Value Added Wheat CRC Ltd

North Ryde, Australia

Value Added Wheat CRC Ltd

North Ryde, Australia
SEARCH FILTERS
Time filter
Source Type

Wu M.J.,Elizabeth Macarthur Agricultural Institute | Wu M.J.,Value Added Wheat CRC Ltd | Giersch T.,Elizabeth Macarthur Agricultural Institute | Giersch T.,Value Added Wheat CRC Ltd | And 11 more authors.
Journal of Cereal Science | Year: 2012

Starch granule (SG)-associated proteins are involved in starch synthesis and the interaction between SGs and the endosperm protein matrix. In this study, SG proteins were sequentially extracted with the chaotropic reagent, urea from 1M to 4M, and then profiled using an integrated proteomic approach including one- and two-dimensional electrophoresis, mass spectrometry and antibody-based enzyme-linked immunosorbent assay (ELISA). The results demonstrated that the SG-associated proteins were dominated by granule-bound starch synthase (GBSS), gliadin, low molecular weight glutenin subunits (LMW-GS), serine protease inhibitors, α-amylase inhibitors and puroindolines. A protein with an apparent molecular mass of 50kDa, expressed in cultivar hard wheat Kukri but not in soft wheat Triller was identified as a novel member of the 'S' group of LMW-GS, designated as LMW-GS-'S'. Further characterization using a broad wheat population revealed that LMW-GS-'S' was selectively expressed in hard wheat cultivars while deleted in all soft wheats tested. Its relationship with hardness was confirmed by its expression in tetraploid durum wheats, which are among the hardest wheats around the world. Monoclonal antibody (MAb) F8-14E6 against LMW-GS-'S' was developed and used in an ELISA to screen 90 Glu allele-defined doubled haploid Janz/Kukri wheat lines. The allele that encodes LMW-GS-'S' was mapped to GluB3h (p<0.001). © 2011 Elsevier Ltd.


Wu M.J.,Elizabeth Macarthur Agricultural Institute | Wu M.J.,Value Added Wheat CRC Ltd | McKay S.,Elizabeth Macarthur Agricultural Institute | McKay S.,Value Added Wheat CRC Ltd | And 3 more authors.
Journal of Cereal Science | Year: 2012

Specificity of monoclonal antibodies (MAbs) is required in diagnostic immunoassays. However, structural homology between a target antigen and other cellular proteins often causes promiscuous cross-reactivity of a MAb to proteins other than its target antigen, thereby hindering specificity. It is proposed in this study that specific proteolytic enzymes could be useful in the reduction or elimination of cross-reactive epitopes while retaining target epitopes for certain MAbs. The hypothesis was tested using a novel 'S'-type low molecular weight glutenin subunit (LMW-GS-'S') and its antibodies. The results demonstrated that the protease, chymotrypsin, markedly reduced the cross-reactivity in samples that would otherwise obscure the specific detection of LMW-GS-'S' by MAb F8-14E6 in an enzyme-linked immunosorbent assay (ELISA). Further investigations revealed that the working mechanism for this proteolytic treatment was indeed due to the selective cleavage that destroyed cross-reactive epitopes whilst retaining the intact specific epitope. As more proteases are being discovered or engineered, the successful utilization of protease in immunoassays as reported here will benefit general immunodiagnostic assay development in both medicine and agriculture by targeting specific epitopes with tailored proteolytic treatment of antigens. © 2011 Elsevier Ltd.


Wu M.J.,Elizabeth Macarthur Agricultural Institute | Wu M.J.,Value Added Wheat CRC Ltd | McKay S.,Elizabeth Macarthur Agricultural Institute | McKay S.,Value Added Wheat CRC Ltd | And 5 more authors.
Journal of Cereal Science | Year: 2012

Serpins constitute a large family of related proteins, the majority of which are serine protease inhibitors (from which their name is derived). They are known to be abundantly and polymorphically expressed in individual wheat cultivars. However, to date there lacks a detailed investigation into their inter-genotypic polymorphisms in larger wheat collections. In this study, a systematic proteomic approach, combining native polyacrylamide gel electrophoresis (PAGE), SDS-PAGE, two-dimensional gel electrophoresis, tandem mass spectrometry (MS/MS) and immunoblotting, was taken to characterize serpin polymorphisms among 177 Australian and 19 foreign hexaploid as well as 6 tetraploid wheat varieties. A total of seven serpin isoforms were identified and five unique expression patterns of these serpins were clearly revealed among varieties. In particular, the novel serpin isoforms 3a and 3b were highly polymorphic in the collection of wheat cultivars and are structurally diverse from the other serpin isoforms. The findings and methodology established in this study are not only immediately applicable in wheat breeding and varietal differentiation but may also be useful for delineation of serpin function in bread or pasta making in the future. © 2011 Elsevier Ltd.


Wu M.J.,Elizabeth Macarthur Agricultural Institute | Wu M.J.,Value Added Wheat CRC Ltd | McKay S.,Elizabeth Macarthur Agricultural Institute | McKay S.,Value Added Wheat CRC Ltd | And 5 more authors.
Journal of Cereal Science | Year: 2011

β-Amylase encoded by multiple loci on chromosomes of triticeae crop plants is a starch metabolic enzyme with a significant physiological role in flour functionality. In this study, wheat grain β-amylase isozymes were characterized using an extensive Australian wheat population and genetic breeding/mapping lines. β-amylase isozymes were fractionated by high-resolution native polyacrylamide gel electrophoresis (PAGE) and validated by zymogram and peptide sequencing using electrospray ionization tandem mass spectrometry (ESI-MS/MS). Specific antibodies were generated against the β-amylase purified through protein chromatography and used for β-amylase identification. Further characterization of wheat β-amylases was carried out with immunoblotting and 2-dimensional electrophoresis (2-DE). The enzyme was found to be polymorphic among the Australian and foreign wheat cultivars, with a total of six isozymes: two common and four varying isozymes, the latter group including one novel β-amylase isozyme of fast-mobility in native PAGE, According to pedigree analysis, the origin of this particular isozyme was from the ancestral genotype, Ciano F 67. © 2011 Elsevier Ltd.

Loading Value Added Wheat CRC Ltd collaborators
Loading Value Added Wheat CRC Ltd collaborators