Plant Cell, Tissue and Organ Culture

, Volume 57, Issue 2, pp 113–119 | Cite as

Analysis of somaclonal variation through tissue culture and chromosomal localization of rDNA sites by fluorescent in situ hybridization in wild Allium tuberosum and a regenerated variant

  • Geum-Sook Do
  • Bong-Bo Seo
  • Jong-Min Ko
  • Seon-Hee Lee
  • Jae-Hong Pak
  • In-Sun Kim
  • Seung-Dal Song
Article

Abstract

The effects of basal media and growth regulators on callus initiation and shoot regeneration have been investigated in wild Allium tuberosum (2n = 4x = 32). Callus initiation was greatest from flower bud explants cultured on MS medium supplemented with 2,4-D and BA at 1 mg l−1 each. Maximum number of shoots was obtained from callus grown on MS medium supplemented with NAA and BA at 0.2 and 2 mg l−1, respectively. The chromosome analysis of regenerants derived from callus revealed variation in ploidy, such as 2n = 28, 29, 30, 31, 33 as well as normal tetraploid. During the culture period for two generations, one aneuploid regenerant with 2n = 30 (named At30) showed better viability and growth than tetraploid plants and other aneuploid variants. In a karyotypic analysis of At30, the chromosomal positions of 5S and 18S-5.8S-26S rDNA were physically mapped by fluorescent in situ hybridization and compared to chromosomes of wild type A. tuberosum. Both wild type A. tuberosum and At30 exhibited two sets of 5S rDNA sites, one on the proximal position of the short arm of chromosome 3, and the other on the intercalary region on the long arm of chromosome 6. There was one 18S-5.8S-26S rDNA site in the secondary constriction including flanking short chromosomal segments of satellite and terminal regions on the short arm of chromosome 8 in wild type A. tuberosum. However, At30 showed only three labelled chromosome 8 indicating that this was one of the lost chromosomes of At30.

