Skip to main content
SpringerLink
Log in

Enzyme fingerprint analyses in tissue regenerated from anther culture of maize

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

Abstract

The objectives of our studies were to investigate the effect of cold pre-treatment duration and the effect of two different culture media (YP and N6) on maize anther culture response in two maize genotypes (A 18 and A 19) and to identify the gametic origin of the maize regenerants. Androgenic induction and callus formation was compared in anther cultures following pre-treatment applied to both media tested and with both maize genotypes. Higher plant regeneration was observed in case of YP media independently of the genotype used. The best results were achieved when 12 days (genotype A 18) or 14 days (in case of genotype A 19) cold pre-treatment at 10°C was applied. We have tested the possibility of using enzyme isoform analyses to identify the microspore origin of calli and plants derived from anther cultures. The 11 enzymes tested in our experiments were acid phosphatase, alcohol dehydrogenase, catalase, diaphorase, β-glucosidase, glutamate oxaloacetate transaminase, isocitrate dehydrogenase, malate dehydrogenase, 6-phosphogluconate dehydrogenase, phosphoglucomutase and phosphoglucoisomerase. Analysis of malate dehydrogenase proved the gametic origin of the calli initiated and of the DH plants regenerated from anther culture, when the coleoptile of the donor plant material showed two forms of enzyme 3/6 and the analysed calli showed only one of the two forms (3 or 6).

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

Similar content being viewed by others

References

  • Ambrus H, Darkó E, Szabó L, Bakos F, Király Z, Barnabás B (2006) In vitro microspore selection in maize anther culture with oxidative stress stimulators. Protoplasma 228:87–94

    Article  PubMed  CAS  Google Scholar 

  • Aulinger IE, Peter SO, Schmid JE, Stamp P (2003) Rapid attainment of a doubled haploid line from transgenic maize (Zea mays L.) plants by means of anther culture. In Vitro Cell Dev Plant 39:165–170

    Article  Google Scholar 

  • Barloy D, Denis L, Beckert M (1989) Comparison of the aptitude for anther culture in some androgenic double haploid maize lines. Maydica 34:303–308

    Google Scholar 

  • Barnabás B, Fransz PF, Schel JHN (1987) Ultrastructural studies on pollen embryogenesis in maize (Zea mays L.). Plant Cell Rep 6:212–215

    Article  Google Scholar 

  • Barnabás B, Obert B, Kovács G (1999) Colchicine, an efficient genome-doubling agent for maize (Zea mays L.) microspores cultured in anthero. Plant Cell Rep 18:858–862

    Article  Google Scholar 

  • Barnabás B, Pónya Z, Szakács É, Timár I, Obert B, Preťová A (2001) Biotechnology and micromanipulation of sexual processes in flowering plants. Biologia 56:7–12

    Google Scholar 

  • Barnabás B (2003) Anther culture of maize (Zea mays L.). In: Maluszynski M, Kasha KJ, Forster BP, Szarejko I (eds) Doubled haploid production in crop plants a manual. Kluwer Academic Publishers, Dordrecht, p 428

    Google Scholar 

  • Barret P, Brinkman M, Dufour P, Murigneux A, Beckert M (2004) Identification of candidate genes for in vitro androgenesis induction in maize. Theor Appl Genet 109:1660–1668

    Article  PubMed  CAS  Google Scholar 

  • Bartošová Z, Obert B, Takáč T, Kormuťák A, Preťová A (2005) Using enzyme polymorphism to identify the gametic origin of flax regenerants. Acta Biol Cracov Bot 47:173–178

    Google Scholar 

  • Bourgoin-Greneche M, Lallemand J (1993) Electrophoresis and its application to the description of varieties. A presentation of the techniques used by GEVES. GEVES, La Miniere, France, p 63

  • Bourgoin-Greneche M, Lallemand J, Pouget R (1998) Technical reference manual for the isoenzymatic analysis of maize. F 78285, GEVES, Guyancourt, France, p 73

