Summary
Genomic in situ hybridization was used to identify alien chromatin in chromosome spreads of wheat, Triticum aestivum L., lines incorporating chromosomes from Leymus multicaulis (Kar. and Kir.) Tzvelev and Thinopyrum bessarabicum (Savul. and Rayss) Löve, and chromosome arms from Hordeum chilense Roem. and Schult, H. vulgare L. and Secale cereale L. Total genomic DNA from the introgressed alien species was used as a probe, together with excess amounts of unlabelled blocking DNA from wheat, for DNA:DNA in-situ hybridization. The method labelled the alien chromatin yellow-green, while the wheat chromosomes showed only the orange-red fluorescence of the DNA counterstain. Nuclei were screened from seedling root-tips (including those from half-grains) and anther wall tissue. The genomic probing method identified alien chromosomes and chromosome arms and allowed counting in nuclei at all stages of the cell cycle, so complete metaphases were not needed. At prophase or interphase, two labelled domains were visible in most nuclei from disomic lines, while only one labelled domain was visible in monosomic lines. At metaphase, direct visualization of the morphology of the alien chromosome or chromosome segment was possible and allowed identification of the relationship of the alien chromatin to the wheat chromosomes. The genomic in-situ hybridization method is fast, sensitive, accurate and informative. Hence it is likely to be of great value for both cytogenetic analysis and in plant breeding programmes.
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Anamthawat-Jónsson K, Schwarzacher T, Leitch AR, Bennett MD, Heslop-Harrison JS (1990) Discrimination between closely related Triticeae species using genomic DNA as a probe. Theor Appl Genet 79:721–728
Cremer T, Lichter P, Borden J, Ward DC, Manuelidis L (1988) Detection of chromosome aberrations in metaphase and interphase tumor cells by in-situ hybridization using chromosome-specific library probes. Hum Genet 80:235–246
Dong Y, Zhouzhou Y, Ganyuan Z (1986) Study on hybridization of Triticum aestivum with Leymus multicaulis and Leymus racemosus. In: Li Z, Swaminathan MS (eds) Proc 1st Int Symp on Chromosome Engineering in Plants. Xian, China, pp 185–187
Fatih AM (1983) Analysis of the breeding potential of wheat-Agropyron and-wheat-Leymus derivatives. Hereditas 98:287–295
Flavell RB, O'Dell M, Hutchinson J (1981) Nucleotide sequence organization in plant chromosomes and evidence for sequence translocation during evolution. Cold Spring Harbor Symp Quant Biol 45:501–508
Forster BP, Gorham J, Miller TE (1987) Salt tolerance of an amphiploid between Triticum aestivum and Agropyron junceum. Plant Breed 98:1–8
Forster BP, Miller TE, Law CN (1988) Salt tolerance of two wheat-Agropyron junceum disomic addition lines. Genome 30:559–564
Friebe B, Heun M, Bushuk W (1989) Cytological characterization, powdery mildew resistance and storage protein composition of tetraploid and hexaploid 1BL/1RS wheat rye translocation lines. Theor Appl Genet 78:425–432
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 and in-situ hybridization. Theor Appl Genet 81:381–389
Gale MD, Miller TE (1987) The introduction of alien genetic variation in wheat. In: Lupton FGH (ed) Wheat breeding: Its scientific basis. Chapman and Hall, London, pp 173–210
Gale MD, Sharp PJ, Chao S, Law CN (1989) Applications of genetic markers in cytogenetic manipulation of the wheat genome. Genome 31:137–142
Guidet F, Rogowsky P, Taylor C, Song W, Langridge P (1991) Cloning and characterisation of a new rye-specific repeated sequence. Genome 34:81–87
Hart GE, Islam AKMR, Shepherd KW (1980) Use of isozymes as chromosome markers in the isolation and characterization of wheat-barley chromosome addition lines. Genet Res Cambridge 36:311–325
Heslop-Harrison JS (1991) The molecular cytogenetics of plants. J Cell Sci 100:15–21
Heslop-Harrison JS, Bennett MD (1990) Nuclear architecture in plants. Trends Genet 6:401–405
Heslop-Harrison JS, Schwarzacher T, Leitch AR, Anamthawat-Jónsson K, Bennett MD (1988) A method of identifying DNA sequences in chromosomes of plants. European Patent Application Number 8828130.8. December 2
Heslop-Harrison JS, Leitch AR, Schwarzacher T, Anamthawat-Jónsson K (1990) Detection and characterization of 1B/1R translocations in hexaploid wheat. Heredity 65:385–392
Islam AKMR, Shepherd KW (1991a) Alien genetic variation in wheat improvement. In: Gupta PK, Tsuchiya T (eds) Chromosome engineering in plants: genetics, breeding, evolution, part A. Elsevier, Amsterdam, pp 291–312
Islam AKMR, Shepherd KW (1991b) Production of wheat-barley recombinant chromosomes through induced homoeologous pairing. 1. Isolation of recombinants involving barley arm 3HL and 6HL. Theor Appl Genet 83:489–494
