Food Chemistry Advances

Protein characteristics, starch properties and end-use quality of soft and hard wheat - a
comparative review
--Manuscript Draft--

Manuscript Number: FOCHA-D-23-00383R2

Article Type: Review Article

Keywords: Hard vs soft wheat; protein secondary structure; Polymeric vs monomeric proteins;
Rheology; Starch properties

Abstract: Hardness of wheat endosperm is a physical property that depends on the genetic
makeup of the grain and varies amongst different types/varieties. It is associated with
proteins, particularly 14-15 kDa friabilins, which exist in a higher amount at the surface
of starch in soft wheat grains while either absent or occur in smaller quantities on
granules of hard wheat varieties. The storage proteins in hard wheat flours exhibit
higher proportion of polymeric proteins, β-sheets and β-turns, while soft wheat starches
are characterized by a lower crystallinity, lesser amount of amylose-lipid complexes, a
particle size distribution with smaller proportion of A-type granules (disk or lenticular-
shaped granules with diameter >10 µm) and higher-gelatinisation temperatures,
swelling, breakdown susceptibility and retrogradation than hard wheat starches. Flours
from hard and soft wheat also differ for processing and end-use quality based on
differences in dough rheological properties, solvent retention capacities and particle
size.

Powered by Editorial Manager® and ProduXion Manager® from Aries Systems Corporation
Response to Reviewers

Response to reviewers’ comments

We thank the Reviewer for valuable comments/suggestions on our manuscript (MS).
As recommended, we have revised the MS. Changes made in the revised MS are
shown as red text and the point-wise responses to reviewer’s comments/suggestions
are tabulated below for kind consideration, please.

Sr. no. Reviewer’s comment/suggestion Response

Overall, the paper is well-executed, and it addresses
a pertinent topic in line with its intended purpose.
However, concerning the title "Soft and hard wheat:
differences from protein and starch perspective - a
concise review," it implies that the paper will
provide a concise yet comprehensive review of the
disparities between soft and hard wheats from the We have modified the title
1 protein and starch perspective. However, upon of the MS as per the
reviewing the citations, it is evident that only 70 recommendation.
references are included, suggesting that the review
may not be as exhaustive as implied by such an
ambitious title. It is recommended to consider
modifying the title to more accurately represent the
intended scope of the paper, avoiding overly
definitive statements.

With sincere thanks and regards.

Khetan Shevkani
Highlights (for review)

Highlights

 Wheat with different hardness differ for dough functionality and end-use suitability
 Hard wheats have high unextractable polymeric proteins and produce more elastic
dough
 Soft wheat starches show low crystallinity while higher swelling and retrogradation
REVISED Manuscript (text with changes Marked) Click here to view linked References