  • Büter B, Pescitelli SM, Berger K, Schmid JE, Stamp P (1993) Autoclaved and filter sterilized liquid media in maize anther culture—significance of activated charcoal. Plant Cell Rep 13:79–82

    Article  Google Scholar 

  • Cardy BJ, Stuber CW, Goodman MM (1980) Techniques for starch gel electrophoresis of enzymes from maize (Zea mays L.). Institute of Statistics Mimeograph Series No. 1317, North Carolina State University, Raleigh, p 87

  • Coumans MP, Sohota S, Swanson EB (1989) Plant development from isolated microspores of Zea mays L. Plant Cell Rep 7:618–621

    Google Scholar 

  • Čurn V (1995) Acid phosphatase and leucine aminopeptidase isozymes as biochemical markers of homogeneity in oil seed rape androgenetic lines. Plant Growth Regul 16:59–63

    Article  Google Scholar 

  • Chu CC (1981) The N6 medium and its applications to anther culture of cereal crops. In: Proc. sym. plant tissue culture, Beijing 1978, Pitman, Boston, pp 43–50

  • Dieu P, Beckert M (1986) Further studies of androgenic embryo production and plant regeneration from in vitro cultured anthers in maize (Zea mays L). Maydica 31:245–259

    Google Scholar 

  • Elmaghrabi A, Ochatt S (2006) Isoenzymes and flow cytometry for the assessment of true-to-typeness of calluses and cell suspensions of barrel medic prior to regeneration. Plant Cell Tissue Organ 85:31–43

    Article  CAS  Google Scholar 

  • Forster BP, Thomas WTB (2004) Doubled haploids in genetics and plant breeding. Plant Breed Rev 25:57–88

    Google Scholar 

  • Frova C (1990) Analysis of gene expression in microspores, pollen, and silks of Zea mays L. Sex Plant Reprod 3:200–206

    Article  Google Scholar 

  • Frova C, Binelli G, Ottaviano E (1987) Isozyme and HSP gene expression during male gametophyte development in maize. In: Isozymes: current topics in biological and medical research, vol 15: genetics, development, and evolution, pp 97–120

  • Genovesi AD, Collins GB (1982) In vitro production of haploid plants of corn via anther culture. Crop Sci 22:1137–1144

    Google Scholar 

  • Gorla MS, Frova C, Binelli G, Ottaviano E (1986) The extent of gametophytic-sporophytic gene expression in maize. Theor Appl Genet 72:42–47

    Article  Google Scholar 

  • Greneche M, Giraud G (1989) Manuel technique de reference pour l’analyse isoenzymatique du Mais. GEVES, Surgeres, France, p 78

    Google Scholar 

  • Jähne A, Lörz H (1995) Cereal microspore culture. Plant Sci 109:1–12

    Article  Google Scholar 

  • Jäger K, Kószegi DV, Barnabás B (2005) Regeneration capacity of microspore—derived structures produced in anther cultures of maize (Zea mays L.). Acta Physiol Plant 27:621–629

    Article  Google Scholar 

  • Ku MK, Cheng WC, You LC, Kuan HP, Huang CH (1978) Induction factors and morphocytological characteristic of pollen derived plants in maize (Zea mays L.). In: Proc. symp. plant tissue culture, 1978, Science Press, Peking, pp 35–45

  • Kuo CS, Sun AC, Wang YY, Gui YL, Gu SR, Miao SH (1978) Studies on induction of pollen plants and androgenesis in maize. Acta Bot Sin 20:204–209

    Google Scholar 

  • Maraschin SF, de Priester W, Spaink HP, Wang M (2005) Androgenic switch: an example of plant embryogenesis from the male gametophyte perspective. J Exp Bot 56:1711–1726

    Article  PubMed  CAS  Google Scholar 

  • Markert CL, Moller F (1959) Multiple forms of enzymes: tissues, cytogenic and species specific patterns. Proc Natl Acad Sci USA 45:753–763

