New possibilities in micro-scale wheat     quality characterisation: micro-gluten      determination and starch isolationS...
New possibilities in micro-scale wheat quality characterisation                                       Why micro-tests?    ...
New possibilities in micro-scale wheat quality characterisation       Instrumental and methodological background of       ...
New possibilities in micro-scale wheat quality characterisation    Available micro-scale tools today    (most frequently u...
New possibilities in micro-scale wheat quality characterisation  Successful longterm ongoing Hungarian-Australian   collab...
New possibilities in micro-scale wheat quality characterisationTradition 1: Supporting the birth of wheat chemistry with H...
New possibilities in micro-scale wheat quality characterisation   Continuation of our tradition: Sample preparation with m...
New possibilities in micro-scale wheat quality characterisation                                 Continuation of our tradit...
New possibilities in micro-scale wheat quality characterisation   Tradition 2: Development of routine tests for determinat...
New possibilities in micro-scale wheat quality characterisation     Continuation of our tradition: development the prototy...
New possibilities in micro-scale wheat quality characterisation           Applications: Comparison of mixing properties   ...
New possibilities in micro-scale wheat quality characterisation                 Applications in research activities (examp...
New possibilities in micro-scale wheat quality characterisation    Application in routine analysis – Micro-doughLab by Per...
New possibilities in micro-scale wheat quality characterisation                      Tradition 3: Sedimentation tests     ...
New possibilities in micro-scale wheat quality characterisation    Principle and interpretation of sedimentation tests   T...
New possibilities in micro-scale wheat quality characterisation Recent developments: Semi-automatic instrument and methods...
New possibilities in micro-scale wheat quality characterisation     Applications: method validation, routine analysis   Su...
New possibilities in micro-scale wheat quality characterisation Application in research activities: QTL mapping for Zeleny...
New possibilities in micro-scale wheat quality characterisation  New member of the small-scale family: Development of GluS...
New possibilities in micro-scale wheat quality characterisation                           Applications: method validation,...
New possibilities in micro-scale wheat quality characterisation            Application in research activities: An example ...
New possibilities in micro-scale wheat quality characterisation       Application in research activities: Investigation of...
New possibilities in micro-scale wheat quality characterisation     Application in research activities: Investigation the ...
New possibilities in micro-scale wheat quality characterisation         Application in research activities: Relatinoship b...
New possibilities in micro-scale wheat quality characterisation  Instead of summary … a completted       picture of micro-...
New possibilities in micro-scale wheat quality characterisation                         Acknowledgement Many thanks all of...
Thank you for your kind      attention!
New possibilities in micro-scale wheat quality characterisation                    Instead of summary…             The who...
New possibilities in micro-scale wheat quality characterisation     Applications in research activities: measured relation...
Budapest University of Technology and Economics            Invitation to the next MoniQA Conference….
New possibilities in micro-scale wheat quality characterisation       Requirements for high throughput methods            ...
New possibilities in micro-scale wheat quality characterisationInstead of summary: the available micro-scale          inst...
New possibilities in micro-scale wheat quality characterisation      Case 2: Sample preparation with micro-mill for       ...
New possibilities in micro-scale wheat quality characterisation   Bekes, F. and Gras, P.W. Demonstration of the 2 g Mixogr...
New possibilities in micro-scale wheat quality characterisation  Dobraszczyk, B.J. and Roberts C.A. Strain hardening and  ...
New possibilities in micro-scale wheat quality characterisation  R. Kieffer, H. Wieser, M.H. Henderson, A. Graveland:  Cor...
New possibilities in micro-scale wheat quality characterisation   Ide kellene ugyanaz, ami előbb első RVA cikk és abstract...
Budapest University of Technology and Economics                                    Summary…       -Functional data and the...
Budapest University of Technology and Economics
Budapest University of Technology and Economics           Result 3: Relationship between different functional             ...
Budapest University of Technology and Economics                                                    But… Effects of locatio...
But… Effects of treatment             (Pannon-wheat quality program)                                      Location        ...
Budapest University of Technology and Economics                    Pictures from the beginning…
Budapest University of Technology and Economics                    Pictures from the beginning…
Budapest University of Technology and Economics                    Pictures from the beginning…                           ...
Budapest University of Technology and Economics                    Pictures from the beginning…
Budapest University of Technology and Economics             Application fields and results of micro tests
Budapest University of Technology and Economics                       Method(s) 2: Mixing properties        Summary:      ...
Budapest University of Technology and Economics                       Method(s) 2: Mixing properties
Budapest University of Technology and Economics                                     Case 2: Effect of Glu/Gli ratio on the...
