Skip to main content
SpringerLink
Log in

Multiple shoot induction in zygotic embryos: a strategy for acceleration of banana breeding

  • Original Article
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

The presence of residual female fertility in most of the parthenocarpic banana accessions encourages the banana breeder to develop new hybrids through conventional breeding. Desirable trait can be fixed in the first generation of hybrid progenies, but the evaluation of these hybrids in field is the time-consuming process owing to non-availability of uniform suckers/planting material. This can be overcome by developing multiple shoots from single embryo in a short period of time through embryo culture. A protocol for in vitro multiplication of plantlets from zygotic embryos was standardized in seeded accessions. Multiple shoots from zygotic embryos were achieved up to 55.2% and 64.1% in seeded accessions of Musa acuminata and M. velutina respectively in medium supplemented with 17.76 µM of BAP. The Single shoot derived (only germination) from zygotic embryos was decapitated and the apical meristem were disturbed for further multiple shoot formation in media supplemented with 17.76 µM of BAP. Present studies revealed that in total 75% and 91% of the M. acuminata and M.velutina embryos were able to produce multiple shoot from single embryo by manipulating the media composition and decortications technique. The above protocol was applied for zygotic embryos obtained from controlled pollination (18 cross combinations) and open pollination (nine accessions) of various genomic groups (ABB, AAB, AA). The multiple shoots derived from zygotic embryos and plantlet germinated from zygotic embryos was examined for genetic fidelity analysis by SSR markers.

Key message

The protocol for multiple shoot formation from single zygotic embryo under in vitro culture developed in this study will accelerate the banana breeding program.

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

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Ahmed KZ, Remy S, Sagi L, Swennen R (2006) Germination of Musa balbisiana seeds and embryos. XVII International ACROBAT Meeting, Joinville, Santa Catarina State, Brasil, pp 510–512

  • Ali N, Skirvin R, Splittstoesser WE, George WL (1991) Germination and regeneration of plants from old cucumber seed. HortScience 26(7):917–918

    Article  Google Scholar 

  • Allam EK, Othman BA, Sawy EL, Thabet SD (2000) Establishment of an aseptic culture of banana micropropagation in vitro. Ann Agric Sci 38:1121–1136

    Google Scholar 

  • Andrus L, Ding Z, Jiang F, Jin B, Ding X, Sun J, Guiyuan L (1971) Induction and identification of hexadecaploid of Pinellia ternate. J Euphytica 186(2):479–488

    Google Scholar 

  • Arun K, Uma S, Saraswathi MS, Backiyarani S, Durai P (2013) Effects of whole seed priming on the in vitro germination of hybrid banana embryos (Musa spp.). Seed Sci Technol 41:439–451

    Article  Google Scholar 

  • Asif MJ, Mak C, Othman RY (2001) In vitro zygotic embryo culture of wild Musa accuminata ssp. Malaccensis and factors affecting germination and seedling growth. Plant Cell Tiss Org Cult 67:267–270

    Article  CAS  Google Scholar 

  • Backiyarani S, Uma S, Maria Doss A, Saraswathi MS, Selvaraj V, Arun K, Durai P (2016) Enhancing the seed set in banana using antitoxins in National Conference on Fruit Breeding in Tropics and subtropics-An Indian Perspective. ICAR-Indian Institute of horticultural Research, Bengaluru, p 116

  • Backiyarani S, Chandrasekar A, Uma S, Saraswathi MS (2019) MusatransSSRDB (a transcriptome derived SSR database)—an advanced tool for banana improvement. J Biosci 43:110–116

    Google Scholar 

  • Batte M, Swennen R, Uwimana B, Akech V, Brown A, Tumuhimbise R, Hovmalm HP, Geleta M, Ortiz R (2019) Crossbreeding East African highland bananas: lessons learnt relevant to the botany of the crop after 21 years of genetic enhancement. Front Plant Sci 10:81

    Article  PubMed  PubMed Central  Google Scholar 

  • Bhattacharya J, Renukdas NN, Khuspe SS, Rawal SK (2003) Multiple shoot regeneration from immature embryo explants of papaya. Biol Plant 47(3):327–331

    Article  Google Scholar 

  • Burgos-Hernández M, Castillo-Campos G, Mata-Rosas M, González D, Vovides AP, Murguía-González J (2014) Seed germination of wild banana Musa ornata (Musaceae). Seed Sci Technol 42:16–27

    Article  Google Scholar 

  • Chang FC, Yang CM (1992) Allelopathic potential of purple nutsedge (Cyperusrotundus L.) and barnyardgrass (Echinochloa crus-galli (L.) Beavu.) on corn (Zea mays L.) weed residues effect on corn emergence and seedling growth. J Agric Res China 41:9–23

