Conservation, ex vitro direct regeneration, and genetic uniformity assessment of alginate-encapsulated nodal cuttings of Sphagneticola calendulacea (L.) Pruski

  • Suprabuddha Kundu
  • Umme Salma
  • Md. Nasim Ali
  • Nirmal Mandal
Original Article
  • 3 Downloads

Abstract

A well-organized procedure was established for the conservation and distribution of Sphagneticola calendulacea (L.) Pruski [synonym Wedelia chinensis (Osbeck) Merrill] for the first time, using alginate-encapsulated nodal segments (NSs) as synthetic seeds. The ideal beads were obtained through a combination of 2.5% sodium alginate and 75 mM calcium chloride with 84.40 ± 2.20% rate of shoot emergence. The maximum regeneration (88.84 ± 2.24%) from synthetic seeds was achieved on liquid 1/2Murashige and Skoog (MS) medium in comparison to its other formulations. Furthermore, superior frequency (91.09 ± 2.24%) of complete plantlet (having both shoots and roots) formation was achieved when synthetic seeds were cultured on liquid 1/2MS (1.5% sucrose) fortified with 1.0 mg L−1 N6-benzyladenine plus 0.25 mg L−1 α-naphthalene acetic acid. Synthetic seeds could be effectively stored at low temperature (8 °C) up to 90 days with a survival rate of 52.38 ± 3.06%, whereas higher temperature (25 °C) did not support regeneration after 75 days of storage. The plantlets were successfully acclimatized to natural conditions with ~ 89% survival frequency. To by-pass the time-consuming in vitro culture step after encapsulation, synthetic seeds were directly regrown into complete plantlets ex vitro on sand, soil, and vermicompost (1:1:1; w/w). Regeneration rate of 42.22 ± 2.22% was attained when NSs were pretreated on 1/2MS medium containing 4.0 mg L−1 indole-3-acetic acid for 24 h in dark, prior to encapsulation. The random amplified polymorphic DNA and intersimple sequence repeat fingerprinting profiles demonstrated genetic uniformity amongst the regenerated plantlets, in vitro mother plant, as well as in vivo wild plant.

Keywords

Encapsulation Germplasm exchange Sodium alginate Calcium chloride Genetic uniformity 

Abbreviations

BA

N6-Benzyladenine

CC

Calcium chloride

IBA

Indole-3-butyric acid

LAF

Laminar airflow

MS

Murashige and Skoog medium

NAA

α-Naphthalene acetic acid

NS

Nodal segment

PGR

Plant growth regulator

SA

Sodium alginate

Notes

Acknowledgements

Authors acknowledge the laboratory as well as library assistance from the Bidhan Chandra Krishi Viswavidyalaya, West Bengal, India. We further are thankful to the anonymous reviewers and the editor of this article for their critical comments and suggestions on the manuscript. The authors declare that there are no conflicts of interest. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

