Plant regeneration through somatic embryogenesis from immature and mature zygotic embryos of Musa acuminata ssp. burmannica

  • S. Uma
  • S. Lakshmi
  • M. S. Saraswathi
  • A. Akbar
  • M. M. Mustaffa
Plant Tissue Culture


A simple and efficient protocol has been developed for in vitro regeneration of M. acuminata ssp. burmannica (AA) plants. Somatic embryos were produced when immature and mature zygotic embryo explants were cultured on Murashige and Skoog medium supplemented with plant growth regulators 2,4-dichlorophenoxyacetic acid; (2,4-D), picloram or benzyl adenine and indole acetic acid. In general, immature embryos responded better than mature embryos. Callus proliferation was highest in medium supplemented with 2,4-D (4.5 μM). Subsequent transfer of callus to fresh medium produced rapidly proliferating embryogenic calli. Embryogenic calli were maintained in complete darkness for 15 d followed by cycles of 8 h dark and 16 h light, under white fluorescent lamps with a light intensity of 3,000 lm/m2 and at temperature of 28 ± 2°C. Regeneration of embryogenic calli into plantlets was higher for immature embryos (76.6%) than for mature embryos (50.6%). This plant regeneration protocol using mature or immature zygotic embryos, via somatic embryogenesis, has significant potential to improve germination efficiencies of hybrid progenies used in conventional breeding strategies. Furthermore, tests on seed storage showed that seed viability rapidly decline after harvesting and was negligible after 9 mo of storage. This indicates using freshly harvested seeds as explant material is necessary for maximizing the tissue culture response.


Banana and plantains Seed storage Musa acuminata ssp. burmannica Plant regeneration Somatic embryogenesis Zygotic embryo 


  1. Afele JC, De Langhe E (1991) Increasing in vitro germination of Musa balbisiana seed. Plant Cell Tiss Organ Cult 27:33–36CrossRefGoogle Scholar
  2. Aslam M, Zafar AR, Siddiqi TO (2010) Standardization of seed viability protocol for Pinus wallichiana A. B. Jackson in Kashmir, India. Int J Pharm Sci Rev Res 4:93–98Google Scholar
  3. Bakry F, Horry JP (1992) Tetraploid hybrids from interploid 3x/2x crosses in cooking bananas. Fruits 47:641–655Google Scholar
  4. Botti C, Vasil IK (1983) Plant regeneration by somatic embryogenesis from parts of cultured mature embryos of Pennisetum americanum (L.) K. Schum. Z. Pflanzenphysiol 111:319–325Google Scholar
  5. Brown DCW, Thorpe A (1995) Crop improvement through tissue culture. World J Microbiol Biotechnol 11:409–415CrossRefGoogle Scholar
  6. Côte F, Domergue R, Monmarson S, Schwendiman J, Teisson C, Escalant J (1996) Embryogenic cell suspension from the male flower of Musa AAA cv. Grande Naine (AAA). Physiol Plant 97:285–290CrossRefGoogle Scholar
  7. Cox EA, Stotzky G, Goos RD (1960) In-vitro culture of Musa balbisiana Colla embryos. Nature 185:403–404CrossRefGoogle Scholar
  8. Cronauer-Mitra SS, Krikorian AD (1988) Plant regeneration via somatic embryogenesis in the seeded diploid banana Musa ornata Roxb. Plant Cell Rep 7:23–25PubMedCrossRefGoogle Scholar
  9. de Oliveira e Silva S, da Silva KM, de F Borges M, de Oliveira RP (1999) Pollination and culture of banana embryos. Infomusa 8:24–26Google Scholar
  10. Escalant JV, Teisson C (1988) Somatic embryogenesis in Musa sp. C.R. Seances-Acad Sci Ser 306:277–281Google Scholar
  11. Escalant JV, Teisson C (1989) Somatic embryogenesis and plants from immature zygotic embryos of species Musa acuminata and Musa balbisiana. Plant Cell Rep 7:665–668Google Scholar
  12. Fortescue JA, Turner DW (2011) Reproductive biology. In: Pillay M, Tenkouano A (eds) Banana Breeding Progress and Challenges. CSRC, Taylor and Francis Group, Florida, pp 145–180CrossRefGoogle Scholar
  13. Johri BM, Rao PS (1984) Experimental embryology. In: Johri BM (ed) Embryology of Angiosperms. Springer, Berlin, pp 744–802CrossRefGoogle Scholar
  14. Krishnaraj S, Vasil IK (1995) Somatic embryogenesis in herbaceous monocots. In: Thorpe TA (ed) In vitro embryogenesis in plants. Kluwer, Dordrecht, pp 417–470CrossRefGoogle Scholar
  15. Morel G (1950) Sur la culture des tissus de deux monocotyl’edones. C R Acad Sci Paris 230:1099–1101Google Scholar
  16. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  17. NRCB—National Research Centre for Banana, (2011) Annual report. NRCB, Tiruchirappalli, pp 16–17Google Scholar
  18. Pancholi N, Wetten A, Caligari (1995) Germination of Musa velutina seeds: comparison of in vivo and in vitro systems. In Vitro Cell Dev Biol Plant 31:127–130CrossRefGoogle Scholar
  19. Pillay M, Cullis CA, Talengera D, Tripathi L (2011) Propagation methods in Musa. In: Pillay M, Tenkouano A (eds) Banana Breeding—Progress and challenges. Taylor and Francis Group, vol II. pp 285–303Google Scholar
  20. Simmonds NW (1952) The germination of banana seeds. Trop Agric Trin 37:211–221Google Scholar
  21. Simmonds NW (1959) Experiments on the germination of banana seeds. Trop Agric Trin 36:259–273Google Scholar
  22. Simmonds NW (1966) Bananas, 2nd edn. Longman, LondonGoogle Scholar
  23. Uma S, Lakshmi S, Saraswathi MS, Akbar A, Mustaffa MM (2011) Embryo rescue and plant regeneration in banana (Musa spp.). Plant Cell Tiss Organ Cult 105:105–111CrossRefGoogle Scholar
  24. Uma S, Mustaffa MM, Saraswathi MS, Durai P (2009) Exploitation of diploids in Indian banana breeding programmes. Acta Hort (ISHS) 897:215–223Google Scholar
  25. Uma S, Saraswathi MS, Sevarajan R (2010) Banana and plantains. In: Singh HP, Parthasarathy VA, Nirmal Babu K (eds) Advances in Horticulture Biotechnology—regeneration systems—fruit crops, plantation crops and spices, vol I. Westville, New Delhi, pp 15–72Google Scholar
  26. Walker KA, Sato SJ (1981) Morphogenesis in callus tissue of Medicago sativa: The role of ammonium ion in somatic embryogenesis. Plant Cell Tiss Organ Cult 1:109–121CrossRefGoogle Scholar
  27. Walker KA, Yu PC, Sato SJ, Jaworski EG (1978) The hormonal control of organ formation in callus of Medicago sativa L. cultured in vitro. Am J Bot 65:654–659CrossRefGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2012

Authors and Affiliations

  • S. Uma
    • 1
  • S. Lakshmi
    • 1
  • M. S. Saraswathi
    • 1
  • A. Akbar
    • 1
  • M. M. Mustaffa
    • 1
  1. 1.National Research Centre for BananaTrichyIndia

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