    Google Scholar 

  • Crouch JH, Crouch HK, Tenkouano A, Ortiz R (1999) VNTR-based diversity analysis of 2x and 4x full-sib Musa hybrids. Electron J Biotechnol 2:130–139

    Article  Google Scholar 

  • Dayarani M, Dhanarajan MS, Arun K, Uma S, Narayani Padma (2014) Embryo culture and embryo rescue studies in wild Musa spp. (Musa ornata). J Appl Hort 16(2):126–130

    Google Scholar 

  • Diro M, Van Staden J (2003) In vitro regeneration of Ensete ventricosum from zygotic embryos of stored seeds. S Afr J Bot 69:364–369

    Article  Google Scholar 

  • Dissanayaka NP, Kodikara KAS, Vithanage DS, Krishnarajah SA, Rubasinghe MK, Dayananda TG (2015) Effects of 6-benzylaminopurine (BAP) treatment on seed germination and seedling vigour of endemic herb exacumtrinervium L. in Sri Lanka: conservation strategy. J Univ Ruhuna 1:14–20

    Article  Google Scholar 

  • Ferreira CF, Silva SO, Damasceno Sobrinho LP, Damascena SCS, Alves FSAA, Paz OP (2004) Molecular characterization of banana (AA) diploid with contrasting levels of black and yellow sigatoka resistance. Am J Appl Sci 1:276–278

    Article  CAS  Google Scholar 

  • Franklin G, Jeyachandran R, Ignicimuthu S (2000) Factors affecting regeneration of pigeon pea (Cajanuscajan L. Millsp) from mature embryonal axes. Plant Growth Regul 30:31–36

    Article  CAS  Google Scholar 

  • Ganeshan S, Baga M, Harvey BL, Rossnagel BG, Scoles GJ, Chibbar RN (2003) Production of multiple shoots from thidiazuron-treated mature embryos and leaf-base/apical meristems of barley (Hordeum vulgare). Plant Cell Tiss Org Cult 73:57–64

    Article  CAS  Google Scholar 

  • Gawel NL, Jarret RL (1991) A modified CTAB DNA extraction procedure of Musa and Ipomoea. Plant Mol Biol Report 9:262–266

    Article  CAS  Google Scholar 

  • González de León D, Fauré S (1993) Genetic mapping of the banana diploid genome: toward an integrated approach to the study of the Musa genome and the use of molecular marker technologies in Musa breeding. In: Proceedings of the workshop on “Biotechnology applications for banana and plantain improvement”, San Jose, Costa Rica (edited by INIBAP, Montpellier, France) pp 29

  • Grafi G, Barak S (2014) Stress induces cell dedifferentiation in plants. BBA-Gene Regul Mech 1849(4):378–384

    Google Scholar 

  • Guan KL (1991) Studies on complex dormancy of Cornus officinalis seed. Chin J of Bot 3:145–150

    Google Scholar 

  • Heslop-Harrison TS (2007) Domestication, genomics and the future for banana. Ann Bot 100:1073–1084

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Johri BM, Rao PS (1984) Experimental embryology. In: Johri BM (ed) Embryology of angiosperms. Springer-Verlag, Berlin, pp 744–802

  • Kadota M, Niimi Y (2003) Effects of cytokinin types and their concentrations on shoot proliferation and hyperhydricity in in vitro pear cultivar shoots. Plant Cell Tiss Org Cult 72:261–265

    Article  CAS  Google Scholar 

  • Klem M, Balla J, Machackova I, Eder J, Prochazka S (2004) The uptake and metabolism of benzyl amino purine in tobacco (Nicotiana tabacum L.) and cucumber (Cucumissativus L.) explants. Plant Growth Regul 31:135–142

    Article  Google Scholar 

  • Köhler C, Hennig L, Spillane C, Stephane P, Gruissem W, Grossniklaus U (2003) The polycomb-group protein MEDEA regulates seed development by controlling expression of the MADS-box gene PHERES. Genes Dev 17:1540–1553

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Laliberté B (2016) Global strategy for the conservation and use of Musa (banana) genetic resources: a consultative document prepared by the Global Musa Genetic Resources Network (Musa Net). Bioversity International, Rome

    Google Scholar 

  • Mamidala P, Nanna RS (2011) Effect of genotype, explants source and medium on in vitro regeneration of tomato. Int J Genet Mol Biol 3:45–50

    CAS  Google Scholar 

  • Manchanda P, Gosal SS (2011) Molecular assessment of genetic fidelity of micropropagated banana (Musa acuminata L.) using SSR markers. Int J Plant Res 24(2):91–101

    Google Scholar 

  • Mensuali-Sodi A, Panizza M, Tognoni F (1995) Endogenous ethylene requirement for adventitious root induction and growth in tomato cotyledons and lavandin microcuttings in vitro. Plant Growth Regul 17:205–212