References

  1. Adhikari S, Bandyopadhyay TK, Ghosh P (2014) Assessment of genetic stability of Cucumis sativus L. regenerated from encapsulated shoot tips. Sci Hortic 170:115–122CrossRefGoogle Scholar
  2. Ahmed MR, Anis M, Al-Etta HA (2015) Encapsulation technology for short-term storage and germplasm exchange of Vitex trifolia L. Rend Lincei 26(2):133–139CrossRefGoogle Scholar
  3. Alatar AA, Ahmad N, Javed SB, Abdel-Salam EM, Basahi R, Faisal M (2017) Two-way germination system of encapsulated clonal propagules of Vitex trifolia L.: an important medicinal plant. J Hortic Sci Biotechnol 92(2):175–182CrossRefGoogle Scholar
  4. Bianchetti RE, de Resende CF, Pacheco VS, Dornellas FF, de Oliveira AMS, Freitas JCE, Peixoto PHP (2017) An improved protocol for in vitro propagation of the medicinal plant Mimosa pudica L. Afr J Biotechnol 16(9):418–428CrossRefGoogle Scholar
  5. Brar DS, Jain SM (1998) Somaclonal variation: mechanism and applications in crop improvement. In: Jain SM, Brar DS, Ahloowalia BS (eds) Somaclonal variation and induced mutations in crop improvement. Kluwer, Dordrecht, pp 15–24CrossRefGoogle Scholar
  6. Chand S, Singh AK (2004) Plant regeneration from encapsulated nodal segments of Dalbergia sissoo Roxb., a timber-yielding leguminous tree species. J Plant Physiol 161:237–243CrossRefPubMedGoogle Scholar
  7. Cuesta C, Ordás RJ, Rodríguez A, Fernández B (2010) PCR-based molecular markers for assessment of somaclonal variation in Pinus pinea clones micro-propagated in vitro. Biol Plant 54:435–442CrossRefGoogle Scholar
  8. Duncan DB (1955) Multiple range and multiple F test. Biometrics 11:1–42CrossRefGoogle Scholar
  9. Faisal M, Anis M (2007) Regeneration of plants from alginate-encapsulated shoots of Tylophora indica—an endangered medicinal plant. J Hortic Sci Biotechnol 82:351–354CrossRefGoogle Scholar
  10. Gantait S, Kundu S (2017) Artificial seed technology for storage and exchange of plant genetic resources. In: Malik CP, Wani SH, Kushwaha HB, Kaur R (eds) Advanced technologies for crop improvement and agricultural productivity. Agrobios International, Jodhpur, pp 135–159Google Scholar
  11. Gantait S, Sinniah UR (2013) Storability, post-storage conversion and genetic stability assessment of alginate-encapsulated shoot tips of monopodial orchid hybrid Aranda Wan Chark Kuan ‘Blue’ × Vanda coerulea Grifft. ex. Lindl. Plant Biotechnol Rep 7:257–266CrossRefGoogle Scholar
  12. Gantait S, Bustam S, Sinniah UR (2012) Alginate-encapsulation, short-term storage and plant regeneration from protocorm-like bodies of Aranda Wan Chark Kuan ‘Blue’ × Vanda coerulea Grifft. ex. Lindl. (Orchidaceae). Plant Growth Regul 68:303–311CrossRefGoogle Scholar
  13. Gantait S, Kundu S, Ali MN (2015a) Influence of encapsulating agent and matrix levels on synseed production of Bacopa monnieri (L.) Pennell. Med Plants 7:182–187Google Scholar
  14. Gantait S, Kundu S, Ali N, Sahu NC (2015b) Synthetic seed production of medicinal plants: a review on influence of explants, encapsulation agent and matrix. Acta Physiol Plant 37(5):1–12CrossRefGoogle Scholar
  15. Gantait S, Kundu S, Yeasmin L, Ali MN (2017) Impact of differential levels of sodium alginate, calcium chloride and basal media on germination frequency of genetically true artificial seeds of Rauvolfia serpentina (L.) Benth. ex Kurz. J Appl Res Med Aromat Plants 4:75–81Google Scholar
  16. Ghanbarali S, Abdollahi MR, Zolnorian H, Moosavi SS, Seguí-Simarro JM (2016) Optimization of the conditions for production of synthetic seeds by encapsulation of axillary buds derived from minituber sprouts in potato (Solanum tuberosum). Plant Cell Tissue Organ Cult 126(3):449–458CrossRefGoogle Scholar
  17. Hung CD, Trueman SJ (2012) Alginate encapsulation of shoot tips and nodal segments for short-term storage and distribution of the eucalypt Corymbia torelliana × C. citriodora. Acta Physiol Plant 34(1):117–128CrossRefGoogle Scholar
  18. Islam S, Banik H, Alam S, Tarek M, Rahman M (2009) In vitro Propagation of Holarrhena antidysenterica Wall., Wedelia chinensis (Osb.) Merr. And Woodfordia fruticosa (L.) Kurz. Plant Tissue Cult Biotechnol 19:253–255Google Scholar
  19. Koul S, Pandurangan A, Khosa RL (2012) Wedelia chinensis (Asteraceae)—an overview. Asian Pac J Trop Biomed 2:S1169–S1175CrossRefGoogle Scholar
  20. Kundu S, Salma U, Ali MN, Mandal N (2017) Factors influencing large scale micropropagation of Sphagneticola calendulacea (L.) Pruski and clonality assessment using RAPD and ISSR markers. In Vitro Cell Dev Biol Plant 53:167–177CrossRefGoogle Scholar
