pp 1–10 | Cite as

Efficient and rapid in-vitro plantlet regeneration via somatic embryogenesis in ornamental bananas (Musa spp.)

  • Nandhakumar NatarajanEmail author
  • Sathish Sundararajan
  • Sathishkumar Ramalingam
  • Perinba Suresh Chellakan
Original Article


A simple and efficient protocol developed for plantlet regeneration through in vitro somatic embryogenesis in three ornamental bananas. In vitro propagation, especially somatic embryogenesis has been a method of choice to obtain homogenous planting material in ornamental bananas. In the present investigation, true-to-type plants via somatic embryogenesis were achieved in three ornamental bananas viz. Musa laterita, Musa beccarii and Musa velutina using immature male flower bud as explants. The evaluation was carried out at two stages of somatic embryogenesis, induction, and maturation of somatic embryos. Embryogenic calli desiccated up to 2 h at 25 ± 1 °C resulted in higher frequencies of embryo induction and maturation compared to non-desiccated embryos. The plantlets generated were hardened, and the genetic fidelity of the plantlets was confirmed using SRAP markers. It was observed that banana plants derived via somatic embryogenesis displayed normal phenotype as that of plants grown from a single elite mother plant. In the present study, the successful establishment of plants from male flower bud was achieved in 6 to 7 months which is a relatively short duration compared to other reports involving different species of banana. The protocol developed could be useful for large-scale micropropagation studies in these commercially important ornamental bananas.


Ornamental banana Male flower bud Genetic fidelity Somatic embryogenesis 



Author Sathish S acknowledges Indian Council for Medical Research, New Delhi, India (No.3/1/2/102/2018-Nut.) for fellowship support.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest.