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Ortiz R (1997) Occurrence and inheritance of 2n pollen in Musa. Ann Bot 79:449–453

    Article  Google Scholar 

  • Ortiz R, Vuylsteke D (1995) Factors influencing seed set in triploid musa spp. L. and production of euploid hybrids. Ann Bot 75(2):151–155

    Article  Google Scholar 

  • Palanivel S, Parvathi S, Jayabalan N (2002) Callus induction and plantlet regeneration from mature cotyledonaury segments of groundnut (Arachis hypogaea L.). J Plant Biol 45(1):22–27

    Article  Google Scholar 

  • Pandey H, Nandi SK, Nadeem M, Palni LM (2000) Chemical stimulation of seed germination in Aconitum heterophyllum Wall, and A. balfourii Stapf.: important Himalayan species of medicinal value. International Seed Testing Association. Seed Sci Technol 28:39–48

    Google Scholar 

  • Peschke VM, Phillips RL (1992) Genetic implications of somaclonal variation in plants. Adv Genet 30:41–75

    Article  CAS  Google Scholar 

  • Punyasingh K (1947) Chromosome numbers in crosses of diploid, triploid and tetraploid maize. Genetics 32:541–554

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ramírez-Mosqueda MA, Iglesias-Andreu LG (2015) Indirect organogenesis and assessment of somaclonal variation in plantlets of Vanilla planifolia Jacks. Plant Cell Tiss Org Cult 123(3):657–664

    Article  Google Scholar 

  • Rao MS, Purohit SD (2006) In vitro shoot bud differentiation and plantlet regeneration in Celastrus paniculatus Wild. Biol Plant 50:501–506

    Article  CAS  Google Scholar 

  • Ravishankar KV, Raghavendra KP, Athani V, Rekha A, Sudeepa K, Bhavya D, Srinivar V, Ananad L (2013) Development and characterisation of microsatellite markers for wild banana (Musa balbisiana). J Hort Sci Biotechnol 88:605–609

    Article  CAS  Google Scholar 

  • Rotchanapreeda T, Wongniam S, Swangpol SC, Chareonsap PP, Sukkaewmanee N (2014) Development of SSR markers from Musa balbisiana for genetic diversity analysis among thai bananas. Plant Syst Evol 302:739–761

    Article  CAS  Google Scholar 

  • Rowe PR, Rosales F (1993) Diploid breeding at FHIA and the development of Goldfinger (FHIA-01). InfoMusa 2:9–11

    Google Scholar 

  • Scott RJ, Spielman M, Bailey J, Dickinson HG (1998) Parent-of-origin effects on seed development in Arabidopsis thaliana. Development 125:3329–3341

    Article  CAS  PubMed  Google Scholar 

  • Shepherd K (1987) Banana breeding—past and present. Acta Hortic 196:37–43

    Article  Google Scholar 

  • Simmonds NW (1962) The evolution of the bananas. Tropical science series. Longmans, London, p 170

    Google Scholar 

  • Sivanesan I (2007) Shoot regeneration and somaclonal variation from leaf callus cultures of Plumbago zeylanica. Asian J Plant Sci 6:83–86

    Article  CAS  Google Scholar 

  • Sobhakumari VPP, Lalithakumari D (2003) Direct plant regeneration from shoot tip cultures of Capsicum annuum L.cv.PLR-1. Phytomorphology 53:235–242

    Google Scholar 

  • Suman S (2017) Plant tissue culture: a promising tool of quality material production with special reference to micropropagation of banana. Biochem Cell Arch 17:1–26

    Google Scholar 

  • Swennen R, Vuylsteke D, Hahn SK (1992) The use of simple biotechnological tools to facilitate plantain breeding. In: Thottappilly G, Monti LM, Mohan Raj DR, Moore AW (eds) Biotechnology: enhancing research on tropical crops in Africa. IITA, Nigeria, pp 69–74

    Google Scholar 

  • Talengera D, Vuylsteke D, Karamura E (1996) In vitro germination of Ugandan banana hybrids. Musa Africa 10:14

    Google Scholar 

  • Tiryaki I, Korkmaz A, Nas MN, Ozbay N (2009) Priming combined with plant growth regulators promotes germination and emergence of dormant Amaranthus cruentus L.seeds. International Seed Testing Association. Seed Sci Technol 33(3):571–579

    Article  Google Scholar 

  • Uma S, Mustaffa MM, Saraswathi MS, Durai P (2011) Exploitation of diploids in Indian breeding programmes. Acta Hortic 897:215–223