1
2
3
4
5 1 Protein characteristics, starch properties and end-use quality of soft and hard
6
7 2 wheat - a comparative review
8
9
10 3 Khetan Shevkani1, Mehak Katyal2, Narpinder Singh3
11
12
4
13
14
15 5 1 Department of Applied Agriculture, Central University of Punjab, Bathinda-151401, India
16
17 6 (shevkani@gmail.com; khetan.shevkani@cup.edu.in)
18
19
20 7 2 Department of Nutrition and Dietetics (Food Technology), Manav Rachna International Institute
21
22 8 of Research & Studies, Faridabad, India (mehak.katyal3@gmail.com)
23
24
25 9 3 Department of Food Science and Technology, Graphic Era Deemed to be University, Dehradun-
26
27 10 248002, India (narpinders@yahoo.com)
28
29
30 11
31
32 12 Corresponding authors: KS (shevkani@gmail.com)
33
34
13
35
36
37 14 Running title: Hard and soft wheat: a comparison
38
39 15
40
41
42 16 Abstract
43
44 17 Hardness of wheat endosperm is a physical property that depends on the genetic makeup of the
45
46
47 18 grain and varies amongst different types/varieties. It is associated with proteins, particularly 14-15
48
49 19 kDa friabilins, which exist in a higher amount at the surface of starch in soft wheat grains while
50
51
20 either absent or occur in smaller quantities on granules of hard wheat varieties. The storage
52
53
54 21 proteins in hard wheat flours exhibit higher proportion of polymeric proteins, β-sheets and β-turns,
55
56 22 while soft wheat starches are characterized by a lower crystallinity, lesser amount of amylose-lipid
57
58
59 23 complexes, a particle size distribution with smaller proportion of A-type granules (disk or
60 1
61
62
63
64
65
 1
2
3
4 24 lenticular-shaped granules with diameter >10 µm) and higher-gelatinisation temperatures,
5
6
7 25 swelling, breakdown susceptibility and retrogradation than hard wheat starches. Flours from hard
8
9 26 and soft wheat also differ for processing and end-use quality based on differences in dough
10
11
12 27 rheological properties, solvent retention capacities and particle size.
13
14 28
15
16
17 29 Keywords: Hard vs soft wheat; Protein secondary structure; Polymeric vs monomeric proteins;
18
19 30 Rheology; Starch properties
20
21
31
22
23
24 32 1. Introduction
25
26 33 Wheat is considered unique amongst cereals because of its ability to form a viscoelastic dough
27
28
29 34 when mixed with water. Wheat flour is used to prepare a number of palatable and satisfying
30
31 35 foods. It is grown and utilized in most parts of the world for food, feed and industrial purposes.
32
33
34 36 Thousands of cultivars/varieties of wheat varying in grain and flour properties are grown. They
35
36 37 have been categorized based on planting season (winter or spring), colour (white or red) and
37
38
38 endosperm characteristics (vitreous or mealy and soft or hard). However, wheat is primarily
39
40
41 39 classed on the basis of grain hardness/softness as it has strong association with the finished-
42
43 40 product quality (Shewry, 2023). Hard wheat grains, generally, yield strong flour with high
44
45
46 41 protein content which is suitable for bread making while soft wheat mill into flour with lesser
47
48 42 protein and damaged starch content suitable for manufacturing cookies, biscuits and pastries
49
50
51 43 (Quayson et al., 2016ab; Shang et al., 2020; Sharma et al., 2022).
52
53 44 Grain hardness is a genetic property of wheat and it depends on the degree of adhesion
54
55
56
45 between starch granules and protein matrix within the endosperm, the continuity of the protein
57
58 46 matrix and the absence of cracks/fissures. The adhesion between starch and proteins in the wheat
59
60 2
61
62
63
64
65
 1
2
3
4 47 endosperm is governed primarily by the level of specific membrane-bound proteins of 14-15 kDa
5
6
7 48 present on the surface of starch granules known as friabilins. These proteins can soften wheat
8
9 49 endosperm by hindering the adhesion between starch granules and protein matrix. Soft wheat
10
11
12 50 grains contain higher levels of friabilins than hard wheat while these proteins are absent in
13
14 51 durum varieties, considered the hardest type of wheat (Shevkani et al., 2017). Friabilins are made
15
16
17 52 up of two protein subunits, puroindoline-A and puroindoline-B, which are gene products of
18
19 53 alleles Pina-D1a and Pinb-D1a, respectively, and are located at the Ha locus on the short arm of
20
21 54 the 5D chromosome in soft wheats (Morris et al., 2021). Hard wheat varieties are characterized
22
23
24 55 by specific mutations in either Pina-D1 or Pinb-D1 genes or might lack these genes (Morris,
25
26 56 2021; Nadolska-Orczyk et al, 2009). The level of friabilins on starch granules also depends on
27
28
29 57 the level of polar lipids including glycol- and phospholipids which were essential for the binding
30
31 58 of friabilins to the granule surface in the endosperm (Greenblatt et al., 1995). Puroindolines can
32
33
34 59 bind with lipids on the surface of starch granules through a hydrophobic tryptophan-rich domain
35
36 60 (Clifton et al., 2007). A negative association between starch surface lipids and grain hardness has
37
38
61 been reported (Kim et al., 2012). Environment conditions also affect the hardness of wheat
39
40
41 62 grains. Singh et al. (2021) reported that heat stressed wheat had grains with higher hardness and
42
43 63 lower weight and diameter caused by lower starch synthesis.
44
45
46 64 The hardness of wheat grains is commonly measured as grain hardness index (GHI) using
47
48 65 a Single Kernel Characterization System (SKCS) of Perten Instruments (Australia). This
49
50
51 66 instrument records the force required to crush a grain between a toothed and a narrowing
52
53 67 crescent gap. GHI is an indicator of the resistance of the grain to fracture. It also translates the
54
55
56
68 ability to produce flour of variable quality as hard and soft wheat varieties differ widely for their
57
58 69 protein content/composition, starch characteristics and dough rheology (Issarny et al., 2017;
59
60 3
61
62
63
64
65
 1
2
3
4 70 Katyal et al., 2017; 2019; Shang et al., 2020; Sharma et al., 2022). In recent years, hard and soft
5
6
7 71 wheat varieties have been investigated to study the basis of differences in their properties and
8
9 72 applications (Cao et al., 2017; Issarny et al., 2017; Katyal et al., 2017; 2019; Quayson et al.,
10
11
12 73 2016ab; Sharma et al., 2022; Shang et al., 2020; Singh et al., 2017; 2019; Tao et al., 2018).
13
14 74 However, a review comparing hard and soft wheat from the perspective of starch and protein
15
16
17 75 properties is not available. Therefore, this article attempted to provide a concise overview of the
18
19 76 differences in hard and soft wheat varieties in the context of the structure and functionality of
20
21 77 proteins and starches.
22
23
24 78
25
26 79 2. Protein characteristics
27
28
29 80 Gliadins and glutenins are major storage proteins in wheat endosperm, constituting up to 85% of
30
31 81 total proteins (Guo et al., 2021). Glutenins refer to proteins soluble in dilute alkali or acidic
32
33
34 82 solutions while gliadins include aqueous alcohol soluble proteins. Figure 1 depicts the typical size
35
36 83 exclusion chromatographic peaks of wheat flour glutenins and gliadins. These proteins are
37
38
84 responsible for the typical viscoelastic behaviour (characterised as the presence of both elasticity
39
40
41 85 and extensibility) of wheat flour dough which is pivotal in determining the end-use quality of
42
43 86 wheat varieties. Elasticity in dough results from the formation of a strong three-dimensional gluten
44
45
46 87 network desirable for making a good quality bread while extensibility/fluidity is due to a mellow
47
48 88 network formation suitable for the preparation of cookies, cakes and wheat tortillas (Dizlek et al.,
49
50
51 89 2022; Mondal et al., 2009).
52
53 90 Amongst wheat proteins, glutenins are polymeric proteins and they play a crucial role in
54
55
56
91 gluten strength and the suitability of wheat flour for bread making. The hydration of flour proteins
57
58 92 during dough kneading results in the formation of a three-dimensional gluten network. This
59
60 4
61
62
63
64
65
 1
2
3
4 93 involves the polymerization of glutenins through interchain disulphide linkages (Shewry and
5
6
7 94 Lafiandra, 2022) and the aggregation of the polymeric glutenin molecules with gliadins via
8
9 95 hydrogen bonds, hydrophobic and electrostatic interactions (Morel et al., 2020). Although both
10
11
12 96 these phenomena contribute to stabilise the gluten network and dough formation, glutenins endow
13
14 97 the dough with the strength/elasticity needed for good quality bread making owing to the ability to
15
16
17 98 form intermolecular covalent disulphide bonds, whereas the presence of gliadins promotes
18
19 99 extensibility by weakening the gluten network after being incorporated into the network structure
20
21 100 (Schopf et al., 2021). Shewry (2023) showed that dough extensibility results from the slippage of
22
23
24 101 gliadins and glutenins linked non-covalently. Therefore, hard wheat flour with higher glutenin
25
26 102 content than soft wheat flour (Figure 1) generally forms more elastic dough exhibiting stronger
27
28
29 103 gluten network required for bread-making.
30
31 104 Based on the differences in molecular weight, glutenins can be categorised as low and
32
33
34 105 high-molecular-weight glutenin subunits (LMW-GS and HMW-GS, respectively) held together by
35
36 106 intra and inter-chain disulphide bonds (Figure 2). HMW-GS are associated with improved bread-
37
38
107 making properties while flours with lower levels of HMW-GS could be useful for making
39
40
41 108 products where a relatively lower gluten strength is required. Hard and soft wheat differ for the
42
43 109 relative levels of LMW-GS and HMW-GS fractions (Figure 2). The glutenins from hard and soft
44
45
46 110 wheat varieties showed the presence of 19 significant polypeptides (PPs) ranging in MW between
47
48 111 26 kDa and 98 kDa which were categorized as HMW-GS (98-79 kDa), LMW-GS-B (49-40 kDa)
49
50
51 112 and LMW-GS-C (35-26 kDa). Hard wheat varieties with GHI of 97 to 100 showed specific
52
53 113 HMW-GS composition with 91, 80, 78, and 74 kDa PPs, which was not the property of soft wheat
54
55
56
114 flours (Katyal et al., 2017). In addition, hard wheat flours exhibited glutenins with GluD1 locus
57
58
59
60 5
61
62
63
64
65
 1
2
3
4 115 with 5+10 subunits whereas soft wheat HMW-GS with allelic composition of 2*, 7 and 2+12 were
5
6
7 116 linked with low dough stability and improved cookie-making properties (Sharma et al., 2022).
8
9 117 Wheat flour proteins have also been evaluated as the proportion of unextractable and
10
11
12 118 extractable polymeric and monomeric proteins (UnEx-PP, Ex-PP, UnEx-MP and Ex-MP,
13
14 119 respectively) based on their extractability in sodium dodecyl sulphate buffer. In addition to the
15
16
17 120 ratio of gliadin/glutenin, the quantity of both extractable and unextractable proteins affects dough
18
19 121 strength and end-use quality. Loaf volume and dough/gluten strength correlated positively with the
20
21 122 amount of glutenins and negatively with gliadins (Tsilo et al., 2010), while polymeric to
22
23
24 123 monomeric protein ratio was associated with superior baking qualities (Labuschagne et al., 2004).
25
26 124 Hard and soft wheat also differ for the proportion of polymeric and monomeric proteins. Flours
27
28
29 125 from harder wheat varieties contained polymeric proteins (both extractable and unextractable) in
30
31 126 higher proportions than soft wheat (Baasandorj et al., 2016; Katyal et al., 2017). The mean
32
33
34 127 proportion of Ex-PP and UnEx-PP for flours from hard and medium hard wheat varieties was 33-
35
36 128 46% and 50-62%, respectively, against 27% and 42%, respectively, for soft wheat flours, whereas
37
38
129 soft wheat flours contained monomeric proteins in higher proportions than hard wheat flours
39
40
41 130 (Table 1).
42
43 131 The secondary structure of gluten proteins is also related to dough functionality and it has
44
45
46 132 also been linked with grain hardness. The elasticity of gluten proteins was attributed to β-spiral
47
48 133 structures in HMW-GS which were formed of regularly repeated β-turn structures (Shewry and
49
50
51 134 Halford, 2002; Tatham et al., 2001). Belton (2005) showed the formation of hydrogen-bonded
52
53 135 inter-chain β-sheet structures in gluten proteins by the orientation of β-turns in adjacent β-spirals
54
55
56
136 after hydration. The dough with a higher proportion of β-sheets and lower α-helix exhibited higher
