Summary
Wheat lines carrying homologous pairs of complete or telocentric rye chromosomes have been used to investigate the detection of rye chromosomes or chromosome fragments in a wheat genetic background, by nucleic acid hybridisation. Three different radioactive rye probes were used: (1) the most highly repeated DNA sequences made radioactive in vitro by “nick translation”, (2) the whole complement of repeated sequences radioactively labelled in vivo and (3) 25s and 18s ribosomal RNAs radioactively labelled in vivo. Radioactive highly repeated sequence DNA was hybridised to unlabelled DNAs from wheat-rye chromosome addition lines immobilised on nitrocellulose filters. Rye-specific highly repeated sequences were detected in all the lines, illustrating that rye-specific highly repeated sequences reside on all chromosome arms. The effect of the size of the radioactive probe DNA was investigated by hybridising labelled rye repeated sequences to unlabelled DNAs from wheat-rye chromosome addition lines in solution. Short fragments round 300 nucleotides long detected rye-specific repeated sequences in all the unlabelled DNAs. Longer fragments, greater than 2000 nucleotides, were used to show that some of these rye-specific repeated sequences are in chromosomal regions which lack repeated sequences related to repeated sequences in wheat. The regions are probably considerably longer than 10,000 base pairs.
Saturating concentrations of 25s and 18s ribosomal RNAs were hybridised to DNAs from Holdfast wheat-King II rye chromosome addition lines bound to nitrocellulose filters. The results showed that chromosome 1R of King II possesses the major cluster of ribosomal RNA genes of rye. However with DNAs from Chinese Spring wheat-Imperial rye chromosome addition lines, those with rye chromosome 1R and chromosome 5R had considerably more ribosomal RNA genes than Chinese Spring wheat.
Using labelled barley repeated sequences as probe DNA it was shown in wheat-barley DNA mixtures that it was possible to detect 0.05 per cent barley DNA, i.e. 5 parts in 10,000, by incubating the DNAs to C0t 100 at 70°C in 0.12-M phosphate buffer.
It is concluded that these sorts of experiments should be of considerable use for detecting the presence of alien chromosome segments in wheat and other plant species.
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Barber, H N, Driscoll, C J, and Vickery, R S. 1968. Enzymic markers for wheat and rye chromosomes. Proc 3rd Int Wheat Genet Symp, 116.
Bennett, M D. 1974. Meiotic, gametophytic and early endosperm development in Triticale. In R. Mclntyre and M. Campbell (eds), Triticale; Proc Int Symp El Batan, Mexico.
Bennett, M D, and Smith, J B. 1975. Confirmation of the identification of the rye chromosomes in 1B/IR wheat-rye chromosome substitution and translocation lines. Can J Genet Cytol, 17, 117–120.
Bennett, M D, and Smith, J B. 1976. Nuclear DNA amounts in angiosperms. Phil, trans of Royal Soc (Lond), 274, 227–274.
Bhatia, C R, and Smith, H H. 1966. Variation associated with an Aegilops umbellulata chromosome segment incorporated into wheat. Nature, 211, 1425–1426.
Birnstiel, M L, Chipchase, M, and Spiers, J. 1971. The ribosomal RNA cistrons. Progress in Nucleic Acid and Molecular Biology, 11, 351–389.
Brown, D D, and Weber, C S. 1968. Gene linkage by RNA-DNA hybridisation. Unique DNA sequences homologous to 4s RNA, 5s RNA and ribosomal RNA. J Mol Biol, 34, 661–680.
Burgi, E, and Hershey, A D. 1963. Sedimentation rate as a measure of molecular weight of DNA. Biophys J, 3, 309–321.
Darvey, N L, and Gustafson, J P. 1975. Identification of rye chromosomes in wheat-rye addition lines and triticale by heterochromatin bands. Crop Sci, 15, 239–243.
Driscoll, C J. 1975. First compendium of wheat-alien chromosome lines. Annual Wheat Newsletter, 21, 16–32.
Driscoll, C J, and Jensen, N F. 1963. A genetic method for detecting intergeneric translocation. Genetics, 48, 459–468.
Driscoll, C, and Sears, E R. 1971. Individual addition of the chromosomes of “Imperial” rye to wheat. Agron Abst, 6.
Dvorak, J, and Knott, D R. 1974. Disomic and ditelosomic additions of diploid Agropyron elongatum chromosomes to Triticum aestivum. Can J Gen Cytol, 16, 399–417.
Flavell, R B, and Rimpau, J. 1975. Ribosomal RNA genes and supernumerary B chromosomes of rye. Heredity, 35, 127–131.
Flavell, R B, Rimpau, J, and Smith, D B. 1977. Repeated sequence DNA relationships in four cereal genomes. Chromosoma (Berl), 63, 205–222.
Flavell, R B, and Smith, D B. 1974. Variation in nucleolar organiser ribosomal RNA gene multiplicity in wheat and rye. Chromosoma (Berl), 47, 327–334.
Flavell, R B, and Smith, D B. 1976. Nucleotide sequence organisation in the wheat genome. Heredity, 37, 231–252.
Gustafson, J P, and Bennett, M D. 1976. Preferential selection for wheat-rye substitutions in 42-chromosome Triticale. Crop Science, 16, 688–693.
Hart, G E, McMillin, D E, and Sears, E R. 1976. Determination of the chromosomal location of a glutamate oxaloacetate transaminase structural gene using Triticum agropyron translocations. Genetics, 83, 49–61.