Knott DR (1961) The inheritance of rust resistance. VI. The transfer of stem rust resistance from Agropyron elongatum to common wheat. Can J Plant Sci 41:109–123
Koebner RMD, Shepherd KW, Appels R (1986) Controlled introgression to wheat of genes from rye chromosome arm 1RS by induction of allosyndesis. 2. Characterisation of recombinants. Theor Appl Genet 73:209–217
Lapitan NLV, Sears RG, Rayburn AL, Gill BS (1986) Wheatrye translocations. J Hered 77:415–419
Law CN (1981) Chromosome manipulation in wheat. Chromosomes Today 7:194–205
Le HT, Armstrong KC (1991) In-situ hybridization as a rapid means to assess meiotic pairing and detection of alien DNA transfers in interphase cells of wide crosses involving wheat and rye. Mol Gen Genet 225:33–37
Le HT, Armstrong KC, Miki B (1989) Detection of rye DNA in wheat-rye hybrids and wheat translocation stocks using total genomic DNA as a probe. Plant Mol Biol Reporter 7:150–158
Leitch AR, Mosgöller W, Schwarzacher T, Bennett MD, Heslop-Harrison JS (1990) Genomic in-situ hybridization to sectioned nuclei shows chromosome domains in grass hybrids. J Cell Sci 95:335–341
Leitch IJ, Leitch AR, Heslop-Harrison JS (1991) Physical mapping of plant DNA sequences by simultaneous in-situ hybridization of two differently labelled fluorescent probes. Genome 34:329–333
Maluszynska J, Heslop-Harrison JS (1991) Localization of tandemly repeated DNA sequences in Arabidopsis thaliana. The Plant Jour 1:159–166
McIntosh RA (1988) Catalogue of gene symbols for wheat. In: Miller TE, Koebner RMD (eds) Proc 7th Int Wheat Genetics Symp. Institute of Plant Science Research, Cambridge, pp 1225–1323
Meinkoth J, Wahl G (1984) Hybridization of nucleic acids immobilized on solid supports. Anal Biochem 138:267–284
Miller TE, Reader SM (1987) A guide to the homoeology of chromosomes within the Triticeae. Theor Appl Genet 74:214–217 (Addendum to Table 1: Theor Appl Genet 74:680)
Mukai Y, Gill BS (1991) Detection of barley chromatin added to wheat by genomic in-situ hybridization. Genome 34:448–452
Porceddu E, Ceoloni C, Lafiandra D, Tanzarella OA, Scarascia Mugnozza GT (1988) Genetic resources and plant breeding: problems and prospects. In: Miller TE, Koebner RMD (eds) Proc 7th Int Wheat Genetics Symp. Institute of Plant Science Research, Cambridge, pp 7–21
Rayburn AL, Gill BS (1987) Molecular analysis of the D-genome of the Triticeae. Theor Appl Genet 73:385–388
Rogowsky PM, Guidet FLY, Langridge P, Shepherd KW, Koebner RMD (1991) Isolation and characterization of wheat-rye recombinants involving chromosome arm 1DS of wheat. Theor Appl Genet 82:537–544
Schwarzacher T, Leitch AR, Bennett MD, Heslop-Harrison JS (1989) In-situ localization of parental genomes in a wide hybrid. Ann Bot 64:315–324
Schwarzacher T, Heslop-Harrison JS, Anamthawat-Jónsson K, Finch RA, Bennett MD (1992) Parental genome separation in reconstructions of somatic and premeiotic metaphases of Hordeum vulgare x H. bulbosum. J Cell Sci 101:13–24
Sears EJ (1956) The transfer of leaf-rust resistance from Aegilops umbellulata into wheat. Brookhaven Symp Biol 9:1–22
Sears EJ (1977) An induced mutant with homoeologous pairing in common wheat. Can J Genet Cytol 19:585–593
Sharp PJ, Desai S, Chao S, Gale MD (1988) Isolation, characterization and application of a set of 14 RFLP probes for identifying each homoeologous chromosome in the Triticeae. In: Miller TE, Koebner RMD (eds) Proc 7th Int Wheat Genetics Symp. Institute of Plant Science Research, Cambridge, pp 639–646
Shepherd KW, Islam AKMR (1988) Fourth compendium of wheat-alien chromosome lines. In: Miller TE, Koebner RMD (eds) Proc 7th Int Wheat Genetics Symp. Institute of Plant Science Research, Cambridge, pp 1373–1398
Swanson CP, Webster PL (1975) The cell, 4th edn. Prentice-Hall, Englewood Cliffs, New Jersey
Tkachuk DC, Pinkel D, Kuo WL, Weier HU, Gray JW (1991) Clinical applications of fluorescence in-situ hybridization. Genet Anal Techn Appl 8:67–74
Zeller FJ (1973) 1B/1R wheat-rye chromosome substitutions and translocations. In: Sears ER, Sears LMS (eds) Proc 4th Int Wheat Genetics Symp. Agricultural Experimental Station, College of Agriculture, University of Missouri, Columbia, pp 209–222
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Communicated by J. W. Snape
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Schwarzacher, T., Anamthawat-Jónsson, K., Harrison, G.E. et al. Genomic in situ hybridization to identify alien chromosomes and chromosome segments in wheat. Theoret. Appl. Genetics 84, 778–786 (1992). https://doi.org/10.1007/BF00227384
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DOI: https://doi.org/10.1007/BF00227384