Abstract
A rapidly growingTriticum aestivum L. (wheat) derived long term suspension culture (named TaKB1), that is probably not regenerable, was analysed for karyotype rearrangements, stability and changes in repetitive DNA. The cell line has an average chromosome number of 21 and the DNA amount of unreplicated cells of TaKB1 measured by flow cytometry is about 30% lower than an unreplicated (1C) bread wheat genome.In situ hybridization of a repetitive DNA sequence (pSc119.2), which occurs as tandemly repeated blocks (heterochromatin) in wheat, shows that chromosomes from the TakB1 line have fewer and weaker subtelomeric locations of the sequence than wheat, suggesting deletions of distal chromosome segments and a reduction in the sites and copy number of the sequence. Thein situ hybridization pattern and chromosome morphology allowed 27 chromosome types to be identified in the cell line. No two analysed cells contained the same chromosome complement, although some chromosome types were present in every cell. Using Southern hybridization the structure and copy number of a retroelement (Wis-2) and its flanking sequence was shown to be the same in the TaKB1 cell line and wheat. Anin situ analysis of rDNA in the TaKB1 cell line (using the probe pTa71) showed a reduction in number of sites and rRNA genes in each cell from that in wheat. Interphase cells of the cell line showed dispersed signal throughout the nucleolus with no evidence for clusters of condensed and inactive rRNA genes.
Similar content being viewed by others
References
Ambros PE, Matzke MA & Matzke AJM (1986) Detection of a 17 kb unique sequence (T-DNA) in plant chromosomes byin situ hybridization. Chromosoma 94: 11–18
Anamthawat-Jónsson K, Schwarzacher T, Leitch AR, Bennett MD & Heslop-Harrison JS (1990) Discrimination between closely relatedTriticeae species using genomic DNA as a probe. Theor. Appl. Genet. 79: 721–728
Arumuganathan K, Slattery JP, Tanksley SD & Earle ED (1991) Preparation and flow cytometric analysis of metaphase chromosomes of tomato. Theor. Appl. Genet. 82: 101–111
Bayliss MW (1980) Chromosomal variation in plant tissues in culture. Int. Rev. Cytol. 11A: 113–144
Bedbrook JR, Jones J, O'Dell M, Thompson RD & Flavell RB (1980) A molecular description of telomeric heterochromatin inSecale species. Cell 19: 545–560
Bennett MD & Smith JB (1976) Nuclear DNA amounts in angiosperms. Philos. Trans. R. Soc. Lond. Ser. B 274: 227–274
Brettell RIS & Pallotta MA, Gustafson JP & Appels R (1986) Variation at theNor loci in triticale derived from tissue culture. Theor. Appl. Genet. 71: 637–643
Brown PTH, Göbel E & Lörz H (1991) RFLP analysis ofZea mays callus cultures and their regenerated plants. Theor. Appl. Genet. 81: 227–232
Eriksson T (1966) Partial synchronization of cell division in suspension cultures ofHaplopappus gracilis. Physiol. Plant. 19: 900–910
Flavell RB & O'Dell M (1979) The genetic control of nucleolus formation in wheat. Chromosoma 71: 135–152
Frearson EM, Power JB & Cocking EC (1973) The isolation, culture and regeneration ofPetunia leaf protoplasts. Dev. Biol. 33: 130–137
Friebe B, Mukai Y, Dhaliwal HS, Martin TJ & Gill BS (1991) Identification of alien chromatin specifying resistance to wheat streak mosaic and greenbug in wheat germ plasm by C-banding andin situ hybridization. Theor. Appl. Genet. 81: 381–389
Gerlach WL & Bedbrook JR (1979) Cloning and characterization of ribosomal RNA genes from wheat and barley. Nucleic Acids Res. 7: 1869–1885
Hadlaczky Gy, Bisztray Gy, Praznovszky T & Dudits D (1983) Mass isolation of plant chromosomes and nuclei. Planta 157: 278–285
Heslop-Harrison JS, Schwarzacher T, Anamthawat-Jónsson K, Leitch AR, Shi M & Leitch IJ (1992)In situ hybridization with automated chromosome denaturation. Technique 3: 104–115
Kao KN (1977) Chromosomal behaviour in somatic hybrids of soybean —Nicotiana glauca. Mol. Gen. Genet. 150: 225–230
Kao KN, Miller RA, Gamborg OL & Harvey BL (1970) Variations in chromosome number and structure in plant cells grown in suspension culture. Can. J. Genet. Cytol. 12: 297–301
