biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 51:22-26, 2007 | DOI: 10.1007/s10535-007-0005-0

Assessment of genetic fidelity of micropropagated Swertia chirayita plantlets by ISSR marker assay

P. Joshi1,*, V. Dhawan1
1 TERI School of Advanced Studies, Darbari Seth Block, Habitat Place, New Delhi, India

Inter simple sequence repeat (ISSR) marker assay was employed to validate the genetic fidelity of Swertia chirayita plantlets multiplied in vitro by axillary multiplication upto forty-two passages. Sixteen ISSR primers generated a total of 102 amplicons among the tissue-cultured plants. Forty-eight amplicons were amplified in the outlier (a Swertia species). The outlier (negative control) was employed to rule out the possibility that the invariant fingerprint was due to chance alone and that the ISSR technique employed was not discriminatory enough to detect the off-types. A homogenous amplification profile was observed for all the micropropagated plants. The results confirmed the clonal fidelity of the tissue culture-raised S. chirayita plantlets and corroborated the fact that axillary multiplication is the safest mode for multiplication of true to type plants.

Keywords: axillary multiplication; conservation; endangered medicinal plant; molecular markers; somaclonal variations
Subjects: amplified fragment length polymorphism (AFLP); auxins; cytokinins; endangered plants; in vitro culture, micropropagation, propagation; in vitro culture, regeneration, proliferation, differentiation; medicinal plants; polymerase chain reaction (PCR); somaclonal variation; Swertia chirayita

Received: June 9, 2005; Accepted: November 11, 2005; Published: March 1, 2007  Show citation

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Joshi, P., & Dhawan, V. (2007). Assessment of genetic fidelity of micropropagated Swertia chirayita plantlets by ISSR marker assay. Biologia plantarum, 51(1), 22-26. doi: 10.1007/s10535-007-0005-0
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    References

