Hostname: page-component-78c5997874-xbtfd Total loading time: 0 Render date: 2024-11-11T01:16:55.794Z Has data issue: false hasContentIssue false
  • English
  • Français

Identification of biotinylated proteins in soybean [Glycine max (L.) Merrill] seeds and their characterization during germination and seedling growth 1

Published online by Cambridge University Press:  19 September 2008

Robert G. Shatters Jr ,
Soon P. Boo ,
José B. França Neto  and
S. H. West
Robert G. Shatters Jr*
Affiliation:
USDA/ARS at the University of Florida, Agronomy Seed Laboratory, P.O. Box 110770, Gainesville, FL 32611–0770, USA
Soon P. Boo
Affiliation:
Embrapa-National Center for Soybean Research, Caixa Postal 231, 86001–970 Londrina, PR, Brazil
José B. França Neto
Affiliation:
Graduate Student, Agronomy Department, University of Florida, Agronomy Seed Laboratory, P.O. Box 110770, Gainesville, FL 32611–0770, USA
S. H. West
Affiliation:
USDA/ARS at the University of Florida, Agronomy Seed Laboratory, P.O. Box 110770, Gainesville, FL 32611–0770, USA
*
*Correspondence
Get access Rights & Permissions [Opens in a new window]

Abstract

Biotin is an important vitamin. It is biologically active as a protein prosthetic group, where it functions in enzymatically catalysed carboxylation reactions. It has previously been shown that the ability to synthesize biotin is not necessary for germination of Arabidopsis thaliana seeds, but that this process is required for early seedling growth. This research was conducted to determine if changes in the detection of biotinylated proteins could be observed that reflect changes in the need for biotin-mediated enzyme reactions observed during early soybean seedling growth. A seed specific 75-kDa biotinylated protein present in the embryonic axes and the cotyledons was lost during the first 3 d of germination. Seed specificity, and pattern of expression during germination suggest that this protein is a homologue of the seed specific 65-kDa biotinylated protein previously identified in pea (Pisum sativum). If samples were not treated with 2-mercaptoethanol, three equally spaced proteins at approx. 85 kDa were visible. In the presence of 2-ME these proteins appeared as a single 85-kDa band. This triplet was distinct only in the embryonic axes of dry seeds and not in imbibed seeds or in other plant parts. This demonstrates that imbibitional changes do occur in the pool of biotinylated proteins present in dry soybean seeds, and that 2-ME treatment can inhibit complete identification of the biotinylated proteins present in seed tissues.

Keywords

SoybeanGlycine maxbiotinbiotinylated proteinsgermination
Type
Physiology and Biochemistry
Information
Seed Science Research , Volume 7 , Issue 4 , December 1997 , pp. 373 - 376
Copyright
Copyright © Cambridge University Press 1997

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

1

Research funded by USDA-ARS and Embrapa-Brazihan Corporation for Agricultural Research

References

Alban, C., Jallian, J., Job, D. and Douce, R. (1995) Isolation and characterization of biotin carboxylase from pea chloroplasts. Plant Physiology 109, 927–935.CrossRefGoogle ScholarPubMed
AOSA (1983) Rules for testing seed. Springfield, Illinois. Association of Official Seed Analysts.Google Scholar
Duval, M., DeRose, R.T., Job, C., Faucher, D., Douce, R. and Job, D. (1994a) The major biotinyl protein from Pisum sativum seeds covalently binds biotin at a novel site. Plant Molecular Biology 26, 265–273.CrossRefGoogle Scholar
Duval, M., Job, C., Alban, C., Douce, R. and Job, D. (1994b) Development patterns of free and protein-bound biotin during maturation and germination of seeds of Pisum sativum: Characterization of a novel seed-specific biotinylated protein. Biochemical Journal 299, 141–150.CrossRefGoogle ScholarPubMed
Fling, S.P. and Gregerson, D.S. (1986) Peptide and protein molecular weight determination by electrophoresis using a high-molarity tris buffer system without urea. Analytical Biochemistry 155, 83–88.CrossRefGoogle ScholarPubMed
França Neto, J.B., West, S.H., Shatters, R.G. and Boo, S.P. (1997) Developmental pattern of biotinylated proteins during embryogenesis and maturation of soybean seed. Seed Science Research 7, 377–384.CrossRefGoogle Scholar
Schneider, T., Dinkins, R., Robinson, K., Shellhammer, J. and Meinke, D.W. (1989) An embryo-lethal mutant of Arabidopsis thaliana is a biotin auxotroph. Developmental Biology 131, 161–167.CrossRefGoogle ScholarPubMed
Shu, T.F., Hsieh, K.L., Hsing, Y.I., Chen, Z.Y. and Chow, T.Y. (1996) Glycine max 68 kDa LEA protein mRNA, complete cds. Direct sequence submission to Genbank. Accessiou 59626.Google Scholar
Weston, S.A., Crossett, B., Tuckwell, D.S. and Humphries, M.J. (1995) Effects of β-mercaptoethanol on the detection of biotinylated proteins. Analytical Biochemistry 225, 28–33.CrossRefGoogle ScholarPubMed
Wurtele, E.S. and Nikolau, B.J. (1990) Plants contain multiple biotin enzymes: Discovery of 3-methylcrotonyl-CoA carboxylase propionyl-CoA carboxylase and pyruvate carboxylase in the plant kingdom. Archives of Biochemistry and Biophysics 278, 179–186.CrossRefGoogle ScholarPubMed