Advertisement

Biologia

, Volume 74, Issue 1, pp 25–33 | Cite as

Influence of auxins on somatic embryogenesis in Haworthia retusa Duval

  • Doo Hwan Kim
  • Kyung Won Kang
  • Iyyakkannu SivanesanEmail author
Original Article

Abstract

An efficient in vitro plant regeneration method through somatic embryogenesis has been established in Haworthia retusa. Somatic embryos were induced from leaf explants on Murashige and Skoog (MS) medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and α-naphthaleneacetic acid (NAA) either alone or in combination with 4 μM thidiazuron (TDZ). Of the four auxins studied, IBA was found to be the most promising in terms of somatic embryo induction, followed in decreasing frequency by 2,4-D, IAA, and NAA. The highest somatic embryo induction (60.7%), with a mean of 20.7 embryos per leaf explant, was observed on MS medium amended with 20 μM IBA. The inclusion of 4 μM TDZ to the auxin-containing medium significantly (p < 0.0001) increased the somatic embryo induction frequency as well as the number of somatic embryos. The best combination for somatic embryogenesis was IBA + TDZ. The highest incidence of somatic embryo induction (100%), with a mean of 55.8 somatic embryos, was obtained on a culture medium containing 16 μM IBA + 4 μM TDZ. Somatic embryos germinated best on MS medium supplemented with 2 μM gibberellic acid. Morphological variations were observed among the regenerated plantlets. Well-developed plantlets obtained from germination media were acclimatized in the greenhouse.

Keywords

Auxin Leaf explants Gibberellic acid Somatic embryos Thidiazuron 

Notes

Acknowledgments

This article was supported by the KU Research Professor Program of Konkuk University.

Author contribution

DHK, KWK, and IS conceived and designed the experiments and wrote the paper. IS performed the somatic embryogenesis studies, and KWK performed the greenhouse experiments. All authors have seen and agreed to the submitted manuscript.

Compliance with ethical standards

Conflicts of interest

The authors declare that there are no conflicts of interest.