57
58 137 gluten strength (Kaur et al., 2014). The secondary structure of gluten/dough can be evaluated
59
60 6
61
62
63
64
65
 1
2
3
4 138 qualitatively and quantitatively using Fourier transform infrared (FTIR) spectroscopy which is a
5
6
7 139 vibrational spectroscopic analytical technique useful for studying the structure of proteins under
8
9 140 diverse environments. Amongst different regions observed when proteins are studied using FTIR,
10
11
12 141 amide I and II regions (attributing to stretching of C=O bond and bending vibrations at N-H bond
13
14 142 in infrared spectra) are very sensitive to protein secondary structure (Shevkani et al., 2019). The
15
16
17 143 analysis of wheat proteins using FTIR generally reveals the presence of three predominant bands
18
19 144 in the amide-I region at wavenumbers between ≈1660 and ≈1690 cm-1 while two bands in the
20
21 145 amide-II region at ≈1570 and ≈1580 cm-1. The amide-I bands at around 1629-1632 cm-1, 1650-
22
23
24 146 1660 cm-1, 1665-1670 cm-1 and 1687-1690 cm-1 were ascribed to antiparallel-β-sheets, α-helical
25
26 147 structures, β-turns and β-sheets, respectively (Wang et al., 2014). The proteins from hard and soft
27
28
29 148 wheat differed for their secondary structure. Hard wheat proteins contained a lesser proportion of
30
31 149 α-helices and higher β-sheets and β-turns than soft wheat flours (Jazaeri et al., 2015; Quayson et
32
33
34 150 al., 2016a). Katyal et al. (2017) also reported that the dough from soft wheat flours had higher
35
36 151 proportion of intermolecular-β-sheets than medium-hard and hard wheat varieties. It has also been
37
38
39
152 reported that β-sheets are formed in soft wheat dough by hydrophobic interactions while in hard
40
41 153 wheat dough, disulfide bonds stabilise the secondary structure (Jazaeri et al. 2015).
42
43 154
44
45
46 155 3. Starch properties
47
48 156 Starch is the most abundant storage carbohydrate in wheat, comprising about 60-75% of grain and
49
50
51 157 70-80% of flour. The content and characteristics of starch in flour affect the quality of wheat-based
52
53 158 products (Cao et al., 2019; Shang et al., 2021). The pattern of breakage of wheat during milling
54
55
56
159 affects the amount of damaged starch in flour. Hard wheat varieties yield flours with higher
57
58 160 damaged starch content having greater affinity to water than intact granules. Li et al. (2013)
59
60 7
61
62
63
64
65
 1
2
3
4 161 attributed lower water absorption of soft wheat flour to higher content of intact starch granules
5
6
7 162 (lesser damaged granules) together with lower protein content. Similarly, the higher water
8
9 163 absorption capacity of hard wheat flour was ascribed to higher content of damaged starch
10
11
12 164 (Baasandorj et al., 2020; Katyal et al., 2019; Singh et al., 2019). Shewry (2023) reported that
13
14 165 damaged starch can absorb 3-4 times its own weight of water in comparison to ≈ 0.5 times for
15
16
17 166 intact starch granules.
18
19 167 Hard and soft wheat varieties also differ for amylose content, morphological properties,
20
21 168 crystallinity, gelatinisation properties and pasting properties of their starches. Starches from soft
22
23
24 169 wheat were less crystalline and contained a lower proportion of ‘A’ granules (disk or lenticular
25
26 170 shaped granules with diameter > 10 µm) while higher of ‘B’ granules (spherical granules with
27
28
29 171 diameters < 10 µm) than starch of hard wheat varieties (Katyal et al., 2019; Singh et al., 2017;
30
31 172 Shang et al., 2020). In general, higher gelatinisation temperatures and enthalpy of starches are
32
33
34 173 attributed to a higher degree of crystallinity (associated with lower content of amorphous amylose
35
36 174 and a greater proportion of long amylopectin chains) which provides structural stability to
37
38
175 granules and makes them more resistant towards gelatinisation (Shevkani et al., 2017). However,
39
40
41 176 soft wheat starches, despite lower crystallinity, showed higher gelatinisation temperatures and
42
43 177 enthalpy (mean onset-, peak-, endset-temperature and enthalpy = 59.96-61.17 °C, 63.25-63.84 °C,
44
45
46 178 66.67-66.81 °C and 7.55-9.22 J/g respectively) than medium-hard (58.15-60.96 °C, 62.08-63.58
47
48 179 °C, 66.09-66.46 °C and 7.05-8.45 J/g, respectively) and hard varieties (57.55-60.27 °C, 61.72-
49
50
51 180 63.08 °C, 65.85-66.12 °C and 6.84-8.31 J/g, respectively) (Katyal et al., 2019; Singh et al., 2017).
52
53 181 Such behaviour of soft wheat starches was likely due to higher puroindolines content and lesser
54
55
56
182 damage to the granules during milling which made them more resistant to gelatinise by inhibiting
57
58
59
60 8
61
62
63
64
65
 1
2
3
4 183 the availability of water (Brites et al., 2008; Quayson et al., 2016b) and suppressing starch
5
6
7 184 hydration and dispersion in water (Han et al., 2022; Leon et al., 2006), respectively.
8
9 185 However, varying results were observed for amylose content in starches of hard and soft
10
11
12 186 wheat grains. Singh et al. (2017), Katyal et al. (2019) and Shang et al. (2020) reported lower
13
14 187 amylose in starch from softer wheat varieties (mean amylose content = 12.1-22.8%) than harder
15
16
17 188 varieties (mean amylose content = 15.0-25.2%). In contrast, Kumar et al. (2016) observed higher
18
19 189 amylose content for starch from soft wheat (25.78%) than hard wheat varieties (24.11%), whereas
20
21 190 Biduski et al. (2022) reported no difference in amylose content of starch from soft (25.62%) and
22
23
24 191 hard wheat (25.53%), but for medium hard wheat (23.3%).
25
26 192 Pasting properties represent changes in the viscosity of starch suspensions during and after
27
28
29 193 gelatinisation. Pasting properties are considered an important index of starch functionality and
30
31 194 depend not only on starch morphology and structure but also on the state of granules
32
33
34 195 (damaged/intact) and the presence/absence of non-starch constituents (Han et al., 2022; Singh et
35
36 196 al., 2014; Shevkani et al., 2021). Starches from soft wheat showed higher peak viscosity
37
38
197 (maximum viscosity attained during heating representing the point of maximum swelling of
39
40
41 198 granules), breakdown viscosity (the fall in viscosity during heating and shearing representing the
42
43 199 susceptibility of swollen granules to rupture at high temperatures), final and setback viscosity
44
45
46 200 (increase in the viscosity on cooling due to retrogradation) than hard wheat starches (Katyal et al.,
47
48 201 2019; Shang et al., 2020). These results were possibly due to the presence of lower levels of
49
50
51 202 amylose-lipid complexes, starch-associated proteins and damaged granules in soft wheat starches,
52
53 203 which modified the behaviour of starch suspensions during pasting by affecting the potential of
54
55
56
204 granules to swell and disintegrate during heating and shearing. The swelling of granules during
57
58 205 heating in water has been shown to occur more quickly after the exudation of amylose (Tester &
59
60 9
61
62
63
64
65
 1
2
3
4 206 Karkalas, 2001) while amylose-lipid complexes and proteins suppress granular swelling and
5
6
7 207 subsequent disintegration by increasing granule integrity/rigidity and preventing amylose leaching
8
9 208 (Shang et al., 2020; Shevkani et al., 2011). Mechanical damage to starch, in contrast, could
10
11
12 209 promote the formation of less viscous paste by distorting granules, damaging their ordered
13
14 210 structure, reducing crystallinity and facilitating water absorption leading to greater disintegration
15
16
17 211 of the gelatinized starch and the production of more leachate materials (Barrera et al., 2013; Han et
18
19 212 al., 2022; Wu et al., 2018).
20
21 213 Differences in retrogradation tendencies were also reported for starches from soft and hard
22
23
24 214 wheat varieties (Shang et al., 2020). Soft starch gels showed higher retrogradation than starches
25
26 215 from hard wheat varieties. Gels of soft wheat starches exhibited higher syneresis (separation of
27
28
29 216 water from starch gels during freezing-thawing cycles due to the reassociation of dissociated
30
31 217 starch chains during cooling/storage of gels), gel hardness and fracturability in comparison to
32
33
34 218 starches from hard wheat varieties. These phenomena were linked with the presence of more
35
36 219 granular remnants, higher amylose content and lower levels of proteins and lipids associated with
37
38
220 starches. Amylose has been shown to enhance the rate of retrogradation while the presence of
39
40
41 221 proteins and amylose-lipid complexes could reduce retrogradation by making interactions with
42
43 222 starch chains (Shevkani et al., 2017; 2021). However, more work is required to be conducted on
44
45
46 223 the retrogradation tendencies of hard and soft wheat starches as only limited information is
47
48 224 available in the literature.
49
50
51 225
52
53 226 4. Finished product suitability
54
55
56
227 Wheat is consumed in almost all countries of the world. It is typically milled into flour and then
57
58 228 used as a fundamental ingredient for preparing a wide range of products. Flours from different
59
60 10
61
62
63
64
65
 1
2
3
4 229 wheat varieties differ for their physicochemical and functional characteristics, hence for finished
5
6
7 230 product suitability. Usually, the suitability of wheat flour for producing different types of products
8
9 231 is assessed by evaluating dough rheology, solvent retention capacity (SRC) and flour particle size.
10
11
12 232 The rheological properties of wheat flour dough determine baking performance,
13
14 233 machinability and processing quality (Ktenioudaki et al., 2010). Farinograph and dynamic
15
16
17 234 rheometer are commonly used to determine the suitability of wheat flour for different baked
18
19 235 foods. Farinograph is a widely employed empirical rheological measurement technique, whereas
20
21 236 dynamic rheometers are used to measure the fundamental rheological aspects of dough.
22
23
24 237 Empirical rheological techniques are more common than dynamic rheometry, and they record the
25
26 238 changes in dough consistency during mixing while making use of large deformations. The most
27
28
29 239 useful parameters obtained from the farinographic analysis include water absorption (WA; a
30
31 240 measure of the amount of water required for the formation of an optimally viscoelastic dough),
32
33
34 241 dough development time (DDT; an indicative of optimum mixing/kneading time to have a well-
35
36 242 developed dough), dough stability (DS; an indicator of tolerance of dough towards mixing) and
37
38
243 degree of softening (DOS; decrease in the consistency of dough with extended mixing). Flours
39
40
41 244 with higher WA are generally desirable for bread making. A flour exhibiting high WA along
42
43 245 with high DS and low DOS results in the production of bread with desirable quality attributes
44
45
46 246 (high volume, soft and porous crumb, springy texture, etc.); however, a high WA combined with
47
48 247 a high DOS indicates poor quality flour. Hard wheat flours generally show higher WA, DDT and
49
50
51 248 DS in comparison to flours from soft wheat varieties (Table 1; Figure 3). Higher values of DDT
52
53 249 and DS for hard wheat were associated with a stronger gluten network and higher protein content
54
55
56
250 as compared to soft wheat (Tian et al., 2007). The DOS has been shown to vary widely between
57
58
59
60 11
61
62
63
64
65
 1
2
3
4 251 14 and 136 BU amongst different wheat varieties, while soft varieties were generally
5
6
7 252 characterized by exceptionally high DOS (Katyal et al., 2017).
8
9 253 Dynamic rheometer, in comparison, evaluates elastic and viscous properties of dough
10
11
12 254 which are expressed as storage modulus (G′), loss modulus (G″) and loss tangent (tanδ = G″/G′).
13
14 255 The elastic dough with greater DS exhibits higher G' and G" and lower tanδ values (Singh et al.
15
16
17 256 2011). Kaur et al. (2013) observed positive relations of both G′ and G″ with protein content.
18
19 257 Hard wheat varieties showed higher G' and G″ than soft wheat varieties, indicating a more elastic
20
21 258 nature of hard wheat flour dough. Both G' and G'' also related positively to DS and DDT. Wheat
22
23
24 259 varieties with HMW-GS of 91, 80, 78 and 74 kDa had lower tanδ (higher elasticity) than those
25
26 260 which did not show these proteins (Katyal et al., 2017).
27
28
29 261 Solvent retention capacity is another useful method to determine end-use quality of wheat
