Alteration of media enables efficient in vitro cloning of mature Elaeocarpus serratus L. (Ceylon olive): a commercially important fruit tree

Abstract

Elaeocarpus serratus is a fruit tree able to propagate through conventional vegetative means to a limited extent restricts its wide cultivation by the farmers. In the present report, we have developed an efficient in vitro propagation protocol using mature nodal explants from a 17-year-old tree for the first time with 6.6 shoots/culture. Explants cultured on agar (0.8%) gelled standard Murashige and Skoog (MS) medium, ½ MS, ¾ MS, White’s, Gamborg’s B5 or woody plant medium (WPM) supplemented with 2.5 µM benzyl adenine (BA) and 0.1 µM α-naphthalene acetic acid (NAA) showed the superiority of ½ MS medium in terms of explant response and number shoots (6.6). Further optimization of ½ MS medium by altering nutrient elements (macros, micros, vitamins and Fe EDTA) were undertaken, and MS medium composed of half-strength major salts, original strength of minor salts and vitamins were supplemented with BA (2.5 µM) and NAA (0.1 µM), produced enhanced axillary bud proliferation (8.88/explant) and shoot elongation (3.83 cm). Reculturing of original explant on this medium after IV passages produced more than 16 healthy shoots per culture which attained a length of 4.13 cm. Microshoots raised through this way were rooted (86.11%) ex vitro by pulse treatment with 2 mM indole-3-butyric acid (IBA) for 5 min followed by planting in nursery pots containing a 1:1:1 (v/v/v) mix of sand, soil, and farmyard manure. The hardened plants were successfully planted in the fruit tree garden of the Department. Genetic fidelity of micropropagated and mother plants were tested using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) markers which showed a high degree of monomorphism thus supported morphological uniformity of micropropagated plants.

This is a preview of subscription content, access via your institution.

We’re sorry, something doesn't seem to be working properly.

Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

References

  1. Ahmad A, Anis M (2019) Meta-topolin improves in vitro morphogenesis, rhizogenesis and biochemical Analysis in Pterocarpus marsupium Roxb.: a potential drug-yielding tree. Plant Growth Regul 38:1007–1016

    CAS  Google Scholar 

  2. Ahmed MR, Anis M, Alatar AA, Faisal M (2017) In vitro clonal propagation and evaluation of genetic fidelity using RAPD and ISSR marker in micropropagated plants of Cassia alata L.: a potential medicinal plant. Agro Forest Syst 91:637–647

    Google Scholar 

  3. Amirchakhmaghi N, Hosseinpour B, Yousefzadeh H (2019) Development of a micropropagation protocol for Malus orientalis using axillary buds. In Vitro Cell Dev Biol Plant. https://doi.org/10.1007/s11627-019-09992-4

    Article  Google Scholar 

  4. Arshad SM, Kumar A (2006) Tissue culture investigation of Elaeocarpus tuberculatus a highly valued rudraksha. Vegetos-An Int J Plant Res 19:111–114

    Google Scholar 

  5. Awodoyin RO, Olubode OS, Ogbu JU, Balogun RB, Nwawuii JU, Orji KO (2015) Indigenous fruit trees of tropical Africa: Status, opportunity for development and biodiversity management. Agri Sci 6:31–41

    Google Scholar 

  6. Baruah PS, Deka K, Lahkar L, Sarma B, Borthakur SK, Tanti B (2019) Habitat distribution modelling and reinforcement of Elaeocarpus serratus L.: A threatened tree species of Assam, India for improvement of its conservation status. Act Ecol Sinica. https://doi.org/10.1016/j.chnaes.2018.06.002

    Article  Google Scholar 

  7. Bell RL, Reed BM (2002) In vitro tissue culture of pear: advances in techniques for micropropagation and germplasm preservation. Acta Hortic 596:412–418

    Google Scholar 

  8. Borkowska B (2001) Morphological and physiological characteristics of micropropagated strawberry plants rooted in vitro or ex vitro. Sci Hort 89:195–206

