Czech J. Genet. Plant Breed., 2014, 50(2):123-129 | DOI: 10.17221/137/2013-CJGPB

Obtaining calli and regenerated plants in anther cultures of peaOriginal Paper

Sergey BOBKOV
All-Russia Research Institute of Legumes and Groat Crops, Orel, Russia

Pea (Pisum sativum L.) is a species for which there is no efficient method for the recovery of haploid plants yet. This research investigated the influence of various genotypes, nutrient media, and stress treatments on callus formation, embryogenesis and plant regeneration in anther cultures of pea. A wide range of pea genotypes and nutrient media was studied. Morphogenic calli were initiated on media supplemented with α-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and 2,4-dichlorophenoxyacetic acid (2,4-D) without application of stress treatments. Embryogenic calli and embryos were regenerated on media with low sucrose content in the presence of 2,4-D or indole-3-butyric acid (IBA) after cold stress (4°C) of isolated buds, alone or in combination with in vitro treatment of isolated anthers at higher temperatures (35-38°C). The efficiency of regeneration via shoot morphogenesis on different nutrient media and the peculiarities of regeneration from embryogenic calli were investigated. Green embryogenic calli initiated on 2,4-D were able to develop through shoot morphogenesis on a medium supplemented with BA and NAA. This process led to regeneration of hypertrophic embryos at various developmental stages. The origin of regenerated plants (i.e. from microspores or somatic anther cells) was estimated using marker alleles determining morphological traits. Almost all R0 regenerants derived from morphogenic calli originated from anther somatic cells.

Keywords: callus; embryo; haploid plant; microspore; regeneration; stress treatment

