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Differential, temporal and spatial expression of genes involved in storage oil and oleosin accumulation in developing rapeseed embryos: implications for the role of oleosins and the mechanisms of oil-body formation

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Abstract

The temporal and spatial expression of oleosin and Δ9-stearoyl-ACP desaturase genes and their products has been examined in developing embryos of rapeseed, Brassica napus L. var. Topas. Expression of oleosin and stearate desaturase genes was measured by in situ hybridisation at five different stages of development ranging from the torpedo stage to a mature-desiccating embryo. The temporal pattern of gene expression varied dramatically between the two classes of gene. Stearate desaturase gene expression was relatively high, even at the torpedo stage, whereas oleosin gene expression was barely detectable at this stage. By the stage of maximum embryo fresh weight, stearate desaturase gene expression had declined considerably while oleosin gene expression was at its height.

In contrast to their differential temporal expression, the in situ labelling of both classes of embryo-specific gene showed similar, relatively uniform patterns of spatial expression throughout the embryo sections. Immunogold labelling of ultra-thin sections from radicle tissue with anti-oleosin antibodies showed similar patterns to sections from cotyledon tissue. However, whereas at least three oleosin isoforms were detectable on western blots of homogenates from cotyledons, only one isoform was found in radicles. This suggests that some of the oleosin isoforms may be expressed differentially in the various types of embryo tissue. The differential timing of stearate desaturase and oleosin gene expression was mirrored by similar differences in the timing of the accumulation of their ultimate products, i.e. storage oil and oleosin proteins. Oil-body fractions prepared from young (2.5 mg) embryos contained very little oleosin protein, as examined by SDS-PAGE and western blotting, whereas identically prepared fractions from dry seeds contained over 10% (w/w) oleosin. Dehydration of oil bodies from young embryos resulted in their breakdown and coalescence into large clumps of oil which could not be re-emulsified, even after rehydration. In contrast, the oleosin-rich oil bodies from mature embryos were stable to dehydration and subsequent rehydration. It is suggested that, in developing rapeseed embryos, the accumulation of storage oil and oleosins is not concomitant but that the eventual deposition of oleosins onto the surfaces of storage oil bodies is essential for their stability during seed desiccation.

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Abbreviations

ABA:

abscisic acid

ACP:

acyl carrier protein

GLC:

gas-liquid chromatography

PBS:

phosphate-buffered saline

References

  1. Brockman, HL: General features of lipolysis: reaction scheme, interfacial structure and experimental approaches. In: Brockman, HL, Borgstrom, B (eds) Lipolytic Enzymes, pp. 4–46. Elsevier Press, Amsterdam (1984).

    Google Scholar 

  2. Coen, ES, Romero, JM, Doyle, S, Elliot, R, Murphy, G, Carpenter, R: Floricaula: a homeotic gene required for flower development in Anthirrhinum majus. Cell 63: 1311–1322 (1990).

    Article  PubMed  Google Scholar 

  3. Cummins, I, Murphy, DJ: Mechanism of oil body synthesis and maturation in developing seeds. In: Harwood, JL, Quin, PJ (eds) Plant Lipid Biochemistry, Structure and Function pp. 231–233. Portland Press, London (1990).

    Google Scholar 

  4. Davidowicz-Grzegorzewsha, A, Podstolski, A: Age-related changes in the ultrastructure and membrane properties of Brassica napus L. seeds. Ann Bot 69: 39–46 (1992).

    Google Scholar 

  5. deOlivera Franco, LO, Simoens, C, Seurinck, J, Coppieters, J, Botterman, J, VanMontagu, M: Inflorescence-specific genes from Arabidopsis thaliana encoding glycine-rich proteins. Plant J 3: 495–507 (1993).

    Article  PubMed  Google Scholar 

  6. Dure, L, Greenway, SC, Galau, GA: Developmental biochemistry of cottonseed embryogenesis and germination. XIV. Changing mRNA populations as shown by in vitro and in vivo protein synthesis. Biochemistry 20: 4162–4168 (1981).

    PubMed  Google Scholar 

  7. Espelund, M, Saeboe-Larssen, S, Hughes, DW, Galau, GA, Larsen, F, Jakobsen, KS: Late embryogenesis-abundant genes encoding proteins with different numbers of hydrophilic repeats are regulated differently by abscisic acid and osmotic stress. Plant J 2: 241–252 (1992).

    Article  PubMed  Google Scholar 

  8. Evans, DE, Rothnie, NE, Sang, JP, Palmer, MV, Mulcahy, DL, Singh, MB, Knox, RB: Correlations between gametophytic (pollen) and sporophytic (seed) generations for polyunsaturated fatty acids in oilseed rape Brassica napus L. Theor Appl Genet 76: 411–419 (1988).

    Article  Google Scholar 

  9. Evans, DE, Taylor, PE, Singh, MB, Knox, RB: The interrelationship between the accumulation of lipids, protein and the level of acyl carrier protein during the development of Brassica napus L. pollen. Planta 186: 343–354 (1992).