Budapest University of Technology and Economics                      Case 2: Effect of Glu/Gli ratio on the mixing        ...
Budapest University of Technology and Economics           Case 3: Effect of protein subunit incorporation on              ...
Budapest University of Technology and Economics         Method 1: Sample preparation with micro mill
Budapest University of Technology and Economics        Method 1: Sample preparation with micro mill                       ...
Budapest University of Technology and Economics                                  Standardization         • ICC Standar dNo...
Budapest University of Technology and Economics          Measured relationships among Zeleny values and                  d...
Budapest University of Technology and Economics      Case 4 - Molecular backcround: Relationship between         allelic c...
Budapest University of Technology and Economics                                               Case 4 - Kazakh wheat variet...
Budapest University of Technology and Economics          Kazakh wheat varieties – Micro-Zeleny values
Budapest University of Technology and Economics                                     Case 5 - Preliminary QTL analysis     ...
Budapest University of Technology and Economics                        Case 5 - Preliminary QTL analysis                  ...
Budapest University of Technology and Economics             A tészta-jellemzők (erősség, nyújthatóság) szignifikáns       ...
Budapest University of Technology and Economics         Case 6 - NIR-prediction for Zeleny based on Kazakh                ...
Budapest University of Technology and Economics                    Case 6 - NIR-prediction for Zeleny based on Kazakh     ...
Budapest University of Technology and Economics          Case +1: Evaluation of NIR spectras for Hungarian                ...
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New possibilities in micro-scale wheat quality characterisation: micro-gluten determination and starch isolation

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International Gluten Workshop, 11th; Beijing (China); 12-15 Aug 2012

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New possibilities in micro-scale wheat quality characterisation: micro-gluten determination and starch isolation

  1. 1. New possibilities in micro-scale wheat quality characterisation: micro-gluten determination and starch isolationS. TÖMÖSKÖZI1, SZ. SZENDI1, A. BAGDI1, A. HARASZTOS1, G. BALÁZS1, D. DIEPEVEEN2, R. APPELS3 and F. BÉKÉS4 1 Budapest University of Technology and Economics (BUTE), Department of Applied Biotechnology and Food Science, Budapest, Hungary; 2Department of Agriculture and Food Western Australia 3 Centre for Comparative Genomics, Murdoch University, Western Australia, Australia; 4 FBFD PTY LTD, Beecroft, NSW, Australia 11th International Gluten Workshop August 12-15, 2012 Beijing, China
  2. 2. New possibilities in micro-scale wheat quality characterisation Why micro-tests? Where the amount of sample is limited… Earlier quality-based selection of wheat lines Studying the roles of protein subunits in modified wheat varieties Investigation of the effects of different types of additives (protein subunits, carbohydrates, lipoproteins, etc.) on the quality parameters. Application in routine analytical work. However, the causes of the differences between standard and small-scale procedures have to be identified.
  3. 3. New possibilities in micro-scale wheat quality characterisation Instrumental and methodological background of micro-scale research
  4. 4. New possibilities in micro-scale wheat quality characterisation Available micro-scale tools today (most frequently used examples)
  5. 5. New possibilities in micro-scale wheat quality characterisation Successful longterm ongoing Hungarian-Australian collaboration with CSIRO Plant Industry, Newport Scientific Ltd (Australia), Metefém Ltd and Labintern Ltd, Hungary
  6. 6. New possibilities in micro-scale wheat quality characterisationTradition 1: Supporting the birth of wheat chemistry with Hungarian micro-mill in Australia from 1870s F.B. Gurthi drafted the philosophy of the wheat quality measurement and research, namely: We have to mimic the industrial processes in the laboratory. Hungarian Ganz-type micro-mill
  7. 7. New possibilities in micro-scale wheat quality characterisation Continuation of our tradition: Sample preparation with micro mill and sieve Technical parameters Optimal moisture content : durum wheat 16 % aestivum wheat 15 % Milling yields : 51-70 % Weight : 17.5 kg Dimensions : 270×210×350 mm Sample size : >3g
  8. 8. New possibilities in micro-scale wheat quality characterisation Continuation of our tradition: Comparison with standard laboratory milling procedures 80 Correlation between milling Summary: Milling Yield on Micro Mill 75 yields measured with micro- mill vs. Quadromat-Junior 70 type labmill - Milling yields are slightly lower in 65 case of micro-mill. 60 - The mixing curves are similar and the 55 calculated parameters are in good correlation. 50 50 55 60 65 70 75 80 Milling Yield on Traditional Test Mills Effect on Mixing Time Effect on Loaf Height 350 36Comparison studies of 300 35 Mixing Time (sec) 34 dough properties 250 Loaf Height (mm) 33 with 200 32 BUHLER and 150 31 FQC-2000 flours 100 30 50 29 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 28 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Flour Samples Flour Samples
  9. 9. New possibilities in micro-scale wheat quality characterisation Tradition 2: Development of routine tests for determination of wheat qulaity Jenő Hankóczy (1869 – 1939) Hungarian inventor The prototype of the Farinograph of the Farinograph (1912- 1928) and Extensograph
  10. 10. New possibilities in micro-scale wheat quality characterisation Continuation of our tradition: development the prototype of micro-scale Z-arm mixer in cooperation with CSIRO and Newport Scientific, Australia Magvas Macro Valorigraph Magvas Micro Z-arm Mixer
  11. 11. New possibilities in micro-scale wheat quality characterisation Applications: Comparison of mixing properties Summary: - Characteristics of curves registered with micro- and macro methods are similar. - The correlations between macro and micro parameters are highly significant. - Some alterations can be identified – for example in case of dough development time. We have to learn more about the causes of identified differences.