References

  1. Bairu MW, Aremu AO, Van Staden J (2011) Somaclonal variation in plants: causes and detection methods. Plant Growth Regul 63:147–173.  https://doi.org/10.1007/s10725-010-9554-x CrossRefGoogle Scholar
  2. Beyl CA, Sharma GC (1983) Picloram induced somatic embryogenesis in Gasteria and Haworthia. Plant Cell Tissue Organ Cult 2:23–132.  https://doi.org/10.1007/BF00043357 CrossRefGoogle Scholar
  3. Bradaï F, Pliego-Alfaro F, Sánchez-Romero C (2016) Somaclonal variation in olive (Olea europaea L.) plants regenerated via somatic embryogenesis: influence of genotype and culture age on phenotypic stability. Sci Hortic 213:208–215.  https://doi.org/10.1016/j.scienta.2016.10.031 CrossRefGoogle Scholar
  4. Devi K, Sharma M, Ahuja PS (2014) Direct somatic embryogenesis with high frequency plantlet regeneration and successive cormlet production in saffron (Crocus sativus L.). S Afr J Bot 93:207–216.  https://doi.org/10.1016/j.sajb.2014.04.006 CrossRefGoogle Scholar
  5. Hatzilazarou S, Kostas S, Economou A, Scaltsoyiannes A (2017) Efficient propagation of Nerium oleander L. through tissue culture. Propag Ornam Plants 17:64–74Google Scholar
  6. Kaul K, Sabharwal PS (1972) Morphogenetic studies on Haworthia: establishment of tissue culture and control of differentiation. Am J Bot 59:377–385.  https://doi.org/10.2307/2441548 CrossRefGoogle Scholar
  7. Kaul K, Sabharwal PS (1975) Morphogenetic studies on Haworthia: effects of inositol on growth and differentiation. Am J Bot 62:655–659.  https://doi.org/10.1002/j.1537-2197.1975.tb14098.x CrossRefGoogle Scholar
  8. Kim DH, Kang KW, Sivanesan I (2017a) In vitro propagation of Cymbidium hybrid. Propag Ornam Plants 17:48–54Google Scholar
  9. Kim DH, Kang KW, Sivanesan I (2017b) Micropropagation of Haworthia retusa Duval. Propag Ornam Plants 17:77–82Google Scholar
  10. Kumar V, Moyo M, Van Staden J (2016) Enhancing plant regeneration of Lachenalia viridiflora, a critically endangered ornamental geophyte with high floricultural potential. Sci Hortic 211:263–268.  https://doi.org/10.1016/j.scienta.2016.08.024
  11. Lincy KA, Remashree AB, Sasikumar B (2009) Indirect and direct somatic embryogenesis from aerial stem explants of ginger (Zingiber officinale Rosc.). Acta Bot Croat 68:93–103Google Scholar
  12. Liu B, Fang H, Meng C, Chen M, Chai Q, Zhang K, Liu S (2017) Establishment of a rapid and efficient micropropagation system for succulent plant Haworthia turgida haw. HortScience 52:1278–1282.  https://doi.org/10.21273/HORTSCI12056-17
  13. Mahendran G, Narmatha Bai V (2016) Direct somatic embryogenesis of Malaxis densiflora (A. Rich.) Kuntze. J Genet Eng Biotechnol 14:77–81.  https://doi.org/10.1016/j.jgeb.2015.11.003 CrossRefGoogle Scholar
  14. Moradi Z, Farahani F, Sheidai M, Satari TN (2017) Somaclonal variation in banana (Musa acuminate colla cv. Valery) regenerated plantlets from somatic embryogenesis: histological and cytogenetic approaches. Caryologia 70:1–6.  https://doi.org/10.1080/00087114.2016.1198665 CrossRefGoogle Scholar
  15. Mujib A, Ali M, Tonk D, Zafar N (2017) Nuclear 2C DNA and genome size analysis in somatic embryo regenerated gladiolus plants using flow cytometry. Adv Hortic Sci 31:165–174.  https://doi.org/10.13128/ahs-21956 Google Scholar
  16. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497.  https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  17. Mycock DJ, Watt MP, Hannweg KF, Naicker K, Makwarela M, Berjak P (1997) Somatic embryogenesis of two indigenous south African Haworthia spp. (H. limifolia and H. koelmaniorum). South Afr J Bot 63:345–350.  https://doi.org/10.1016/S0254-6299(15)30784-5 CrossRefGoogle Scholar
  18. Ogihara Y (1981) Tissue culture in Haworthia. Theor Appl Genet 60:353–363.  https://doi.org/10.1007/BF00264330 CrossRefGoogle Scholar
  19. Ogihara Y, Tsunewaki K (1978) Tissue culture in Haworthia I. effects of auxins and kinetin on callus growth. Bot Mag Tokyo 91:83–91.  https://doi.org/10.1007/BF02489105
  20. Ouyang Y, Chen Y, Lü J, Teixeira da Silva JA, Zhang X, Ma G (2016) Somatic embryogenesis and enhanced shoot organogenesis in Metabriggsia ovalifolia W. T. Wang. Sci Rep 19:6–24.  https://doi.org/10.1038/srep24662 Google Scholar
  21. Pandey KN, Sabharwal PS, Calkins J (1979) Effects of ionizing radiation (60Co gamma rays) on growth and morphogenesis of Haworthia mirabilis, haw. callus tissues. In Vitro 15:246–251.  https://doi.org/10.1007/BF02618947 CrossRefGoogle Scholar