    Article  PubMed  CAS  Google Scholar 

  • Mejza SJ, Morgant V, DiBona DE, Wong JR (1993) Plant regeneration from isolated microspores of Triticum aestivum. Plant Cell Rep 12:149–153

    Article  Google Scholar 

  • Miao SH, Kuo CS, Kwei YL, Sun AT, Ku SY, Lu WL, Wang YY (1978) Introduction of pollen plants of maize and observation of their progeny. In: Proc. symp. plant tissue culture, 1978, Science Press, Peking, pp 23–24

  • Múdry P, Dragúň M (2005a) Above standard description and evaluation of local populations of maize (Zea mays L.). In: Proceedings of 4th seminar: evaluation of plant genetic resources, VÚRV, Piešťany, pp 175–176

  • Múdry P, Dragúň M (2005b) Proteomic classification of self-pollinated lines of maize (Zea mays L.) and their single crosses—reality and perspectives of utilization. In: New achievements of genetics and breeding in agricultural crops 12, VÚRV, Piešťany, 23.–24.11.2005

  • Múdry P, Juráček L’ (1998) Biochemical and genetic biodiversity of maize lines on the base of polymorphism of selected types of enzymes. In: Short communications volume II, fifth congress 28 June–2 July 1998, European Society for Agronomy, Nitra, The Slovac Republic, pp 279–280

  • Múdry P, Juráček L’ (2001) Modifikovaná štandardizovaná metodika analýzy polymorfizmu jedenástich druhov enzýmov—molekulárnych značkovačov kukurice siatej (Zea mays L.). In: Biotechnologické metódy v šľachtení rastlín BIOS 2001, Zborník referátov zo VII. Vedeckej konferencie s medzinárodnou účasťou, SPU, Nitra

  • Nageli M, Schmid JE, Stamp P, Büter B (1999) Improved formation of regenerable callus in isolated microspore culture of maize: impact of carbohydrates, plating density and time transfer. Plant Cell Rep 19:177–184

    Article  CAS  Google Scholar 

  • Obert B, Orosz A, Kovács G, Barnabás B (1998) Study of the androgenic capacity in crosses between highly androgenic exotic DH lines and recalcitrant commercial inbreeds in maize anther culture. Novenytermeles 47:473–481

    Google Scholar 

  • Obert B, Preťová A, Büter B, Schmid JE (2000) Effect of different saccharides on viability of isolated microspores and androgenic induction in Zea mays. Biol Plant 43:125–128

    Article  CAS  Google Scholar 

  • Obert B, Barnabás B (2004) Colchicine induced embryogenesis in maize. Plant Cell Tissue Organ 77:283–285

    Article  CAS  Google Scholar 

  • Obert B, Szabó L, Mitykó J, Preťová A, Barnabás B (2005) Morphological events undergoing in cultures of mechanically isolated maize microspores. In Vitro Cell Dev Plant 41:775–782

    Article  Google Scholar 

  • Pescitelli SM, Johnson CD, Petolino JF (1990) Isolated microspore culture of maize: effect of isolation technique reduced temperature and sucrose level. Plant Cell Rep 8:628–631

    Article  Google Scholar 

  • Petolino JF, Jones AM (1986) Anther culture of elite genotypes of maize. Crop Sci 26:1072–1074

    Google Scholar 

  • Petolino JF, Jones AM, Thompson SA (1988) Selection for increased anther culture response in maize. Theor Appl Genet 76:157–159

    Article  Google Scholar 

  • Preťová A, de Ruijter NCA, van Lammeren AAM, Schel JHN (1993) Structural observations during androgenic microspore culture of the 4c1 genotype of Zea mays L. Euphytica 65:61–69

    Article  Google Scholar 

  • Preťová A, Obert B, Bartošová Z (2006) Haploid formation in maize, barley, flax and potato. Protoplasma 228:107–114