  12. 12. New possibilities in micro-scale wheat quality characterisation Applications in research activities (examples) 250 250 Z-arm mixer 2g Mixograph 1 , 17+18,5+10 Research on protein - , 17+18,5+10D 200 200 subunit composition 1 , - ,5+10D 150 150 (Békés et al) 1 , 17+18, -T 100 Mixes of flour prepared from wheat with deletions of glutenin subunits,5+10 100 -, - genes show much better discrimination between poor and good wheats - - , 17+18, 50 50 than the Mixograph probably because the mechanism of dough - , - 1, 0 development in the Z-arm mixer is not as dependant on dough strength ,as 0 -, - - in pin mixers. Research on glutenin subunit incorporation into rice dough (Oszvald et al) Please visit our poster, also: Balázs, G. et al: Complex quality characterisation…
  13. 13. New possibilities in micro-scale wheat quality characterisation Application in routine analysis – Micro-doughLab by Perten Instuments Features and benefits - Small (4 gram) Sample Size - Correlates with Standard Tests - Programmable Mixing Speed (Source: www.perten.com. 01 08 2012)
  14. 14. New possibilities in micro-scale wheat quality characterisation Tradition 3: Sedimentation tests History 1916. First experimental results for correlation between collodial swelling properties and the baking quality of wheat flours by Upson és Calvin 1918. In diluted acid solulion, the rate and extent of the swelling of „strong” wheat flours are higher than the same value of weak flours (Gortner és Doherty) …. 1946. …relationship between water-retention capacity and loaf volume for hard ret winter wheat (Finney és Yamazaki). 1947. Higher gluten content and better gluten quality gives slower sedimentation and higher Zeleny value. (Lawrence Zeleny) 1950- Standardization of Zeleny- and other sedimentation tests...
  15. 15. New possibilities in micro-scale wheat quality characterisation Principle and interpretation of sedimentation tests The Zeleny value means the degree of sedimentation of suspended flour in lactic acid in a given time. The method is characterising the swelling properties of wheat proteins and insoluble components. First experiments Corr. between prot. cont. and Corr. between loaf valume and sedimentation value sedimentation value Zeleny, L, november, 1947
  16. 16. New possibilities in micro-scale wheat quality characterisation Recent developments: Semi-automatic instrument and methods for determination of Zeleny value on micro- and micro-scale in cooperation with Labintern Ltd, Hungary Detector Easy sample handling Micro-test tubes Macro-test tubes Shaking unit Operational sequence SediCom Tester
  17. 17. New possibilities in micro-scale wheat quality characterisation Applications: method validation, routine analysis Summary: - The new instrument is an automated version of the traditional equipment, applicable also for macro (standard)- and micro-scale measurements, where the sample requirements are 3.2g and 0,4 g of flour, respectively. - Using the equipment, the measurement is simpler, faster, and more accurate. The daily throughput can be increased by 50% per day.
