Planta

, Volume 222, Issue 3, pp 512–520 | Cite as

Two naturally occurring deletion mutants of 12S seed storage proteins in Arabidopsis thaliana

  • Anfu Hou
  • Kede Liu
  • Niramol Catawatcharakul
  • Xurong Tang
  • Vi Nguyen
  • Wilfred A. Keller
  • Edward W. T. Tsang
  • Yuhai Cui
Original Article

Abstract

Two naturally occurring Arabidopsis mutants, Cape Verde Islands and Monte (Mr-0), with aberrant 12S seed storage protein (SSP) profiles have been identified by SDS-PAGE. In both mutants, one of the 12S globulin bands is missing while a new band of lower molecular mass is present. Tandem mass spectrometry-mass spectrometry (MS/MS) analyses of the mutant peptides have revealed that both are shorter variants of 12S globulin with deletion sites detected within the α-subunits of 12S globulin cruciferin B (CRB) and C (CRC), respectively. Sequence analyses of the genomic DNA flanking the deletion sites have demonstrated that both deletions occurred at the genomic level. These two mutants are referred to as CRBΔ12 and CRCΔ13 with the delta sign indicating a deletion and the number indicating amino acids deleted. Alignment of these two mutant sequences with that of soybean A3B4 subunit, for which the crystal structure was determined recently, have revealed that the CRCΔ13 deletion is located in a hypervariable/disordered region, and will probably not affect the structure of the hexameric globulin. The CRBΔ12 deletion, however, is located in a binding region that is thought to be important for the hexamer formation. However, CRBΔ12 appears to accumulate normally as judged by its band intensity relative to the other SSP subunits on the protein gels. Thus it seems that the seed can, to a certain extent, tolerate some mutations in its storage proteins.

Keywords

Arabidopsis Deletion mutants Mass spectrometry Natural variations Seed storage proteins 

Abbreviations

cM

Centimorgan

CRB

Cruciferin B

CRC

Cruciferin C

MS

Mass spectrometry

MS/MS

Tandem mass spectrometry-mass spectrometry

QTL

Quantitative trait loci

RIL

Recombinant inbred line

SSP

Seed storage protein

Notes

Acknowledgements

We thank Sangeeta Dhaubhadel, Frederic Marsolais, and Brian McGarvey for critical reading of the manuscript; Jeremy Murray for help with genetic mapping; Cunjie Zhang, Biological Mass Spectrometry Laboratory of the University of Western Ontario, for her expert peptide sequence service; Patrick Chapman for bioinformatics help with Fig. 5; Alex Molnar for help in preparing the figures; Ida van Grinsven for DNA sequencing; the Arabidopsis Biological Resource Center (Columbus, Ohio, USA) for Arabidopsis accessions; Agriculture and Agri-Food Canada’s Canadian Crop Genomics Initiative and Genome Canada/Genome Prairie for funding.

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Anfu Hou
    • 1
  • Kede Liu
    • 1
    • 2
  • Niramol Catawatcharakul
    • 1
  • Xurong Tang
    • 1
    • 3
  • Vi Nguyen
    • 1
  • Wilfred A. Keller
    • 3
  • Edward W. T. Tsang
    • 3
  • Yuhai Cui
    • 1
  1. 1.Southern Crop Protection and Food Research CentreAgriculture and Agri-Food CanadaLondonCanada
  2. 2.National Key Laboratory of Crop Genetic ImprovementHuazhong Agriculture UniversityWuhanChina
  3. 3.Plant Biotechnology InstituteNational Research Council of CanadaSaskatoonCanada

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