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The effect of different alleles at the r locus on the synthesis of seed storage proteins in Pisum sativum

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Abstract

Rocket immunoelectrophoresis was used to measure the accumulation of storage proteins in developing cotyledons of two Pisum sativum (pea) genotypes, that were close to isogenic except for the nature of the allele at the r locus. There was a marked decrease in legumin accumulation in the rr (wrinkled-seeded) genotype compared to the RR (round-seeded) genotype. The accumulation of vicilin did not differ greatly between the two genotypes. Pulse-labelling studies indicated that the differences in rates of accumulation of legumin between the rr and RR genotypes were a consequence of differences in rates of protein synthesis.

Measurements of relative amounts of specific mRNAs, using cDNA clones as probes, showed lower amounts of legumin mRNA in developing cotyledons of the rr, compared to the RR, genotype. Both vicilin mRNAs and convicilin mRNA, the latter of which shows a similar temporal pattern of expression to those of the major legumin species, are relatively unaffected by the nature of the allele at the r locus. Nuclear run-on transcription experiments indicated no differences in the rate of synthesis of legumin transcripts in the rr and RR near-isolines. The consequences of homozygosity for the r allele on storage protein mRNA levels in vitro may be mimicked by manipulating the sucrose concentration of the culture medium.

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References

  1. Ambrose MJ, Wang TL, Cook SK, Hedley CL: An analysis of seed development in Pisum sativum. IV. Cotyledon cell populations in vivo and in vitro. J Exp Bot 38: 1909–1920 (1987).

    Google Scholar 

  2. Bain JM, Mercer FV: Subcellular organization of the developing cotyledons of Pisum sativum L. Aust J Biol Sci 19: 49–67 (1966).

    Google Scholar 

  3. Barratt DHP: Regulation of storage protein accumulation by abscisic acid in Vicia faba L. cotyledons cultured in vitro. Ann Bot 57: 245–256 (1986).

    Google Scholar 

  4. Beach LR, Spencer D, Randall PJ, Higgins TJV: Transcriptional and post-transcriptional regulation of storage protein gene expression in sulfur-deficient pea seeds. Nucl Acids Res 13: 999–1013 (1985).

    PubMed  Google Scholar 

  5. Bown D, Ellis THN, Gatehouse JA: The sequence of a gene encoding convicilin from pea (Pisum sativum L.) shows that convicilin differs from vicilin by an insertion near the N-terminus. Biochem J 251: 717–726 (1988).

    PubMed  Google Scholar 

  6. Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254 (1976).

    Article  PubMed  Google Scholar 

  7. Casey R: Immunoaffinity chromatography as a means of purifying legumin from Pisum (pea) seeds. Biochem J 177: 509–520 (1979).

    PubMed  Google Scholar 

  8. Casey R, Domoney C, Nielsen NC: Isolation of a cDNA clone for pea (Pisum sativum) seed lipoxygenase. Biochem J 232: 79–85 (1985).

    PubMed  Google Scholar 

  9. Casey R, Domoney C, Ellis N: Legume storage proteins and their genes. Oxford Surv Plant Mol Cell Biol 3: 1–95 (1986).

    Google Scholar 

  10. Chandler PM, Spencer D, Randall PJ, Higgins TJV: Influence of sulfur nutrition on developmental patterns of some major pea seed proteins and their mRNAs. Plant Physiol 75: 651–657 (1984).

    Google Scholar 

  11. Chappell J, Hahlbrock K: Transcription of plant defence genes in response to UV light or fungal elicitor. Nature 311: 76–78 (1984).

    Google Scholar 

  12. Chappell J, Chrispeels MJ: Transcriptional and posttranscriptional control of phaseolin and phytohemag-glutinin gene expression in developing cotyledons of Phaseolus vulgaris. Plant Physiol 81: 50–54 (1986).

    Google Scholar 

  13. Corke FMK, Hedley CL, Shaw PJ, Wang TL: An analysis of seed development in Pisum sativum V. Fluorescence triple staining for investigating cotyledon cell development. Protoplasma 140: 164–172 (1987).

    Google Scholar 

  14. Coxon DT, Davies DR: The effect of the ra and rb loci on the lipid content of the seed of Pisum sativum. Theor Appl Genet 64: 47–50 (1982).

    Article  Google Scholar 

  15. Craig S, Goodchild DJ, Hardham AR: Structural aspects of protein accumulation in developing pea cotyledons. I. Qualitative and quantitative changes in parenchyma cell vacuoles. Aust J Plant Physiol 6: 81–98 (1979).

