet. 79: 609-617

    Google Scholar 

  • Rhodes CA, Green CE & Phillips RL (1986) Factors affecting culture initiation from maize tassels. Plant Sci. 46: 225-232

    Google Scholar 

  • Rout JR & Lucas WJ (1996) Characterization and manipulation of embryogenic response from in vitro cultured immature inflorescences of rice (Oryza sativa L.). Planta 198: 127-138

    Google Scholar 

  • Ruiz ML, Rueda J, Peláez MI, Espino FJ, Candela M, Sendino AM & Vásques AM (1992) Somatic embryogenesis, plant regeneration and somaclonal variation in barley. Plant Cell Tiss. Org. Cult. 28: 97-101

    Google Scholar 

  • Sangwan RS, Bourgeois Y, Brown S, Vasseur G & Sangwan-Nor-reel BS (1992) Characterization of competent cells and early events of Agrobacterium-mediated genetic transformation in Arabidopsis thaliana. Planta 188: 439-456

    Google Scholar 

  • Sears RG & Deckard EL (1982) Tissue culture variability in wheat: Callus induction and plant regeneration. Crop Sci. 22: 546-550

    Google Scholar 

  • Shimada T (1978) Plant regeneration from the callus induced from wheat embryos. Jpn. J. Genet. 53: 371-374

    Google Scholar 

  • Songstad DD, Petersen WL & Armstrong CL (1992) Establishment of friable embryogenic (type II) callus from immature tassels of Zea mays (Poaceae). Am. J. Bot. 79: 761-764

    Google Scholar 

  • Stolarz A & Lörz H (1986) Somatic embryogenesis, in vitro multiplication and plant regeneration from immature embryo explants of hexaploid Triticale (× Triticosecale Wittmack). Z. Pflanzenzuchtung 96: 353-362

    Google Scholar 

  • Thomas MR & Scott KJ (1985) Plant regeneration by somatic embryogenesis from callus initiated from immature embryos and immature inflorescences of Hordeum vulgare L. J. Plant Physiol. 121: 159-169

    Google Scholar 

  • Tingay S, McElroy D, Kalla R, Fieg S, Wang M, Thornton S & Brettell R (1997) Agrobacterium tumefaciens-mediated barley transformation. Plant J. 11: 1369-1376

    Google Scholar 

  • Varshney A, Kant T, Sharma VK, Rao A & Kothari SL (1996) High frequency plant regeneration from immature embryo cultures of Triticum aestivum and T. durum. Cereal Res. Comm. 24: 409-416

    Google Scholar 

  • Wan Y & Leumax PG (1994) Generation of large numbers of independently transformed fertile barley plants. Plant Physiol. 104: 37-48

    Google Scholar 

  • Weeks JT, Anderson OD & Blechl AE (1993) Rapid production of multiple independent lines of fertile transgenic wheat (Triticum aestivum L.). Plant Physiol. 102: 1077-1084

    Google Scholar 

  • Zhang LJ & Seilleur P (1987) A simple and fast method to obtain high frequency of plant regeneration from mature and immature wheat embryos. Bull. Rech. Agron. Gembloux 22: 187-189

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to A. Nadolska-Orczyk.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Przetakiewicz, A., Orczyk, W. & Nadolska-Orczyk, A. The effect of auxin on plant regeneration of wheat, barley and triticale. Plant Cell, Tissue and Organ Culture 73, 245–256 (2003). https://doi.org/10.1023/A:1023030511800

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1023030511800