Irani, B N, and Bhatia, C R. 1972. Chromosomal location of alcohol dehydrogenase gene(s) in rye using wheat-rye addition lines. Genetica, 43, 195–200.
Islam, A K M R, Shepherd, K W, and Sparrow, D H B. 1975. Addition of individual barley chromosomes to wheat. Proc 3rd Int Barley Genetics Symp, 260–270.
Knott, D R. 1961. The inheritance of rust resistance. VI. The transfer of stem rust resistance from Agropyron elongatum to common wheat. Canad J Plant Sci, 41, 109–123.
Koller, O L, and Zeller, F J. 1976. The homoeologous relationships of rye chromosomes 4R and 7R with wheat chromosomes. Genet Res, Camb, 28, 177–188.
Lurquin, P F. 1977. Integration versus degradation of exogenous DNA in plants: an open question. Progress in Nucleic Acid Research and Molecular Biology, 20, 161–207.
Macdonald, T, and Smith, H H. 1972. Variation associated with an Aegilops umbellulata chromosome segment incorporated into wheat. II. Peroxidase and leucine amino-peptidase isozymes. Genetics, 72, 77–86.
Macgregor, H C, and Mizuno, S. 1976. In situ hybridisation of nick-translated 3H ribosomal DNA to chromosomes from salamanders. Chromosoma, 54, 15–25.
Maniatis, T, Jeffrey, A, and Kleid, D G. 1975. Nucleotide sequence of the rightward operator of phage λ. Proc Nat Acad Sci (Wash), 72, 1184–1188.
Mettin, D, Bluthner, W D, and Schlegel, C. 1973. Additional evidence on spontaneous 1B/1R wheat-rye substitutions and translocations. Proc 4th Wheat Genet Symp, 179–184. Mo. Agric. Exp. Stn., Columbia, Mo.
Mohan, J, and Flavell, R B. 1974. Ribosomal RNA cistron multiplicity and nucleolar organisers in hexaploid wheat. Genetics, 76, 33–44.
Pardue, M L, and Gall, J G. 1975. Nucleic acid hybridisation to the DNA of cytological preparations. In: Methods in Cell Biol, 10, 1–16.
Riley, R, and Chapman, V. 1958. The production and phenotypes of wheat-rye chromosome addition lines. Heredity, 12, 301–305.
Riley, R, and Chapman, V. 1960. The meiotic behaviour, fertility and stability of wheat-rye chromosome addition lines. Heredity, 14, 89–100.
Riley, R, Chapman, V, and Johnson, R. 1968. The incorporation of alien disease resistance in wheat by genetic interference with the regulation of meiotic chromosome synapsis. Genet Res (Camb), 12, 199–219.
Riley, R, and Kimber, G. 1966. The transfer of alien genetic variation to wheat. Annual Report of Plant Breeding Institute, Cambridge. 6–36.
Riley, R, and Macer, R C F. 1966. The chromosomal distribution of the genetic resistance of rye to wheat pathogens. Canad J of Genetics and Cytol, 4, 640–653.
Sarma, N P, and Natarajan, A T. 1973. Identification of heterochromatic regions in the chromosomes of rye. Hereditas, 74, 233–238.
Sears, E R. 1956. The transfer of leaf-rust resistance from Aegilops umbellulata to wheat. Brookhaven Symp Biol, 9, 1–22.
Sears, E R. 1972. Chromosome engineering in wheat. Stadler Symposium, 4, 23–38. University of Missouri, Columbia, U.S.A.
Singh, R J, and Röbbelen, G. 1976. Giemsa banding technique reveals deletions within rye chromosomes in addition lines. Z Pflanzenzüchtg, 76, 11–18.
Smith, D B, and Flavell, R B. 1974. The relatedness and evolution of repeated nucleotide sequences in the genomes of some Gramineae species. Biochem Genet, 12, 243–256.
Smith, D B, and Flavell, R B. 1975. Characterisation of the wheat genome by renaturation kinetics. Chromosoma (Berl), 50, 223–242.
Smith, D B, and Flavell, R B. 1977. Nucleotide sequence organisation in the rye genome. Biochim Biophys Acta, 474, 82–97.
Smith, D B, Rimpau, J, and Flavell, R B. 1976. Interspersion of different repeated sequences in the wheat genome revealed by interspecies DNA/DNA hybridisation. Nucleic Acid Res, 3, 2811–2826.
Studier, F W. 1965. Sedimentation studies of the size and shape of DNA. J Molec Biol, 11, 373–390.
Tang, K S, and Hart, G E. 1975. Use of isozymes as chromosome markers in wheat-rye addition lines and in triticale. Genet Res (Camb), 26, 187–201.
Verma, S C, and Rees, H. 1974. Giemsa staining and the distribution of heterochromatin in rye chromosomes. Heredity, 32, 118–122.
Wienhues, A. 1966. Transfer of rust resistance otAgropyron to wheat by addition, substitution and translocation. Proc 2nd Int Wheat Genet Symp Hereditas Suppl: 2, 370–381.
Zeller, F J. 1973. 1B/IR wheat-rye chromosome substitutions and translocations. Proc 4th Wheat Genet Symp, 209–221. Mo. Agric. Exp. Stn., Columbia, Mo.
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Flavell, R., O'Dell, M., Rimpau, J. et al. Biochemical detection of alien DNA incorporated into wheat by chromosome engineering. Heredity 40, 439–455 (1978). https://doi.org/10.1038/hdy.1978.49
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DOI: https://doi.org/10.1038/hdy.1978.49