  18. 18. New possibilities in micro-scale wheat quality characterisation Application in research activities: QTL mapping for Zeleny values Experimental design followed, consisting of blocking for individual days and day order recorded. Population: Chara (Aus) x Glenlea (Can), 180 DH lines. Field and milling effects were also included in the QTL model. Replication at the zeleny testing consisted of double replicates for each genotype (Cavanagh, C. et al) Functional data and their relationships to allelic composition indicated that • Most significant on for Zeleny value while dough strength and partly extensibility are dependQTLthe presence of were detected on chromosomes not HMW glutenin alleles, the allelic composition of LMW glutenins are mostly responsible Glu3B variation of Zeleny indeces of the wheats, investigated. for the harbouring HMW Glutenin loci • Among glutenin loci, significant QTLs wereChrom 6A the regions of both type of found in -LOD 6.1 (possibly Gliadin) • Chrom 1 and 6. gliadin loci on chromosome 5A -LOD 4.5 (protein gene?) . Chrom 3B –LOD 3.5 • 1B Rmax EXT Please visit our poster, also: Balázs, G. et al: Complex quality characterisation…
  19. 19. New possibilities in micro-scale wheat quality characterisation New member of the small-scale family: Development of GluStar system and methods for determination of gluten content and separation of wheat starch (in collaboration with Labintern Ltd, Hungary and CSIRO Plant Industry, Australia) The novelty is the capability for the determination of wet gluten content and for the separation of the starch from flour samples, in same time. The modularity of the measuring system ensures the utilization of basic modules, separately. Filters for separation Vacuum of starch (soluble system The combined measuring system is carbohydrate available with standard (10 g) and polimers) micro-scale (5g) bowls.
  20. 20. New possibilities in micro-scale wheat quality characterisation Applications: method validation, routine analysis Both small scale and macro-methods were validated for determination of wet gluten content. The results are correlated with the generally used standard methods (ISO 21415-2:2006, ICC-Standard No.106/2). Table 1: Wet Gluten content (%) Glutomatic 2200 vs. Micro GluStar System Martonvásár vs. Micro GluStar System A Micro GluStar System = -3,796 + 1,0491 * Glutomatic 2200 B Micro GluStar System = -8,012 + 1,2224 * Martonvásár Correlation: r = ,99281 Correlation: r = ,98815 42 42 40 40 38 38 36 36 Micro GluStar System Micro GluStar System 34 34 32 32 30 30 28 28 26 26 24 24 22 22 24 26 28 30 32 34 36 38 40 42 44 24 26 28 30 32 34 36 38 40 42 Glutomatic 2200 0,95 Conf.Int. Martonvásár 0,95 Conf.Int.
  21. 21. New possibilities in micro-scale wheat quality characterisation Application in research activities: An example for investigation of gluten proteins Rudi Appels et al: Proteome bioinformatics and genetics for associating proteins with grain phenotype •A total of 740 flour samples from lines in a MAGIC population (4-way cross) established in CSIRO Plant Industry were analysed using the GluStar equipment. •Based on control samples (from cv Carnamah) analysed every 10 samples, the %wet gluten ranged from 1.295 - 1.405 g/4.5g flour and hence the reproducibility of the measurement for these 74 control samples was good. •The genetic variation assayed by GluStar for the set of samples from the structured MAGIC population, ranged from 0.95 g wet gluten/4.5g flour to 2.8g wet gluten/4.5 g flour and within this analysis the reproducibility was consistent with that defined by the control Carnamah samples. •A particularly useful aspect of the small scale analyses, in addition to the % wet gluten measurement, was the fast qualitative assessment of the dough formed by the flour samples.
  22. 22. New possibilities in micro-scale wheat quality characterisation Application in research activities: Investigation of the effect of amylose content on the rheological properties Comparison of viscosity (RVA) profiles of the flours and separated starch Comparison of amilosy content measured in flour blends and from their isolates Comparison of viscosity (RVA) profiles of starches separated from different wheat varieties
  23. 23. New possibilities in micro-scale wheat quality characterisation Application in research activities: Investigation the effect of non starchy carbohidrates on the rheological properties Water extractable (WEAX) and unextractable (WUAX) arabinoxilane content in the wheat flours and separated carbohydrates -WUAX remains in the filtrate of the effluent -WEAX can barely be measured in the isolates
  24. 24. New possibilities in micro-scale wheat quality characterisation Application in research activities: Relatinoship between carbohydrate composition and rheological parameters Correlation between carbohydrate constitution and RVA parameters of flours (21 Hungarian wheat varieties) Correlation between carbohydrate constitution and RVA parameters of isolates
  25. 25. New possibilities in micro-scale wheat quality characterisation Instead of summary … a completted picture of micro-scale tools
  26. 26. New possibilities in micro-scale wheat quality characterisation Acknowledgement Many thanks all of our mentioned and non-mentioned colleagues, students, technicians who took part in the mentioned projects. This work was and is supported by - Hungarian Scientific Research Found (OTKA, Projects No.: 80292 and 80334 ) - Hungarian National Project titled ‘Development of breeding, agricultural production and food industrial processing system of Pannon wheat varieties , (NTP, PojectTECH-09-A3-2009-0221) - National Research and Development Programs (NKFP, Project No: 04/35/2001) - Hungarian Economic Competitiveness Operative Programme (GAK, Project No.: ALAP1-00126/2004). - Hungarian Gábor Baross Innocsek Programme (KM-CSEKK-2005-000044) - CIMMYT
  27. 27. Thank you for your kind attention!