    Google Scholar 

  16. Creech RG: Genetic control of carbohydrate synthesis in maize endosperm. Genetics 52: 1175–1186 (1965).

    Google Scholar 

  17. Croy RRD, Gatehouse JA, Evans IM, Boulter D: Characteristion of the storage protein subunits synthesised in vitro by polyribosomes and RNA from developing pea (Pisum sativum L.) I. Legumin. Planta 148: 49–56 (1980).

    Google Scholar 

  18. Croy RRD, Gatehouse JA, Tyler M, Boulter D. The purification and characterization of a third storage protein (convicilin) from the seeds of pea (Pisum sativum L.). Biochem J 191: 509–516 (1980).

    PubMed  Google Scholar 

  19. Croy RRD, Lycett GW, Gatehouse JA, Yarwood JN, Boulter D: Cloning and analysis of cDNAs encoding plant storage protein precursors. Nature 295: 76–79 (1982).

    Google Scholar 

  20. Danielsson CE: Seed globulins of the gramineae and leguminosae. Biochem J 44: 387–400 (1949).

    Google Scholar 

  21. Davies DR: The ra locus and legumin synthesis in Pisum sativum. Biochem Genet 18: 1207–1219 (1980).

    PubMed  Google Scholar 

  22. DiFonzo N, Fornasari E, Salamini F, Reggiani R, Soave C: Interaction of maize mutants floury-2 and opaque-7 with opaque-2 in the synthesis of endosperm proteins. J Heredity 71: 397–402 (1980).

    Google Scholar 

  23. Domoney C, Davies DR, Casey R: The initiation of legumin synthesis in immature embryos of Pisum sativum L. grown in vivo and in vitro. Planta 149: 454–460 (1980).

    Google Scholar 

  24. Domoney C, Casey R: Cloning and characterization of complementary DNA for convicilin, a major seed storage protein in Pisum sativum L. Planta 159: 446–453 (1983).

    Google Scholar 

  25. Domoney C, Casey R: Measurement of gene number for seed storage proteins in Pisum. Nucl Acids Res 13: 687–699 (1985).

    PubMed  Google Scholar 

  26. Domoney C, Ellis THN, Davies DR: Organization and mapping of legumin genes in Pisum. Mol Gen Genet 202: 280–285 (1986).

    Article  Google Scholar 

  27. Domoney C, Casey R: Changes in legumin messenger RNAs throughout seed development in Pisum sativum L. Planta 170: 562–566 (1987).

    Google Scholar 

  28. Edwards J, Ap Rees T: Sucrose partitioning in developing embryos of round and wrinkled varieties of Pisum sativum. Phytochemistry 25: 2027–2032 (1986).

    Google Scholar 

  29. Ellis THN, Domoney C, Castleton J, Cleary W, Davies DR: Vicilin genes of Pisum. Mol Gen Genet 205: 164–169 (1986).

    Google Scholar 

  30. Evans IM, Gatehouse JA, Boulter D: Regulation of storage-protein synthesis in pea (Pisum sativum L.) cotyledons under conditions of sulphur deficiency. Biochem J 232: 261–265 (1985).

    PubMed  Google Scholar 

  31. Gallagher TF, Ellis RJ: Light-stimulated transcription of genes for two chloroplast polypeptides in isolated pea leaf nuclei. EMBO J 1: 1493–1498 (1982).

    Google Scholar 

  32. Gatehouse JA, Evans IM, Bown D, Croy RRD, Boulter D: Control of storage-protein synthesis during seed development in pea (Pisum sativum L.). Biochem J 208: 119–127 (1982).

    PubMed  Google Scholar 

  33. Hartings H, Maddaloni M, Lazzaroni N, DiFonzo N, Motto M, Salamini F, Thompson R: The O2 gene which regulates zein deposition in maize endosperm encodes a protein with structural homologies to transcriptional activators. EMBO J 8: 2795–2801 (1989).

    PubMed  Google Scholar 

  34. Hedley CL, Smith CM, Ambrose MJ, Cook S, Wang TL: An analysis of seed development in Pisum sativum II. The effect of the r-locus on the growth and development of the seed. Ann Bot 58: 371–379 (1986).

    Google Scholar 

  35. Jen G, Thach RE: Inhibition of host translation in encephalomyocarditis virus-infected L cells: a novel mechanism. J Virol 43: 250–261 (1982).