30
31 262 flours. Lu and Seetharaman (2014) and Hammed et al. (2015) reported relation of SRC values
32
33
34 263 with WA, DS, protein content, gluten content and damaged starch content. SRC quantifies the
35
36 264 swelling of gluten networks and the ability to retain various solvents such as water, sucrose, lactic
37
38
265 acid and sodium carbonate (Kweon et al., 2011). Sodium carbonate SRC (NaSRC) was related
39
40
41 266 with the damaged starch content while sucrose SRC (SuSRC) indicated gliadins, pentosan and
42
43 267 arabinoxylans content. Lactic acid SRC (LASRC) was an indicator of gluten strength while water
44
45
46 268 SRC (WSRC) indicated general water retention capacity of various flour constituents. Cao et al.
47
48 269 (2017) observed higher NaSRC for hard wheat (73.6-90.1%) than soft wheat flours (66.3-81.6%).
49
50
51 270 LASRC was related to gluten strength parameters and bread volume (Guttieri et al., 2001; Xiao et
52
53 271 al., 2006). Soft wheat varieties showed lower SuSRC associated with lower arabinoxylans content,
54
55
56
272 while LASRC positively correlated with UnEx-PP reflecting the contribution of glutenins to
57
58
59
60 12
61
62
63
64
65
 1
2
3
4 273 swelling and gluten network formation. Hard wheat flours were characterized by greater LASRC,
5
6
7 274 WSRC, SuSRC and NaSRC than soft wheat flour (Issarny et al., 2017).
8
9 275 Particle size distribution also has a pronounced effect on the baking quality and
10
11
12 276 functionality of wheat flours. In addition, the particle size of flour also influences the glycemic
13
14 277 index, fermentation and bioavailability of nutrients (Hallfrisch & Behall, 2000). Flours from soft
15
16
17 278 and hard wheat vary for particle size distribution. The flour particle size distribution was
18
19 279 correlated with GHI (Devaux et al., 1998). Wheat with higher GHI produced flour with a lesser
20
21 280 proportion of small-size particles and a greater of large-size particles; however, soft wheat
22
23
24 281 varieties showed the opposite results (Kaur et al., 2014; Katyal et al., 2016; 2018). Soft wheat with
25
26 282 higher proportion of fine particles showed lower-DDT, DS, gluten strength and SRC, and
27
28
29 283 produced cookies with greater spread (Kaur et al., 2014; Singh et al., 2019; Sharma et al., 2022).
30
31 284
32
33
34 285 Conclusion
35
36 286 Hard and soft wheat differ for milling and functional characteristics. Hard wheat mill into flours
37
38
287 with a higher proportion of coarse particles, damaged starch content and protein content as
39
40
41 288 compared to soft wheat varieties. Proteins in hard wheat flours yield dough with high gluten
42
43 289 strength attributing to the higher proportion of polymeric glutenins, β-sheets and β-turns than soft
44
45
46 290 wheat flours. Generally, hard wheat flours exhibit strong gluten attributed to high protein content,
47
48 291 HMW-GS and UnEx-PP; however, the differences due to genetics, environmental conditions,
49
50
51 292 duration of crop and cultural practices affect the ultimate suitability of flour for different products.
52
53 293 Soft wheat starches are characterized by a lower proportion of A-type granules (disk or lenticular-
54
55
56
294 shaped granules with diameter > 10 µm), a lower crystallinity, higher gelatinisation temperature,
57
58 295 more swelling, greater breakdown susceptibility and increased retrogradation tendencies than
59
60 13
61
62
63
64
65
 1
2
3
4 296 starches from hard wheat varieties. The lower proportion of damaged granules, amylose-lipid
5
6
7 297 complexes and granule-associated proteins in soft wheat starches contribute to higher swelling,
8
9 298 breakdown and retrogradation. However, more work is required on the comparison of hard and
10
11
12 299 soft wheat starches for amylose and amylopectin fine structure (chain length distribution) owing to
13
14 300 the association of these attributes with the functionality of starches.
15
16
17 301
18
19 302 Acknowledgement
20
21 303 NS acknowledges financial support from CSIR as research project grant and SERB as JC Bose
22
23
24 304 Fellowship grant.
25
26 305
27
28
29 306 Conflict of Interest
30
31 307 The authors have declared no conflict of interest.
32
33
34 308
35
36 309 References
37
38
310 Baasandorj, T., Ohm, J. B., & Simsek, S. (2016). Effects of kernel vitreousness and protein level
39
40 311 on protein molecular weight distribution, milling quality, and breadmaking quality in
41 312 hard red spring wheat. Cereal Chemistry, 93, 426-434.
42
43 313 Baasandorj, T., Ohm, J.B., & Simsek, S. (2020). Comparison of different experimental
44
45 314 breadmaking methods and their associations with flour quality parameters in hard red
46 315 spring wheat. Cereal Chemistry, 97(2), 515-526.
47
48 316 Barrera, G.N., Bustos, M.C., Iturriaga, L., Flores, S.K., León, A.E., & Ribotta, P.D. (2013).
49 317 Effect of damaged starch on the rheological properties of wheat starch suspensions.
50
51 318 Journal of Food Engineering, 116, 233-239.
52
53 319 Belton, P.S. (2005). New approaches to study the molecular basis of the mechanical properties of
54 320 gluten. Journal of Cereal Science,41, 203-211.
55
56 321 Biduski, B., Werlang, S., Colussi, R., Pinto, V. Z., Zavareze, E. D. R., Gutkoski, L. C., &
57 322 Bertolin, T. E. (2022). Starches Properties from Soft, Medium‐ Hard, and Hard Brazilian
58
59 323 Wheat upon Annealing. Starch‐ Stärke, 74(9-10), 2100267.
60 14
61
62
63
64
65
 1
2
3
4 324 Brites, C.M., Santos, C.A.L.D., Bagulho, A.S., & Beirão-da-Costa, M.L. (2008). Effect of wheat
5
6 325 puroindoline alleles on functional properties of starch. European Food Research and
7 326 Technology, 226, 1205-1212.
8
9 327 Cao, W., Falk, D., & Bock, J.E. (2017). Protein structural features in winter wheat:
10
11
328 Benchmarking diversity in Ontario hard and soft winter wheat. Cereal Chemistry, 94, 199-
12 329 206.
13
14 330 Cao, X., Tong, J., Ding, M., Wang, K., Wang, L., Cheng, D., Li, H., Liu, A., Liu, J., Zhao, Z.,
15 331 Wamg, Z, & Gao, X. (2019). Physicochemical properties of starch in relation to
16
17 332 rheological properties of wheat dough (Triticum aestivum L.). Food Chemistry, 297,
18 333 125000.
19
20 334 Clifton, L.A., Lad, M.D., Green, R.J., & Frazier, R.A. (2007). Single amino acid substitutions in
21 335 puroindoline-b mutants influence lipid binding properties. Biochemistry, 46, 2260-2266.
22
23 336 Delcour, J.A., Joye, I.J., Parey, B., Wilderjans, E., Brijs, K., & Lagrain, B. (2012). Wheat gluten
24
25 337 functionality as a quality determinant in cereal-based food products. Annual Review of
26 338 Food Science and Technology, 3, 469-492.
27
28 339 Devaux, M.F., de Monredon, F.L.D., Guibert, D., Novales, B., & Abecassis, J. (1998). Particle
29
340 size distribution of break, sizing and middling wheat flours by laser diffraction. Journal
30
31 341 of the Science of Food and Agriculture, 78, 237-244.
32
33 342 Dizlek, H., Girard, A.L., & Awika, J.M. (2022). High protein and gliadin content improves
34 343 tortilla quality of a weak gluten wheat. LWT-Food Science and Technology, 160, 113320.
35
36 344 Greenblatt, G.A., Bettge, A.D., & Morris, C.F. (1995). Relationship between endosperm texture
37
38
345 and the occurrence of friabilin and bound polar lipids on wheat-starch. Cereal Chemistry,
39 346 72, 172-176.
40
41 347 Gupta, R.B., Khan, K., & Macritchie, F. (1993). Biochemical basis of flour properties in Bread
42 348 wheats. 1. Effects of variation in the quantity and size distribution of polymeric protein.
43
44 349 Journal of Cereal Science, 18, 23-44.
45
46 350 Guo, J., Wang, F., Zhang, Z., Wu, D., & Bao, J. (2021). Characterization of gluten proteins in
47 351 different parts of wheat grain and their effects on the textural quality of steamed bread.
48 352 Journal of Cereal Science, 102, 103368.
49
50 353 Guttieri, M.J., Bowen, D., Gannon, D., O'Brien, K., & Souza, E. (2001). Solvent retention
51
52 354 capacities of irrigated soft white spring wheat flours. Crop Science, 41, 1054-1061.
53
54 355 Hallfrisch, J., & Behall, K.M. (2000). Mechanisms of the effects of grains on insulin and glucose
55 356 responses. Journal of the American College of Nutrition, 19, 320S-325S.
56
57
58
59
60 15
61
62
63
64
65
 1
2
3
4 357 Hammed, A.M., Ozsisli, B., Ohm, J.B., & Simsek, S. (2015). Relationship between solvent
5
6 358 retention capacity and protein molecular weight distribution, quality characteristics, and
7 359 breadmaking functionality of hard red spring wheat flour. Cereal Chemistry, 92, 466-474.
8
9 360 Han, N., Fan, J.L., Chen, N., & Chen, H.Q. (2022). Effect of ball milling treatment on the
10
11
361 structural, physicochemical and digestive properties of wheat starch, A-and B-type starch
12 362 granules. Journal of Cereal Science, 104, 103439.
13
14 363 Issarny, C., Cao, W., Falk, D., Seetharaman, K., & Bock, J.E. (2017). Exploring functionality of
15 364 hard and soft wheat flour blends for improved end‐ use quality prediction. Cereal
16
17 365 Chemistry, 94, 723-732.
18
19 366 Jazaeri, S., Bock, J.E., Bagagli, M.P., Iametti, S., Bonomi, F., & Seetharaman, K. (2015).
20 367 Structural modifications of gluten proteins in strong and weak wheat dough during
21 368 mixing. Cereal Chemistry, 92, 105-113.
22
23 369 Katyal, M., Virdi, A.S., Kaur, A., Singh, N., Kaur, S., Ahlawat, A.K., & Singh, A.M. (2016).
24
25 370 Diversity in quality traits amongst Indian wheat varieties I: flour and protein
26 371 characteristics. Food Chemistry, 194, 337-344.
27
28 372 Katyal, M., Singh, N., Virdi, A.S., Kaur, A., Chopra, N., Ahlawat, A.K., & Singh, A.M. (2017).
29
373 Extraordinarily soft, medium-hard and hard Indian wheat varieties: composition, protein
30
31 374 profile, dough and baking properties. Food Research International, 100, 306-317.
32
33 375 Katyal, M., Virdi, A.S., Singh, N., Chopra, N., Kaur, A., Ahlawat, A.K., & Singh, A.M. (2018).
34 376 Fractionation and grain hardness effect on protein profiling, pasting and rheological
35
36 377 properties of flours from medium-hard and extraordinarily soft wheat varieties. Journal
37 378 of Food Science and Technology, 55, 4661-4674.
38
39 379 Katyal, M., Singh, N., Chopra, N., & Kaur, A. (2019). Hard, medium-hard and extraordinarily
40 380 soft wheat varieties: Comparison and relationship between various starch properties.
41
42 381 International Journal of Biological Macromolecules, 123, 1143-1149.
43
44 382 Kaur, A., Shevkani, K., Katyal, M., Singh, N., Ahlawat, A.K. & Singh, A.M. (2016).
45 383 Physicochemical and rheological properties of starch and flour from diff erent durum
46
384 wheat varieties and their relationships with noodle quality. Journal of Food Science and
47
48 385 Technology, 53, 2127-2138
49
50 386 Kaur, A., Singh, N., Ahlawat, A.K., Kaur, S., Singh, A.M., Chauhan, H., & Singh, G.P. (2013).
51 387 Diversity in grain, flour, dough and gluten properties amongst Indian wheat cultivars
52
53 388 varying in high molecular weight subunits (HMW-GS). Food Research International, 53,
54 389 63-72.
55
56 390 Kaur, A., Singh, N., Kaur, S., Ahlawat, A.K., & Singh, A.M. (2014). Relationships of flour
57 391 solvent retention capacity, secondary structure and rheological properties with the cookie
58
59 392 making characteristics of wheat cultivars. Food Chemistry, 158, 48-55.
60 16
61
62
63
64
65
 1
2
3
4 393 Kim, K.H., Feiz, L., Martin, J.M., & Giroux, M.J. (2012). Puroindolines are associated with
5
6 394 decreased polar lipid breakdown during wheat seed development. Journal of Cereal
7 395 Science, 56, 142-146.
8
9 396 Ktenioudaki, A., Butler, F., & Gallagher, E. (2010). Rheological properties and baking quality of
10
11
397 wheat varieties from various geographical regions. Journal of Cereal Science, 51, 402-
12 398 408.
13
14 399 Kumar, R., Kumar, A., Sharma, N.K., Kaur, N., Chunduri, V., Chawla, M., Sharma, S., Singh,
15 400 K., & Garg, M. (2016). Soft and hard textured wheat differ in starch properties as
16
17 401 indicated by trimodal distribution, morphology, thermal and crystalline properties.
18 402 PloSone, 11, e0147622.
19
20 403 Kweon, M., Slade, L., & Levine, H. (2011). Solvent retention capacity (SRC) testing of wheat
21 404 flour: Principles and value in predicting flour functionality in different wheat‐ based food
22
23 405 processes and in wheat breeding-A review. Cereal Chemistry, 88, 537-552.
24
25 406 Labuschagne, M.T., Koen, E., & Dessalegn, T. (2004). Use of size exclusion high-performance
26 407 liquid chromatography for wheat quality prediction in Ethiopia. Cereal Chemistry, 81, 533-
27
28
408 537.
29
409 León, A.E., Barrera, G.N., Pérez, G.T., Ribotta, P.D., & Rosell, C.M. (2006). Effect of damaged