    Article  Google Scholar 

  • Redha A, Islam SMS, Büter B, Stamp P, Schmid JE (2000) The improvement in regenerated doubled haploids from anther culture of wheat by anther transfer. Plant Cell Tissue Organ 63:167–172

    Article  CAS  Google Scholar 

  • Shariatpanahi ME, Bal U, Heberle-Bors E, Touraev A (2006) Stresses applied for the re-programming of plant microspores towards in vitro embryogenesis. Physiol Plant 127:519–534

    Article  CAS  Google Scholar 

  • Shim YS, Kasha KJ, Simion E, Letarte J (2006) The relationship between induction of embryogenesis and chromosome doubling in microspore cultures. Protoplasma 228:79–86

    Article  PubMed  CAS  Google Scholar 

  • Stuber CW, Wendler JF, Goodman MN, Smith JSC (1988) Techniques and scoring procedures for starch gel electrophoresis of enzymes from maize (Zea mays L.). Technical Bulletin 286, North Carolina Agricultural Research Service, North Carolina State University, Raleigh, pp 1–87

  • Testillano PS, Georgiev S, Mogensen HL, Coronado MJ, Dumas C, Risueno MC, Matthys-Rochon E, Ting YC, Margaret YU, Wan Z (2004) Spontaneous chromosome doubling results from nuclear fusion during in vitro maize induced microspore embryogenesis. Chromosoma 112:342–349

    Article  PubMed  CAS  Google Scholar 

  • Ting YC, Yu M, Zheng WZ (1981) Improved anther culture of maize (Zea mays). Plant Sci Lett 23:139–145

    Article  Google Scholar 

  • Touraev A, Vicente O, Heberle-Bors E (1997) Initiation of microspore embryogenesis by stress. Trends Plant Sci 2:297–302

    Article  Google Scholar 

  • Tsay HS, Miao SH, Widholm JM (1986) Factors affecting haploid plant regeneration from maize anther culture. J Plant Physiol 126:33–40

    Google Scholar 

  • Wang M, Hoekstra S, Van Bergen S, Lamers GEM, Oppedijk BJ, Van der Heiden MW, De Priester W, Shilperoort RA (1999) Apoptosis in developing anthers and the role of ABA in this process during androgenesis in Hordeum vulgare L. Plant Mol Biol 39:489–501

    Article  PubMed  CAS  Google Scholar 

  • Wang M, Van Bergen S, Van Duijn B (2000) Insight into a key developmental switch and its importance for efficient plant breeding. Plant Physiol 124:523–530

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by grant APVT-51-002302 and VEGA 2/5079/5. We would like to thank Professor Beáta Barnabás for providing seeds of maize genotypes used in our studies. B. O. is grateful to Alexander von Humboldt Foundation (Bonn, Germany) for his research fellowship.

Author information

Authors and Affiliations

  1. Institute of Plant Genetics and Biotechnology, Slovak Academy of Sciences, Akademická 2, P. O. Box 39/A, 95007, Nitra, Slovak Republic

    Ľubica Uváčková, Bohuš Obert & Anna Preťová

  2. Faculty of Education, Department of Biology, Trnava University, Priemyselná St. 4, 91843, Trnava, Slovak Republic

    Pavol Múdry

  3. Institute of Cellular and Molecular Botany, University Bonn, Kirschalle 1, 53115, Bonn, Germany

    Bohuš Obert

Authors
  1. Ľubica Uváčková
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pavol Múdry
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bohuš Obert
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Anna Preťová
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bohuš Obert.

Additional information

Communicated by E. Lojkowska.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Uváčková, Ľ., Múdry, P., Obert, B. et al. Enzyme fingerprint analyses in tissue regenerated from anther culture of maize. Acta Physiol Plant 30, 779–785 (2008). https://doi.org/10.1007/s11738-008-0180-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11738-008-0180-4

Keywords

Search

Navigation