    Article  Google Scholar 

  10. Fernandez, DE, Turner, FD, Crouch, ML: In situ localization of storage protein mRNAs in developing meristems of Brassica napus embryos. Development 111: 299–313 (1991).

    PubMed  Google Scholar 

  11. Finkelstein, RR, Tenbarge, KM, Shimway, JE, Crouch, ML: Role of ABA in the maturation of rapeseed embryos. Plant Physiol 78: 630–636 (1985).

    Google Scholar 

  12. Fowler, DB, Downey, RK: Lipid and morphological changes in developing rapeseed, Brassica napus. Can J Plant Sci 50: 233–247 (1970).

    Google Scholar 

  13. Harada, JJ, DeLisle, AJ, Baden, CS, Crouch, ML: Unusual sequence of an abscisic acid induced mRNA which accumulates late in Brassica napus seed development. Plant Mol Biol 12: 395–401 (1989).

    Article  Google Scholar 

  14. Hills, MJ, Watson, MD, Murphy, DJ: Targeting of oleosins to the oil bodies of oilseed rape (Brassica napus L.). Planta 189: 24–29 (1993).

    Article  PubMed  Google Scholar 

  15. Holbrook, LA, vanRooijen, GJH, Wilen, RW, Maloney, MM: Oil body proteins in microspore derived embryos of Brassica napus. Plant Physiol 97: 1051–1058 (1991).

    Google Scholar 

  16. Hsu, RC: Abscisic acid accumulation in developing seeds of Phaseolus vulgaris L. Plant Physiol 117: 211–221 (1979).

    Google Scholar 

  17. Huang, AHC: Oil bodies and oleosins in seeds. Annu Rev Plant Physiol Plant Mol Biol 43: 177–200 (1992).

    Article  Google Scholar 

  18. Hughes, DW, Galau, GA: Temporally modular gene expression during cotyledon development. Genes Devel 3: 358–369 (1989).

    PubMed  Google Scholar 

  19. Hughes, DW, Galau, GA: Cotton (Gossypium hirsutum) Mat P6 and Mat P7 oleosin genes. Plant Physiol 101: 697–698 (1993).

    Article  PubMed  Google Scholar 

  20. Jackson, DP: In situ hybridisation in plants. In: Bowles, DJ, Gurr, SJ, McPherson, M (eds) Molecular Plant Pathology: A Practical Approach, pp. 140–154. Oxford University Press, Oxford (1991).

    Google Scholar 

  21. Kater, MM, Koningstein, GM, Nijkamp, JJ, Stuitje, AR: cDNA cloning and expression of the Brassica napus enoyl-acyl carrier protein reductase in Escherichia coli. Plant Mol Biol 17: 895–909 (1991).

    PubMed  Google Scholar 

  22. Keddie J: The regulation of gene expression during Brassica napus embryogenesis. Ph. D. thesis, University of East Anglia (1993).

  23. Keddie, J, Hubner, G, Slocombe, SP, Jarvis, RP, Cummins, I, Edwards, EW, Shaw, CH, Murphy, DJ: Cloning and characterisation of an oleosin gene from Brassica napus. Plant Mol Biol 19: 443–453 (1992).

    PubMed  Google Scholar 

  24. Keddie, J, Edwards, EW, Gibbons, T, Shaw, CH, Murphy, DJ: Sequence of an oleosin cDNA from Brassica napus. Plant Mol Biol 19: 1079–1083 (1992).

    Article  PubMed  Google Scholar 

  25. Kermode, AR, Bewley, DD: Regulatory processes involved in the switch from seed development to germination. Possible roles for desiccation and ABA. In: Monti, L, Porceddu, E (eds) Drought Resistance in Plants: Physiological and Genetic Aspects, pp. 59–76 EEC Brussels, (1987).

    Google Scholar 

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

    PubMed  Google Scholar 

  27. Li, M, Smith, LJ, Clark, DC, Wilson, R, Murphy, DJ: Secondary structures of a new class of lipid body proteins from oilseeds. J Biol Chem 267: 8245–8253 (1992).

    PubMed  Google Scholar 

  28. Loer, DS, Herman, EM. Cotranslational integration of soybean oil body membrane protein oleosin into microsomal membranes. Plant Physiol 101: 993–998 (1993).

    PubMed  Google Scholar 

  29. Mansfield, SG, Briarty, LG: Cotyledon cell development in Arabidopsis thaliana during reserve deposition. Can J Bot 70: 151–164 (1992).

    Google Scholar 

  30. Murphy, DJ: Storage lipid bodies in plants and other organisms. Prog Lipid Res 29: 299–324 (1990).

    PubMed  Google Scholar 

  31. Murphy, DJ: Biochemical and molecular regulation of storage product formation in oilseeds. In: Current Topics in Plant Physiology. American Society of Plant Physiology, USA, pp. 228–237 (1993).

    Google Scholar 

  32. Murphy, DJ, Cummins, I: Biosynthesis of storage products during embryogenesis in rapeseed. J Plant Physiol 135: 63–69 (1989).

    Google Scholar 

  33. Murphy, DJ, Cummins, I, Kang, AS: Synthesis of the major oil-body membrane protein in developing rapeseed embryos. Biochem J 258: 285–293 (1989).

