Influence of Culture Media and Their Compositions on Haploid Induction in Indian Short Day Onion

  • Gururaj Basaya Mathapati
  • Pritam Kalia
  • Sabina Islam
  • Navinder Saini
  • Arun Kumar
  • Anil Khar
Research Article
  • 15 Downloads

Abstract

Two media, Gamborg’s medium (B5) and Murashige and Skoog’s medium (MS) with various combinations, were used to study their effect on haploid induction efficiency from unpollinated flowers or ovaries in tropical short day onion. In B5 medium, highest induction efficiency (1.9%) was observed on HAP18 followed by HAP17 and HAP05 whereas in MS medium, HAP40 recorded highest induction frequency (5.0%) followed by HAP32. Kinetin, meta-topolin, thidiazuron did not influence embryo induction. Sucrose at 7.5% in B5 medium and at 10% in MS medium was found to be optimum for induction. Days for plantlet induction were 98.9 ± 3.9 days in B5 medium and 83.1 ± 2.7 days in MS medium. Induction rate of 0.51% and survival of 27.8% in B5 medium and induction rate of 0.72% and survival of 33% in MS medium were observed. Three haploids, 1 mixoploid and 1 diploid in B5 medium and 2 haploids, one diploid in MS medium were obtained. Flow cytometry and cytology confirmed the status of haploid plants. MS medium was found to be better than B5 medium and newer combinations for haploid induction were identified. Plant Preservative Mixture (PPM™) at 1 mg/l was found to be efficient for inhibiting contamination from field grown flower buds. These combinations will be helpful in devising new strategies for faster and higher rate of haploid induction in tropical short day Indian cultivars.

Keywords

Allium cepa L. Gynogenesis Tropical onion Short day Murashige and Skoog’s medium Gamborg’s medium 

Notes

Acknowledgements

This study was supported by the Department of Biotechnology, Government of India under the project code BT/PR12181/BPA/118/22/2014. The authors are thankful to the Director, ICAR-IARI and Head, Division of Vegetable Science, ICAR-IARI for providing the laboratory facilities and encouragement to carry out the work.

Compliance with Ethical Standards

Conflict of interest

The authors declare no conflict of interest.

