The feasibility of genetically engineering soybean seed coats to divert metabolism towards the production of novel biochemicals was tested. The genes phbA, phbB, phbC from Ralstonia eutropha each under the control of the seed coat peroxidase promoter were introduced into soybean and the production of polyhydroxybutyrate (PHB) was assayed. The analysis of seed coats arising from 4 independent transformation events demonstrated that PHB was produced at a mean of 0.12% seed coat dried weight with individual values up to 0.36%. These values demonstrate that it is possible to metabolically engineer soybean seed coats.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Bland JM, Altman DG (1995) Multiple significance tests: the Bonferroni method. Br Med J 310:169–170
Bohmert K, Balbo I, Kopka J, Mittendorf V, Nawrath C, Poirier Y, Tischendorf G, Tretheway RN, Willmitzer L (2000) Transgenic Arabidopsis plants can accumulate polyhydroxybutyrate to up to 4% of their fresh weight. Planta 211:841–845
Bohmert K, Balbo I, Steinbuche A, Tischendorf G, Willmitzer L (2002) Constitutive expression of the β-ketothiolase gene in transgenic plants. A major obstacle for obtaining polyhydroxybutyrate-producing plants. Plant Physiol 128:1282–1290
Capell T, Christou P (2004) Progress in plant metabolic engineering. Curr Opin Biotechnol 15:148–154
Chang T-CL, Karr C Jr (1959) Gas-liquid chromatographic analysis of aromatic hydrocarbons boiling up to 218° in a low-temperature coal tar. Anal Chim Acta 21:474–490
Corner EJH (1951) The leguminous seed. Phytomorphology 1:117–150
Gijzen M (1997) A deletion mutation at the Ep locus causes low seed coat peroxidase activity in soybean. Plant J 12:991–998
Gijzen M, van Huystee R, Buzzell RI (1993) Soybean seed coat peroxidase: a comparison of high-activity and low-activity genotypes. Plant Physiol 103:1061–1066
Gijzen M, Miller SS, Bowman LA, Batchelor AK, Boutilier K, Miki BLA (1999) Localization of peroxidase mRNAs in soybean seeds by in situ hybridization. Plant Mol Biol 41:57–63
Gillikan JW, Graham JS (1991) Purification and developmental analysis of the major anionic peroxidase from the seed coat of Glycine max. Plant Physiol 96:214–220
Gray JSS, Montgomery R (2006) Asymmetric glycosylation of soybean seed coat peroxidase. Carbohydr Res 341:198–209
Gudynaite-Savitch L, Johnson DA, Miki BL (2009) Strategies to mitigate transgene–promoter interactions. Plant Biotechnol J 7:472–485
Hahn JJ, Eschenlauer AC, Sleytr UB, Somers DA, Srienc F (1999) Peroxisomes as sites for synthesis of polyhydroxyalkanoates in transgenic plants. Biotechnol Prog 15:1053–1057
Han SY (2009) Development of soybean seed coat as a bioreactor for the production of industrial enzymes. Dissertation, University of Ottawa
John ME, Keller G (1996) Metabolic pathway engineering in cotton: biosynthesis of polyhydroxybutyrate in fiber cells. Proc Natl Acad Sci USA 93:12768–12773
Malik K, Wu K, Li X-Q, Martin-Heller T, Hu M, Foster E, Tian L, Wang C, Ward K, Jordan M, Brown D, Gleddie S, Simmonds D, Zheng S, Simmonds J, Miki B (2002) A constitutive gene expression system derived from the tCUP cryptic promoter elements. Theor Appl Genet 105:505–514
Miller S, Jin Z, Schnell JA, Romero C, Brown D, Johnson DA (2010) Hourglass cell development in the soybean seed coat. Ann Bot 106:235–242
Moïse JA, Han S-Y, Gudynaitę-Savitch L, Johnson DA, Miki BLA (2005) Seed coats: structure, development, composition, and biotechnology. In Vitro Cell Dev Biol Plant 41:620–644
