High-Throughput Extraction and Enzymatic Determination of Sugars and Fructans in Fructan-Accumulating Plants.
Degree of polymerization
Enzymatic assay
Ethanolic extraction
Fructan determination
Sugar determination
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2022
2022
Historique:
entrez:
21
10
2021
pubmed:
22
10
2021
medline:
15
12
2021
Statut:
ppublish
Résumé
Fructans are carbohydrates present in more than 15% of flowering plants. They represent the major pool of carbohydrates in some species, especially when facing cold or drought. However, the functions of fructans with high or low degrees of polymerization (DP), their diurnal use, and the regulation of their synthesis and degradation in response to stresses still remain unclear. Here we present an enzymatic protocol adapted to 96-well microplates that simultaneously allows the determination of fructans and glucose, fructose, and sucrose. Moreover, the protocol allows to estimate the average DP of the fructans in the samples. The protocol is based on the enzymatic degradation of fructans into glucose and fructose and their subsequent conversion into gluconate 6-phosphate concomitant with the formation of NADH in the presence of ATP.
Identifiants
pubmed: 34674172
doi: 10.1007/978-1-0716-1912-4_10
doi:
Substances chimiques
Carbohydrates
0
Fructans
0
Sugars
0
Fructose
30237-26-4
Glucose
IY9XDZ35W2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
107-119Informations de copyright
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
Références
Sicher RC, Kremer DF, Harris WG (1984) Diurnal carbohydrate-metabolism of barley primary leaves. Plant Physiol 76(1):165–169
pubmed: 16663791
pmcid: 1064249
doi: 10.1104/pp.76.1.165
Farrar SC, Farrar JF (1986) Compartmentation and fluxes of sucrose in intact leaf blades of barley. New Phytol 103(4):645–657
doi: 10.1111/j.1469-8137.1986.tb00839.x
Bihmidine S, Julius BT, Dweikat I, Braun DM (2016) Tonoplast Sugar Transporters (SbTSTs) putatively control sucrose accumulation in sweet sorghum stems. Plant Signal Behav 11(1):e1117721
pubmed: 26619184
doi: 10.1080/15592324.2015.1117721
pmcid: 26619184
Cairns AJ, Cookson A, Thomas BJ, Turner LB (2002) Starch metabolism in the fructan-grasses: patterns of starch accumulation in excised leaves of Lolium temulentum L. J Plant Physiol 159(3):293–305
doi: 10.1078/0176-1617-00644
van Arkel J, Sevenier R, Hakkert JC, Bouwmeester HJ, Koops AJ, van der Meer IM (2013) Tailor-made fructan synthesis in plants: a review. Carbohydr Polym 93(1):48–56
pubmed: 23465900
doi: 10.1016/j.carbpol.2012.02.001
pmcid: 23465900
Larry MW (1973) Carbohydrate reserves of grasses: a review. J Range Manag 26(1):13–18
doi: 10.2307/3896873
Lunn JE, Hatch MD (1995) Primary partitioning and storage of photosynthate in sucrose and starch in leaves of C-4 plants. Planta 197(2):385–391
doi: 10.1007/BF00202661
Farrar SC, Farrar JF (1985) Carbon fluxes in leaf blades of barley. New Phytol 100(3):271–283
doi: 10.1111/j.1469-8137.1985.tb02778.x
Meuriot F, Morvan-Bertrand A, Noiraud-Romy N, Decau ML, Escobar-Gutierrez AJ, Gastal F, Prud’homme MP (2018) Short-term effects of defoliation intensity on sugar remobilization and N fluxes in ryegrass. J Exp Bot 69(16):3975–3986
pubmed: 29931373
pmcid: 6054246
doi: 10.1093/jxb/ery211
Yoshida M, Abe J, Moriyama M, Kuwabara T (1998) Carbohydrate levels among winter wheat cultivars varying in freezing tolerance and snow mold resistance during autumn and winter. Physiol Plant 103(1):8–16
doi: 10.1034/j.1399-3054.1998.1030102.x
Peukert M, Thiel J, Peshev D, Weschke W, Van den Ende W, Mock HP, Matros A (2014) Spatio-temporal dynamics of fructan metabolism in developing barley grains. Plant Cell 26(9):3728–3744
pubmed: 25271242
pmcid: 4213166
doi: 10.1105/tpc.114.130211
del Viso F, Puebla AF, Fusari CM, Casabuono AC, Couto AS, Pontis HG, Hopp HE, Heinz RA (2009) Molecular characterization of a putative sucrose: Fructan-6-fructosyltransferase (6-SFT) of the cold-resistant Patagonian grass Bromus pictus associated with fructan accumulation under low temperatures. Plant Cell Physiol 50(3):489–503
