Advertisement

Physiology and Molecular Biology of Plants

, Volume 18, Issue 3, pp 273–280 | Cite as

In vitro propagation of spine gourd (Momordica dioica Roxb.) and assessment of genetic fidelity of micropropagated plants using RAPD analysis

  • Govind Kumar Rai
  • Major Singh
  • Neha Prakash Rai
  • D. R. Bhardwaj
  • Sanjeev Kumar
Short Communication

Abstract

An efficient protocol for rapid in vitro clonal propagation of spine gourd (Momordica dioica Roxb.) genotype RSR/DR15 (female) and DR/NKB-28 (male) was developed through enhanced axillary shoot proliferation from nodal segments. Maximum shoot proliferation of 6.2 shoots per explant with 100 % shoot regeneration frequency was obtained from the female genotype on Murashige and Skoog’s (1962) medium supplemented with 0.9 μM N6-benzyladenine (BA) and 200 mg l-1 casein hydrolysate (CH). While from the male genotype the optimum shoot regeneration frequency (86.6 %) and 6.4 shoots per explant was obtained on MS medium supplemented with 2.2 μM BA. CH induced vigorous shoots, promoted callus formation, and proved inhibitory for shoot differentiation and shoot length, especially in explants from male genotype. Rooting was optimum on half-strength MS medium (male 92.8 %, female 74.6 %) containing 4.9 μM indole-3-butyric acid (IBA). Plantlets were transferred to plastic cups containing a mixture of cocopit and perlite (1:1 ratio) and then to soil after 2–3 weeks. 84 % female and 81 % male regenerated plantlets survived and grew vigorously in the field. Genetic stability of the regenerated plants was assessed using random amplified polymorphic DNA (RAPD). The amplification products were monomorphic in the in vitro propagated plants and similar to those of mother plant. No polymorphism was detected revealing the genetic integrity of in vitro propagated plants. This micropropagation procedure could be useful for raising genetically uniform planting material of known sex for commercial cultivation or build-up of plant material of a specific sex-type.

Keywords

Momordica dioica Dioecious Axillary shoot proliferation Micropropagation Nodal explants Genetic fidelity 