  1. Ali KS, ELhassan A, Ehiweris O, Maki HE (2013) Embryogenesis and plantlet regeneration via immature male flower culture of banana (Musasp.) cv. Grand Nain. J Forest Prod Indus 2:48–52Google Scholar
  2. Arnold S, Sabala I, Bozhkov P, Dyachok J, Filonova L (2002) Developmental pathways of somatic embryogenesis. Plant Cell Tissue Organ Cult 69:233–249. CrossRefGoogle Scholar
  3. Becker DK, Dugdale B, Smith MK, Harding RM, Dale JL (2000) Genetic transformation of Cavendish banana (Musa spp. AAA group) cv grand Nain via microprojectile bombardment. Plant Cell Rep 19:229–234. CrossRefGoogle Scholar
  4. Cote FX, Domergue R, Monmarson S, Schwendiman J, Teisson C, Escalant JV (1996) Embryogenic cell suspensions from the male flower of Musa AAA cv. Grand Naine. Physiol Plant 97:285–290. CrossRefGoogle Scholar
  5. Daniells JW, Geering AD, Bryde NJ, Thomas JE (2001) The effect of Banana streak virus on the growth and yield of dessert bananas in tropical Australia. Ann Appl Biol 139:51–60CrossRefGoogle Scholar
  6. Duangkongsan W, Promtab W (2014) In vitro multiplication of Musa laterita Roxb. J Agric Technol 10:1173–1181Google Scholar
  7. FAO Stat. FAO Stat accessed vides Accessed 12/8/2018
  8. Fonsah EG, Wallace R, Krewer G (2008) Why are there seeds in my banana? A look at ornamental banana. J Food Distrib Res 39:67–71Google Scholar
  9. Ganapathi TR, Higgs NS, Balint-Kurti PJ, Arntzen CJ, May GD, Van Eck JM (2001) Agrobacterium-mediated transformation of embryogenic cell suspensions of the banana cultivar Rasthali (AAB). Plant Cell Rep 20:157–162. CrossRefGoogle Scholar
  10. Giridhar P, Vaddadi S, Matam P, Shreelakshmi SV (2018) TDZ induced diverse in vitro responses in some economically important plants. In: Thidiazuron: from urea derivative to plant growth regulator. Springer, Singapore, pp 329–341. CrossRefGoogle Scholar
  11. Häkkinen M (2001) Musa laterita: an ornamental banana. Fruit Gard 33:6–7Google Scholar
  12. Häkkinen M (2007) Ornamental bananas: focus on Rhodochlamys. Chron Horticult 47:7–12Google Scholar
  13. Häkkinen M, Vare H (2008) Typification and check-list of Musa L. names (Musaceae) with nomenclatural notes. Adansonia 30:63–112Google Scholar
  14. Heslop-Harrison JS, Schwarzacher T (2007) Domestication, genomics and the future for banana. Ann Bot 100:1073–1084. CrossRefGoogle Scholar
  15. Hiraga S, Minakawa H, Takahashi K, Takahashi R, Hajika M, Harad K, Ohtsubo N (2007) Evaluation of somatic embryogenesis from immature cotyledons of Japanese soybean cultivars. Plant Biotechnol 24:435–440. CrossRefGoogle Scholar
  16. Hrahsel L, Basu A, Sahoo L (2014) In vitro propagation and assessment of the genetic Fidelity of Musa acuminata (AAA) cv. Vaibalhla derived from immature male flowers. App Biochem Biotechnol 172:1530. CrossRefGoogle Scholar
  17. Husin N, Jalil M, Othman RY, Khalid N (2014) Enhancement of regeneration efficiency in banana (Musa acuminata cv. Berangan) by using proline and glutamine. Sci Hortic 168:33–37. CrossRefGoogle Scholar
  18. Israeli Y, Ben-Bassat D, Reuveni H (1996) Selection of stable banana clones which do not produce dwarf somaclonal variants during in vitro culture. Sci Hortic 67:197–205. CrossRefGoogle Scholar
  19. Itoh JI, Sato Y, Nagato Y, Matsuoka M (2006) Formation, maintenance and function of the shoot apical meristem in rice. Plant Mol Biol 60:827–842. CrossRefGoogle Scholar
  20. Jin S, Mushke R, Zhu H, Tu L, Lin Z, Zhang Y, Zhang X (2008) Detection of somaclonal variation of cotton (Gossypium hirsutum) using cytogenetics, flow cytometry and molecular markers. Plant Cell Rep 27:1303–1316. CrossRefGoogle Scholar
  21. Joe A, Sabu M, Sreejith PE (2013) A new variety of Musa velutina H.Wendl. & Drude (Musaceae) from Assam, North-East India. Plant Syst Evol 300:13–17. CrossRefGoogle Scholar
  22. Kahia J, Ndaruhutse F, Waweru B, Bonaventure N, Mutaganda A, Sallah PY, Asiimwe T (2015) In vitro propagation of two elite cooking banana cultivars-FHIA 17 and INJAGI. Intl J Biotechnol Mol Biol Res 6:40–47CrossRefGoogle Scholar
  23. Khalil SM, Elbanna AAM (2004) Highly efficient somatic embryogenesis and plant regeneration via suspension cultures of banana (Musa spp.). Arab J Biotechnol 7:99–110Google Scholar
  24. Krewer G, Fonsah EG, Rieger M, Wallace R, Linvill D, Mullinix B (2008) Evaluation of commercial banana cultivars in southern Georgia for ornamental and nursery production. Hort Technol 18:529–535. CrossRefGoogle Scholar
  25. Kulkarni VM, Bapat VA (2013) Somatic embryogenesis and plant regeneration from cell suspension cultures of Rajeli (AAB), an endangered banana cultivar. J Plant Biochem Biotechnol 22:132–137. CrossRefGoogle Scholar
  26. Kumari N, Misra P (2016) Mass in-vitro micropropagation of banana (Musa sp.). Int J Plant Protec 9:204–210CrossRefGoogle Scholar
  27. Martin KP, Pachathundikandi SK, Zhang CL, Slater A, Madassery J (2006) RAPD analysis of a variant of banana (Musa sp.) cultivar Grande Naine and its propagation via shoot tip culture. In vitro Cell Dev Biol-Plant 42:188–192. CrossRefGoogle Scholar
