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

  • Original Papers
  • Published:
Biologia Plantarum

Abstract

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.

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Abbreviations

AFLP:

amplified fragment length polymorphism

dNTPs:

deoxyribonucleotide triphosphates

ISSR:

inter simple sequence repeat

PCR:

polymarase chain reaction

RAPD:

random amplification for polymorphic DNA

RFLP:

restriction fragment length polymorphism

SAMPL:

selectively amplified microsatellite polymorphic locus

SSR:

simple sequence repeat

Taq:

Thermus aquaticus

UBC:

University of British Columbia

References

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • Doyle, J.J., Doyle, J.L.: Isolation of plant DNA from fresh tissue.-Focus 12: 13–15 1990.

    Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • Hirochika, H.: Activation of tobacco transposons during tissue culture.-EMBO J. 12: 2521–2528, 1993.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • Kaeppler, S.M., Phillips, R.L.: Tissue culture-induced DNA methylation variation in maize.-Proc. nat. Acad. Sci. USA 90: 8773–8776, 1993.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • Morgante, M., Vogel, J.: Compound microsatellite primers for detection of genetic polymorphisms.-US Patent Appl. 08/326456, 1994.

  • 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.

    Article  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • Rahman, M., Rajora, O.: Microsatellite DNA somaclonal variation in micropropagated trembling aspen (Populus tremuloides).-Plant Cell Rep. 20: 531–536, 2001.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • Tautz, D.: Hypervariability of simple sequences as a general source of polymorphic DNA markers.-Nucl. Acids Res. 17: 6463–6467, 1989.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • Vasil, I.K.: Developing cell and tissue culture systems for improvement of cereal and grass crops.-J. Plant Physiol. 128: 193–218, 1987.

    Google Scholar 

  • Vendrame, W.A., Kochertm, G., Wetzstein, H.Y.: AFLP analysis of variation in pecan somatic embryos.-Plant Cell Rep. 18: 853–857, 1999.

    Article  CAS  Google Scholar 

  • 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.

    CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Zietekiewicz, E., Rafalski, A., Labuda, D.: Genome fingerprinting by simple sequence repeats (SSR)-anchored polymerase chain reaction amplification.-Genomics 20: 176–183, 1994.

    Article  Google Scholar 

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  1. TERI School of Advanced Studies, Darbari Seth Block, Habitat Place, Lodhi Road, New Delhi, 110 003, India

    P. Joshi & V. Dhawan

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  1. P. Joshi
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  2. V. Dhawan
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Joshi, P., Dhawan, V. Assessment of genetic fidelity of micropropagated Swertia chirayita plantlets by ISSR marker assay. Biol Plant 51, 22–26 (2007). https://doi.org/10.1007/s10535-007-0005-0

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  • DOI: https://doi.org/10.1007/s10535-007-0005-0

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