    PubMed  Google Scholar 

  34. Murphy, DJ, Cummins, I, Ryan, AJ: Biosynthesis and mobilisation of the major seed storage of Brassica napus L. An immunocytochemical and biochemical study. Plant Physiol Biochem 27: 647–657 (1989).

    Google Scholar 

  35. Murphy, DJ, Keen, JN, O'Sullivan, JN, Au, DMY, Edwards, EIW, Jackson, PJ, Cummins, I, Gibbons, T, Shaw, CH, Ryan, AJ: A class of amphipathic plant proteins with structural similarities to human and animal serum apolipoproteins. Biochim Biophys Acta 1088: 86–94 (1991).

    PubMed  Google Scholar 

  36. Norton, G, Harris, JF: Compositional changes in developing rapeseed (Brassica napus L.). Planta 123: 163–174 (1975).

    Google Scholar 

  37. Qu, R, Vance, VB, Huang, AHC: Expression of genes encoding oleosin isoforms in the embryos of maturing maize kernels. Plant Sci 72: 223–232 (1990).

    Article  Google Scholar 

  38. Rakow, G, McGregor, DI: Oil fatty acid and chlorophyll accumulation in developing seeds of two ‘linolenic acid lines’ of low erucic acid rapeseed. Can J Plant Sci 55: 197–203 (1995).

    Google Scholar 

  39. Rest, JA, Vaughan, JG: The development of protein and oil bodies in the seed of Sinapis alba L. Planta 105: 245–262 (1972).

    Google Scholar 

  40. Roberts, MR, Hodge, R, Ross, JHE, Sorensen, A-M, Murphy, DJ, Draper, J, Scott, R. Characterisation of a new class of oleosins indicates a male gametophyte-specific lipid storage pathway. Plant J 3: 629–636 (1993).

    Article  PubMed  Google Scholar 

  41. Ross, JHE, Murphy, DJ: Biosynthesis and localisation of storage proteins, oleosins and lipid during embryo development in Coriandrum sativum and other Umbelliferae. Plant Sci 86: 59–70 (1992).

    Article  Google Scholar 

  42. Ross, JHE, Murphy, DJ: Differential accumulation of oleosins, starch, storage proteins and triacylglycerols in embryos and cell cultures of Daucus carota L. Plant Sci 88: 1–11 (1993).

    Article  Google Scholar 

  43. Schägger, H, vonJagow, G: Tricine-sodium dodecyl sulphate polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166: 368–379 (1987).

    PubMed  Google Scholar 

  44. Senaratna, T, Kott, L, Beversdorf, WD, McKersie, BD: Desiccation of microspore derived embryos of oilseed rape (Brassica napus L.). Plant Cell Rep 10: 342–344 (1991).

    Article  Google Scholar 

  45. Senaratna, T: Artificial seeds. Biotech Adv 10: 379–392 (1992).

    Article  Google Scholar 

  46. Skriver, K, Mundy, J: Gene expression in response to abscisic acid and osmotic stress. Plant Cell 2: 503–512 (1990).

    Article  PubMed  Google Scholar 

  47. Slocombe, SP, Cummins, I, Jarvis, P, Murphy, DJ: Structure and temporal regulation of an embryo-specific Brassica napus cDNA encoding a stearoyl-acyl carrier protein (ACP) desaturase. Plant Mol Biol 20: 151–156 (1992).

    PubMed  Google Scholar 

  48. Smith, CG: The ultrastructural development of sherosomes and oil bodies in the developing embryo of Crambe abyssinica. Planta 119: 125–142 (1974).

    Google Scholar 

  49. Suzuki, Y, Kurogochi, S, Nurofushi, N, Ota, Y, Takahashi, N: Seasonal changes in GA1, GA19 and abscisic acid in three rice cultivars. Plant Cell Physiol 22: 1085–1093 (1981).

    Google Scholar 

  50. Tzen, JTC, Cao, Y-Z, Laurent, P, Ratnayake, C, Huang, AHC: Lipids, proteins and structure of seed oil bodies from diverse species. Plant Physiol 101: 267–276 (1993).

    PubMed  Google Scholar 

  51. vanStanden, J, Gilliland, MG, Brown, NAC: Ultrastructure of dry viable and non-viable Protea compacta embryos. Z Pflanzenphysiol 76: 28–35 (1975).

    Google Scholar 

  52. Vivekananda, J, Drew, MC, Thomas, T: Hormonal and environmental regulation of the carrot lea-class gene Dc3. Plant Physiol 100: 176–581 (1992).

    Google Scholar 

  53. Yatsu, LY, Jacks, TJ: Spherosome membranes. Plant Physiol 49: 937–943 (1972).

    Google Scholar 

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Cummins, I., Hills, M.J., Ross, J.H.E. et al. Differential, temporal and spatial expression of genes involved in storage oil and oleosin accumulation in developing rapeseed embryos: implications for the role of oleosins and the mechanisms of oil-body formation. Plant Mol Biol 23, 1015–1027 (1993). https://doi.org/10.1007/BF00021816

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