    1. Carvalho, L.C., Goulao, L., Oliveira, C., Goncalves, J.C., Amancio, S.: RAPD assessment for clonal and genetic stability of in vitro propagated chestnut hybrids.-Plant Cell Tissue Organ Cult. 77: 23-27, 2004. Go to original source...
    2. Chowdari, K.V., Ramakrishna, W., Tamhankar, S.A., Hendre, R.R., Gupta, V.S., Sahasrabudhe, N.A., Ranjekar, P.K.: Identification of minor DNA variations in rice somaclonal variants.-Plant Cell Rep. 18: 55-58, 1998. Go to original source...
    3. Devarumath, R., Nandy, S., Rani, V., Marimuthu, S., Muraleedharan, N., Raina, S. RAPD, ISSR and RFLP fingerprints as useful markers to evaluate genetic integrity of micropropagated plants of three diploid and triploid elite tea clones representing Camellia sinensis (China type) and C. assamica ssp. assamica (Assam-India type).-Plant Cell Rep. 21: 166-173, 2002. Go to original source...
    4. Doyle, J.J., Doyle, J.L.: Isolation of plant DNA from fresh tissue.-Focus 12: 13-15 1990. Go to original source...
    5. Fang, D.Q., Roose, M.L.: Identification of closely related citrus cultivars with inter-simple sequence repeat markers.-Theor. appl. Genet. 95: 408-417, 1997. Go to original source...
    6. Goto, S., Thakur, R.C., Ishii, K.: Determination of genetic stability in long-term micropropagated shoots of Pinus thunbergii Parl. using RAPD markers.-Plant Cell Rep. 18: 193-197, 1998. Go to original source...
    7. Gupta, M., Chyi, Y.S., Romero-Severson, J., Owen, J.L.: Amplification of DNA markers from evolutionarily diverse genomes using single primers of simple sequence repeats.-Theor. appl. Genet. 89: 998-1006, 1994. Go to original source...
    8. Hirochika, H.: Activation of tobacco transposons during tissue culture.-EMBO J. 12: 2521-2528, 1993. Go to original source...
    9. Hirochika, H., Sugimoto, K., Otsuki, Y., Tsugawa, H., Kanda, M.: Retrotransposon of rice involved in mutations induced by tissue culture.-Proc. nat. Acad. Sci. USA 93: 7783-7788, 1996. Go to original source...
    10. Hofmann, N.E., Raja, A., Nelson, R.L., Korban, S.S.: Mutagenesis of embryogeneic cultures of soybean and detecting polymorphism using RAPD markers.-Biol. Plant. 48: 173-177, 2004. Go to original source...
    11. Kaeppler, S.M., Phillips, R.L.: Tissue culture-induced DNA methylation variation in maize.-Proc. nat. Acad. Sci. USA 90: 8773-8776, 1993. Go to original source...
    12. Leroy, X.J., Leon, K., Charles, G., Branchard, M.: Cauliflower somatic embryogenesis and analysis of regenerant stability by ISSRs.-Plant Cell Rep. 19: 1102-1107, 2000. Go to original source...
    13. Leroy, X.J., Leon, K., Hily, J.M., Chaumeil, P., Branchard, M.: Detection of in vitro culture-induced instability through inter-simple sequence repeat analysis.-Theor. appl. Genet. 102: 885-891, 2001. Go to original source...
    14. Liu, Z.L., Han, F.P., Tan, M., Shan, X.H., Dong, Y.Z., Wang, X.Z., Fedak, G., Hao, S., Liu, B.: Activation of a rice endogenous retrotransposon Tos17 in tissue culture is accompanied by cytosine demethylation and causes heritable alteration in methylation pattern of flanking genomic regions.-Theor. appl. Genet. 109: 200-209, 2004. Go to original source...
    15. Martins, M., Sarmento, D., Oliveira, M.M.: Genetic stability of micropropagated almond plantlets, as assessed by RAPD and ISSR markers.-Plant Cell Rep. 23: 492-496, 2004. Go to original source...
    16. Matthes, M., Singh, R., Cheah, S.C., Karp, A.: Variation in oil palm Elaeis guineensis (Jacq.) tissue culture-derived regenerants revealed by AFLPs with methylation-sensitive enzymes.-Theor. appl. Genet. 102: 971-979, 2001. Go to original source...
    17. Meyer, W., Michell, T.G., Freedman, E.Z., Vilgalys, R.: Hybridization probes for conventional DNA fingerprinting used as single primers in polymerase chain reaction to distinguish strains of Cryptococcus neoformans.-J. clin. Biol. 31: 2274-2280, 1993. Go to original source...
    18. Morgante, M., Vogel, J.: Compound microsatellite primers for detection of genetic polymorphisms.-US Patent Appl. 08/326456, 1994.
    19. Nagaoka, T., Ogihara, Y.: Applicability of inter-simple sequence repeat markers in wheat for use as DNA markers in comparison to RFLP and RAPD markers.-Theor. appl. Genet. 97: 597-602, 1997. Go to original source...
    20. Palombi, M., Damiano, V.: Comparison between RAPD and SSR molecular markers in detecting genetic variation in kiwifruit (Actinidia deliciosa A. Chev).-Plant Cell Rep. 20: 1061-1066, 2002. Go to original source...
    21. Phillips, R.L., Kaeppler, S.M., Olhoft, P.: Genetic instability of plant tissue cultures: Breakdown of normal controls.-Proc. nat. Acad. Sci. USA 91: 5222-5226, 1994. Go to original source...
    22. Powell, W., Morgante, R., Andre, C., Hanafey, M., Vogel, J., Tingey, S., Rafalsky, A.: The comparison of RFLP, RAPD, AFLP and SSR (microsatellite) markers for germplasm analysis.-Mol. Breed. 2: 225-238, 1996. Go to original source...
    23. Rahman, M., Rajora, O.: Microsatellite DNA somaclonal variation in micropropagated trembling aspen (Populus tremuloides).-Plant Cell Rep. 20: 531-536, 2001. Go to original source...
    24. Salvi, N.D., George, L., Eapen, S.: Plant regeneration from the leaf base callus of turmeric and random amplified polymorphic DNA analysis of the regenerated plants.-Plant Cell Tissue Organ Cult. 66: 113-119, 2001. Go to original source...
    25. Shu, Q.Y., Liu, G.S., Qi, D.M., Chu, C.C., Liu, J. Li, H.J.: An effective method for axillary bud culture and RAPD analysis of cloned plants in tetraploid black locust.-Plant Cell Rep. 22: 175-180, 2003. Go to original source...
    26. Singh, A., Negi, M.S., Moses, V.K., Venkateswarlu, B., Srivastava, P.S., Lakshmikumaram, M.: Molecular analysis of micropropagated neem plants using AFLP markers for ascertaining clonal fidelity.-In vitro cell. dev. Biol. Plant 38: 519-521, 2002. Go to original source...
    27. Singh, M., Saroop, J., Dhiman, B.: Detection of intra-clonal genetic variability in vegetatively propagated tea using RAPD markers.-Biol. Plant. 48: 113-115, 2004. Go to original source...
    28. Tautz, D.: Hypervariability of simple sequences as a general source of polymorphic DNA markers.-Nucl. Acids Res. 17: 6463-6467, 1989. Go to original source...
    29. Varshney, A., Lakshmikumaran, M., Srivastava, P.S., Dhawan, V.: Establishment of genetic fidelity of in vitro raised Lillium bulblets through RAPD markers.-In vitro cell. Dev. Biol. Plant 37: 227-231, 2001. Go to original source...
    30. Vasil, I.K.: Developing cell and tissue culture systems for improvement of cereal and grass crops.-J. Plant Physiol. 128: 193-218, 1987. Go to original source...
    31. Vendrame, W.A., Kochertm, G., Wetzstein, H.Y.: AFLP analysis of variation in pecan somatic embryos.-Plant Cell Rep. 18: 853-857, 1999. Go to original source...
    32. Vyskot, B., Reich, J., Fajkus, J., Bedek, M., Soska, J.: Genome modifications in protoplast-derived tobacco plants: Contents of repetitive DNA sequences.-Biol. Plant. 33: 448-454, 1991. Go to original source...
    33. Xu, M., Li. X., Korban, S.S.: DNA-methylation alterations and exchanges in vitro cellular differentiation in rose (Rosa hybrida L.).-Theor. appl. Genet. 109: 899-910, 2004. Go to original source...
    34. Zietekiewicz, E., Rafalski, A., Labuda, D.: Genome fingerprinting by simple sequence repeats (SSR)-anchored polymerase chain reaction amplification.-Genomics 20: 176-183, 1994. Go to original source...

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