Skip to main content

Advertisement

SpringerLink
Log in

Vitrification-Based Cryopreservation of In Vitro-Grown Apical Meristems of Chlorophytum borivilianum Sant et Fernand: A Critically Endangered Species

  • Research Article
  • Published:
Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

This article reports the cryopreservation of apical meristems of Chlorophytum borivilianum, a tropical and IUCN critically endangered species. Initially, in vitro cultured shoots were pre-adapted on 12% (w/v) sucrose for 2-months and were found appropriate stock material for further experimentations. Furthermore, the preculture of meristems excised from pre-adapted in vitro shoots on 12% (w/v) sucrose containing MS medium with 50 mg/l abscisic acid for 48 h, followed by treatment with loading solution (LS), and plant vitrification solution (PVS2) was found crucial for recovery following cryostorage. Thereafter, durations of exposure to the LS and PVS2 were optimized to enhance the regeneration efficiency of apical meristems. Treatment with LS for 20 min followed by 30 min exposure to PVS2 was standardized for the vitrification of the apical meristems before plunging them into liquid nitrogen. Moreover, after cryoexposure thawing was performed for 1 min at 38 °C ± 2 in a water-bath followed by the treatment with unloading solution for 10 min resulted in enhanced recovery up to 33% on 2 mg/l 6-benzyladenine (BA) and 0.2 mg/l α-naphthalene acetic acid containing MS medium.

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

  1. Chauhan R, Keshavkant S, Quraishi A (2018) Enhanced production of diosgenin through elicitation in micro-tubers of Chlorophytum borivilianum Sant et Fernand. Ind Crop Prod 113:234–239

    Article  CAS  Google Scholar 

  2. Tripathi RK, Dethe PD, Bhojne SK, Raut AA, Rege NN (2019) A prospective, randomized, placebo-controlled, double-blind comparative pilot study to evaluate the efficacy of Chlorophytum borivilianum on physical performance. Indian J Pharmacol 51:150–156

    Article  CAS  Google Scholar 

  3. Chauhan R, Quraishi A, Jadhav SK, Keshavkant S (2016) A comprehensive review on pharmacological properties and biotechnological aspects of Genus Chlorophytum. Acta Physiol Plant 38:116

    Article  Google Scholar 

  4. Kaur R, Arora S, Thukral AK (2009) Enhancing seed germination of Chlorophytum borivilianum Sant et Fernand with PGRs, steroidal hormones and zinc. Res J Seed Sci 2:32–39

    Article  Google Scholar 

  5. IUCN (2015): International union for conservation of nature and natural resources. The IUCN red list of threatened species, Version 2015–4. 2015, http://www.iucnredlist.org [Accessed September 2019]

  6. Nayar MP, Sastry ARK (1988) Chlorophytum borivilianum. In: Nayar MP, Sastry ARK (eds) Red data book of Indian plants, vol 2. Botanical Survey of India, Calcutta (India), p 412

    Google Scholar 

  7. Pathirana R, McLachlan A, Hedderley D, Panis B, Carimi F (2016) Pre-treatment with salicylic acid improves plant regeneration after cryopreservation of grapevine (Vitis spp.) by droplet vitrification. Acta Physiol Plant 38:12

    Article  Google Scholar 

  8. Rao NK (2004) Plant genetic resources: advancing conservation and use through biotechnology: a review. Afr J Biotechnol 3:136–145

    Google Scholar 

  9. Villalobos-Olivera A, Martínez J, Quintana N, Zevallos BE, Cejas I, Lorenzo JC et al (2019) Field performance of micropropagated and cryopreserved shoot tips derived pineapple plants grown in the field for 14 months. Acta Physiol Plant 41:34

    Article  Google Scholar 

  10. Tandon P, Kumaria S, Das CM (2012) Plant resources of India: potentials for future development. Proc Natl Acad Sci Ind Sect B Biol Sci 82(S2):283–289

    Google Scholar 

  11. Benelli C, De Carlo A, Engelmann F (2013) Recent advances in the cryopreservation of shoot-derived germplasm of economically important fruit trees of Actinidia, Diospyros, Malus, Olea, Prunus, Pyrus and Vitis. Biotechnol Adv 31:175–185

    Article  CAS  Google Scholar 

  12. Wen B (2019) Seed germination ecology of Alexandra palm (Archontophoenix alexandrae) and its implication on invasiveness. Sci Rep 9:4057

    Article  Google Scholar 

  13. Rahman SM, Ahsan MN, Al-Mamun A, Hossain QZ, Rahman MM, Hasan MM, Chisty MAH (2015) Evaluating the suitability of cryoprotectants and cryopreservation solutions for Olive Barb, Puntius Sarana embryos. Proc Natl Acad Sci Ind Sect B Biol Sci 85(1):175–180

    Article  CAS  Google Scholar 

  14. Chauhan R, Keshavkant S, Jadhav SK, Quraishi A (2016) In vitro slow-growth storage of Chlorophytum borivilianum Sant. et Fernand: a critically endangered herb. Vitro Cell Dev Biol-Plant 52:315–321

