Skip to main content
Log in

Varietal Discrimination and Genetic Variability Analysis of Cymbopogon Using RAPD and ISSR Markers Analysis

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

Cymbopogon is an important genus of family Poaceae, cultivated mainly for its essential oils which possess high medicinal and economical value. Several cultivars of Cymbopogon species are available for commercial cultivation in India and identification of these cultivars was conceded by means of morphological markers and essential oil constitution. Since these parameters are highly influenced by environmental factors, in most of the cases, it is difficult to identify Cymbopogon cultivars. In the present study, Random amplified polymorphic DNA (RAPD) and Inter-simple sequence repeat (ISSR) markers were employed to discriminate nine leading varieties of Cymbopogon since prior genomic information is lacking or very little in the genus. Ninety RAPD and 70 ISSR primers were used which generated 63 and 69 % polymorphic amplicons, respectively. Similarity in the pattern of UPGMA-derived dendrogram of RAPD and ISSR analysis revealed the reliability of the markers chosen for the study. Varietal/cultivar-specific markers generated from the study could be utilised for varietal/cultivar authentication, thus monitoring the quality of the essential oil production in Cymbopogon. These markers can also be utilised for the IPR protection of the cultivars. Moreover, the study provides molecular marker tool kit in both random and simple sequence repeats for diverse molecular research in the same or related genera.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

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

References

  1. Jagadishchandra, K. S. (1975). Recent studies on Cymbopogon Spreng. (aromatic grasses) with special reference to Indian taxa: taxonomy, cytogenetics, chemistry and scope. Journal of Plantation Crops, 3, 43–57.

    Google Scholar 

  2. Pinder, A. R. (1960). Chemistry of terpenes. (pp. 38–40). Chapman and Hall Ltd, London

  3. Subrahmanyan, V., & Bose, S.M. (1955). Bactericidal property of Indian essential oils with special reference to lemongrass oil. In Essential oils and Aromatic chemicals (pp. 1958). A symposium held at Dehra Dun October 6–9, under Joint Auspices of CSIR and Forest Research Institute and College CSIR, New Delhi.

  4. Tyagi, B. K., Shahi, A. K., & Koul, B. L. (1988). Evaluation of repellent activities of Cymbopogon essential oils against mosquito vectors of Malaria, Filariasis and Dengue fever. Phytomedicine, 5, 324–329.

    Article  Google Scholar 

  5. Nybom, H., & Bartish, I. (2000). Effects of life history traits and sampling strategies on genetic diversity estimates obtained with RAPD markers in plants. Perspectives in Plant Ecology Evolution and Systatics, 3, 93–114.

    Article  Google Scholar 

  6. Williams, J. G. K., Kubelik, A. R., Livak, K. J., Rafalski, J. A., & Tingey, S. V. (1990). DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research, 18, 6531–6535.

    Article  CAS  Google Scholar 

  7. Zietkiewicz, E., Rafalski, A., & Labuda, D. (1994). Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics, 20, 176–183.

    Article  CAS  Google Scholar 

  8. Karatash, H., & Agaoglu, S. Y. (2008). Genetic diversity among Turkish local grape accessions (Vitis vinifera L.) using RAPD markers. Hereditas, 145, 58–63.

    Article  Google Scholar 

  9. Ahmad, F., Khan, A. I., Awan, F. S., & Sadia, B. (2010). Genetic diversity of chickpea (Cicer arietinum L.) germplasm in Pakistan as revealed by RAPD analysis. Genetics and Molecular Research, 9, 1414–1420.

    Article  CAS  Google Scholar 

  10. Geetha, K. A., Kawane, A., Bishoyi, A. K., Phurailatpam, A., Ankita, C., Malik, S. K., Srinivasan, R., & Bhat, S. R. (2013). Characterization of mode of reproduction of Commiphora wightii [(Arnott) Bhandari] reveals novel pollen-pistil interaction and occurrence of obligate sexual female plants. Trees Structure and Function, 27, 567–581.

    Article  Google Scholar 

  11. Bishoyi, A. K., Pillai, V. V., Geetha, K. A., & Maiti, S. (2014). Assessment of genetic diversity in Clitoria ternatea populations from different parts of India by RAPD and ISSR markers. Genetic Resources and Crop Evolution, 6, 1597–1609.

    Article  Google Scholar 

  12. Ratnaparkhe, M. B., Santra, D. K., Tullu, A., & Muehlbauer, F. J. (1998). Inheritance of inter-simple-sequence-repeat polymorphisms and linkage with a fusarium wilt resistance gene in chickpea. Theory and Applied Genetics, 96, 348–353.

    Article  CAS  Google Scholar 

  13. Desai, P., Gajera, B., Mankad, M., Shah, S., Patel, A., Patil, G., Narayanan, S., & Kumar, N. (2015). Comparative assessment of genetic diversity among Indian bamboo genotypes using RAPD and ISSR markers. Molecular Biology Reports, 42, 1265–1273.