30
31 410 starch levels on flour-thermal behaviour and bread staling. European Food Research and
32 411 Technology, 224, 187-192.
33
34 412 Li, W., Shan, Y., Xiao, X., Luo, Q., Zheng, J., Ouyang, S., & Zhang, G. (2013). Physicochemical
35
36 413 properties of A-and B-starch granules isolated from hard red and soft red winter wheat.
37 414 Journal of Agricultural and Food Chemistry, 61, 6477-6484.
38
39 415 Lu, Z., & Seetharaman, K. (2014). Suitability of Ontario‐ grown hard and soft wheat flour
40 416 blends for noodle making. Cereal Chemistry, 91, 482-488.
41
42 417 Malalgoda, M., Ohm, J.B., Meinhardt, S., Chao, S., & Simsek, S. (2017). Cluster analysis of
43
44 418 historical and modern hard red spring wheat cultivars based on parentage and HPLC
45 419 analysis of gluten‐ forming proteins. Cereal Chemistry, 94, 560-567.
46
47 420 Marti, A., Augst, E., Cox, S., & Koehler, P. (2015). Correlations between gluten aggregation
48 421 properties defined by the GlutoPeak test and content of quality-related protein fractions
49
50 422 of winter wheat flour. Journal of Cereal Science, 66, 89-95.
51
52 423 Mohler, V., Schmolke, M., Paladey, E., Seling, S., & Hartl, L. (2012). Association analysis of
53 424 Puroindoline-D1 and Puroindoline b-2 loci with 13 quality traits in European winter
54
55
425 wheat (Triticum aestivum L.). Journal of Cereal Science, 56, 623-628.
56
57
58
59
60 17
61
62
63
64
65
 1
2
3
4 426 Mondal, S., Hays, D.B., Alviola, N.J., Mason, R.E., Tilley, M., Waniska, R.D., Bean, S.R., &
5
6 427 Glover, K. D. (2009). Functionality of gliadin proteins in wheat flour tortillas. Journal of
7 428 Agricultural and Food Chemistry, 57, 1600-1605.
8
9 429 Morel, M.H., Pincemaille, J., Chaveau, E., Louhichi, A., Voilleau, F., Menut, P., Ramos, L., &
10
11
430 Banc, A. (2020). Insight into gluten structure in a mild chaotropic solvent by asymmetrical
12 431 flow field flow fractionation (AsF1FFF). Food Hydrocolloids,103, 105676
13
14 432 Morris, C.F., Luna, J., & Caffe‐ Treml, M. (2021). The vromindolines of cv. Hayden oat (Avena
15 433 sativa L.)–A review of the Poeae and Triticeae indolines and a suggested system for
16
17 434 harmonisation of nomenclature. Journal of Cereal Science, 97, 103135.
18
19 435 Nadolska-Orczyk, A., Gasparis, S., & Orczyk, W. (2009). The determinants of grain texture in
20 436 cereals. Journal of Applied Genetics, 50, 185-197.
21
22 437 Quayson, E.T., Marti, A., Bonomi, F., Atwell, W., & Seetharaman, K. (2016a). Structural
23 438 modification of gluten proteins in strong and weak wheat dough as affected by mixing
24
25 439 temperature. Cereal Chemistry, 93, 189-195.
26
27 440 Quayson, E.T., Atwell, W., Morris, C.F., & Marti, A. (2016b). Empirical rheology and pasting
28 441 properties of soft-textured durum wheat (Triticum turgidum ssp. durum) and hard-
29
442 textured common wheat (T. aestivum). Journal of Cereal Science, 69, 252-258.
30
31 443 Schopf, M., Wehrli, M.C., Becker, T., Jekle, M., & Scherf, K.A. (2021). Fundamental
32
33 444 characterization of wheat gluten. European Food Research and Technology, 247, 985-
34 445 997.
35
36 446 Shang, J., Li, L., Liu, C., Hong, J., Liu, M., Zhao, B., & Zheng, X. (2021). Relationships of flour
37
38
447 characteristics with isolated starch properties in different Chinese wheat varieties. Journal
39 448 of Cereal Science, 99, 103210.
40
41 449 Shang, J., Li, L., Zhao, B., Liu, M., & Zheng, X. (2020). Comparative studies on
42 450 physicochemical properties of total, A-and B-type starch from soft and hard wheat
43
44 451 varieties. International Journal of Biological Macromolecules, 154, 714-723.
45
46 452 Sharma, S., Katyal, M., Singh, N., Singh, A.M., & Ahlawat, A.K. (2022). Comparison of effect
47 453 of using hard and soft wheat on the high molecular weight-glutenin subunits profile and the
48 454 quality of produced cookie. Journal of Food Science and Technology, 59, 2545-2561.
49
50 455 Shevkani, K., Singh, N., Singh, S., Ahlawat, A.K. & Singh, A.M. (2011). Relationship between
51
52 456 physicochemical and rheological properties of starches from Indian wheat lines.
53 457 International Journal of Food Science and Technology, 46, 2584-2590.
54
55 458 Shevkani, K., Singh, N., Bajaj, R., & Kaur, A. (2017). Wheat starch production, structure,
56
459 functionality and applications-a review. International Journal of Food Science and
57
58 460 Technology, 52, 38-58.
59
60 18
61
62
63
64
65
 1
2
3
4 461 Shevkani, K., Singh, N., Chen, Y., Kaur, A., & Yu, L. (2019). Pulse proteins: secondary
5
6 462 structure, functionality and applications. Journal of Food Science and Technology, 56,
7 463 2787-2798.
8
9 464 Shevkani, K., Singh, N., Isono, N., & Noda, T. (2021). Structural and functional properties of
10
11
465 amaranth starches from residue obtained during protein extraction. Journal of Food
12 466 Measurement and Characterization, 15, 5087-5096.
13
14 467 Shewry, P. (2023). Wheat grain proteins: Past, present, and future. Cereal chemistry, 100, 9-22.
15
16 468 Shewry, P.R., & Lafiandra, D. (2022). Wheat glutenin polymers 1. Structure, assembly and
17 469 properties. Journal of Cereal Science,106, 104386
18
19 470 Shewry, P.R., & Halford, N.G. (2002). Cereal seed storage proteins: structures, properties and
20
21
471 role in grain utilization. Journal of Experimental Botany, 53, 947-958.
22
472 Singh, M., & Khatkar, B.S. (2005). Structural and functional properties of wheat storage
23
24 473 proteins: a review. Journal of Food Science and Technology, 42, 455-471.
25
26 474 Singh, N., Gujral, H.S., Katyal, M., & Sharma, B. (2019). Relationship of Mixolab
27 475 characteristics with protein, pasting, dynamic and empirical rheological characteristics of
28
29 476 flours from Indian wheat varieties with diverse grain hardness. Journal of Food Science
30 477 and Technology, 56, 2679-2686.
31
32 478 Singh, N., Kaur, N., Katyal, M., Kaur, A., & Shevkani, K. (2017). Characteristics of starch
33 479 separated from coarse and fine flour fractions obtained from hard, medium‐ hard, and
34
35 480 soft Indian wheat cultivars. Starch‐ Stärke, 69, 1600012.
36
37 481 Singh, N., Shevkani, K., Kaur, A., Thakur, S., Parmar, N., & Singh Virdi, A. (2014).
38 482 Characteristics of starch obtained at different stages of purification during commercial wet
39
483 milling of maize. Starch‐ Stärke, 66, 668-677.
40
41 484 Singh, N., Virdi, A.S., Katyal, M., Kaur, A., Kaur, D., Ahlawat, A.K., Singh, A.M., & Sharma,
42
43 485 R.K. (2021). Evaluation of heat stress through delayed sowing on physicochemical and
44 486 functional characteristics of grains, whole meals and flours of India wheat. Food
45
46 487 Chemistry, 344, 128725.
47
488 Tatham, A.S., Hayes, L., Shewry, P.R., & Urry, D.W. (2001). Wheat proteins exhibit a complex
48
49 489 mechanism of protein elasticity. Biochemica et Biophysica Acta, 1548, 187–193.
50
51 490 Tao, H., Huang, J.S., Xie, Q.T., Zou, Y.M., Wang, H.L., Wu, X.Y., & Xu, X.M. (2018). Effect
52 491 of multiple freezing-thawing cycles on structural and functional properties of starch
53
54 492 granules isolated from soft and hard wheat. Food Chemistry, 265, 18-22.
55
56 493 Tester, R.F., & Karkalas, J. (2001). The effects of environmental conditions on the structural
57 494 features and physico‐ chemical properties of starches. Starch‐ Stärke, 53(10), 513-519.
58
59
60 19
61
62
63
64
65
 1
2
3
4 495 Tian, J., Hu, R., Deng, Z., & Wang, Y. (2007). The variation and stability analysis of wheat
5
6 496 dough stability time. Agricultural Sciences in China, 6, 143-149.
7
8 497 Tsilo, T.J., Ohm, J.B., Hareland, G.A., & Anderson, J.A. (2010). Association of size‐ exclusion
9 498 HPLC of endosperm proteins with dough mixing and breadmaking characteristics in a
10
11
499 recombinant inbred population of hard red spring wheat. Cereal Chemistry, 87, 104-111.
12
500 Wang, P., Xu, L., Nikoo, M., Ocen, D., Wu, F., Yang, N., Jin, Z., & Xu, X. (2014). Effect of
13
14 501 frozen storage on the conformational, thermal and microscopic properties of gluten:
15 502 Comparative studies on gluten-, glutenin-and gliadin-rich fractions. Food Hydrocolloids,
16
17 503 35, 238-246.
18
19 504 Wrigley, C.W., Békés, F., Bushuk, W. (2006). Gluten: A balance of gliadin and glutenin. The
20 505 unique balance of wheat quality. C. Wrigley, F. Békés and W. Bushuk, eds. AACC
21 506 International, St. Paul, MN 2006, 3-34.
22
23 507 Wu, F., Li, J., Yang, N., Chen, Y., Jin, Y., & Xu, X. (2018). The roles of starch structures in the
24
25 508 pasting properties of wheat starch with different degrees of damage. Starch‐ Stärke, 70,
26 509 1700190.
27
28 510 Xiao, Z.S., Park, S.H., Chung, O.K., Caley, M.S., & Seib, P.A. (2006). Solvent retention
29
511 capacity values in relation to hard winter wheat and flour properties and straight‐ dough
30
31 512 breadmaking quality. Cereal Chemistry, 83, 465-471.
32
33 513
34 514
35 515
36
516
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60 20
61
62
63
64
65
 1
2
3
4 517 Legends to figures and tables
5
6 518
7 519 Figure 1. Size-exclusion chromatograms of hard and soft wheat (HW and SW, respectively)
8
9 520 flours (Source: Katyal et al., 2017)
10 521 Figure 2. Electrophoretic profile of reduced and alkylated glutenins of flours from soft
11
12
522 (QBP12-9, QBP12-10, QBP12-8 and QBP12-11), medium-hard (HD2967, HI977, UP2672 and
13 523 DBW39) and hard (K307, HD2733, DPW621-50 and HD2932) wheat varieties (Source: Katyal
14 524 et al., 2017)
15
16 525 Figure 3. Farinograms of flours from different soft and hard Indian wheat varieties (Source:
17 526 Katyal et al., 2017)
18
19 527 Table 1. Physicochemical, rheological (farinographic), chromatographic, solvent retention and
20 528 secondary structural properties of soft and hard wheat flours
21
22 529
23 530
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60 21
61
62
63
64
65
 1
2
3
4 531
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20 532
21 533 Figure 1. Size-exclusion chromatograms of hard and soft wheat (HW and SW, respectively)
22 534 flours (Source: Katyal et al., 2017)
23
24
535
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53 536
54 537 Figure 2. Electrophoretic profile of reduced and alkylated glutenins of flours from soft
55 538 (QBP12-9, QBP12-10, QBP12-8 and QBP12-11), medium-hard (HD2967, HI977, UP2672 and
56
57 539 DBW39) and hard (K307, HD2733, DPW621-50 and HD2932) wheat varieties; HMW: high
58 540 molecular weight; LMW: low molecular weight (Source: Katyal et al., 2017)
59
60 22
61
62
63
64
65
 1
2
3
4 541
5
6
7 542
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22 543
23
24 544 Soft wheat (GHI=29) Soft wheat (GHI=19)
25
26 545
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43 546
44
45 547 Hard wheat (GHI=73)
46
Hard wheat (GHI=89)
47
48 548
49
50 549 Figure 3. Farinograms of flours from different soft and hard Indian wheat varieties; GHI: grain
51 550 hardness index (Source: Katyal et al. 2017)
52
53
54 551
55
56 552
57 553
58 554
59
60 23
61
62
63
64
65
 1
2
3
4 555
5
6
7 556 Table 1. Physicochemical, rheological (farinographic), chromatographic, solvent retention
8 557 and secondary structural properties of soft and hard wheat flours
9
10
11 Soft Hard
Parameter Source(s)
12 wheat wheat
13 Katyal et al. (2017); Katyal et al.,
14 Grain hardness index 17-35 71-95
15 (2018); Shang et al. (2020)
16 Katyal et al. (2017); Issarny et al.
17 Protein content (%) 7-10.5 10-13
(2017)
18
19 Damaged starch content (%) 2-3 7-10 Singh et al. (2019)
20
21 Farinographic analysis
22
23 Water absorption (%) 52-56 58-65 Cao et al. (2017); Hammed et al.
24
25 Dough development time (2015); Issarny et al. (2017); Lu
Up to 1.4 1.5-9
26 (min) and Seetharaman (2014);
27
Dough stability (min) Up to 1.3 7-17 Malalgoda et al. (2017)
28
29 Chromatographic characterization of proteins
30
31 Un-extractable polymeric
32 42 50-62
proteins (%)
33
34 Un-extractable monomeric
58 38-50
35 proteins (%)
36 Katyal et al. [2]; Gupta et al. [44]
37 Extractable polymeric proteins
27 33-46
38 (%)
39 Extractable monomeric
40 73 54-67
41 proteins (%)
42 Solvent retention capacity
43
44 Lactic acid (%) 87-102 105-178 Issarny et al. (2017); Cao et al.
45 Sucrose (%) 74-111 99-135
46 (2017); Hammed et al. (2015);
47 Sodium bicarbonate (%) 68-76 79-98 Xiao et al. (2006); Katyal et al.
48 (2018)
49
Water (%) 58-60 65-81
50 Protein secondary structure
51
52 α-helix (%) 3-9 3-10
53 Intermolecular β-sheets (%) 19 15 Cao et al. (2016); Katyal et al.
54
55 β-sheets (%) 32-71 37-71 (2017)
56
57
β-turns (%) 7-42 8-39
58 558 Values shown have been subjected to rounding-off.
59
60 24
61
62
63
64
65
REVISED Manuscript (text UNmarked) Click here to view linked References