    Google Scholar 

  9. Cuenca B, Sánchez C, Aldrey A, Bogo B, Blanco B, Correa B, Vidal N (2017) Micropropagation of axillary shoots of hybrid chestnut (Castanea sativa × C. crenata) in liquid medium in a continuous immersion system. Plant Cell Tiss Organ Cult 131:307–320

    CAS  Google Scholar 

  10. De Klerk GJ (2002) Rooting of microcuttings: theory and practice. In Vitro Cell Dev Biol Plant 38:415–422

    Google Scholar 

  11. Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21

    CAS  Google Scholar 

  12. Dhavala A, Rathore TS (2010) Micropropagation of Embelia ribes Burm F. through proliferation of adult plant axillary shoots. In Vitro Cell Dev Biol Plant 46:180–191

    Google Scholar 

  13. Endres L, Souza BM, Mercier H (2002) In vitro nitrogen nutrition and hormonal pattern in bromeliads. In Vitro Cell Dev Biol Plant 38:481–486

    CAS  Google Scholar 

  14. Esteban R, Ariz I, Cruz C, Moran JF (2016) Review: mechanisms of ammonium toxicity and the quest for tolerance. Plant Sci 248:92–101

    CAS  PubMed  Google Scholar 

  15. Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirement of suspension cultures of soyabean root cells. Exp Cell Res 50:151–158

    CAS  PubMed  Google Scholar 

  16. Ghani A (1998) Monographs: Elaeocarpus serratus Linn. In: Medicinal plants of Bangladesh: chemical constituents and uses. Asiatic Soc Bangla Publ, pp. 167–168

  17. Goyal P, Kachhwaha S, Kothari SL (2012) Micropropagation of Pithecellobium dulce (Roxb.) Benth a multipurpose leguminous tree and assessment of genetic fidelity of micropropagated plants using molecular markers. Physiol Mol Biol Plants 18:169–176

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Goyal AK, Pradhan S, Basistha BC, Sen A (2014) Micropropagation and assessment of genetic fidelity of Dendrocalamus strictus (Roxb.) Nees using RAPD and ISSR markers. 3 Biotech 5:473–482

    PubMed  PubMed Central  Google Scholar 

  19. Greenway MB, Phillips IC, Lloyd MN, Hubstenberger JF, Phillips GC (2012) A nutrient medium for diverse applications and tissue growth of plant species in vitro. In Vitro Cell Dev Biol Plant 48:403–410

    CAS  Google Scholar 

  20. Hand C, Maki S, Reed MB (2014) Modeling optimal mineral nutrition for hazelnut micropropagation. Plant Cell Tiss Organ Cult 119:411–425

    CAS  Google Scholar 

  21. Jayasinghe L, Amarasinghe RN, Arundathie BGS, Rupasinghe GK, Jayatilake NHAN, Fujimoto Y (2011) Antioxidant flavonol glycosides from Elaeocarpus serratus and Filicium decipiens. Nat Prod Res 26:717–721

    PubMed  Google Scholar 

  22. Kabylbekova B, Kovalchuk I, Mukhitdinova Z et al (2020) Reduced major minerals and increased minor nutrients improve micropropagation in three apple cultivars. In Vitro Cell Dev Biol Plant. https://doi.org/10.1007/s11627-019-10019-1

    Article  Google Scholar 

  23. Krishnan SSR, Siril EA (2016) Enhanced in vitro shoot regeneration in Oldenlandia umbellata L. by using quercetin: a naturally occurring auxin-transport inhibitor. Proc Natl Acad Sci India Sect B Biol Sci 21:271–278

    Google Scholar 

  24. Liu Y, Wirén NV (2017) Ammonium as a signal for physiological and morphological responses in plants. J Exp Bot 68:2581–2592

    CAS  PubMed  Google Scholar 

  25. Lloyd GB, McCown BH (1980) Commercially feasible micropropagation of mountain laurel (Kamalia latifolia) by use of shoot tip culture. Comb Proc Int Plant Propagators Soc 30:421–427