Published: June 30, 2014  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
BOBKOV S. Obtaining calli and regenerated plants in anther cultures of pea. Czech J. Genet. Plant Breed. 2014;50(2):123-129. doi: 10.17221/137/2013-CJGPB.
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Bobkov S.V. (2010): Isolated pea anther culture. Russian Agricultural Sciences, 6: 413-416. Go to original source...
    2. Chu C. (1978): The N6 medium and its applications to anther culture of cereal crops. In: Proc. Symp. Plant Tissue Culture. Science Press, Peking, 43-50.
    3. Croser J., Lulsdorf M., Davies P., Clarke H., Wilson J., Sidhu P., Grewal R., Allen K., Dament T., Warketin T., Vandenberg A., Siddique K. (2005): Haploid embryogenesis from chickpea and field pea - progress towards a routine protocol. In: Proc. Australian Branch of the IAPT & B. Perth, 71-82.
    4. Gamborg O., Eveleigh D.E. (1968): Culture methods and detection of gluconases in cultures of wheat and barley. Canadian Journal of Biochemistry, 46: 417-421. Go to original source... Go to PubMed...
    5. Gosal S.S., Bajaj Y.P.S. (1988): Pollen embryogenesis and chromosomal variation in anther culture of three food legumes - Cicer arientum, Pisum sativum and Vigna mungo. SABRAO Journal of Breeding and Genetics, 20: 51-58.
    6. Griga M. (1998): Direct somatic embryogenesis from shoot apical meristems of pea, and thidiazuron-induced high conversion rate of somatic embryos. Biologia Plantarum, 41: 481-495. Go to original source...
    7. Griga M. (2002): Morphology and anatomy of Pisum sativum somatic embryos. Biologia Plantarum, 45: 173-182. Go to original source...
    8. Griga M., Tejklova E., Novak F.J., Kubalakova M. (1986): In vitro clonal propagation of Pisum sativum L. Plant Cell Tissue Organ Culture, 6: 95-104. Go to original source...
    9. Gupta S. (1975): Morphogenetic response of haploid callus tissue of Pisum sativum (var. B22). Indian Agriculturalist, 19: 11-21.
    10. Gupta S., Ghosal K.K., Gadgil V.N. (1972): Haploid tissue culture of Triticum aestivum var. Sonalina and Pisum sativum var. B22. Indian Agriculturalist, 16: 277-278.
    11. Hussey G., Gunn H.V. (1984): Plant production in pea (Pisum sativum L. cvs Puget and Upton) from long-term callus with superficial meristems. Plant Science Letters, 37: 143-148. Go to original source...
    12. Kubaláková M., Tejklová E., Griga M. (1988): Some factors affecting root formation on in vitro regenerated pea shoot. Biologia Plantarum, 30: 179-184. Go to original source...
    13. Kysely W., Myers J.R., Lazzeri P.A., Collins G.B., Jacobsen H.J. (1987): Plant regeneration via somatic embryogenesis in pea. Plant Cell Reports, 6: 305-308. Go to original source... Go to PubMed...
    14. Lazzeri P.A., Hildebrand D.F., Collins D.B. (1987): Soybean somatic embryogenesis: effect of nutritional and сhemical factors. Plant Cell Tissue Organ Culture, 10: 209-220. Go to original source...
    15. Lehminger-Mertens R., Jacobsen H.J. (1989): Plant regeneration from pea protoplast via somatic embryogenesis. Plant Cell Reports, 8: 379-382. Go to original source... Go to PubMed...
    16. Lulsdorf M.M., Croser J.S., Ochatt S. (2011): Androgenesis and doubled-haploid production in food legumes. Chapter 11. In: Pratap A., Kumar J. (eds): Biology and Breeding of Food Legumes. CABI, Oxfordshire, 336-347. Go to original source...
    17. Murashige T., Skoog F. (1962): A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15: 473-497. Go to original source...
    18. Ochatt S.J., Mousset-Declas C., Rancillac M. (2000): Fertile pea plants regenerate from protoplasts when calluses have not undergone endoreduplication. Plant Science, 156: 177-183. Go to original source... Go to PubMed...
    19. Ochatt S.J., Delaitre C., Lionneton E., Hughette O., Patat-Ochatt E.M., Kahane R. (2005): One team, PCMV and one approach, in vitro biotechnology, for one aim, the breeding of quality plants with a wide array of species. In: Ramdane D. (ed.): Crops: Growth, Quality And Biotechnology. WFL, Helsinki, 1038-1067.
    20. Ochatt S., Pech C., Grewal R., Coreux C., Lulsdorf M., Jacas L. (2009): Abiotic stress enhances androgenesis from isolated microspores of some legume species (Fabaceae). Journal of Plant Physiology, 166: 1314-1328. Go to original source... Go to PubMed...
    21. Ribalta F., Croser J., Ochatt S. (2012): Flow cytometry enables identification of sporophytic eliciting stress treatments in gametic cells. Journal of Plant Physiology, 169: 104-110. Go to original source... Go to PubMed...
    22. Sidhu R., Davies P. (2005): Pea anther culture: callus initiation and production of haploid plants. In: Proc. Australian Branch of the IAPT & B. Perth, 180-186.
    23. Stejskal J.H., Griga M. (1992): Somatic embryogenesis and plant regeneration in Pisum sativum L. Biologia Plantarum, 34: 15-22. Go to original source...
    24. Telmer C.A., Simmonds D.H., Newcomb W. (1992): Determination of development stage to obtain high frequencies of embryogenic microspores in Brassica napus. Physiologia Plantarum, 84: 417-424. Go to original source...
    25. Tetu T., Sangwan R.S., Sangwan-Noreel B.S. (1990): Direct somatic embryogenesis and organogenesis in cultured immature zygotic embryos of Pisum sativum L. Journal of Plant Physiology, 137: 102-109. Go to original source...
    26. Uvarov V.N. (1993): Lupinoid - a new type of determinant growth in pea. Selekciya i semenovodstvo, 5-6: 15-20. (in Russian)

    This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY NC 4.0), which permits non-comercial use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.


    Return to the content