1 Protein characteristics, starch properties and end-use quality of soft and hard

2 wheat - a comparative review

3 Khetan Shevkani1, Mehak Katyal2, Narpinder Singh3

5 1 Department of Applied Agriculture, Central University of Punjab, Bathinda-151401, India

6 (shevkani@gmail.com; khetan.shevkani@cup.edu.in)

7 2 Department of Nutrition and Dietetics (Food Technology), Manav Rachna International Institute

8 of Research & Studies, Faridabad, India (mehak.katyal3@gmail.com)

9 3 Department of Food Science and Technology, Graphic Era Deemed to be University, Dehradun-

10 248002, India (narpinders@yahoo.com)

11

12 Corresponding authors: KS (shevkani@gmail.com)

13

14 Running title: Hard and soft wheat: a comparison

15

16 Abstract

17 Hardness of wheat endosperm is a physical property that depends on the genetic makeup of the

18 grain and varies amongst different types/varieties. It is associated with proteins, particularly 14-15

19 kDa friabilins, which exist in a higher amount at the surface of starch in soft wheat grains while

20 either absent or occur in smaller quantities on granules of hard wheat varieties. The storage

21 proteins in hard wheat flours exhibit higher proportion of polymeric proteins, β-sheets and β-turns,

22 while soft wheat starches are characterized by a lower crystallinity, lesser amount of amylose-lipid

23 complexes, a particle size distribution with smaller proportion of A-type granules (disk or
1
24 lenticular-shaped granules with diameter >10 µm) and higher-gelatinisation temperatures,

25 swelling, breakdown susceptibility and retrogradation than hard wheat starches. Flours from hard

26 and soft wheat also differ for processing and end-use quality based on differences in dough

27 rheological properties, solvent retention capacities and particle size.

28

29 Keywords: Hard vs soft wheat; Protein secondary structure; Polymeric vs monomeric proteins;

30 Rheology; Starch properties

31

32 1. Introduction

33 Wheat is considered unique amongst cereals because of its ability to form a viscoelastic dough

34 when mixed with water. Wheat flour is used to prepare a number of palatable and satisfying

35 foods. It is grown and utilized in most parts of the world for food, feed and industrial purposes.

36 Thousands of cultivars/varieties of wheat varying in grain and flour properties are grown. They

37 have been categorized based on planting season (winter or spring), colour (white or red) and

38 endosperm characteristics (vitreous or mealy and soft or hard). However, wheat is primarily

39 classed on the basis of grain hardness/softness as it has strong association with the finished-

40 product quality (Shewry, 2023). Hard wheat grains, generally, yield strong flour with high

41 protein content which is suitable for bread making while soft wheat mill into flour with lesser

42 protein and damaged starch content suitable for manufacturing cookies, biscuits and pastries

43 (Quayson et al., 2016ab; Shang et al., 2020; Sharma et al., 2022).

44 Grain hardness is a genetic property of wheat and it depends on the degree of adhesion

45 between starch granules and protein matrix within the endosperm, the continuity of the protein

46 matrix and the absence of cracks/fissures. The adhesion between starch and proteins in the wheat

2
47 endosperm is governed primarily by the level of specific membrane-bound proteins of 14-15 kDa

48 present on the surface of starch granules known as friabilins. These proteins can soften wheat

49 endosperm by hindering the adhesion between starch granules and protein matrix. Soft wheat

50 grains contain higher levels of friabilins than hard wheat while these proteins are absent in

51 durum varieties, considered the hardest type of wheat (Shevkani et al., 2017). Friabilins are made

52 up of two protein subunits, puroindoline-A and puroindoline-B, which are gene products of

53 alleles Pina-D1a and Pinb-D1a, respectively, and are located at the Ha locus on the short arm of

54 the 5D chromosome in soft wheats (Morris et al., 2021). Hard wheat varieties are characterized

55 by specific mutations in either Pina-D1 or Pinb-D1 genes or might lack these genes (Morris,

56 2021; Nadolska-Orczyk et al, 2009). The level of friabilins on starch granules also depends on

57 the level of polar lipids including glycol- and phospholipids which were essential for the binding

58 of friabilins to the granule surface in the endosperm (Greenblatt et al., 1995). Puroindolines can

59 bind with lipids on the surface of starch granules through a hydrophobic tryptophan-rich domain

60 (Clifton et al., 2007). A negative association between starch surface lipids and grain hardness has

61 been reported (Kim et al., 2012). Environment conditions also affect the hardness of wheat

62 grains. Singh et al. (2021) reported that heat stressed wheat had grains with higher hardness and

63 lower weight and diameter caused by lower starch synthesis.

64 The hardness of wheat grains is commonly measured as grain hardness index (GHI) using

65 a Single Kernel Characterization System (SKCS) of Perten Instruments (Australia). This

66 instrument records the force required to crush a grain between a toothed and a narrowing

67 crescent gap. GHI is an indicator of the resistance of the grain to fracture. It also translates the

68 ability to produce flour of variable quality as hard and soft wheat varieties differ widely for their

69 protein content/composition, starch characteristics and dough rheology (Issarny et al., 2017;

3
70 Katyal et al., 2017; 2019; Shang et al., 2020; Sharma et al., 2022). In recent years, hard and soft

71 wheat varieties have been investigated to study the basis of differences in their properties and

72 applications (Cao et al., 2017; Issarny et al., 2017; Katyal et al., 2017; 2019; Quayson et al.,

73 2016ab; Sharma et al., 2022; Shang et al., 2020; Singh et al., 2017; 2019; Tao et al., 2018).

74 However, a review comparing hard and soft wheat from the perspective of starch and protein

75 properties is not available. Therefore, this article attempted to provide a concise overview of the

76 differences in hard and soft wheat varieties in the context of the structure and functionality of

77 proteins and starches.

78

79 2. Protein characteristics

80 Gliadins and glutenins are major storage proteins in wheat endosperm, constituting up to 85% of

81 total proteins (Guo et al., 2021). Glutenins refer to proteins soluble in dilute alkali or acidic

82 solutions while gliadins include aqueous alcohol soluble proteins. Figure 1 depicts the typical size

83 exclusion chromatographic peaks of wheat flour glutenins and gliadins. These proteins are

84 responsible for the typical viscoelastic behaviour (characterised as the presence of both elasticity

85 and extensibility) of wheat flour dough which is pivotal in determining the end-use quality of

86 wheat varieties. Elasticity in dough results from the formation of a strong three-dimensional gluten

87 network desirable for making a good quality bread while extensibility/fluidity is due to a mellow

88 network formation suitable for the preparation of cookies, cakes and wheat tortillas (Dizlek et al.,

89 2022; Mondal et al., 2009).

90 Amongst wheat proteins, glutenins are polymeric proteins and they play a crucial role in

91 gluten strength and the suitability of wheat flour for bread making. The hydration of flour proteins

92 during dough kneading results in the formation of a three-dimensional gluten network. This

4
 93 involves the polymerization of glutenins through interchain disulphide linkages (Shewry and

94 Lafiandra, 2022) and the aggregation of the polymeric glutenin molecules with gliadins via

95 hydrogen bonds, hydrophobic and electrostatic interactions (Morel et al., 2020). Although both

96 these phenomena contribute to stabilise the gluten network and dough formation, glutenins endow

97 the dough with the strength/elasticity needed for good quality bread making owing to the ability to

98 form intermolecular covalent disulphide bonds, whereas the presence of gliadins promotes

99 extensibility by weakening the gluten network after being incorporated into the network structure

100 (Schopf et al., 2021). Shewry (2023) showed that dough extensibility results from the slippage of

101 gliadins and glutenins linked non-covalently. Therefore, hard wheat flour with higher glutenin

102 content than soft wheat flour (Figure 1) generally forms more elastic dough exhibiting stronger

103 gluten network required for bread-making.