References

  1. 1.
    FAOSTAT (2014) Food and Agriculture Organization of the United Nations—FAO Statistical Database. http://www.fao.org/faostat/en/#data/QC. Assessed 3 June 2017
  2. 2.
    Singh RK, Bhonde SR (2011) Performance studies of exotic onion (Allium cepa L.) hybrids in the Nashik region of Maharashtra. Indian J Hill Farming 24:29–31Google Scholar
  3. 3.
    Nunes RLC, Oliveira ABD, Dutra AS (2014) Agronomic performance of onion hybrids in Baraúna, in the semi-arid region of Brazil. Rev Ciênc Agron 45:606–611CrossRefGoogle Scholar
  4. 4.
    Shigyo M, Kik C (2008) Onion. In: Prohens J, Nuez JF (eds) Vegetables II. Springer, New York, pp 121–159CrossRefGoogle Scholar
  5. 5.
    Muren RC (1989) Haploid plant induction from unpollinated ovaries in onion. HortScience 24:833–834Google Scholar
  6. 6.
    Bohanec B, Jakše M, Ihan A, Javornik B (1995) Studies of gynogenesis in onion (Allium cepa L.): induction procedures and genetic analysis of regenerants. Plant Sci 104:215–224CrossRefGoogle Scholar
  7. 7.
    Alan AR, Brants A, Cobb E, Goldschmied P, Mutschler MA, Earle ED (2004) Fecund gynogenic lines from onion (Allium cepa L.) breeding materials. Plant Sci 167:1055–1066CrossRefGoogle Scholar
  8. 8.
    Fayos O, Vallés MP, Garcés-Claver A, Mallor C, Castillo AM (2015) Doubled haploid production from Spanish onion (Allium cepa L.) germplasm: embryogenesis induction, plant regeneration and chromosome doubling. Front Plant Sci 6:384CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Geoffriau E, Kahane R, Rancillac M (1997) Variation of gynogenesis ability in onion (Allium cepa L.). Euphytica 94:3–44CrossRefGoogle Scholar
  10. 10.
    Jakše M, Hirschegger P, Bohanec B, Havey MJ (2010) Evaluation of gynogenic responsiveness and pollen viability of selfed doubled haploid onion lines and chromosome doubling via somatic regeneration. J Am Soc Hortic Sci 135:67–73Google Scholar
  11. 11.
    Bohanec B, Jakše M (1999) Variations in gynogenic response among long-day onion (Allium cepa L.) accessions. Plant Cell Rep 18:737–742CrossRefGoogle Scholar
  12. 12.
    Dunstan DI, Short KC (1977) Improved growth of tissue cultures of the onion, Allium cepa. Physiol Plant 41:70–72CrossRefGoogle Scholar
  13. 13.
    Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158CrossRefPubMedGoogle Scholar
  14. 14.
    Michalik B, Adamus A, Nowak E (2000) Gynogenesis in Polish onion cultivars. J Plant Physiol 156:211–216CrossRefGoogle Scholar
  15. 15.
    Alan AR, Mutschler MA, Brants A, Cobb E, Earle ED (2003) Production of gynogenic plants from hybrids of Allium cepa L. and A. roylei Stearn. Plant Sci 165:1201–1211CrossRefGoogle Scholar
  16. 16.
    Anandhan S, Chavan AA, Gopal J, Mote SR, Shelke PV, Lawande KE (2014) Variation in gynogenic potential for haploid induction in Indian short-day onions. Indian J Genet 74:526–528Google Scholar
  17. 17.
    Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  18. 18.
    Niedz RP, Bausher MG (2002) Control of in vitro contamination of explants from greenhouse- and field-grown trees. In Vitro Cell Dev Biol Plant 38:468–471CrossRefGoogle Scholar
  19. 19.
    Miyazaki JJ, Tan BH, Errington SG (2010) Eradication of endophytic bacteria via treatment for axillary buds of Petunia hybrida using Plant Preservative Mixture (PPM™). Plant Cell Tissue Organ Cult 102:365–372CrossRefGoogle Scholar
  20. 20.
    Chiancone B, Karasawa MMG, Gianguzzi V, Abdelgalel AM, Bárány I, Testillano PS, Marinoni DT, Botta R, Germanà MA (2015) Early embryo achievement through isolated microspore culture in Citrus clementina Hort. Ex Tan., cvs. ‘Monreal Rosso’ and ‘Nules’. Front Plant Sci 6:413CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Chukwujekwu JC, Fennell CW, Van Staden J (2002) Optimisation of the tissue culture protocol for the endangered Aloe polyphylla. S Afr J Bot 68:424–429CrossRefGoogle Scholar
  22. 22.
    Chen UC, Hsia CN, Yeh MS, Agrawal DC, Tsay HS (2006) In vitro micropropagation and ex vitro acclimation of Bupleurum kaoi—an endangered medicinal plant native to Taiwan. In Vitro Cell Dev Biol Plant 42:128–133CrossRefGoogle Scholar
  23. 23.
    Sulistyaningsih EYA, Tashiro Y (2006) Flower bud culture of shallot (Allium cepa L. Aggregatum group) with cytogenetic analysis of resulting gynogenic plants and somaclones. Plant Cell Tissue Organ Cult 86:249–255CrossRefGoogle Scholar
  24. 24.
    Jakse M, Havey MJ, Bohanec B (2003) Chromosome doubling procedures of onion (Allium cepa L.) gynogenic embryos. Plant Cell Rep 21:905–910PubMedGoogle Scholar
  25. 25.
    Campion B, Alloni C (1990) Induction of haploid plants in onion (Allium cepa L.) by in vitro culture of unpollinated ovules. Plant Cell Tissue Organ Cult 20:1–6CrossRefGoogle Scholar
  26. 26.
    Campion B, Azzimonti MT, Vicini E, Schiavi M, Falavigna A (1992) Advances in haploid plant induction in onion (Allium cepa L.) through in vitro gynogenesis. Plant Sci 86:97–104CrossRefGoogle Scholar
  27. 27.
    Jakše M, Bohanec B, Ihan A (1996) Effect of media components on the gynogenic regeneration of onion (Allium cepa L.) cultivars and analysis of regenerants. Plant Cell Rep 15:934–938CrossRefPubMedGoogle Scholar
  28. 28.
    Bohanec B, Jakše M, Havey MJ (2003) Genetic analyses of gynogenetic haploid production in onion. J Am Soc Hortic Sci 128:571–574Google Scholar

Copyright information

© The National Academy of Sciences, India 2018

Authors and Affiliations

  1. 1.Division of Vegetable ScienceICAR-Indian Agricultural Research Institute (ICAR-IARI)New DelhiIndia
  2. 2.Division of GeneticsICAR-Indian Agricultural Research Institute (ICAR-IARI)New DelhiIndia

Personalised recommendations