Acta Physiologiae Plantarum

, Volume 33, Issue 6, pp 2541–2545 | Cite as

Assessment of genetic fidelity of micropropagated plants of Simmondsia chinensis (Link) Schneider using RAPD and ISSR markers

  • Sunil Kumar
  • Manisha Mangal
  • A. K. Dhawan
  • Narender Singh
Short Communication


RAPD (random amplified polymorphic DNA) and ISSR (inter simple sequence repeat) markers were screened to test the genetic integrity of jojoba (Simmondsia chinensis) plants multiplied through axillary bud multiplication from nodal segments. The in vitro raised plantlets were maintained for up to 12 in vitro subcultures. During the study a total of 48 (32 RAPD and 16 ISSR) primers were screened, out of which 24 RAPD and 13 ISSR primers produced a total of 191 (126 RAPD and 65 ISSR) clear, distinct and reproducible amplicons. The amplified products were monomorphic across all the selected micropropagated plants and were similar to the mother plant. The micropropagation protocol developed by our group for rapid in vitro multiplication is appropriate for clonal propagation of jojoba. The outcome supports the fact that axillary bud multiplication can also be used as one of the safest modes for the production of true-to-type plants.


Axillary bud multiplication Molecular marker Nodal segment Somaclonal variations 



deoxyribonucleotide triphosphates


Polymerase chain reaction




α-Naphthalene acetic acid


  1. Agnihotri RK, Mishra J, Nandi SK (2009) Improved in vitro shoot multiplication and rooting of Dendrocalamus hamiltonii Nees et Arn. Ex Munro: production of genetically uniform plants and field evaluation. Acta Physiol Plant 31:961–967CrossRefGoogle Scholar
  2. Agrawal V, Prakash S, Gupta SC (2002) Effective protocol for in vitro shoot production through nodal explants of Simmondsia chinensis. Biol Plant 45:449–453CrossRefGoogle Scholar
  3. Agrawal V, Sharma K, Sarika G, Kumar R, Prasad M (2007) ISSR marker-assisted selection of male and female plants in a promising dioecious crop: jojoba (Simmondsia chinensis). Plant Biotech Rep 2:239–243Google Scholar
  4. Benzioni A (1992) Flower bud dormancy: ABA concentration and survival during frost of jojoba genotype under water stress. J Am Soc Hortic Sci 117:976–980Google Scholar
  5. Bhattacharya S, Bandopadhyay TK, Ghosh PD (2010) High frequency clonal propagation of Cymbopogon martinii var motia (palmarosa) through rhizome culture and true to type assessment using ISSR marker. J Plant Biochem Biotech 19:27–274Google Scholar
  6. Chaturvedi HC, Sharma M (1989) In vitro production of cloned plants of jojoba (Simmondsia chinensis (Link) Schneider) through shoot proliferation in long term culture. Plant Sci 63:199–207CrossRefGoogle Scholar
  7. Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15Google Scholar
  8. Joshi P, Dhawan V (2007) Assessment of genetic fidelity of micropropagated Swertia chirayita plantlets by ISSR marker assay. Biol Plant 51:22–26CrossRefGoogle Scholar
  9. Kumar S, Singh N, Mangal M (2009) Micropropagation of Simmondsia chinensis (Link) Schneider through enhanced axillary branching from nodal segments. J Plant Biol 36:75–81Google Scholar
  10. Kumar N, Modi AR, Singh AS, Gajera BB, Patel AR, Patel MP, Subhash N (2010a) Assessment of genetic fidelity of micropropagated date palm (Phoenix dactylifera L.) plants by RAPD & ISSR markers assay. Physiol Mol Biol Plants 16:207–213CrossRefGoogle Scholar
  11. Kumar S, Rai MK, Singh N, Mangal M (2010b) Alginate-encapsulation of shoot tips of jojoba [Simmondsia chinensis (Link) Schneider] for germplasm exchange and distribution. Physiol Mol Biol Plants 16:379–382CrossRefGoogle Scholar
  12. Lee CW (1988) Application of plant biotechnology for clonal propagation and yield enhancement in jojoba. In: Baldwin AR (ed) Proceeding of the seventh international conference on jojoba and its uses, Illinois, pp 102–111Google Scholar