Nawrath C, Poirier Y, Somerville C (1994) Targeting of the polyhydroxybutyrate biosynthetic pathway to the plastids of Arabidopsis thaliana results in high levels of polymer accumulation. Proc Natl Acad Sci USA 91:12760–12764
Peoples OP, Sinskey AJ (1989a) Poly-β-hydroxybutyrate biosynthesis in Alcaligenes eutrophus H16: characterization of the genes encoding β-ketothiolase and acetoacetyl-CoA reductase. J Biol Chem 264:15293–15297
Peoples OP, Sinskey AJ (1989b) Poly-β-hydroxybutyrate (PHB) biosynthesis in Alcaligenes eutrophus H16: identification and characterization of the PHB polymerase gene (phbC). J Biol Chem 264:15298–15303
Poirier Y, Dennis D, Klomparens K, Nawrath C, Somerville C (1992a) Perspectives on the production of polyhydroxyalkanoates in plants. FEMS Microbiol Rev 103:237–246
Poirier Y, Dennis D, Klomparens K, Somerville C (1992b) Polyhydroxybutyrate, a biodegradable thermoplastic, produced in transgenic plants. Science 256:520–523
Saruul P, Srienc F, Somers DA, Samac DA (2002) Production of a biodegradable plastic polymer, poly-β-hydroxybutyrate in transgenic alfalfa. Crop Sci 42:919–927
Schnell JA, Han S-Y, Miki BL, Johnson DA (2010) Soybean peroxidase propeptides are functional signal peptides and increase the yield of a foreign protein. Plant Cell Rep 29:987–996
Schubert P, Steinbüchel A, Schlegel GG (1988) Cloning of the Alcaligenes eutrophus genes for synthesis of poly-β-hydroxybutyric acid (PHB) and synthesis of PHB in Escherichia coli. J Bact 170:5837–5847
Sharma AK, Sharma MK (2009) Plants as bioreactors: recent developments and emerging opportunities. Biotechnol Adv 27:811–832
Slater SC, Voige WH, Dennis DE (1988) Cloning and expression in Escherichia coli of the Alcaligenes eutrophus H16 poly-β-hydroxybutyrate biosynthetic pathway. J Bact 170:4431–4436
Snell KD, Peoples OP (2009) PHA bioplastic: a value-added coproduct for biomass biorefineries. Biofuels Bioprod Bioref 3:456–467
Trick HN, Dinkins RD, Santarem ER, Di R, Samoylov V, Meurer CA, Walker DR, Parrott WA, Finer JJ, Collins GB (1997) Recent advances in soybean transformation. Plant Tissue Cultur Biotechnol 3:9–26
United States Department of Agriculture, Foreign Agricultural Service. (2010). http://www.fas.usda.gov/psdonline/psdreport.aspx?hidReportRetrievalName=BVS&hidReportRetrievalID=906&hidReportRetrievalTemplateID=1. Accessed February 24, 2010
Welinder KG, Larsen YB (2004) Covalent structure of soybean seed coat peroxidase. Biochim Biophys Acta 1698:121–126
Wróbel M, Zebrowski J, Szopa J (2004) Polyhydroxybutyrate synthesis in transgenic flax. J Biotechnol 107:41–54
Yang J, Barr LA, Fahnestock SR, Liu Z-B (2005) High yield recombinant silk-like protein production in transgenic plants through protein targeting. Transgenic Res 14:313–324
We thank Dr. D.C.W. Brown and Tara Rintoul of the Southern Crop Protection and Food Research Centre, London, Ontario N5V 4T3 for soybean transformation, and referees for helpful suggestions. This research was supported, in part, by a strategic grant from the Natural Sciences and Engineering Research Council of Canada to DAJ.
Electronic supplementary material
Below is the link to the electronic supplementary material.
About this article
Cite this article
Schnell, J.A., Treyvaud-Amiguet, V., Arnason, J.T. et al. Expression of polyhydroxybutyric acid as a model for metabolic engineering of soybean seed coats. Transgenic Res 21, 895–899 (2012). https://doi.org/10.1007/s11248-011-9575-x
- Soybean seed coat
- Metabolic engineering