104 Based on the differences in molecular weight, glutenins can be categorised as low and

105 high-molecular-weight glutenin subunits (LMW-GS and HMW-GS, respectively) held together by

106 intra and inter-chain disulphide bonds (Figure 2). HMW-GS are associated with improved bread-

107 making properties while flours with lower levels of HMW-GS could be useful for making

108 products where a relatively lower gluten strength is required. Hard and soft wheat differ for the

109 relative levels of LMW-GS and HMW-GS fractions (Figure 2). The glutenins from hard and soft

110 wheat varieties showed the presence of 19 significant polypeptides (PPs) ranging in MW between

111 26 kDa and 98 kDa which were categorized as HMW-GS (98-79 kDa), LMW-GS-B (49-40 kDa)

112 and LMW-GS-C (35-26 kDa). Hard wheat varieties with GHI of 97 to 100 showed specific

113 HMW-GS composition with 91, 80, 78, and 74 kDa PPs, which was not the property of soft wheat

114 flours (Katyal et al., 2017). In addition, hard wheat flours exhibited glutenins with GluD1 locus

5
115 with 5+10 subunits whereas soft wheat HMW-GS with allelic composition of 2*, 7 and 2+12 were

116 linked with low dough stability and improved cookie-making properties (Sharma et al., 2022).

117 Wheat flour proteins have also been evaluated as the proportion of unextractable and

118 extractable polymeric and monomeric proteins (UnEx-PP, Ex-PP, UnEx-MP and Ex-MP,

119 respectively) based on their extractability in sodium dodecyl sulphate buffer. In addition to the

120 ratio of gliadin/glutenin, the quantity of both extractable and unextractable proteins affects dough

121 strength and end-use quality. Loaf volume and dough/gluten strength correlated positively with the

122 amount of glutenins and negatively with gliadins (Tsilo et al., 2010), while polymeric to

123 monomeric protein ratio was associated with superior baking qualities (Labuschagne et al., 2004).

124 Hard and soft wheat also differ for the proportion of polymeric and monomeric proteins. Flours

125 from harder wheat varieties contained polymeric proteins (both extractable and unextractable) in

126 higher proportions than soft wheat (Baasandorj et al., 2016; Katyal et al., 2017). The mean

127 proportion of Ex-PP and UnEx-PP for flours from hard and medium hard wheat varieties was 33-

128 46% and 50-62%, respectively, against 27% and 42%, respectively, for soft wheat flours, whereas

129 soft wheat flours contained monomeric proteins in higher proportions than hard wheat flours

130 (Table 1).

131 The secondary structure of gluten proteins is also related to dough functionality and it has

132 also been linked with grain hardness. The elasticity of gluten proteins was attributed to β-spiral

133 structures in HMW-GS which were formed of regularly repeated β-turn structures (Shewry and

134 Halford, 2002; Tatham et al., 2001). Belton (2005) showed the formation of hydrogen-bonded

135 inter-chain β-sheet structures in gluten proteins by the orientation of β-turns in adjacent β-spirals

136 after hydration. The dough with a higher proportion of β-sheets and lower α-helix exhibited higher

137 gluten strength (Kaur et al., 2014). The secondary structure of gluten/dough can be evaluated

6
138 qualitatively and quantitatively using Fourier transform infrared (FTIR) spectroscopy which is a

139 vibrational spectroscopic analytical technique useful for studying the structure of proteins under

140 diverse environments. Amongst different regions observed when proteins are studied using FTIR,

141 amide I and II regions (attributing to stretching of C=O bond and bending vibrations at N-H bond

142 in infrared spectra) are very sensitive to protein secondary structure (Shevkani et al., 2019). The

143 analysis of wheat proteins using FTIR generally reveals the presence of three predominant bands

144 in the amide-I region at wavenumbers between ≈1660 and ≈1690 cm-1 while two bands in the

145 amide-II region at ≈1570 and ≈1580 cm-1. The amide-I bands at around 1629-1632 cm-1, 1650-

146 1660 cm-1, 1665-1670 cm-1 and 1687-1690 cm-1 were ascribed to antiparallel-β-sheets, α-helical

147 structures, β-turns and β-sheets, respectively (Wang et al., 2014). The proteins from hard and soft

148 wheat differed for their secondary structure. Hard wheat proteins contained a lesser proportion of

149 α-helices and higher β-sheets and β-turns than soft wheat flours (Jazaeri et al., 2015; Quayson et

150 al., 2016a). Katyal et al. (2017) also reported that the dough from soft wheat flours had higher

151 proportion of intermolecular-β-sheets than medium-hard and hard wheat varieties. It has also been

152 reported that β-sheets are formed in soft wheat dough by hydrophobic interactions while in hard

153 wheat dough, disulfide bonds stabilise the secondary structure (Jazaeri et al. 2015).

154

155 3. Starch properties

156 Starch is the most abundant storage carbohydrate in wheat, comprising about 60-75% of grain and

157 70-80% of flour. The content and characteristics of starch in flour affect the quality of wheat-based

158 products (Cao et al., 2019; Shang et al., 2021). The pattern of breakage of wheat during milling

159 affects the amount of damaged starch in flour. Hard wheat varieties yield flours with higher

160 damaged starch content having greater affinity to water than intact granules. Li et al. (2013)

7
161 attributed lower water absorption of soft wheat flour to higher content of intact starch granules

162 (lesser damaged granules) together with lower protein content. Similarly, the higher water

163 absorption capacity of hard wheat flour was ascribed to higher content of damaged starch

164 (Baasandorj et al., 2020; Katyal et al., 2019; Singh et al., 2019). Shewry (2023) reported that

165 damaged starch can absorb 3-4 times its own weight of water in comparison to ≈ 0.5 times for

166 intact starch granules.

167 Hard and soft wheat varieties also differ for amylose content, morphological properties,

168 crystallinity, gelatinisation properties and pasting properties of their starches. Starches from soft

169 wheat were less crystalline and contained a lower proportion of ‘A’ granules (disk or lenticular

170 shaped granules with diameter > 10 µm) while higher of ‘B’ granules (spherical granules with

171 diameters < 10 µm) than starch of hard wheat varieties (Katyal et al., 2019; Singh et al., 2017;

172 Shang et al., 2020). In general, higher gelatinisation temperatures and enthalpy of starches are

173 attributed to a higher degree of crystallinity (associated with lower content of amorphous amylose

174 and a greater proportion of long amylopectin chains) which provides structural stability to

175 granules and makes them more resistant towards gelatinisation (Shevkani et al., 2017). However,

176 soft wheat starches, despite lower crystallinity, showed higher gelatinisation temperatures and

177 enthalpy (mean onset-, peak-, endset-temperature and enthalpy = 59.96-61.17 °C, 63.25-63.84 °C,

178 66.67-66.81 °C and 7.55-9.22 J/g respectively) than medium-hard (58.15-60.96 °C, 62.08-63.58

179 °C, 66.09-66.46 °C and 7.05-8.45 J/g, respectively) and hard varieties (57.55-60.27 °C, 61.72-

180 63.08 °C, 65.85-66.12 °C and 6.84-8.31 J/g, respectively) (Katyal et al., 2019; Singh et al., 2017).

181 Such behaviour of soft wheat starches was likely due to higher puroindolines content and lesser

182 damage to the granules during milling which made them more resistant to gelatinise by inhibiting

8
183 the availability of water (Brites et al., 2008; Quayson et al., 2016b) and suppressing starch

184 hydration and dispersion in water (Han et al., 2022; Leon et al., 2006), respectively.

185 However, varying results were observed for amylose content in starches of hard and soft

186 wheat grains. Singh et al. (2017), Katyal et al. (2019) and Shang et al. (2020) reported lower

187 amylose in starch from softer wheat varieties (mean amylose content = 12.1-22.8%) than harder

188 varieties (mean amylose content = 15.0-25.2%). In contrast, Kumar et al. (2016) observed higher

189 amylose content for starch from soft wheat (25.78%) than hard wheat varieties (24.11%), whereas

190 Biduski et al. (2022) reported no difference in amylose content of starch from soft (25.62%) and

191 hard wheat (25.53%), but for medium hard wheat (23.3%).

192 Pasting properties represent changes in the viscosity of starch suspensions during and after

193 gelatinisation. Pasting properties are considered an important index of starch functionality and

194 depend not only on starch morphology and structure but also on the state of granules

195 (damaged/intact) and the presence/absence of non-starch constituents (Han et al., 2022; Singh et

196 al., 2014; Shevkani et al., 2021). Starches from soft wheat showed higher peak viscosity

197 (maximum viscosity attained during heating representing the point of maximum swelling of

198 granules), breakdown viscosity (the fall in viscosity during heating and shearing representing the

199 susceptibility of swollen granules to rupture at high temperatures), final and setback viscosity

200 (increase in the viscosity on cooling due to retrogradation) than hard wheat starches (Katyal et al.,

201 2019; Shang et al., 2020). These results were possibly due to the presence of lower levels of

202 amylose-lipid complexes, starch-associated proteins and damaged granules in soft wheat starches,

203 which modified the behaviour of starch suspensions during pasting by affecting the potential of

204 granules to swell and disintegrate during heating and shearing. The swelling of granules during

205 heating in water has been shown to occur more quickly after the exudation of amylose (Tester &

9
206 Karkalas, 2001) while amylose-lipid complexes and proteins suppress granular swelling and

207 subsequent disintegration by increasing granule integrity/rigidity and preventing amylose leaching

208 (Shang et al., 2020; Shevkani et al., 2011). Mechanical damage to starch, in contrast, could

209 promote the formation of less viscous paste by distorting granules, damaging their ordered

210 structure, reducing crystallinity and facilitating water absorption leading to greater disintegration

211 of the gelatinized starch and the production of more leachate materials (Barrera et al., 2013; Han et

212 al., 2022; Wu et al., 2018).