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  16. Sakai A, Kobayashi S, Oiyama I (1990) Cryopreservation of nucellar cells of navel orange (Citrus sinensis Osb. var. Brasiliensis Tanaka) by vitrification. Plant Cell Rep 9:30–33

    Article  CAS  Google Scholar 

  17. Chen X-L, Li J-H, Xin X, Zhang Z-E, Xin P-P, Lu X-X (2011) Cryopreservation of in vitro-grown apical meristems of Lilium by droplet-vitrification. S Afr J Bot 77:397–403

    Article  Google Scholar 

  18. Suzuki M, Ishikawa M, Okuda H, Noda K, Kishimoto T, Nakamura T et al (2006) Physiological changes in gentian axillary buds during two-step preculturing with sucrose that conferred high levels of tolerance to desiccation and cryopreservation. Ann Bot 97:1073–1081

    Article  CAS  Google Scholar 

  19. Engelmann F (2004) Plant cryopreservation: progress and prospects. Vitro Cell Dev Biol-Plant 40:427–433

    Article  Google Scholar 

  20. Fang JY, Wetten A, Hadley P (2004) Cryopreservation of cocoa (Theobroma cacao L.) somatic embryos for long-term germplasm storage. Plant Sci 166:669–675

    Article  CAS  Google Scholar 

  21. Popova EV, Lee EJ, Wu CH, Hahn EJ, Paek KY (2009) A simple method for cryopreservation of Ginkgo biloba callus. Plant Cell Tissue Organ Cult 97:337–343

    Article  CAS  Google Scholar 

  22. Chang Y, Reed BM (2001) Preculture conditions influence cold hardiness and regrowth of Pyrus cordata shoot tips after cryopreservation. HortScience 36:1329–1333

    Article  CAS  Google Scholar 

  23. Benson EE (2008) Cryopreservation of phytodiversity: a critical appraisal of theory and practice. Crit Rev Plant Sci 27:141–219

    Article  CAS  Google Scholar 

  24. Santos PAA, Paiva R, Silva LC, Souza AC, Gallo CM, Herrera RC (2013) Effect of load solution and growth media on cryopreservation by droplet vitrification of Hancornia speciose shoot tips. In: VIII International Symposium on in vitro Culture and Horticultural Breeding. ISHS Acta Hortic, 1083: 507–512

  25. Matsumoto T, Sakai A, Yamanda K (1994) Cryopreservation of in vitro grown apical meristems of wasabi (Wasabia japonica) by vitrification and subsequent high plant regeneration. Plant Cell Rep 13:442–446

    Article  CAS  Google Scholar 

  26. Yin Z-F, Bi W-L, Chen L, Zhao B, Volk GM, Wang Q-C (2014) An efficient, widely applicable cryopreservation of Lilium shoot tips by droplet vitrification. Acta Physiol Plant 36:1683–1692

    Article  CAS  Google Scholar 

  27. Panis B, Piette B, Swennen R (2005) Droplet vitrification of apical meristems: a cryopreservation protocol applicable to all Musaceae. Plant Sci 168:45–55

    Article  CAS  Google Scholar 

  28. Sant R, Taylor M, Tyagi A (2006) Cryopreservation of in vitro-grown shoot-tips of tropical taro (Colocasia esculenta var. esculenta) by vitrification. Cryo Letters 27:133–142

    CAS  PubMed  Google Scholar 

  29. Zamecnik J, Faltus M, Kotkova RF, Hejnak V (2011) Glass transition determination in Allium shoot tips after dehydration. Acta Hortic 908:33–38

    Article  Google Scholar 

  30. Berjak P, Pammenter NW (2014) Cryostorage of germplasm of tropical recalcitrant seeded species: approaches and problems. Int J Plant Sci 175:29–39

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur, 492 010, India

    Ravishankar Chauhan, S. Keshavkant & Afaque Quraishi

  2. National Center for Natural Resources, Pt. Ravishankar Shukla University, Raipur, 492 010, India

    Ravishankar Chauhan

  3. School of Life Sciences, Pt. Ravishankar Shukla University, Raipur, 492 010, India

    Vikram Singh

Authors
  1. Ravishankar Chauhan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Vikram Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. Keshavkant
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Afaque Quraishi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Afaque Quraishi.

Ethics declarations

Conflict of interest

The authors declare that no conflict of interest.

Additional information

Publisher's Note

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

Significance Statement: For the cryopreservation of Chlorophytum borivilianum abscisic acid was found crucial that helps in freeze tolerance. Moreover, the exposure times to cryoprotectants were optimized which minimizes the toxicity and helps in regeneration of meristems.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chauhan, R., Singh, V., Keshavkant, S. et al. Vitrification-Based Cryopreservation of In Vitro-Grown Apical Meristems of Chlorophytum borivilianum Sant et Fernand: A Critically Endangered Species. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 91, 471–476 (2021). https://doi.org/10.1007/s40011-021-01260-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s40011-021-01260-z

Keywords

Search

Navigation