  13. Low CB, Hackett WP (1981) Vegetative propagation of jojoba. California Agri 35:12–13Google Scholar
  14. Mills D, Wenkart S, Benzioni A (1997) Micropropagation of Simmondsia chinensis (jojoba). In: Bajaj YPS (ed) Biotechnology in agriculture and forestry. Springer, Berlin, pp 370–393Google Scholar
  15. Murashige T, Skoog FA (1962) Revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  16. Narula A, Kumar S, Srivastava PS (2007) Genetic fidelity of in vitro regenerants encapsulation of shoot tips and high diosgenin content in Dioscorea bulbifera L., a potential alternative source of diosgenin. Biotech Lett 29:623–629CrossRefGoogle Scholar
  17. Phillips RL, Kaeppler SM, Olhoft P (1994) Genetic instability of plant tissue cultures: breakdown of normal controls. Proc Natl Acad Sci USA 91:5222–5226PubMedCrossRefGoogle Scholar
  18. Rahman MH, Rajora OP (2001) Microsatellite DNA somaclonal variation in micropropagated trembling aspen (Populus tremuloides). Plant Cell Rep 20:531–536CrossRefGoogle Scholar
  19. Ray T, Dutta I, Saha P, Das S, Roy SC (2006) Genetic stability of three economically important micropropagated banana (Musa spp.) cultivars of lower Indo-Gangetic plains, as assessed by RAPD and ISSR markers. Plant Cell Tissue Organ Cult 85:11–21CrossRefGoogle Scholar
  20. Reddy MP, Chikara J (2010) Biotechnology advances in jojoba (Simmondsia chinensis). In: Ramawat KG (ed) Desert plants. Springer, Berlin, pp 407–422CrossRefGoogle Scholar
  21. Reddy MP, Sarla N, Siddiq EA (2002) Inter simple sequence repeat (ISSR) polymorphism and its application in plant breeding. Euphytica 128:9–17CrossRefGoogle Scholar
  22. Roussos PA, Marioli TM, Pontikis CA, Kotsias D (1999) Rapid multiplication of jojoba seedling by in vitro culture. Plant Cell Tissue Organ Cult 57:133–137CrossRefGoogle Scholar
  23. Sharma K, Agrawal V, Sarika G, Kumar R, Prasad M (2008) ISSR marker-assisted selection of male and female plants in a promising dioecious crop: jojoba (Simmondsia chinensis). Plant Biotech Rep 2:239–243CrossRefGoogle Scholar
  24. Sharma M, Rai SK, Purshottam DK, Jain M, Chakrabarty D, Awasthi A, Nair KN, Kumar A (2009) In vitro clonal propagation of Clerodendrum serratum (Linn.) Moon (barangi): a rare and threatened medicinal plant. Acta Physiol Plant 31:379–383CrossRefGoogle Scholar
  25. Singh A, Reddy MP, Patolia JS (2008) An improved protocol for micropropagation of elite genotypes of Simmondsia chinensis (Link) Schneider. Biol Plant 52:538–542CrossRefGoogle Scholar
  26. Tawar PN, Sawant RA, Dalvi SG, Nikam AA, Kawar PG, Devarumath RM (2008) An assessment of somaclonal variation in micropropagated plants of sugarcane by RAPD markers. Sugar Tech 10:124–127CrossRefGoogle Scholar
  27. Tyagi RK, Prakash S (2004) Genotype and sex-specific protocol for in vitro micropropagation and medium-term conservation of Jojoba. Biol Plant 48:119–123Google Scholar
  28. Tyagi P, Khanduja S, Kothari SL (2010) In vitro culture of Capparis decidua and assessment of clonal fidelity of the regenerated plants. Biol Plant 54:126–130CrossRefGoogle Scholar
  29. Zietkiewicz E, Rafalski A, Labuda D (1994) Genomic fingerprinting by simple sequence repeat (SSR) anchored polymerase chain reaction amplification. Genomics 20:176–183PubMedCrossRefGoogle Scholar

Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2011

Authors and Affiliations

  • Sunil Kumar
    • 1
    • 2
  • Manisha Mangal
    • 1
  • A. K. Dhawan
    • 1
  • Narender Singh
    • 2
  1. 1.Centre for Plant Biotechnology, CCS Haryana Agricultural University New CampusHisarIndia
  2. 2.Department of BotanyKurukshetra UniversityKurukshetraIndia

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