Abstract
The Li locus in white clover controls the presence of cyanogenic β-glucosidase (linamarase) activity in leaf tissue, such that plants homozygous for the ‘null’ allele (li) have no linamarase activity in this tissue. The isolation of a cDNA clone from linamarase mRNA is described. The cDNA clone is used to further characterise alleles of the Li locus. Northern blot analysis shows that plants homozygous for the ‘null’ allele (li li) produce very reduced levels of mRNA which hybridises to the cDNA. Heterozygous plants (Li li), which have intermediate levels of enzyme activity, produce intermediate levels of mRNA. Southern blot analysis of Hind III digested genomic DNA shows that the white clover genome contains three genes with homology to the linamarase cDNA and that at least two of these genes segregate independently. Analysis of the cosegregation of linamarase activity and the presence of genomic restriction fragments identifies the genomic sequence specifying linamarase structure and indicates either a structural or cis acting control function of the Li locus.
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References
Aviv H, Leder P: Purification of biologically active globin mRNA by chromatography on oligothymidylic acid-cellulose Proc Natl Acad Sci USA 69: 1408–1412 (1972).
Carvalho FJP de C: Studies on aliphatic ketone cyanohydrin hydroxynitile lyase from Manihot esculenta Crantz. Ph.D. dissertation, University of California, Davis, CA (1981).
Collinge DB, Hughes MA: Developmental and physiological studies on the cyanogenic glucosides of white clover J Exp Bot 33: 154–165 (1982).
Collinge DB, Hughes MA: In vitro characterisation of the Ac locus in white clover (Trifolium repens L.) Arch Biochem Biophys 218: 38–45 (1982).
Conn EE: Cyanogenic glycosides. In: Stumpf PK, Conn EE (eds) Biochemistry of Plants, vol 7, pp. 479–500. Academic Press, New York (1981).
Corkill L: Cyanogenesis in white clover (Trifolium repens L.) New Zeal J Sci Tech B 22: 65–67 (1940).
Corkill L: Cyanogenesis in white clover (Trifolium repens L.) 2. Inheritance of cyanogenesis New Zeal J Sci Tech B 23: 178–193 (1942).
Daday H: Gene frequencies in wild populations of Trifolium repens L. IV. Mechanism of natural selection Heredity 20: 355–365 (1965).
Dellaporta SL, Wood J, Hicks JB: A plant DNA mini preparation: version II. Plant Mol Biol Rep 1: 19–21 (1983).
Dunn MA, Hughes MA, Sharif AL: Synthesis of the cyanogenic β-glucosidase linamarase in white clover Arch Biochem Biophys 260: 561–568 (1988).
Ennos RA: Manifold effect of the cyanogenic loci in white clover Heredity 46: 127–132 (1981).
Graham DE: The isolation of high molecular weight DNA from whole organisms or large tissue masses Anal Biochem 85: 609–613 (1978).
Grime JP, Mowforth MA: Variation in genome size—an ecological interpretation Nature 299: 151–153 (1982).
Grunstein M, Hogness DS: Colony hybridisation method for isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci USA 72: 3961–3965 (1975).
Hughes MA, Conn EE: Cyanoglucoside biosynthesis in white clover (Trifolium repens L.) Phytochemistry 15: 697–701 (1976).
Hughes MA, Dunn MA: Biochemical characterisation of the Li locus which controls the activity of the cyanogenic β-glucosidase in Trifolium repens L. Plant Mol Biol 1: 169–181 (1982).
Hughes MA, Stirling JD: A study of dominance at the locus controlling cyanoglucoside production in Trifolium repens L. Euphytica 31: 477–483 (1982).
Hughes MA, Dunn MA, Pearson JR: A regulatory element controlling the synthesis of the cyanogenic β-glucosidase (linamarase) of white clover. Heredity 55: 387–391 (1985).
Kakes P: Linamarase and other β-glucosidases are present in the cell walls of Trifolium repens L. leaves. Planta 166: 156–160 (1985).
Kakes P, Eeltink H: The presence of a specialised β-glucosidase, linamarase, in the leaves of Trifolium repens L. is controlled by the gene Li. Z Naturforsch 40c: 509–513 (1985).
Lesk AM: Protein structure and evolution: similar amino acid sequences sometimes produce strikingly different three-dimensional structures. BioEssays 2: 213–214 (1985).
Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).
Nahrstedt A: Cyanogenic compounds as protecting agents for organisms. Pl Syst Evol 150: 35–47 (1985).
Pritchard RH, Holland IB: Basic Cloning Techniques. Blackwell, Oxford (1985).
Rogers SO, Bendich AJ: Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues Plant Mol Biol 5: 69–76 (1985).
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Hughes, M.A., Sharif, A.L., Dunn, M.A. et al. Restriction fragment length polymorphism segregation analysis of the Li locus in Trifolium repens L.. Plant Mol Biol 14, 407–414 (1990). https://doi.org/10.1007/BF00028776
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DOI: https://doi.org/10.1007/BF00028776