    Article  Google Scholar 

  • Uma S, Lakshmi S, Saraswathi MS, Akbar A, Mustaffa MM (2012) Embryo rescue and plant regeneration in banana (Musa spp.). Plant Cell Tiss Org Cult 105:105–111

    Article  CAS  Google Scholar 

  • Uma S, Saraswathi MS, Backiyarani S, Durai P (2015) Banana breeding—a brief review. Int J Innov Hortic 4(1):11–19

    Google Scholar 

  • Varshney A, Lakshmikumaran M, Srivastava PS, Dhawan V (2001) Establishment of genetic fidelity of in vitro-raised lilium bulblets through rapd markers. Vitro Cell Dev Biol Plant 37:227–231

    Article  CAS  Google Scholar 

  • Vasane SR, Kothari RM (2006) Optimization of secondary hardening process of banana plantlets (Musa paradisiaca L. var. Grand Naine). Indian J Biotechnol 5:394–399

    Google Scholar 

  • Venkatachalam P, Kavikishor PB, Geetha N, Thangavelu M, Jayabalan N (1999) A rapid protocol for somatic embryogenesis from immature leaflets of groundnut (Arachishy pogaea L.). Vitro Cell Dev Biol Plant 35:409–412

    Article  Google Scholar 

  • Venkataramana RK, Sampangi-Ramaiah MH, Ajitha R, Khadke GN, Chellam V (2015) Insights into Musa balbisiana and Musa acuminate species divergence and development of genic microsatellites by transcriptomics approach. Plant Gene 4:78–82

    Article  CAS  Google Scholar 

  • Wattanachaiyingcharoen D (1990) Viability, germination and dormancy of banana seed (Musa acuminata subsp.). Unpublished Thesis. The Faculty of Natural and Agriculture Science, The University of Western Australia

  • Zapata C, Srivatanakul M, Park SH, Lee BM, Salas MG, Smith RH (1999) Improvements in shoot apex regeneration of two fiber crops: cotton and kenaf. Plant Cell Tiss Org Cult 56:185–191

    Article  Google Scholar 

  • Zhang J, Maun MA (1990) Seed size variation and its effects on seedling growth in Agropyron psammophilum. Bot Gaz 151:106–113

    Article  Google Scholar 

  • Zhang S, Zhang H, Zhang MB (1996) Production of multiple shoots from shoot apical meristems of oat (Avena sativa L.). J Plant Physiol 148:667–671

    Article  CAS  Google Scholar 

  • Zhang S, WilliamsCarrier R, Jackson D, Lemaux PG (1998) Expression of CDC2Zm and KNOTTED1 during in vitro axillary shoot meristem proliferation and adventitious shoot meristem formation in maize (Zea mays L.) and barley (Hordeumvulgare L.). Planta 204:542–549

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We express our sincere gratitude to the Director, ICAR-National Research Centre for Banana, India for her support in writing this Research article.

Funding

This study was supported by Indian Council of Agricultural Research (ICAR), under the project entitled “Improvement of Banana through conventional breeding—IXX02905” India.

Author information

Author notes

  1. S. Backiyarani and S. Uma have contributed equally to this work.

Authors and Affiliations

  1. ICAR-National Research Centre for Banana, Thogamalai Road, Thayanur Post, Tiruchirappalli, Tamil Nadu, 620102, India

    Suthanthiram Backiyarani, Subbaraya Uma, Swaminathan Saranya, Palani Durai, Selvaraj Eugin Perianayagaraj, Vadivel Selvaraj, Marimuthu Somasundaram Saraswathi, Raju Karthic & Sathiamoorthy Kalpana

Authors
  1. Suthanthiram Backiyarani
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Subbaraya Uma
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Swaminathan Saranya
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Palani Durai
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Selvaraj Eugin Perianayagaraj
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Vadivel Selvaraj
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Marimuthu Somasundaram Saraswathi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Raju Karthic
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Sathiamoorthy Kalpana
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

SBR and SU conceived the study, designed and managed the experiments. SS, SE and RK performed in vitro trials and collected data at laboratory. PD and VS performed hybridization and collected data at field. SBR and SK completed statistical analyses of phenotypic data. SBR wrote the manuscript and SU participated in correcting the manuscript. All authors contributed to writing the manuscript.

Corresponding author

Correspondence to Suthanthiram Backiyarani.

Ethics declarations

Conflict of interest

The author’s declare that they have no competing interests.

Additional information

Communicated by Bart Panis.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (PDF 35 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Backiyarani, S., Uma, S., Saranya, S. et al. Multiple shoot induction in zygotic embryos: a strategy for acceleration of banana breeding. Plant Cell Tiss Organ Cult 147, 339–350 (2021). https://doi.org/10.1007/s11240-021-02127-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-021-02127-x

Keywords

Search

Navigation