  28. 28. New possibilities in micro-scale wheat quality characterisation Instead of summary… The whole picture of wheat characterisation
  29. 29. New possibilities in micro-scale wheat quality characterisation Applications in research activities: measured relationships among Zeleny values and different functional parameters
  30. 30. Budapest University of Technology and Economics Invitation to the next MoniQA Conference….
  31. 31. New possibilities in micro-scale wheat quality characterisation Requirements for high throughput methods Fordí !!! tás
  32. 32. New possibilities in micro-scale wheat quality characterisationInstead of summary: the available micro-scale instruments and methods Kiegészí teni a sikérmosóval
  33. 33. New possibilities in micro-scale wheat quality characterisation Case 2: Sample preparation with micro-mill for micro-analytical methods Summary: Improved repeability for the determination of glutenin subunit composition with LOC
  34. 34. New possibilities in micro-scale wheat quality characterisation Bekes, F. and Gras, P.W. Demonstration of the 2 g Mixograph as a research tool. Cereal Chemistry 69 (1992) 229–230. 2 g mixograph (National/TMCO,USA)
  35. 35. New possibilities in micro-scale wheat quality characterisation Dobraszczyk, B.J. and Roberts C.A. Strain hardening and dough gas cell-wall failure in biaxial extension. J. Cereal Sci. 20:265, 1994. Dough rheological properties were measured in biaxial extension using a modified Chopin Alveograph under conditions close to those of baking expansion. The resulting stress-strain curves showed power-law strain hardening, indicating an increase in stiffness with increasing inflation. This strain hardening was shown to be critical in determining the limit of bubble expansion, both in the Alveograph and in baking. A criterion for gas cell-wall instability and failure has been derived, which takes into account the strain and strain-rate hardening properties of dough during large extensional deformation. Differences in bubble stability were observed between different wheat cultivars at low rates of inflation close to those of baking expansion, but these differences disappeared at the higher rates normally associated with rheological testing. TA.XTplus Texture Analyser Stable Micro System, UK
  36. 36. New possibilities in micro-scale wheat quality characterisation R. Kieffer, H. Wieser, M.H. Henderson, A. Graveland: Correlations of the Breadmaking Performance of Wheat Flour with Rheological Measurements on a Micro-scale(1998) Cereal Sci 27:53:60 For the characterization of wheat quality, micro-extension tests for dough and gluten and a micro-baking test were developed using comparable dough compositions, the same mixing temperature and cultivar-specific mixing times. By means of these methods, the flours of 26 wheat samples were studied for dough development time, maximum resistance and extensibility of dough and gluten and loaf volume of the baked products. Standard methods (rapid-mix-test, gluten index determination) were used for comparison. The results indicated that the rheological properties of dough and gluten as well as the gluten index are correlated higher with the optimised micro-baking test than with the standard baking test. If flour protein or wet gluten content is included in the correlations, the extension test of gluten, which can be performed easily and reproducibly, allows a reliable prediction of the loaf volume obtained by the micro-baking test. TA.XTplus Texture Analyser Stable Micro System, UK
  37. 37. New possibilities in micro-scale wheat quality characterisation Ide kellene ugyanaz, ami előbb első RVA cikk és abstract vagy eredmény, Feri tudnál segíteni, nem találom? Rapid Visco Analyser Perten Instruments
  38. 38. Budapest University of Technology and Economics Summary… -Functional data and their relationships to allelic composition indicated that while dough strength and partly extensibility are depend on the presence of HMW glutenin alleles, the allelic composition of LMW glutenins are mostly responsible for the variation of Zeleny indeces of the wheats, investigated. Among glutenin loci, significant QTLs were found in the regions of both type of gliadin loci on chromosome 1 and 6. -Infrared spectroscopy can be a useful tool for prediction the sedimentation values, but the results are highly depend on the calibration procedure - The Sedicom System® can be a valuable tool both for basic and applied research and also quality evaluation in the whole wheat verticum, including breeding, grain handling, milling and baking.