  28. Meenakshi S, Shinda BN, Suprasanna P (2011) Somatic embryogenesis from immature male flowers and molecular analysis of regenerated plants in banana, LAL KELA (AAA). Electronic J Fruit Ornam Plant Res 19:15–30Google Scholar
  29. Mizukami M, Takeda T, Satonaka H, Matsuoka H (2008) Improvement of propagation frequency with two-step direct somatic embryogenesis from carrot hypocotyls. Biochem Eng J 38:55–60. CrossRefGoogle Scholar
  30. Morel G, Wetmore RH (1951) Fern callus tissue culture. Am J Bot 38:141-143Google Scholar
  31. Morais-Lino LS, Santos-Serejo JAL, Amorim EP, de Santana JRF, Pasqual M, de Oliveira e Silva S (2015) Somatic embryogenesis, cell suspension, and genetic stability of banana cultivars. In Vitro Cell Dev Biol – Plant 52:99–106. CrossRefGoogle Scholar
  32. Nandhakumar N, Kumar KK, Sudhakar D, Soorianathasundaram K (2018) Plant regeneration, developmental pattern and genetic fidelity of somatic embryogenesis derived Musa spp. J Genet Eng Biotechnol 16:587–598. CrossRefGoogle Scholar
  33. Navarro C, Escobedo RM, Mayo A (1997) In vitro plant regeneration from embryogenic cultures of a diploid and a triploid. Cavendish banana. Plant Cell Tissue Organ Cult 51:17–25. CrossRefGoogle Scholar
  34. Rajoriya P, Singh VK, Lall NJR (2018) Optimizing the effect of plant growth regulators on in vitro micro propagation of Indian red banana (Musa acuminata). J Pharmaco Phytochem 8:628–634Google Scholar
  35. Rashid K, Nezhadahmadi A, Othman RY, Ismail NA, Azhar S, Efzueni S (2012) Micropropagation of ornamental plant Musa Beccarii through tissue culture technique using suckers and male buds as explants. Life Sci J 9:2046–2053Google Scholar
  36. Ray T, Dutta I, Saha P, Das S, Roy SC (2006) Genetic stability of three economically important micropropagated banana (Musa spp.) cultivars of lower Indo-Gangetic plains, as assessed by RAPD and ISSR markers. Plant Cell Tissue Organ Cult 85:11–18. CrossRefGoogle Scholar
  37. Resmi L, Nair AS (2007) Plantlet production from the male inflorescence tips of Musa accuminata cultivars from South India. Plant Cell Tissue Organ Cult 83:333–338. CrossRefGoogle Scholar
  38. Sabu M, Joe A, Sreejith PE (2013) Musa velutina subsp. markkuana (Musaceae): a new subspecies from northeastern India. Phytotaxa 92:49–54CrossRefGoogle Scholar
  39. Santos-Serejo JA, Souza EH, Souza FVD, Soares TL, Silva SO (2007) Morphological characterization of ornamental banana. Magistra 19:326–332Google Scholar
  40. Shepherd K (1999) Cytogenetics of the genus Musa. International network for the improvement of banana and plantain [INIBAP], France: MontpellierGoogle Scholar
  41. Soumya SL, Nair BR (2018) Genetic fingerprinting of two species of Averrhoa using RAPD and SRAP markers. Agric Res 7:402. CrossRefGoogle Scholar
  42. Srinivas I, Ganapathy TR, Suprasanna P, Bapat VA (2006) Desiccation and ABA improves conversion of somatic embryos of banana cv. Rasthali (AAB). Indian J Biotechnol 5:521–526Google Scholar
  43. Styer DJ (1985) Bioreactor technology for plant propagation. In Tissue culture in forestry and agri:117–130.
  44. Takayama S, Akita M (1994) The types of bioreactors used for shoots and embryos. Plant Cell Tissue Organ Cult 39:147–156. CrossRefGoogle Scholar
  45. Tripathi JN, Oduor RO, Tripathi L (2015) A high-throughput regeneration and transformation platform for production of genetically modified banana. Front Plant Sci 6:1025. Google Scholar
  46. Uma S, Sathiamoorthy S, Durai P (2005) Banana Indian genetic resources and catalogue. National Research Centre for Banana (ICAR), Tiruchirapalli, India. 474Google Scholar
  47. Uma S, Saraswathi MS, Durai P, Sathiamoorthy S (2006) Diversity and distribution of section Rhodochlamys (genus Musa, Musaceae) in India and breeding potential for banana improvement programmes. Plant Genetic Resources Newsletter 146:17–23Google Scholar
  48. Wallace R, Häkkinen M (2009) Musa x georgiana, a new intersectional hybrid banana with edible banana breeding relevance and ornamental potential. Nord J Bot 27:182–185. CrossRefGoogle Scholar
  49. Youssef M, James A, Mayo-Mosqueda A, Ku-Cauich JR, Grijalva-Arango R, Escobedo-GM RM (2010) Influence of genotype and age of explant source on the capacity for somatic embryogenesis of two Cavendish banana cultivars (Musa acuminate Colla, AAA). Afr J Biotechnol 9:2216–2223Google Scholar
  50. Yuan ZC, Liu P, Saenkham P, Kerr K, Nester EW (2008) Transcriptome profiling and functional analysis of agrobacterium tumefaciens reveals a general conserved response to acidic conditions (pH 5.5) and a complex acid-mediated signaling involved in agrobacterium-plant interactions. J Bacteriol 190:494–507. CrossRefGoogle Scholar

Copyright information

© Institute of Molecular Biology, Slovak Academy of Sciences 2019

Authors and Affiliations

  • Nandhakumar Natarajan
    • 1
    Email author
  • Sathish Sundararajan
    • 2
  • Sathishkumar Ramalingam
    • 2
  • Perinba Suresh Chellakan
    • 1
  1. 1.Department of HorticultureNorth Eastern Hill UniversityWest Garo HillsIndia
  2. 2.Plant Genetic Engineering Laboratory, Department of BiotechnologyBharathiar UniversityCoimbatoreIndia

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