    Article  CAS  Google Scholar 

  14. Long, Y., Cheng, J., Mei, Z., Zhao, L., Wei, C., Fu, S., Khan, M. A., & Fu, J. (2015). Genetic analysis of litchi (Litchi chinensis Sonn.) in southern China by improved random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR). Molecular Biology Reports, 42, 159–166.

    Article  CAS  Google Scholar 

  15. Sangwan, N. S., Yadav, U., & Sangwan, R. S. (2001). Molecular analysis of genetic diversity in elite Indian cultivars of essential oil trade types of aromatic grasses (Cymbopogon species). Plant Cell Reports, 20, 437–444.

    Article  CAS  Google Scholar 

  16. Shasany, A. K., Lal, R. K., Patra, N. K., Darokar, M. P., Garg, A., Kumar, S., & Khanuja, S. P. S. (2000). Phenotypic and RAPD diversity among Cymbopogon winterianus Jowitt accessions in relation to Cymbopogon nardus Rendle. Genetic Resources and Crop Evolution, 47, 553–559.

    Article  Google Scholar 

  17. Khanuja, S. P. S., Shasany, A. K., Pawar, A., Lal, R. K., Darokar, M. P., Naqvi, A. A., Rajkumar, S., Sundaresan, V., Lal, N., & Kumar, S. (2005). Essential oil constituents and RAPD markers to establish species relationship in Cymbopogon Spreng. (Poaceae). Biochemical Systematic and Ecology, 33, 171–186.

    Article  CAS  Google Scholar 

  18. Ganjewala, D. (2008). RAPD characterization of three selected cultivars OD-19, GRL-1 and Krishna of East Indian Lemongrass (Cymbopogon flexuosus Nees ex Steud) Wats. American Eurasian Journal of Botany, 1, 53–57.

    Google Scholar 

  19. Adhikari, S., Bandopadhyay, T. K., & Ghosh, P. D. (2013). Assessment of genetic diversity in certain Indian elite clones of Cymbopogon species through RAPD analysis. Indian Journal of Biotechnology, 12, 109–114.

    CAS  Google Scholar 

  20. Kumar, J., Verma, V., Shahi, A. K., Qazi, G. N., & Balyan, H. S. (2007). Development of simple sequeme repeats markers in Cymbopogon species. Planta Medica, 73, 262–266.

    Article  CAS  Google Scholar 

  21. Kumar, J., Verma, V., Goyal, A., Shahi, A. K., Sparoo, R., Sangwan, R. S., & Qazi, G. N. (2009). Genetic diversity analysis in Cymbopogon species using DNA markers. Plant Omics Journal, 2, 20–29.

    CAS  Google Scholar 

  22. Shiran, B., Amirbakhtiar, N., Kiani, S., Mohammadi, S., Sayed-Tabatabaei, B. E., & Moradi, H. (2007). Molecular characterization and genetic relationship among almond cultivars assessed by RAPD and SSR markers. Scientia Horticulture, 111, 280–290.

    Article  CAS  Google Scholar 

  23. Rohlf, F. J. (1998). NTSYS-PC, numerical taxonomy and multivariate analysis system, version 2.02a. Setauket, New York: Exeter Publications.

    Google Scholar 

  24. Jaccard, P. (1908). Nouvelles recherches sur la distribution florale. Bulletin de la Societe vaudoise des sciences naturelles, 44, 223–270.

    Google Scholar 

  25. Sneath, P. H. A., & Sokal, R. R. (1973). Numerical taxonomy—the principles and practice of numerical classification. W. H., San Francisco: Freeman.

    Google Scholar 

  26. Prevost, A., & Wilkinson, M. J. (1999). A new system of comparing PCR primers applied to ISSR fingerprinting. Theory and Applied Genetics, 98, 107–112.

    Article  CAS  Google Scholar 

  27. Powell, W., Morgante, M., Andre, C., Hanafey, M., Vogel, J., Tingey, S., & Rafalski, A. (1996). The unity of RFLP, RAPD, AFLP and SSR (microsatellite) markers for germplasm analysis. Molecular Breeding, 2, 25–238.

    Article  Google Scholar 

  28. Anderson, J. A., Churchill, G. A., Autrique, J. E., Tanksley, S. D., & Sorrells, M. E. (1993). Optimizing parental selection for genetic linkage maps. Genome, 36, 181–186.

    Article  CAS  Google Scholar 

  29. Manly, B. F. J. (1997). Randomization, bootstrap and Monte Carlo methods in biology. London: Chapman & Hall.

    Google Scholar 

  30. Divaret, I., Margale, E., & Thomas, G. (1999). RAPD markers on seed bulks efficiently assess the genetic diversity of Brassica oleracea L. collection. Theory and Applied Genetics, 98, 1029–1035.