  22. Pikulthong V, Teerakathiti T, Thamchaipenet A, Peyachoknagul S (2016) Development of somatic embryos for genetic transformation in Curcuma longa L. and Curcuma manga Valeton & Zijp. Agric Nat Resour 50:276–285.  https://doi.org/10.1016/j.anres.2015.08.004 Google Scholar
  23. Pinheiro MVM, Martins FB, da Cruz ACF, de Carvalho ACPP, Ventrella MC, Otoni WC (2014) Somatic embryogenesis in anthurium (Anthurium andraeanum cv. Eidibel) as affected by different explants. Acta Sci Agron 36:87–98.  https://doi.org/10.4025/actasciagron.v36i1.16557 CrossRefGoogle Scholar
  24. Raju CS, Aslam A, Shajahan A (2015) High-efficiency direct somatic embryogenesis and plant regeneration from leaf base explants of turmeric (Curcuma longa L.). Plant Cell Tissue Organ Cult 122:79–87.  https://doi.org/10.1007/s11240-015-0751-1 CrossRefGoogle Scholar
  25. Richwine AM, Tipton JL, Thompson GA (1995) Establishment of Aloe, Gasteria and Haworthia shoot cultures from inflorescence explants. HortScience 30:1443–1444.Google Scholar
  26. Rogers SMD (1993a) Optimization of plant regeneration and rooting from leaf explants of five rare Haworthia. Sci Hortic 56:157–161.  https://doi.org/10.1016/0304-4238(93)90016-J CrossRefGoogle Scholar
  27. Rogers SMD (1993b) Culture phenotype affects on regeneration capacity in the monocot Haworthia comptoniana. In Vitro Cell Dev Biol Plant 26:9–12.  https://doi.org/10.1007/BF02632232 CrossRefGoogle Scholar
  28. Sherif NA, Benjamin JHF, Kumar TS, Rao MV (2018) Somatic embryogenesis, acclimatization and genetic homogeneity assessment of regenerated plantlets of Anoectochilus elatus Lindl., an endangered terrestrial jewel orchid. Plant Cell Tissue Organ Cult 132:303–316.  https://doi.org/10.1007/s11240-017-1330-4 CrossRefGoogle Scholar
  29. Sivanesan I, Jeong BR (2012) Identification of somaclonal variants in proliferating shoot cultures of Senecio cruentus cv. Tokyo Daruma. Plant Cell Tissue Organ Cult 111:247–253.  https://doi.org/10.1007/s11240-012-0186-x CrossRefGoogle Scholar
  30. Sivanesan I, Son MS, Jana S, Jeong BR (2012) Secondary somatic embryogenesis in Crocus vernus (L.) hill. Propag Ornam Plants 12:163–170Google Scholar
  31. Standifer LC, O’Rourke EN, Porche-Sorbet R (1984) Propagation of Haworthia from floral scapes. Plant Prop 30:4–6Google Scholar
  32. Stanišić M, Raspor M, Ninković S, Milošević S, Ćalić D, Bohanec B, Trifunović M, Petrić M, Subotić A, Jevremović S (2015) Clonal fidelity of Iris sibirica plants regenerated by somatic embryogenesis and organogenesis in leaf-base culture-RAPD and flow cytometer analyses. South Afr J Bot 96:42–52.  https://doi.org/10.1016/j.sajb.2014.10.014 CrossRefGoogle Scholar
  33. Sun Y, Heil BM, Kahl G, Kohlenbach HW (1987) Plant regeneration from protoplasts of the monocotyledonous Haworthia magnifica v. Poelln. Plant Cell Tissue Organ Cult 8:91–100.  https://doi.org/10.1007/BF00040736 CrossRefGoogle Scholar
  34. Takamori LM, Neto NBM, Vieira LGE, Ribas AF (2015) Optimization of somatic embryogenesis and in vitro plant regeneration of Urochloa species using picloram. In Vitro Cell Dev Biol Plant 51:554–563.  https://doi.org/10.1007/s11627-015-9701-1
  35. Verma VM (2017) Direct somatic embryogenesis and organogenesis from axillary meristem in taro (Colocasia esculenta var. esculenta). Am J BioSci 5:114–122.  https://doi.org/10.11648/j.ajbio.20170506.13 CrossRefGoogle Scholar
  36. Wang X, Li Y, Zeng H, Cai N, Qiao Z, Wang X (2017) Micropropagation of Weigela florida ‘tango’ through in vitro shoot culture. HortScience 52:274–277.  https://doi.org/10.21273/HORTSCI11413-16 CrossRefGoogle Scholar
  37. Wessels DC, Groenewald EG, Koeleman A (1976) Callus formation and subsequent shoot and root development from leaf tissue of Haworthia planifolia cf. var. setulifera v. Poelln. Z Pflanzenphysiol 78:141–145.  https://doi.org/10.1016/S0044-328X(78)80185-8 CrossRefGoogle Scholar
  38. Zhong L, Liu E, Yang C, Jin S, Diao Y, Hu Z (2017) High embryogenic ability and regeneration from floral axis of Amorphophallus konjac (Araceae). Open Life Sci 12:34–41.  https://doi.org/10.1515/biol-2017-0004 Google Scholar

Copyright information

© Plant Science and Biodiversity Centre, Slovak Academy of Sciences 2018

Authors and Affiliations

  1. 1.Department of Bioresources and Food ScienceKonkuk UniversitySeoulSouth Korea
  2. 2.Babo Orchid FarmNamyangju-siSouth Korea

Personalised recommendations