    PubMed  Google Scholar 

  36. Kodrzycki R, Boston RS, Larkins BA: The opaque-2 mutation of maize differentially reduces zein gene transcription. The Plant Cell 1: 105–114 (1989).

    PubMed  Google Scholar 

  37. Kooistra E: On the difference between smooth and three types of wrinkled peas. Euphytica 11: 357–373 (1962).

    Google Scholar 

  38. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).

    PubMed  Google Scholar 

  39. Lee L, Tsai CY: Zein synthesis in the embryo and endosperm of maize mutants. Biochem Genet 22: 729–737 (1984).

    PubMed  Google Scholar 

  40. Lee L, Tsai CY: The effect of sucrose accumulation on zein synthesis in maize starch deficient mutants. Phytochemistry 24: 225–229 (1985).

    Article  Google Scholar 

  41. Lehrach H, Diamond D, Wozney JM, Boedtker H: RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry 16: 4743–4751 (1977).

    PubMed  Google Scholar 

  42. Muesing MA, Smith DH, Capon DJ: Regulation of mRNA accumulation by a human immunodeficiency virus trans-activator protein. Cell 48: 691–701 (1987).

    Article  PubMed  Google Scholar 

  43. Osborne TB: The Vegetable Proteins. Longmans, Green and Co., London (1924).

    Google Scholar 

  44. Pelham HRB, Jackson RJ: An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem 67: 247–256 (1976).

    PubMed  Google Scholar 

  45. Santiago TC, Purvis IJ, Bettany AJE, Brown AJP: The relationship between mRNA stability and length in Saccharomyces cerevisiae. Nucl Acids Res 14: 8347–8360 (1986).

    PubMed  Google Scholar 

  46. Scharpé A, VanParijs R: The formation of polyploid cells in ripening cotyledons of Pisum sativum L. in relation to ribosome and protein synthesis. J Exp Bot 24: 216–222 (1973).

    Google Scholar 

  47. Schmidt RJ, Burr FA, Burr B: Transposon tagging and molecular analysis of the maize regulatory locus opaque-2. Science 238: 960–963 (1987).

    PubMed  Google Scholar 

  48. Shaw G, Kamen R: A conserved AU sequence from the 3′ untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell 46: 659–667 (1986).

    Article  PubMed  Google Scholar 

  49. Shewry PR, Pratt HM, Finch RA, Miflin BJ: Protein metabolism in developing endosperms of high lysine and normal barley. Cereal Chem 56: 110–117 (1979).

    Google Scholar 

  50. Smith A: Major differences in isoforms of starch-branching enzyme between developing embryos of round- and wrinkled-seeded peas (Pisum sativum L.). Planta 175: 270–279 (1988).

    Google Scholar 

  51. Spencer D, Higgins TJV, Button SC, Davey RA: Pulselabeling studies on protein synthesis in developing pea seeds and evidence of a precursor form of legumin small subunit. Plant Physiol 66: 510–515 (1980).

    Google Scholar 

  52. Spencer D, Higgins TJV: The biosynthesis of legumin in maturing pea seeds. Biochem Int 1: 502–509 (1980).

    Google Scholar 

  53. Stickland RG, Wilson KE: Sugars and starch in developing round and wrinkled pea seeds. Ann Bot 52: 919–921 (1983).

    Google Scholar 

  54. Tsai CY, Larkins BA, Glover DV: Interaction of the Opaque-2 gene with starch-forming mutant genes on the synthesis of zein in maize endosperm. Biochem Genet 16: 883–896 (1978).

    PubMed  Google Scholar 

  55. Walling L, Drews GN, Goldberg RB: Transcriptional and post-transcriptional regulation of soybean seed protein mRNA levels. Proc Natl Acad Sci USA 83: 2123–2127 (1986).

    Google Scholar 

  56. Wang TL, Smith CM, Cook SK, Ambrose MJ, Hedley CL. An analysis of seed development in Pisum sativum III. The relationship between the r locus, the water content and the osmotic potential of seed tissues in vivo and in vitro. Ann Bot 59: 73–80 (1987).

    Google Scholar 

  57. Weeke B: Rocket electrophoresis. In: Axelsen NW, Kroll J, Weeke B (eds) A manual of Quantitative Immunoelectrophoresis-Methods and Applications. Universitetsforlaget, Oslo (1973).

    Google Scholar 

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Turner, S.R., Barratt, D.H.P. & Casey, R. The effect of different alleles at the r locus on the synthesis of seed storage proteins in Pisum sativum . Plant Mol Biol 14, 793–803 (1990). https://doi.org/10.1007/BF00016512

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