A modified Megazyme fructan assay for rapidly screening wheat starch synthase IIa mutation populations reveals high fructan accumulation in mature grains of triple null lines |
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| Affiliation: | 1. Division of Glycoscience, Department of Chemistry, School of Engineering Sciences in Chemistry, Biotechnology and Health, Royal Institute of Technology (KTH), AlbaNova University Centre, Stockholm SE106 91, Sweden;2. Adelaide Glycomics, School of Agriculture, Food and Wine, University of Adelaide, Waite Campus, Urrbrae, SA 5064, Australia;3. Genomics Research Centre, Academia Sinica, Sec. 2, 128 Academia Road, Nankang, Taipei 115, Taiwan;4. School of Pharmacy, College of Medicine, National Taiwan University, Taipei 100, Taiwan;5. College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, 08826, Republic of Korea;6. Wallenberg Wood Science Center, Royal Institute of Technology (KTH), Stockholm SE100 44, Sweden;1. Institute of Food Science and Biotechnology, University of Hohenheim, Garbenstraße 25, D-70599 Stuttgart, Germany;2. State Plant Breeding Institute, University of Hohenheim, Fruwirthstraße 21, D-70599, Stuttgart, Germany;3. Biological Science Department, Faculty of Science, King Abdulaziz University, P.O. Box 80257, Jeddah 21589, Saudi Arabia |
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| Abstract: | Cereal grains are the major source of fructan intake with potential health benefits for human. Measurement of fructan levels in cereal breeding populations is a great challenge and time-consuming. In this study, K-FRUCHK and K-FRUC kits commercialized by Megazyme International Limited (Bray, Ireland) were modified and optimized in a 96 deep well plate and a thermal block (BioShake iQ, Q.Instruments, Jena, Germany). The modified assays are able to measure up to 91 samples per day with a range of fructan concentrations (0.4–10.8% in this experiment). Of the two assays, the modified K-FRUC assay is more desirable for measuring fructan levels in cereal grains with high starch content. This assay was successfully used to screen three wheat starch synthase IIa (SSIIa) mutation populations with a total of 466 lines. All 21 SSIIa triple null mutants showed high fructan levels (3.1–10.8%) in wholemeal flours; while all single and double nulls were similar to wild types with ∼1% of fructans. This result indicates that lack of entire SSIIa activity changes carbon flux from starch synthesis into fructan pathway. The identified 21 triple null wheat lines may have potential use for the production of high fructan staple foods. |
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| Keywords: | Cereal grains Fructans Megazyme |
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