  39. 39. Budapest University of Technology and Economics
  40. 40. Budapest University of Technology and Economics Result 3: Relationship between different functional properties Summary: - Strong relationships can be identified in numerous cases between different quality parameters using our database. - However, the validity of these relationships cannot be generalized, mainly because the different sensitivity of alternative methodologies/parameters on different types of wheat. Marked correlations are significant at p<0,05 level, n=23
  41. 41. Budapest University of Technology and Economics But… Effects of location and treatment Comparison of location and agricultural treatment 80 70 60 Zeleny value (ml) 50 1.treatment 2.treatment 40 3.treatment 30 4.treatment 20 5.treatment 10 0 Which value belongs to a given variety 1 2 for evaluation of6 compositional treatment and location 3 4 5 Location 7 8 9 Comparison of agricultural background? 80 70 60 1.location Zeleny value (ml) 2. location 50 3. location 40 4.location 30 5.location 6. location 20 7. location 10 8. location 0 9. location 1 2 3 4 5 Agriculture treatment
  42. 42. But… Effects of treatment (Pannon-wheat quality program) Location Treatments• Complex qualification for Állampuszta Hungarian varieties Pécs- Reménypuszta •2 varieties (MV Suba and Balatonkeresztúr 100 +*100 kg N/ha GK Kalász), 9 agricultural Balatonkenese 100 + *50 kg N/ha areas, 5 agrotechnological Egyházashetye 100 kg N/ha treatment Csorvás 50 kg N/ha Törökszentmiklós 0 kg N/ha Jászboldogháza Szentistván
  43. 43. Budapest University of Technology and Economics Pictures from the beginning…
  44. 44. Budapest University of Technology and Economics Pictures from the beginning…
  45. 45. Budapest University of Technology and Economics Pictures from the beginning… Comparison of Farinograms and micro-Valorigrams
  46. 46. Budapest University of Technology and Economics Pictures from the beginning…
  47. 47. Budapest University of Technology and Economics Application fields and results of micro tests
  48. 48. Budapest University of Technology and Economics Method(s) 2: Mixing properties Summary: - Characteristics of curves registered with micro- and macro methods are similar. - Some alteration can be identified – for example in case of dough development time. - In most cases the correlations between macro and micro parameters are significant. - More or less similar dough systems are characterized with both types of methods and instruments. - We can learn more about the causes of identified differences.
  49. 49. Budapest University of Technology and Economics Method(s) 2: Mixing properties
  50. 50. Budapest University of Technology and Economics Case 2: Effect of Glu/Gli ratio on the mixing properties of wheat dough ERADU 400 [s] Dough development time control flour 64 Water Absorption 300 gliadin gluten 63 glutenin 200 62 100 61 0 60 Control +10% +20% Control +10% +20% flour flour
  51. 51. Budapest University of Technology and Economics Case 2: Effect of Glu/Gli ratio on the mixing properties of wheat dough Research on protein subunit composition Mixes of flour prepared from wheats with deletions of glutenin subunits genes show TM much better discrimination between poor and good wheats than the Mixograph , probably because the mechanism of dough development in the Z-arm mixer is not as dependant on dough strength as in pin mixers. 250 250 Z-arm mixer 2g Mixograph 1 , 17+18,5+10 - , 17+18,5+10 200 200 1 , - ,5+10 150 150 1 , 17+18, - 100 100 -, - ,5+10 - , 17+18, - 50 50 1, - , - 0 0 -, - , -
  52. 52. Budapest University of Technology and Economics Case 3: Effect of protein subunit incorporation on the mixing properties of rice dough Summary: • Application of micro Z-arm mixer is suitable for investigation of rice functionality • Incorporated glutenin subunits are able to alter dough mixing properties of rice flour (WA, DDT, stability). • Carefully controlled reduction/oxidation procedure can be applied on protein functionality studies in reconstituted rice dough.