alliaceae aneuploid autotetraploid FISH Chinese chives 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Brown PTH, Lange KG, Kranz E & Lörz H (1993) Analysis of single protoplasts and regenerated plants by PCR and RAPD technology. Mol. Gen. Genet. 237: 311–317PubMedGoogle Scholar
  2. Dahleen LS & Eizenga GC (1990) Meiotic and isozymic characterization of plants regenerated from euploid and selfed monosomic tall fescue embryo. Theor. Appl. Genet.79: 39–44CrossRefGoogle Scholar
  3. D'Amato F (1977) Cytogenetics of differentiation in tissue and cell cultures. In: Reinert J & Bajaj YPS (eds) Applied and Fundamental Aspects of Plant Cell, Tissue and Organ Culture (pp. 343–357). Springer-Verlag, BerlinGoogle Scholar
  4. Deumling B & Clermont L (1989) Changes in DNA content and chromosomal size during cell culture and plant regeneration of Scilla siberica: selective chromatin diminution in response to environmental conditions. Chromosoma 97: 439–448CrossRefGoogle Scholar
  5. DeVerno LL, Charest PJ & Bonen L (1994) Mitochondrial DNA variation in somatic embryogenic cultures of Larix. Theor. Appl. Genet. 88: 727–732CrossRefGoogle Scholar
  6. Gerlach WL & Bedbrook JR (1979) Cloning and characterization of ribosomal RNA genes from wheat and barley. Nucleic Acids Res. 7: 1869–1885PubMedGoogle Scholar
  7. Hizume M (1994) Chromosomal localization of 18S and 5S rRNA genes in garlic, Allium sativum. Chromosome Information Service 56: 6–8Google Scholar
  8. Jiang J & Gill BS (1994) New 18S-26S ribosomal RNA gene loci: chromosomal landmarks for the evolution of polyploid wheats. Chromosoma103: 179–185PubMedGoogle Scholar
  9. Karp A, Owen PG, Steele SH, Bebeli PJ & Kaltskies PJ (1992) Variation in the telomeric heterochromatin in somaclones of rye. Genome 35: 590–593Google Scholar
  10. Kidwell KK & Osborn TC (1993) Variation among alfalfa somaclones in copy number of repeated DNA sequences. Genome 36: 906–912PubMedGoogle Scholar
  11. Lapitan NLV, Ganal MW & Tanksley SD (1989) Somatic chromosome karyotype of tomato based on in situ hybridization of the TAGI satellite repeat. Genome 32: 992–998Google Scholar
  12. Lee SH & Seo BB (1997) Chromosomal localization of 5S and 18S-26S rRNA genes using fluorescence in situ hybridization in Allium wakegi. Kor. J. Genet. 19: 19–26Google Scholar
  13. Lee SH, Ryu JA, Do GS, Seo BB, Pak JH, Kim IS & Song SD (1998) Chromosome analysis by fluorescence in situ hybridization of callus-derived regenerants in Allium cyaneum R.Plant Cell Rep. 18: 209–213CrossRefGoogle Scholar
  14. Leitch IJ & Heslop-Harrison JS (1993) Physical mapping of four sites of 5S ribosomal DNA sequences and one site of the x-amylase 2 gene in barley (Hordeum vulgare). Genome 36: 517–523PubMedGoogle Scholar
  15. Leitch AR, Schwarzacher T, Wang ML, Leitch IJ, Surlan-Momirovich G, Moore G & Heslop-Harrison JS (1993) Molecular cytogenetic analysis of repeated sequences in a long term wheat suspension culture. Plant Cell Tiss. Org. Cult. 33: 287–296CrossRefGoogle Scholar
  16. Linacero R & Vazquez AW (1992)Cytogenetic variation in rye regenerated plants and their progenies. Genome 35: 428–430Google Scholar
  17. Maluszynska J & Heslop-Harrison JS (1991) Localization of tandemly-repeated DNA sequences in Arabidopsis thaliana. Plant J. 1: 159–166Google Scholar
  18. Mukai Y, Endo TR & Gill B S (1990) Physical mapping of the 5S rRNA multigene family in common wheat. J. Hered. 81: 290–295Google Scholar
  19. Mukai Y, Endo TR & Gill BS (1991) Physical mapping of the 18S-26S rRNA multigene family in common wheat: identification of a new locus. Chromosoma 100: 71–78CrossRefGoogle Scholar
  20. Müller E, Brown PTH, Hartke S & Lörz H (1990) DNA variation in tissue-culture derived rice plants. Theor. Appl. Genet.80: 673–679CrossRefGoogle Scholar
  21. Munthali MT, Newbury HJ & Ford-Lloyd BV (1996) The detection of somaclonal variants of beet using RAPD. Plant Cell Rep. 15: 474–478CrossRefGoogle Scholar
  22. Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497CrossRefGoogle Scholar
  23. Nair AS & Seo BB (1995) Hormonal effects on chromosomal variability among the regenerated plants in Allium senescens L. var. Minor. Indian J. Exp. Biol. 33: 533–536Google Scholar
  24. Pandey R, Chandel KPS & Rao SR (1992) In vitro propagation of Allium tuberosum Rottl. ex. Spreng. by shoot proliferation. Plant Cell Rep. 11: 375–378Google Scholar
  25. Ricroch A, Peffley EB & Baker RJ (1992) Chromosomal location of rDNA in Allium: in situ hybridization using biotin-and fluorescein-labelled probe. Theor. Appl. Genet. 83: 413–418CrossRefGoogle Scholar
  26. Roy SC (1980) Chromosomal variations in the callus tissues of Allium tuberosum and A. cepa. Protoplasma 102: 171–176CrossRefGoogle Scholar
  27. Sekerka V (1977) Cytological characteristics of a tissue culture of Allium cepa L. Acta F. R. N. Comen. Physiol. Plant. 13: 35–42Google Scholar
  28. Seo BB (1977) Cytogenetic studies of some tetraploids in Allium. Kor. J. Bot. 20: 71–76Google Scholar
  29. Seo BB, Kim HH & Kim JH (1989) Giemsa C-banded karyotypes and their relationship of four diploid taxa in Allium. Kor. J. Bot. 32: 173–180Google Scholar
  30. Seo BB, Lee SH & Mukai Y (1997) Physical mapping of 5S and 18S-26S ribosomal RNA gene families in Allium victorialis var. platyphyllum. Kor. J. Plant Biol. 40: 132–137CrossRefGoogle Scholar
  31. Shoyama Y, Zhu XX, Nakai R, Shiraishi S & Kohda H (1997) Micropropagation of Panix notoginseng by somatic embryogenesis and RAPD analysis of regenerated plantlets. Plant Cell Rep. 16: 450–453Google Scholar
  32. Zee SY, Fung A & Yue SB (1977) Tissue culture and differentiation of chinese chive. HortScience 12: 264Google Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Geum-Sook Do
    • 1
  • Bong-Bo Seo
    • 1
  • Jong-Min Ko
    • 1
  • Seon-Hee Lee
    • 1
  • Jae-Hong Pak
    • 1
  • In-Sun Kim
    • 1
    • 2
  • Seung-Dal Song
    • 1
  1. 1.Department of BiologyKyungpook National UniversityTaeguRepublic of Korea
  2. 2.Department of BiologyKeimyung UniversityTaeguRepublic of Korea

Personalised recommendations