    Google Scholar 

  26. Lodha D, Patel AK, Rai MK, Shekhawat NS (2014) In vitro plantlet regeneration and assessment of alkaloid contents from callus cultures of Ephedra foliata (Unth phog), a source of anti-asthmatic drugs. Acta Physiol Plant 36:3071–3079

    CAS  Google Scholar 

  27. Lozzi A, Abdelwahd R, Mentag R, Abousalim A (2019) Development of a new culture medium and efficient protocol for in vitro micropropagation of Ceratonia siliqua L. In Vitro Cell Dev Biol Plant. https://doi.org/10.1007/s11627-019-09990-6

    Article  Google Scholar 

  28. Lymperopoulos P, Msanne J, Rabara R (2018) Phytochrome and phytohormones: working in tandem for plant growth and development. Front Plant Sci. https://doi.org/10.3389/fpls.2018.01037

    Article  PubMed  PubMed Central  Google Scholar 

  29. Martin KP (2003) Rapid in vitro multiplication and ex vitro rooting of Rotula aquatica Lour., a rare rhoeophytic woody medicinal plant. Plant Cell Rep 21:415–420

    CAS  PubMed  Google Scholar 

  30. Morgan MJ, Lehmann M, Schwarzländer M et al (2008) Decrease in manganese superoxide dismutase leads to reduced root growth and affects tricarboxylic acid cycle flux and mitochondrial redox homeostasis. Plant Physiol 147:101–114

    CAS  PubMed  PubMed Central  Google Scholar 

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

    CAS  Google Scholar 

  32. Nas MN, Read PE (2004) A hypothesis for the development of a defined tissue culture medium of higher plants and micropropagation of hazelnuts. Sci Hortic 101:189–200

    CAS  Google Scholar 

  33. Newell C, Growns DJ, Mc Comb JA (2005) A novel in vitro rooting method employing an aerobic medium. Aust J Bot 53:81–89

    Google Scholar 

  34. Niedz RP, Hyndman SE, Evens TJ, Weathersbee AA (2014) Mineral nutrition and in vitro growth of Gerbera hybrida (Asteraceae). In Vitro Cell Dev Biol Plant 50:458–470

    CAS  Google Scholar 

  35. Oberschelp GPJ, Gonçalves AN (2016) Assessing the effects of basal media on the in vitro propagation and nutritional status of Eucalyptus dunnii Maiden. In Vitro Cell Dev Biol Plant 52:28–37

    CAS  Google Scholar 

  36. Patel AK, Lodha D, Ram K, Shekhawat S, Shekhawat NS (2016) Evaluation of physiochemical factors affecting high-frequency plant regeneration of Blyttia spiralis (synonym: Pentatropis spiralis), a threatened climber of medicinal value. In Vitro Cell Dev Biol Plant 52:10–19

    Google Scholar 

  37. Patel AK, Phulwaria M, Rai MK, Gupta AK, Shekhawat S, Shekhawat NS (2014) In vitro propagation and ex vitro rooting of Carallumaedulis (Edgew.) Benth. and Hook. f.: an endemic and endangered edible plant species of the Thar Desert. Sci Hortic 165:175–180

    CAS  Google Scholar 

  38. Peiris KHS (2007) Veralu, Elaeocarpus serratus L. In: Pushpakumara DKNG, Gunasena HPM and Singh VP (eds), Underutilized fruit in Sri Lanka, vol I, Asian Center for underutilized fruit crops, Sri Lanka

  39. Phillips GC, Garda M (2019) Plant tissue culture media and practices: an overview. In Vitro Cell Dev Biol Plant. https://doi.org/10.1007/s11627-019-09983-5

    Article  Google Scholar 

  40. Phulwaria M, Ram K, Gahlot P, Shekhawat NS (2011) Micropropagation of Salvadora persica: a tree of arid horticulture and forestry. New For 42:317–327