  53. 53. Budapest University of Technology and Economics Method 1: Sample preparation with micro mill
  54. 54. Budapest University of Technology and Economics Method 1: Sample preparation with micro mill Effect on Mixing Time Effect on Peak Resistance 350 100 Comparison of milling yields: 400 BUHLER test and FQC-2000 300 75 Mixing Time (sec) 300 Peak Resistance 250 200 50 200 150 100 25 100 50 0 1 0 2 31 4 2 6 3 7 4 9 510 6 12 13 814 15 5 8 11 7 9 1610 11 17 18 12 13 14 15 16 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 17 18 Flour Samples sample number Flour Samples Effect on Resistance Breakdown Effect on Loaf Height 40 36 35 Resistance Breakdown 30 34 Loaf Height (mm) 33 20 32 31 10 30 29 0 28 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Flour Samples Flour Samples
  55. 55. Budapest University of Technology and Economics Standardization • ICC Standar dNo.116: Determination of the Sedimentation Value • AACC method 56-60, • AACC method 56-61A • AACC method 56-62, • AACC method 56-63, • AACC method 56-70 • Magyar szabvány- MSZ ISO 5529 Quality parameter for European Wheat Intervention System
  56. 56. Budapest University of Technology and Economics Measured relationships among Zeleny values and different functional parameters
  57. 57. Budapest University of Technology and Economics Case 4 - Molecular backcround: Relationship between allelic composition and sedimentation values selection from local 1891 1929 Noe (Volga region) variety selection from local 1919 1938 Smena (Siberia) variety selection from local 1919 1939 Lutescens 956 (Siberia) variety -Historical 10 Irtyshanka sets Skala/Saratovskaya 36 1964 1981 Lade (Norway) / FKN-25 1970 1984 Omskaya 12 (USA) Spontaneous hybrid 1986 ---- -Biodiversity (Canada)/Novosibirskaya Omskaya 21 22 Novosibirskaya 22 / 1986 1998 Omskaya 26 -Biofortification W.W.16151 (Sweden) Saratovskaya 29 / 1987 2000 Pamyaty Azyeva Lutescens 99-80-1 Lutescens 162-84-1 / 1989 2001 -Kazakhstan-Hungary-Australia-Turkey Omskaya 32 Chris (USA) cooperation Omsraya 20 / Lutescens 1992 2004 204-80-1//Lutescens 3-86- Kazanskaya yubileynaya 6 -See also the lecture titled „The effects of genetic and Biryusinka / Bezostaya 1 1970 * 1989 Tulunskaya 12 environmental factors on the 67 / Novosibirskaya quality of Kazakhstan and 1981 * 1995 Siberian wheat” by Dr. Alexey Morgounov Altayskaya 92 Lutescens 4029 Physical mutant from 1979 1997 Rosinka Sibakovskaya 3, Buryatscaya 34 / 1978 * 1994 Selenga Buryatscaya 79 OmSHI 6 / ANK 17 // 1985 2000 Shernyava 13 OmSHI 6
  58. 58. Budapest University of Technology and Economics Case 4 - Kazakh wheat varieties 2007.05.11 Contribution % Contribution % Input file: all.csv Glu1-A Glu1-B Glu1-D Glu3-A Glu3-B Glu3-D RMAX H L HH LL HL EXT H L HH LL HL Aktobe 32 B-001 c g a b c d 247 51,35 25,10 0,77 2,32 20,46 15 17,15 61,05 1,74 6,10 13,95 Stepnaya 1 B-004 b c a c g a 349 Contribution % 2,95 4,18 18 32,08 32,08 4,87 9,51 21,46 67,32 7,13 18,43 GVK 1369-2B-007 b c a RMAX j Hb e 368L 62,84 HH80 17,20 LL0,00 12,16HL 19 29,53 35,64 4,48 1,83 28,51 7, 247 51,35 25,10 0,77 2,32 20,46 Lutescens 53-95 b B-010 c d 349 b 67,32 4347,1379,30 18,43 14,54 2,95 3,74 4,18 27,43 31,13 3,31 4,28 33,85 c b 0,22 2,20 19 Astana B-013 b c a 368 g 62,84 3857,8064,47 17,20 17,65 0,00 12,7112,16 21,17 37,23 3,21 5,99 32,41 434 e b 79,30 0,22 4,71 14,54 0,47 2,20 23 3,74 Shortandinskaya 95 b B-016 c a 385 g 64,47 3854,7164,47 17,65 17,65 0,47 12,7112,71 21,17 37,23 3,21 5,99 32,41 e b 4,71 0,47 23 Contribution % Chelyaba B-019 a d a 385 j 64,47 3814,7146,46 17,65 24,93 0,47L24,6712,71 60,18 19,17LL12,09 3,2HL 5,31 381 c a 46,46 EXT 24,93 3,94 3,94 H 0,00 24,670,00 14HH 4 Lutescens 148-97-16 b B-022 c a 368 j 62,84 3687,8062,81517,20 17,20 0,00 12,1612,16 