References

  1. Agrawal V, Prakash S, Gupta SC (1999) Differential hormonal requirements for clonal propagation of male and female jojoba plants. In: Altman A, Ziv M, Izhar S (eds) Current science and biotechnology in agriculture: plant biotechnology and in vitro biology in the 21st century. Kluwer Academic Publishers, Dordrecht, pp 23–26Google Scholar
  2. Ali M, Okubo H, Fujii T, Fujieda K (1991) Techniques for propagation and breeding of kakrol (Momordica dioica Roxb.). Sci Hortic 47(3–4):335–343CrossRefGoogle Scholar
  3. Ali M, Shrivastava V (1998) Characterization of phytoconstitutents of the fruits of Momordica dioica. J Pharamaceutical Sci 60:287–289Google Scholar
  4. Ahmad N, Anis M (2005) In vitro mass propagation of Cucumis sativus L. from nodal segments. Turk J Bot 29:237–240Google Scholar
  5. Andrade LB, Echeverrigaray S, Fracaro F, Pauletti GF, Rota L (1999) The effect of growth regulators on shoot propagation and rooting of common lavender (Lavandula vera DC). Plant Cell Tiss Org Cult 56(2):79–83CrossRefGoogle Scholar
  6. Banerjee S, Zehra M, Kumar S (1999) In vitro multiplication of Centella asiatica, a medicinal herb from leaf explants. Curr Sci 76:147–148Google Scholar
  7. Bharathi LK, Naik G, Singh HS, Dora DK (2007) Spine gourd. In: Peter KV (ed) Underutilized and underexploited horticultural crops. New India Publishing, New Delhi, pp 289–295Google Scholar
  8. Bopana N, Saxena S (2008) In vitro propagation of a high value medicinal plant: Asparagus racemosus Willd. In Vitro Cell Dev Biol—Plant 44:525–532CrossRefGoogle Scholar
  9. Carvalho LC, Goulao L, Oliveira C, Goncalves JC, Amancio S (2004) RAPD assessment for identification of clonal identity and genetic stability of in vitro propagated chestnut hybrids. Plant Cell Tiss Org Cult 77:23–27CrossRefGoogle Scholar
  10. Ceasar SA, Maxwell SL, Prasad KB, Karthigan M, Ignacimuthu S (2010) Highly efficient shoot regeneration of Bacopa monnieri L. using a two-stage culture procedure and assessment of genetic integrity of micropropagated plants by RAPD. Acta Physiol Plant 32:443–452CrossRefGoogle Scholar
  11. Driver JA, Suttle GR (1987) Nursery handling of propagules. In: Bonga JM, Durzan DJ (eds) Cell and tissue culture in forestry. The Netherlands, Dordrecht, pp 320–335CrossRefGoogle Scholar
  12. Frabetti M, Gutiérrez-Pesce P, Mendoza-de Gyves E, Rugini E (2009) Micropropagation of Teucrium fruticans L., an ornamental and medicinal plant. In Vitro Cell Dev Biol—Plant 45:129–134CrossRefGoogle Scholar
  13. Hoque A, Islam R, Joarder OI (1995) In vitro plantlets differentiation in kakrol (Momordica dioica Roxb.). Plant Tiss Cult 5(2):119–124Google Scholar
  14. Hoque A, Islam R, Arima S (2000) High frequency plant regeneration from cotyledon-derived callus of Momordica dioica (Roxb.) Willd. Phytomorphology 50:267–272Google Scholar
  15. Hoque A, Hossain M, Alam S, Arima S, Islam R (2007) Adventitious shoot regeneration from immature embryo explant obtained from female × female Momordica dioica. Plant Tiss Cult Biotech 17(1):29–36Google Scholar
  16. Karim MA, Ahmed SU (2010) Somatic embryogenesis and micropropagation in teasle gourd. Int J of Environ Sci Dev 1(1):10–14CrossRefGoogle Scholar
  17. Martins M, Sarmento D, Oliveira MM (2004) Genetic stability of micropropagated almond plantlets as assessed by RAPD and ISSR markers. Plant Cell Rep 23:492–496CrossRefPubMedGoogle Scholar
  18. Mondal A, Ghosh GP, Zuberi MI (2006) Phylogenetic relationship in different kakrol collections of Bangladesh. Pakistan J Biol Sci 9(8):1516–1524CrossRefGoogle Scholar
  19. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  20. Nabi SA, Rashid MM, Al-Amin M, Rasul MG (2002) Organogenesis in teasle gourd (Momordica dioica Roxb.). Plant Tiss Cult 12(2):173–180Google Scholar
  21. Prakash S, Agrawal V, Gupta SC (2003) Influence of some adjuvants on in vitro clonal propagation of male and female jojoba plants. In Vitro Cell Dev Biol—Plant 39:217–222CrossRefGoogle Scholar
  22. Purohit SD, Dave A, Kukda G (1994) Micropropagation of safed musli (Chlorophytum borivilianum), a rare Indian medicinal herb. Plant Cell Tiss Org Cult 39(1):93–96CrossRefGoogle Scholar
  23. Ram D, Banerjee MK, Pandey S, Srivastava U (2001) Collection and evaluation of Kartoli (Momordica dioica Roxb. Ex. Willd.). Indian J Plant Genet Resour 14:114–116Google Scholar
  24. Rasul MG, Hiramatsu M, Okubo H (2007) Genetic relatedness (diversity) and cultivar identification by randomly amplified polymorphic DNA (RAPD) markers in teasle gourd (Momordica dioica Roxb.). Sci Hortic 111:271–279CrossRefGoogle Scholar
  25. Shu QY, Liu GS, Qi DM, Chu CC, Liu J, Li HJ (2003) An effective method for axillary bud culture and RAPD analysis of cloned plants in tetraploid black locust. Plant Cell Rep 22:175–180CrossRefPubMedGoogle Scholar
  26. Sultana RS, Bari Miah MA (2003) In vitro propagation of karalla (Momordica charantea Linn.) from nodal segment and shoot tip. J Biol Sci 3(12):1134–1139CrossRefGoogle Scholar
  27. Tiwari KN, Sharma NC, Tiwari V, Singh BD (2000) Micropropagation of Centella asiatica (L.), a valuable medicinal herb. Plant Cell Tiss Org Cult 63:179–185CrossRefGoogle Scholar
  28. Trivedi RN, Roy RP (1972) Cytological studies in some species of Momordica. Genetica 43:282–291CrossRefGoogle Scholar
  29. Venkatachalam L, Sreedhar RV, Bhagyalakshmi N (2007) Micropropagation in banana using high levels of cytokinins does not involve any genetic changes as revealed by RAPD and ISSR markers. Plant Growth Regul 51:193–205CrossRefGoogle Scholar

Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2012

Authors and Affiliations

  • Govind Kumar Rai
    • 1
  • Major Singh
    • 1
  • Neha Prakash Rai
    • 1
  • D. R. Bhardwaj
    • 1
  • Sanjeev Kumar
    • 1
  1. 1.Indian Institute of Vegetable ResearchVaranasiIndia

Personalised recommendations