pubmed: 19153157
doi: 10.1093/pcp/pcp008
pmcid: 19153157
Vijn I, Smeekens S (1999) Fructan: more than a reserve carbohydrate? Plant Physiol 120(2):351–359
pubmed: 10364386
pmcid: 1539216
doi: 10.1104/pp.120.2.351
Muir JG, Shepherd SJ, Rosella O, Rose R, Barrett JS, Gibson PR (2007) Fructan and free fructose content of common Australian vegetables and fruit. J Agric Food Chem 55(16):6619–6627
pubmed: 17625872
doi: 10.1021/jf070623x
pmcid: 17625872
Livingston DP (1990) Fructan precipitation from a water/ethanol extract of oats and barley. Plant Physiol 92(3):767–769
pubmed: 16667347
pmcid: 1062366
doi: 10.1104/pp.92.3.767
Liu Z, Mouradov A, Smith KF, Spangenberg G (2011) An improved method for quantitative analysis of total fructans in plant tissues. Anal Biochem 418(2):253–259
pubmed: 21867673
doi: 10.1016/j.ab.2011.08.004
pmcid: 21867673
Ku Y, Jansen O, Oles CJ, Lazar EZ, Rader JI (2003) Precipitation of inulins and oligoglucoses by ethanol and other solvents. Food Chem 81(1):125–132
doi: 10.1016/S0308-8146(02)00393-X
Peshev D, Vergauwen R, Moglia A, Hideg E, Van den Ende W (2013) Towards understanding vacuolar antioxidant mechanisms: a role for fructans? J Exp Bot 64(4):1025–1038
pubmed: 23349141
pmcid: 3580814
doi: 10.1093/jxb/ers377
Hincha DK, Livingston DP, Premakumar R, Zuther E, Obel N, Cacela C, Heyer AG (2007) Fructans from oat and rye: composition and effects on membrane stability during drying. Biochim Biophys Acta Biomembr 1768(6):1611–1619
doi: 10.1016/j.bbamem.2007.03.011
Van den Ende W, Valluru R (2008) Sucrose, sucrosyl oligosaccharides, and oxidative stress: scavenging and salvaging? J Exp Bot 60(1):9–18
pubmed: 19036839
doi: 10.1093/jxb/ern297
pmcid: 19036839
Matros A, Peshev D, Peukert M, Mock H-P, Van den Ende W (2015) Sugars as hydroxyl radical scavengers: proof-of-concept by studying the fate of sucralose in Arabidopsis. Plant J 82(5):822–839
pubmed: 25891826
doi: 10.1111/tpj.12853
pmcid: 25891826
Roth A, Luscher N, Sprenger N, Boller T, Wiemken A (1997) Fructan and fructan-metabolizing enzymes in the growth zone of barley leaves. New Phytol 136(1):73–79
doi: 10.1111/j.1469-8137.1997.tb04732.x
Cimini S, Locato V, Vergauwen R, Paradiso A, Cecchini C, Vandenpoel L, Verspreet J, Courtin CM, D’Egidio MG, Van den Ende W, De Gara L (2015) Fructan biosynthesis and degradation as part of plant metabolism controlling sugar fluxes during durum wheat kernel maturation. Front Plant Sci 6:89
pubmed: 25750648
pmcid: 4335405
doi: 10.3389/fpls.2015.00089
Versluys M, Kirtel O, Toksoy Oner E, Van den Ende W (2018) The fructan syndrome: evolutionary aspects and common themes among plants and microbes. Plant Cell Environ 41(1):16–38
pubmed: 28925070
doi: 10.1111/pce.13070
pmcid: 28925070
Schnyder H (1993) The role of carbohydrate storage and redistribution in the source-sink relations of wheat and barley during grain filling — a review. New Phytol 123(2):233–245
doi: 10.1111/j.1469-8137.1993.tb03731.x
Nagaraj VJ, Altenbach D, Galati V, Luscher M, Meyer AD, Boller T, Wiemken A (2004) Distinct regulation of sucrose: sucrose-1-fructosyltransferase (1-SST) and sucrose: fructan-6-fructosyltransferase (6-SFT), the key enzymes of fructan synthesis in barley leaves: 1-SST as the pacemaker. New Phytol 161(3):735–748
pubmed: 33873712
doi: 10.1111/j.1469-8137.2004.00995.x
pmcid: 33873712
Nagaraj VJ, Riedl R, Boller T, Wiemken A, Meyer AD (2001) Light and sugar regulation of the barley sucrose: fructan 6-fructosyltransferase promoter. J Plant Physiol 158(12):1601–1607
doi: 10.1078/0176-1617-00592
Ritsema T, Brodmann D, Diks SH, Bos CL, Nagaraj V, Pieterse CM, Boller T, Wiemken A, Peppelenbosch MP (2009) Are small GTPases signal hubs in sugar-mediated induction of fructan biosynthesis? PLoS One 4(8):e6605
pubmed: 19672308
pmcid: 2720452
doi: 10.1371/journal.pone.0006605