  21. Lata H, Chandra S, Khan IA, ElSohly MA (2009) Propagation through alginate encapsulation of axillary buds of Cannabis sativa L.—an important medicinal plant. Physiol Mol Biol Plants 15:79–86CrossRefPubMedPubMedCentralGoogle Scholar
  22. Meena AK, Rao MM, Meena RP, Panda P (2011) Pharmacological and phytochemical evidences for the plants of Wedelia genus—a review. Asian J Pharm Res 1:7–12Google Scholar
  23. Mehrotra S, Khwaja O, Kukreja AK, Rahman L (2012) ISSR and RAPD based evaluation of genetic stability of encapsulated micro shoots of Glycyrrhiza glabra following 6 months of storage. Mol Biotechnol 52(3):262–268CrossRefPubMedGoogle Scholar
  24. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–495CrossRefGoogle Scholar
  25. Murray M, Thopson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acid Res 8:4321–4325CrossRefPubMedPubMedCentralGoogle Scholar
  26. Nower AA (2014) In vitro propagation and synthetic seeds production: an efficient method for Stevia rebaudiana Bertoni. Sugar Tech 16(1):100–108CrossRefGoogle Scholar
  27. Palanyandy SR, Gantait S, Suranthran P, Sinniah UR, Subramaniam S (2015) Storage of encapsulated oil palm polyembryoids: influence of temperature and duration. Vitro Cell Dev Biol Plant 51(1):118–124CrossRefGoogle Scholar
  28. Parveen S, Shahzad A (2014) Encapsulation of nodal segments of Cassia angustifolia Vahl. for short-term storage and germplasm exchange. Acta Physiol Plant 36(3):635–640CrossRefGoogle Scholar
  29. Rahman MM, Bhadra SK (2011) Development of protocol for in vitro culture and rapid propagation of Wedelia chinensis (Osbeek) Merr. J Med Plants Res 5:2387–2392Google Scholar
  30. Rai MK, Jaiswal VS, Jaiswal U (2008) Encapsulation of shoot tips of guava (Psidium guajava L.) for short-term storage and germplasm exchange. Sci Hortic 118:33–38CrossRefGoogle Scholar
  31. Saha S, Sengupta C, Ghosh P (2014) Molecular and phytochemical analyses to assess genetic stability in alginate-encapsulated microshoots of Ocimum gratissimum L. following in vitro storage. Nucleus 57(1):33–43CrossRefGoogle Scholar
  32. Saha S, Sengupta C, Ghosh P (2015) Encapsulation, short-term storage, conservation and molecular analysis to assess genetic stability in alginate-encapsulated microshoots of Ocimum kilimandscharicum Guerke. Plant Cell Tissue Organ Cult 120(2):519–530CrossRefGoogle Scholar
  33. Sharma S, Shahzad A (2012) Encapsulation technology for short-term storage and conservation of a woody climber, Decalepis hamiltonii Wight and Arn. Plant Cell Tissue Organ Cult 111(2):191–198CrossRefGoogle Scholar
  34. Sharma S, Shahzad A, Kumar J, Anis M (2014) In vitro propagation and synseed production of scarlet salvia (Salvia splendens). Rend Fis Acc Lincei 25:359–368CrossRefGoogle Scholar
  35. Siddique I, Anis M (2009) Morphogenic response of the alginate encapsulated nodal segment and antioxidative enzymes analysis during acclimatization of Ocimum basilicum L. J Crop Sci Biotechnol 12(4):233–238CrossRefGoogle Scholar
  36. Singh AK, Sharma M, Varshney R, Agarwal SS, Bansal KC (2006) Plant regeneration from alginate to encapsulated shoot tips of Phyllanthus amarus Schum and Thonn, a medicinally important plant species. In Vitro Cell Dev Biol Plant 42:109–113CrossRefGoogle Scholar
  37. Singh SK, Rai MK, Asthana P, Pandey S, Jaiswal VS, Jaiswal U (2009) Plant regeneration from alginate encapsulated shoot tips of Spilanthes acmella L. Murr. A medicinally important and herbal pesticidal plant species. Acta Physiol Plant 31:649–653CrossRefGoogle Scholar
  38. Sundararaj SG, Agrawal A, Tyagi RK (2010) Encapsulation for in vitro short-term storage and exchange of ginger (Zingiber officinale Rosc.) germplasm. Sci Hortic 125(4):761–766CrossRefGoogle Scholar
  39. Suresh V, Kumar RM, Suresh A, Kumar NS, Arunachalam G, Umasankar K (2010) CNS activity of ethanol extract of Wedelia chinensis in experimental animals. Int J Pharm Sci Nanotechnol 3:881–886Google Scholar
  40. Verma SK, Rai MK, Asthana P, Jaiswal VS, Jaiswal U (2010) In vitro plantlets from alginate-encapsulated shoot tips of Solanum nigrum L. Sci Hort 124:517–521CrossRefGoogle Scholar

Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2018

Authors and Affiliations

  1. 1.Department of Agricultural Biotechnology, Faculty of AgricultureBidhan Chandra Krishi ViswavidyalayaNadiaIndia

Personalised recommendations