1,74 35,64 6,10 1,813,951 e b 4 7,80 17,15 0,0061,05 19 29,53 4,48 3 28,5 447 76,43 6,16 1814,01 32,08 2,55 32,08 0,85 4,87 9,51 21,46 Fora B-025 b c d 400 g 66,50 447 76,41918,20 14,01 1,46 0,85 3,64 4,48 36,25 1,83 10,728,510 c a 10,19 3 6,16 29,53 2,5535,64 16 35,25 4,25 5 13,5 Golubkovskaya b B-026 c a c g c 400 66,519 10,19 27,43 1,4631,13 18 3,31 32,58 4,28 7,633,855 0 18,20 3,64 32,58 4,94 4 22,2 23 21,17 37,23 3,21 5,99 32,41 Iren B-029 b u a c ? b 247 72,623 10,81 21,17 2,1037,23 17 3,21 20,80 5,99 2,632,417 7 12,31 2,10 32,39 5,44 0 38,7 Eritrospermum 78 a B-032 c d e b a 819 37,014 6,59 60,18 11,6019,17 2212,09 35,33 3,24 2,37 5,316 0 19,90 24,91 31,39 3,15 27,7 19 29,53 35,64 4,48 1,83 28,51 Akmola 2 D-001 b c a e g e 448 61,116 7,81 35,25 0,4536,25 22 4,25 37,2510,75 6,313,508 6 16,74 13,84 23,69 3,59 7 29,0 Lutescens 25/93-3-5 b D-002 c d c g a 447 76,418 6,16 32,58 2,5532,58 16 4,94 36,25 7,64 10,722,250 3 14,01 0,85 35,25 4,25 5 13,5 *Rmax and Ext was predicted with PQI
  59. 59. Budapest University of Technology and Economics Kazakh wheat varieties – Micro-Zeleny values
  60. 60. Budapest University of Technology and Economics Case 5 - Preliminary QTL analysis Histogram of zeleny phenotype 45 40 35 Frequency 30 25 Frequency 20 15 10 5 0 5 4 9 5 6 3. 3. 4. 5. • Most significant QTL for zeleny were detected on chromosomes not Zeleny units harbouring HMW Glutenin loci • Chrom 6A -LOD 6.1 (possibly Gliadin) • Chrom 5A -LOD 4.5 (protein gene?) • Chrom 3B –LOD 3.5
  61. 61. Budapest University of Technology and Economics Case 5 - Preliminary QTL analysis Zeleny szedimentációs teszt: szignifikáns QTL a LMW glutenin alegységek génjeinek környezetében Glu3B
  62. 62. Budapest University of Technology and Economics A tészta-jellemzők (erősség, nyújthatóság) szignifikáns Case 5 - Preliminary QTL analysis QTL-lel rendelkeznek a HMW glutenin alegységek génjeinek környezetében 1B Rmax EXT
  63. 63. Budapest University of Technology and Economics Case 6 - NIR-prediction for Zeleny based on Kazakh wheat varieties Correlation between Zeleny values obtained with micro- methods and NIR prediction using roboust calibration Micro-Zeleny values Low correlation coefficient… Reasons? NIR values
  64. 64. Budapest University of Technology and Economics Case 6 - NIR-prediction for Zeleny based on Kazakh wheat varieties Comparison of calibrations based on Scatterplot (Spreadsheet - AE_1+2 +.stw 15v*352c) North European (), (KAZ) = 9,0905+0,8157*x; 0,95 Conf.Int. () samples #1 2221 Hungarian () and Kazah referencia:2221 (KAZ): r2 = 0,8157; r = 0,9031; p = 0,0000; y = 9,0905 + 0,8157*x 90 Scatterplot (Spreadsheet - AE_1+2 +.stw 15v*352c) WH371523 WH612196 0011 (HUN) 2221 (HUN) 0011 (KAZ) 2221 (KAZ) 2 referencia:WH371523: r = 0,2495; r = 0,4995; p = 0,0010; y = 31,9414 + 0,4517*x 80 referencia:WH612196: r2 = 0,1445; r = 0,3801; p = 0,0155; y = 39,533 + 0,2166*x referencia:0011 (HUN): r2 = 0,0790; r = 0,2810; p = 0,0790; y = 56,3925 + 0,1952*x referencia:2221 (HUN): r2 = 0,2189; r = 0,4678; p = 0,0023; y = 49,4463 + 0,3765*x 70 referencia:0011 (KAZ): r2 = 0,5867; r = 0,7660; p = 0,000000008; y = 16,8756 + 0,6746*x referencia:2221 (KAZ): r2 = 0,8157; r = 0,9031; p = 0,0000; y = 9,0905 + 0,8157*x 100 60 90 2221 (KAZ) 80 50 70 40 60 50 30 40 30 Reference values are based on Kazakh variation 20 20 r2 = 0,8157 10 10 10 20 30 40 50 60 70 80 90 100 10 20 30 40 50 60 70 80 90 referencia referencia
  65. 65. Budapest University of Technology and Economics Case +1: Evaluation of NIR spectras for Hungarian and Kasakh wheat varieties Principal component analysis (PCA) of NIT spectra Hungarian samples (●) vs. Kazah samples – B (●), D (●), H (●), K (●) groups)

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