    Google Scholar 

  41. Purohit S, Jugran AK, Bhatt ID, Palni LMS, Bhatt A, Nandi SK (2017) In vitro approaches for conservation and reducing juvenility of Zanthoxylum armatum DC: an endangered medicinal plant of Himalayan region. Trees Struct Funct 31:1101–1108

    CAS  Google Scholar 

  42. Rahman MM, Amin MN, Ishiguri F, Yokota S, Sultana RS, Takashima Y, Iizuka K, Yoshizawa N (2009) In vitro plantlet regeneration of ‘“dwarf”’ Indian olive (Elaeocarpus robustus Roxb.): a fruit plant of Bangladesh. Plant Biotechnol Rep 3:259–266

    Google Scholar 

  43. Raji R, Siril EA (2018) Assessment of different pretreatments to breakage dormancy and improve the seed germination in Elaeocarpus serratus L.: an underutilized multipurpose fruit tree from South India. For Sci Technol 14:160–168

    Google Scholar 

  44. Raji R, Siril EA (2016) Cloning of Ceylon olive (Elaeocarpus serratus L.) using conventional methods. J Hortic Sci Biotech 91:292–298

    Google Scholar 

  45. Ramage CM, Williams RR (2002) Mineral nutrition and plant morphogenesis. In Vitro Cell Dev Biol Plant 38:116–124

    CAS  Google Scholar 

  46. Rathore JS, Rathore MS, Shekhawat NS (2005) Micropropagation of Maerua oblongifolia: a liana of arid areas. Phytomorphology 55:241–247

    Google Scholar 

  47. Siril EA, Joseph N (2013) Micropropagation of annatto (Bixa orellana L.) from mature tree and assessment of genetic fidelity of micropropagated plants with RAPD markers. Physiol Mol Biol Plants 1:147–155

    Google Scholar 

  48. Sreeranjini S, Siril EA (2014) Field performance and genetic fidelity evaluation of micropropagated Morinda citrifolia L. Indian J Biotech 13:121–130

    CAS  Google Scholar 

  49. Timofeeva SN, Elkonin LA, Tyrnov VS (2014) Micropropagation of Laburnum anagyroides Medic. through axillary shoot regeneration. In Vitro Cell Dev Biol Plant 50:561–567

    Google Scholar 

  50. Tiwari SK, Tiwari KP, Siril EA (2002) An improved micropropagation protocol for teak. Plant Cell Tiss Organ Cult 71:1–6

    CAS  Google Scholar 

  51. Tripathi M, Kumari N (2010) Micropropagation of a tropical fruit tree Spondias mangifera Willd. through direct organogenesis. Acta Physiol Plant 32:1011–1015

    Google Scholar 

  52. White PR (1939) Potentially unlimited growth of excised plant callus in an artificial medium. Am J Bot 26:59–64

    Google Scholar 

  53. Wang Y, Jiao Y (2018) Auxin and above-ground meristems. J Exp Bot 69:147–154

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors are grateful to Dr. Suhara Beevy S, Professor and Head, Department of Botany for providing the facilities and Kerala State Council for Science Technology and Environment (KSCSTE), Government of Kerala, Thiruvananthapuram, India, for the financial assistance (P.1409/2014/KSCSTE) through Junior Research Fellowship.

Author information

Affiliations

Authors

Contributions

RR and EAS designed the experiments. RR performed the experiments and data collection. EAS and RR analyzed data. RR prepared the first draft of the manuscript and EAS edited and communicated for publishing.

Corresponding author

Correspondence to E. A. Siril.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants performed by any of the authors.

Informed consent

Informed consent not obtained as the article does not contain any studies with human participants performed by any of the authors.

Additional information

Publisher's Note

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

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Raji, R., Siril, E.A. Alteration of media enables efficient in vitro cloning of mature Elaeocarpus serratus L. (Ceylon olive): a commercially important fruit tree. Physiol Mol Biol Plants 27, 429–443 (2021). https://doi.org/10.1007/s12298-021-00955-x

Download citation

Keywords

  • Culture media
  • Elaeocarpus serratus
  • Ex vitro rooting
  • In vitro culture
  • Micropropagation