Karp A, Wu QS, Steele SH & Jones MGK (1987) Chromosome variation in dividing protopasts and cell suspensions of wheat. Theor. Appl. Genet. 74: 140–146
Karp A (1988) Origins and causes of chromosome instability in plant tissue culture and regeneration. In: Brandham PE (Ed) Kew Chromosome Conference III (pp 185–192). HMSO, London
Kihlman BA, Eriksson T & Odmark G (1966) Effects of hydroxyurea on chromosomes, cell division and nucleic acid synthesis inVicia faba. Hereditas 55: 386–397
Landsmann J & Uhrig H (1985) Somaclonal variation inSolanum tuberosum detected at the molecular level. Theor. Appl. Genet. 71: 500–505
Lapitan NLV, Sears RG & Gill BS (1988) Amplification of repeated DNA sequences in wheat × rye hybrids regenerated from tissue culture. Theor. Appl. Genet. 75: 381–388
Leitch IJ, Leitch AR & Heslop-Harrison JS (1991) Physical mapping of plant DNA sequences by simultaneousin situ hybridization of two differently labelled fluorescent probes. Genome 34: 329–333
Leitch AR, Mosgöller W, Shi M & Heslop-Harrison JS (1992) Different patterns of rDNA organization at interphase in nuclei of wheat and rye. J. Cell Sci. 101: (in press)
McIntyre CL, Pereira S, Moral LB & Appels R (1990) NewSecale cereale (rye) DNA derivatives for the detection of rye chromosome segments in wheat. Genome 33: 635–640
Moore G, Lucas H, Batty N & Flavell R (1991) A family of retrotransposons and associated genomic variation in wheat. Genomics 10: 461–468
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: 71–78
Murashige T & Skoog F (1962) A revised mediumn for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497
Nagata T, Okada K & Takebe I (1982) Mitotic protoplasts and their infection with Tobacco Mosaic Virus RNA encapsulated in liposomes. Plant Cell Rep. 1: 250–252
Rayburn AL & Gill BS (1985) Use of biotin-labelled probes to map specific DNA sequences on wheat chromosomes. The Journal of Heredity 76: 78–81
Rayburn AL & Gill BS (1987) Molecular analysis of the D-genome of the Triticeae. Theor. Appl. Genet. 73: 385–388
Schwarzacher HG (1976) Chromosomes in mitosis and interphase. Springer-Verlag Berlin, Heidelberg, New York
Schwarzacher-Robinson T, Kraemer PM & Cram LS (1988) Spontaneousin vitro neoplastic evolution of cultured Chinese Hamster cells. Cancer Genet. Cytogenet. 35: 119–128
Schwarzacher T, Leitch AR, Bennett MD & Heslop-Harrison IS (1989)In situ localization of parental genomes in a wide hybrid. Ann. Bot. 64: 315–324
Schweizer D (1980) Fluorescent chromosome banding in plants: Applications, mechanisms, and implications for chromosome structure. In: Davies DR & Hopwood DA (Eds) The Plant Genome (pp 61–72). The John Innes Charity, Norwich, UK
Sharp PJ, Kreis M, Shewry PR & Gale MD (1988) Location of beta-amylase sequences in wheat and its relatives. Theor. Appl. Genet. 75: 286–290
Sinclair WK (1965) Hydroxyurea: Differential lethal effects on cultured mammalian cells during the cell cycle. Science 150: 1729–1731
Szabados L, Hadlaczky Gy & Dudits D (1981) Uptake of isolated plant chromosomes by plant protoplasts. Planta 151: 141–145
Tiersch TR, Chandler RW, Wachtel SS & Elias S (1989) Reference standards for flow cytometry and application in comparative studies of nuclear DNA content. Cytometry 5: 108–117
Vasil IK (1988) Progress in the regeneration and genetic manipulation of cereal crops. Bio/technology 6: 397–402
Wang ML, Leitch AR, Schwarzacher T, Heslop-Harrison IS & Moore G (1992) Construction of a chromosome-enriched Hpall library from flow-sorted wheat chromosomes. Nucl. Acids Res. 20: 1897–1901
Woolston CJ, Reynolds HV, Stacey NJ & Mullineaux PM (1989) Replication of wheat dwarf virus DNA in protoplasts and analysis of coat protein mutants in protoplasts and plants. Nucl. Acids Res. 17: 6029–6041
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Leitch, A.R., Schwarzacher, T., Wang, M.L. et al. Molecular cytogenetic analysis of repeated sequences in a long term wheat suspension culture. Plant Cell Tiss Organ Cult 33, 287–296 (1993). https://doi.org/10.1007/BF02319014
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02319014