Gao L.,Capital Normal University |
Ma W.,Western Australian Department of Agriculture and Food |
Chen J.,CAS Chengdu Institute of Biology |
Wang K.E.,Capital Normal University |
And 5 more authors.
Journal of Agricultural and Food Chemistry | Year: 2010
High molecular weight glutenin subunits (HMW-GS) from 60 germplasms including 30 common wheat cultivars and 30 related species were separated and characterized by a suite of separation methods including sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), reversed-phase high-performance liquid chromatography (RP-HPLC), high-performance capillary electrophoresis (HPCE), and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Comparative analysis demonstrated that each methodology has its own advantages and disadvantages. The main drawback of SDS-PAGE was its overestimation of molecular mass and incorrect identification of HMW-GS due to its low resolution. However, it had the advantages of technical simplicity and low requirements of equipment; thus, it is suitable for large-scale and high-throughput HMW-GS screening for breeding programs, especially when the glutenin composition is clear in the breeding material. MALDI-TOF-MS clearly expressed many technical advantages among the four methods evaluated, including high throughput, high resolution, and accuracy; it was, however, associated with high equipment cost, thus preventing many breeding companies from accessing the technology. RP-HPLC and HPCE were found to be intermediate between SDS-PAGE and MALDI-TOF-MS. Both RP-HPLC and HPCE demonstrated higher resolution and reproducibility over SDS-PAGE but lower detection power than MALDI-TOF-MS. Results demonstrated that MALDI-TOF-MS is suitable for analyzing HMW-GS for routine breeding line screening and for identifying new genotypes. © 2010 American Chemical Society.
Vanhercke T.,Food Futures National Research Flagship |
El Tahchy A.,Food Futures National Research Flagship |
Shrestha P.,Food Futures National Research Flagship |
Zhou X.-R.,CSIRO |
And 2 more authors.
FEBS Letters | Year: 2013
Metabolic engineering approaches to increase plant oil levels can generally be divided into categories which increase fatty acid biosynthesis ('Push'), are involved in TAG assembly ('Pull') or increase TAG storage/decrease breakdown ('Accumulation'). In this study, we describe the surprising synergy when Push (WRI1) and Pull (DGAT1) approaches are combined. Co-expression of these genes in the Nicotiana benthamiana transient leaf expression system resulted in TAG levels exceeding those expected from an additive effect and biochemical tracer studies confirmed increased flux of carbon through fatty acid and TAG synthesis pathways. Leaf fatty acid profile also synergistically shifts from polyunsaturated to monounsaturated fatty acids. © 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
Netzel M.,Food Futures National Research Flagship |
Netzel M.,CSIRO |
Netzel G.,Food Futures National Research Flagship |
Netzel G.,CSIRO |
And 12 more authors.
Food Research International | Year: 2011
The release and absorption (bioavailability) of carotenoids is a prerequisite for their nutritional impact. This can be strongly affected by the processing conditions used to prepare the food matrix that contains them. To determine the effect of processing on carotenoid bioavailability, homogenized, raw, blanched and cooked carrots were exposed to an in vitro gastric and intestinal digestion model. Final digest samples were placed onto a Caco-2 cell trans-well monolayer culture to mimic intestinal absorption. The results show that the cooked carrot puree consisting of primarily single plant cell particles had the highest release of carotenes, followed by blanched consisting primarily of plant cell clusters and raw carrot puree consisting of larger plant cell clusters. Absorption through the Caco-2 cell layer was the highest from the digesta of cooked carrot puree followed by the digesta of blanched carrot puree. This study demonstrates that thermal processing and/or mechanical homogenization to disrupt plant cell wall matrix enhances the in vitro bioavailability of carotenes from carrots. © 2011.