Jin Y, Fei M, Rosenquist S, Jin L, Gohil S, Sandstrom C, Olsson H, Persson C, Hoglund AS, Fransson G, Ruan Y, Aman P, Jansson C, Liu C, Andersson R, Sun C (2017) A dual-promoter gene orchestrates the sucrose-coordinated synthesis of starch and fructan in barley. Mol Plant 10(12):1556–1570
pubmed: 29126994
doi: 10.1016/j.molp.2017.10.013
pmcid: 29126994
Arkel V (2014) Fructan biosynthesis regulation and the production of tailor-made fructan in plants. In: Polysaccharides: natural fibers in food and nutrition. CRC Press, Boca Raton, FL, pp 1–29
Cairns AJ (2003) Fructan biosynthesis in transgenic plants. J Exp Bot 54(382):549–567
pubmed: 12508066
doi: 10.1093/jxb/erg056
pmcid: 12508066
Chalmers J, Lidgett A, Cummings N, Cao Y, Forster J, Spangenberg G (2005) Molecular genetics of fructan metabolism in perennial ryegrass. Plant Biotechnol J 3(5):459–474
pubmed: 17173633
doi: 10.1111/j.1467-7652.2005.00148.x
pmcid: 17173633
Apolinario AC, de Lima Damasceno BP, de Macedo Beltrao NE, Pessoa A, Converti A, da Silva JA (2014) Inulin-type fructans: a review on different aspects of biochemical and pharmaceutical technology. Carbohydr Polym 101:368–378
pubmed: 24299785
doi: 10.1016/j.carbpol.2013.09.081
pmcid: 24299785
Xue GP, Drenth J, Glassop D, Kooiker M, McIntyre CL (2013) Dissecting the molecular basis of the contribution of source strength to high fructan accumulation in wheat. Plant Mol Biol 81(1–2):71–92
pubmed: 23114999
doi: 10.1007/s11103-012-9983-1
pmcid: 23114999
Obenland DM, Simmen U, Boller T, Wiemken A (1991) Regulation of sucrose-sucrose-fructosyltransferase in barley leaves. Plant Physiol 97(2):811–813
pubmed: 16668471
pmcid: 1081079
doi: 10.1104/pp.97.2.811
Suarez-Gonzalez EM, Lopez MG, Delano-Frier JP, Gomez-Leyva JF (2014) Expression of the 1-SST and 1-FFT genes and consequent fructan accumulation in Agave tequilana and A. inaequidens is differentially induced by diverse (a)biotic-stress related elicitors. J Plant Physiol 171(3–4):359–372
pubmed: 23988562
doi: 10.1016/j.jplph.2013.08.002
pmcid: 23988562
Wagner W, Wiemken A (1987) Enzymology of fructan synthesis in grasses: properties of sucrose-sucrose-fructosyltransferase in barley leaves (Hordeum-vulgare-L cv Gerbel). Plant Physiol 85(3):706–710
pubmed: 16665764
pmcid: 1054326
doi: 10.1104/pp.85.3.706
Wagner W, Wiemken A, Matile P (1986) Regulation of fructan metabolism in leaves of barley (Hordeum vulgare L cv Gerbel). Plant Physiol 81(2):444–447
pubmed: 16664835
pmcid: 1075355
doi: 10.1104/pp.81.2.444
Bohacenko J (2014) Fructan content determination by HPLC method with refractometric detection. Listy Cukrov Repar 130(1):28–32
Longland AC, Dhanoa MS, Harris PA (2012) Comparison of a colorimetric and a high-performance liquid chromatography method for the determination of fructan in pasture grasses for horses. J Sci Food Agric 92(9):1878–1885
pubmed: 22297902
doi: 10.1002/jsfa.5555
pmcid: 22297902
Verspreet J, Hansen AH, Dornez E, Courtin CM, Harrison SJ (2014) A new high-throughput LC-MS method for the analysis of complex fructan mixtures. Anal Bioanal Chem 406(19):4785–4788
pubmed: 24825765
doi: 10.1007/s00216-014-7861-1
pmcid: 24825765
Rao RSP, Andersen JR, Dionisio G, Boelt B (2011) Fructan accumulation and transcription of candidate genes during cold acclimation in three varieties of Poa pratensis. J Plant Physiol 168(4):344–351
pubmed: 20880605
doi: 10.1016/j.jplph.2010.07.019
pmcid: 20880605
Stitt M, Lilley RM, Gerhardt R, Heldt HW (1989) Metabolite levels in specific cells and subcellular compartments of plant leaves. Methods Enzymol 174:518–552
doi: 10.1016/0076-6879(89)74035-0
Cross JM, von Korff M, Altmann T, Bartzetko L, Sulpice R, Gibon Y, Palacios N, Stitt M (2006) Variation of enzyme activities and metabolite levels in 24 Arabidopsis accessions growing in carbon-limited conditions. Plant Physiol 142(4):1574
pubmed: 17085515
pmcid: 1676042
doi: 10.1104/pp.106.086629
McComb RB, Bond LW, Burnett RW, Keech RC, Bowers GN Jr (1976) Determination of the molar absorptivity of NADH. Clin Chem 22(2):141–150
pubmed: 2388
doi: 10.1093/clinchem/22.2.141
pmcid: 2388