    Article  CAS  Google Scholar 

  31. Kumar, L. S., Sawant, A. S., Gupta, V. S., & Ranjekar, P. K. (2001). Comparative analysis of genetic diversity among Indian populations of Scirpophaga incertulas by ISSR-PCR and RAPD-PCR. Biochemical Genetics, 39, 297–309.

    Article  CAS  Google Scholar 

  32. Ngezahayo, F., Dong, Y., & Liu, B. (2007). Somaclonal variation at the nucleotide sequence level in rice (Oryza sativa L.) as revealed by RAPD and ISSR markers, and by pairwise sequence analysis. Journal of Applied Genetics, 48, 329–336.

    Article  Google Scholar 

  33. Maslova, E. V. (2008). Differentiation of two hemp nettle species (Galeopsis bifida Boenn. and G. tetrahit L.) inferred from morphological characters and DNA markers. Russian Journal of Genetics, 44, 312–319.

    Article  CAS  Google Scholar 

  34. Mohanty, S., Das, A. B., Ghosh, N., Panda, B. B., & Smith, D. W. (2010). Genetic diversity of 28 wild species of fodder legume Cassia using RAPD, ISSR, and SSR markers: a novel breeding strategy. Journal of Biotechnology Research, 2, 44–55.

    CAS  Google Scholar 

  35. Chandrashekar, R., Prasad, B., Vankudothu, N., Jyothi, C. P., Lakshmi, B. N., Rama, R. Y., & Kudle, K. R. (2015). A comparative genetic diversity study among elite Bt and Non-Bt cotton varieties of Andhra Pradesh by RAPD markers. Discover Biotechnology, 16, 30–37.

    Google Scholar 

  36. Khaled, K. A., El-Demardash, I. S., & Amer, E. A. M. (2015). Genetic polymorphism among some sugarcane germplasm collections as revealed by RAPD and ISSR analyses. Life Science Journal, 12, 159–167.

    CAS  Google Scholar 

  37. Thangjam, R. (2014). Inter-simple sequence repeat (ISSR) marker analysis in Parkia timoriana (DC.) Merr. populations from Northeast India. Applied Biochemistry and Biotechnology, 172, 1727–1734.

    Article  CAS  Google Scholar 

  38. Mishra, P., Kumar, L. D., Kumar, A., Gokul, S., Ravikumar, K., Shukla, A. K., & Sundaresan, V. (2015). Population dynamics and conservation implications of Decalepis arayalpathra (J. Joseph and V. Chandras.) Venter., a steno endemic species of Western Ghats, India. Applied Biochemistry and Biotechnology, 176, 1413–1430.

    Article  CAS  Google Scholar 

  39. Mattioni, C., Casasoli, M., Gonzalez, M., & Ipinza, R. (2002). Comparison of ISSR and RAPD markers to characterize three Chilean Nothofagus species. Theory and Applied Genetics, 104, 1064–1070.

    Article  CAS  Google Scholar 

  40. Mukherjee, A., Sikdar, B., Ghosh, B., Banerjee, A., Ghosh, E., Bhattacharya, M., & Roy, S. C. (2013). RAPD and ISSR analysis of some economically important species, varieties, and cultivars of the genus Allium (Alliaceae). Turkish Journal of Botany, 37, 605–618.

    CAS  Google Scholar 

  41. Blair, M. W., Panaud, O., & McCouch, S. R. (1999). Inter-simple sequence repeat (ISSR) amplification for analysis of microsatellite motif frequency and fingerprinting in rice (Oryza sativa L.). Theory and Applied Genetics, 98, 780–792.

    Article  CAS  Google Scholar 

Download references

Acknowledgment

The authors are thankful to the ICAR-Directorate of Medicinal and Aromatic Plants Research (ICAR-DMAPR) for providing facilities for this study. We also acknowledge the encouragement and support provided by Dr. Satyabrata Maiti, former Director, ICAR-DMAPR, for conducting the study.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to K.A. Geetha.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflict of interest.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Table 1

(DOC 17 kb)

Supplementary Table 2

(DOC 85 kb)

Supplementary Table 3

(DOC 93 kb)

Supplementary Table 4

(DOC 45 kb)

Supplementary Table 5

(DOC 30 kb)

Supplementary Table 6

(DOC 20 kb)

Supplementary Table 7

(DOC 20 kb)

Supplementary Table 8

(DOC 19 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bishoyi, A.K., Sharma, A., Kavane, A. et al. Varietal Discrimination and Genetic Variability Analysis of Cymbopogon Using RAPD and ISSR Markers Analysis. Appl Biochem Biotechnol 179, 659–670 (2016). https://doi.org/10.1007/s12010-016-2022-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-016-2022-y

Keywords

Navigation