213 Differences in retrogradation tendencies were also reported for starches from soft and hard

214 wheat varieties (Shang et al., 2020). Soft starch gels showed higher retrogradation than starches

215 from hard wheat varieties. Gels of soft wheat starches exhibited higher syneresis (separation of

216 water from starch gels during freezing-thawing cycles due to the reassociation of dissociated

217 starch chains during cooling/storage of gels), gel hardness and fracturability in comparison to

218 starches from hard wheat varieties. These phenomena were linked with the presence of more

219 granular remnants, higher amylose content and lower levels of proteins and lipids associated with

220 starches. Amylose has been shown to enhance the rate of retrogradation while the presence of

221 proteins and amylose-lipid complexes could reduce retrogradation by making interactions with

222 starch chains (Shevkani et al., 2017; 2021). However, more work is required to be conducted on

223 the retrogradation tendencies of hard and soft wheat starches as only limited information is

224 available in the literature.

225

226 4. Finished product suitability

227 Wheat is consumed in almost all countries of the world. It is typically milled into flour and then

228 used as a fundamental ingredient for preparing a wide range of products. Flours from different

10
229 wheat varieties differ for their physicochemical and functional characteristics, hence for finished

230 product suitability. Usually, the suitability of wheat flour for producing different types of products

231 is assessed by evaluating dough rheology, solvent retention capacity (SRC) and flour particle size.

232 The rheological properties of wheat flour dough determine baking performance,

233 machinability and processing quality (Ktenioudaki et al., 2010). Farinograph and dynamic

234 rheometer are commonly used to determine the suitability of wheat flour for different baked

235 foods. Farinograph is a widely employed empirical rheological measurement technique, whereas

236 dynamic rheometers are used to measure the fundamental rheological aspects of dough.

237 Empirical rheological techniques are more common than dynamic rheometry, and they record the

238 changes in dough consistency during mixing while making use of large deformations. The most

239 useful parameters obtained from the farinographic analysis include water absorption (WA; a

240 measure of the amount of water required for the formation of an optimally viscoelastic dough),

241 dough development time (DDT; an indicative of optimum mixing/kneading time to have a well-

242 developed dough), dough stability (DS; an indicator of tolerance of dough towards mixing) and

243 degree of softening (DOS; decrease in the consistency of dough with extended mixing). Flours

244 with higher WA are generally desirable for bread making. A flour exhibiting high WA along

245 with high DS and low DOS results in the production of bread with desirable quality attributes

246 (high volume, soft and porous crumb, springy texture, etc.); however, a high WA combined with

247 a high DOS indicates poor quality flour. Hard wheat flours generally show higher WA, DDT and

248 DS in comparison to flours from soft wheat varieties (Table 1; Figure 3). Higher values of DDT

249 and DS for hard wheat were associated with a stronger gluten network and higher protein content

250 as compared to soft wheat (Tian et al., 2007). The DOS has been shown to vary widely between

11
251 14 and 136 BU amongst different wheat varieties, while soft varieties were generally

252 characterized by exceptionally high DOS (Katyal et al., 2017).

253 Dynamic rheometer, in comparison, evaluates elastic and viscous properties of dough

254 which are expressed as storage modulus (G′), loss modulus (G″) and loss tangent (tanδ = G″/G′).

255 The elastic dough with greater DS exhibits higher G' and G" and lower tanδ values (Singh et al.

256 2011). Kaur et al. (2013) observed positive relations of both G′ and G″ with protein content.

257 Hard wheat varieties showed higher G' and G″ than soft wheat varieties, indicating a more elastic

258 nature of hard wheat flour dough. Both G' and G'' also related positively to DS and DDT. Wheat

259 varieties with HMW-GS of 91, 80, 78 and 74 kDa had lower tanδ (higher elasticity) than those

260 which did not show these proteins (Katyal et al., 2017).

261 Solvent retention capacity is another useful method to determine end-use quality of wheat

262 flours. Lu and Seetharaman (2014) and Hammed et al. (2015) reported relation of SRC values

263 with WA, DS, protein content, gluten content and damaged starch content. SRC quantifies the

264 swelling of gluten networks and the ability to retain various solvents such as water, sucrose, lactic

265 acid and sodium carbonate (Kweon et al., 2011). Sodium carbonate SRC (NaSRC) was related

266 with the damaged starch content while sucrose SRC (SuSRC) indicated gliadins, pentosan and

267 arabinoxylans content. Lactic acid SRC (LASRC) was an indicator of gluten strength while water

268 SRC (WSRC) indicated general water retention capacity of various flour constituents. Cao et al.

269 (2017) observed higher NaSRC for hard wheat (73.6-90.1%) than soft wheat flours (66.3-81.6%).

270 LASRC was related to gluten strength parameters and bread volume (Guttieri et al., 2001; Xiao et

271 al., 2006). Soft wheat varieties showed lower SuSRC associated with lower arabinoxylans content,

272 while LASRC positively correlated with UnEx-PP reflecting the contribution of glutenins to

12
273 swelling and gluten network formation. Hard wheat flours were characterized by greater LASRC,

274 WSRC, SuSRC and NaSRC than soft wheat flour (Issarny et al., 2017).

275 Particle size distribution also has a pronounced effect on the baking quality and

276 functionality of wheat flours. In addition, the particle size of flour also influences the glycemic

277 index, fermentation and bioavailability of nutrients (Hallfrisch & Behall, 2000). Flours from soft

278 and hard wheat vary for particle size distribution. The flour particle size distribution was

279 correlated with GHI (Devaux et al., 1998). Wheat with higher GHI produced flour with a lesser

280 proportion of small-size particles and a greater of large-size particles; however, soft wheat

281 varieties showed the opposite results (Kaur et al., 2014; Katyal et al., 2016; 2018). Soft wheat with

282 higher proportion of fine particles showed lower-DDT, DS, gluten strength and SRC, and

283 produced cookies with greater spread (Kaur et al., 2014; Singh et al., 2019; Sharma et al., 2022).

284

285 Conclusion

286 Hard and soft wheat differ for milling and functional characteristics. Hard wheat mill into flours

287 with a higher proportion of coarse particles, damaged starch content and protein content as

288 compared to soft wheat varieties. Proteins in hard wheat flours yield dough with high gluten

289 strength attributing to the higher proportion of polymeric glutenins, β-sheets and β-turns than soft

290 wheat flours. Generally, hard wheat flours exhibit strong gluten attributed to high protein content,

291 HMW-GS and UnEx-PP; however, the differences due to genetics, environmental conditions,

292 duration of crop and cultural practices affect the ultimate suitability of flour for different products.

293 Soft wheat starches are characterized by a lower proportion of A-type granules (disk or lenticular-

294 shaped granules with diameter > 10 µm), a lower crystallinity, higher gelatinisation temperature,

295 more swelling, greater breakdown susceptibility and increased retrogradation tendencies than

13
296 starches from hard wheat varieties. The lower proportion of damaged granules, amylose-lipid

297 complexes and granule-associated proteins in soft wheat starches contribute to higher swelling,

298 breakdown and retrogradation. However, more work is required on the comparison of hard and

299 soft wheat starches for amylose and amylopectin fine structure (chain length distribution) owing to

300 the association of these attributes with the functionality of starches.

301

302 Acknowledgement

303 NS acknowledges financial support from CSIR as research project grant and SERB as JC Bose

304 Fellowship grant.

305

306 Conflict of Interest

307 The authors have declared no conflict of interest.

308

309 References

310 Baasandorj, T., Ohm, J. B., & Simsek, S. (2016). Effects of kernel vitreousness and protein level
311 on protein molecular weight distribution, milling quality, and breadmaking quality in
312 hard red spring wheat. Cereal Chemistry, 93, 426-434.
313 Baasandorj, T., Ohm, J.B., & Simsek, S. (2020). Comparison of different experimental
314 breadmaking methods and their associations with flour quality parameters in hard red
315 spring wheat. Cereal Chemistry, 97(2), 515-526.
316 Barrera, G.N., Bustos, M.C., Iturriaga, L., Flores, S.K., León, A.E., & Ribotta, P.D. (2013).
317 Effect of damaged starch on the rheological properties of wheat starch suspensions.
318 Journal of Food Engineering, 116, 233-239.
319 Belton, P.S. (2005). New approaches to study the molecular basis of the mechanical properties of
320 gluten. Journal of Cereal Science,41, 203-211.
321 Biduski, B., Werlang, S., Colussi, R., Pinto, V. Z., Zavareze, E. D. R., Gutkoski, L. C., &
322 Bertolin, T. E. (2022). Starches Properties from Soft, Medium‐ Hard, and Hard Brazilian
323 Wheat upon Annealing. Starch‐ Stärke, 74(9-10), 2100267.
14
324 Brites, C.M., Santos, C.A.L.D., Bagulho, A.S., & Beirão-da-Costa, M.L. (2008). Effect of wheat
325 puroindoline alleles on functional properties of starch. European Food Research and
326 Technology, 226, 1205-1212.
327 Cao, W., Falk, D., & Bock, J.E. (2017). Protein structural features in winter wheat:
328 Benchmarking diversity in Ontario hard and soft winter wheat. Cereal Chemistry, 94, 199-
329 206.
330 Cao, X., Tong, J., Ding, M., Wang, K., Wang, L., Cheng, D., Li, H., Liu, A., Liu, J., Zhao, Z.,
331 Wamg, Z, & Gao, X. (2019). Physicochemical properties of starch in relation to
332 rheological properties of wheat dough (Triticum aestivum L.). Food Chemistry, 297,
333 125000.
334 Clifton, L.A., Lad, M.D., Green, R.J., & Frazier, R.A. (2007). Single amino acid substitutions in
335 puroindoline-b mutants influence lipid binding properties. Biochemistry, 46, 2260-2266.
336 Delcour, J.A., Joye, I.J., Parey, B., Wilderjans, E., Brijs, K., & Lagrain, B. (2012). Wheat gluten
337 functionality as a quality determinant in cereal-based food products. Annual Review of
338 Food Science and Technology, 3, 469-492.
339 Devaux, M.F., de Monredon, F.L.D., Guibert, D., Novales, B., & Abecassis, J. (1998). Particle
340 size distribution of break, sizing and middling wheat flours by laser diffraction. Journal
341 of the Science of Food and Agriculture, 78, 237-244.
342 Dizlek, H., Girard, A.L., & Awika, J.M. (2022). High protein and gliadin content improves
343 tortilla quality of a weak gluten wheat. LWT-Food Science and Technology, 160, 113320.
344 Greenblatt, G.A., Bettge, A.D., & Morris, C.F. (1995). Relationship between endosperm texture
345 and the occurrence of friabilin and bound polar lipids on wheat-starch. Cereal Chemistry,
346 72, 172-176.
347 Gupta, R.B., Khan, K., & Macritchie, F. (1993). Biochemical basis of flour properties in Bread
348 wheats. 1. Effects of variation in the quantity and size distribution of polymeric protein.
349 Journal of Cereal Science, 18, 23-44.
350 Guo, J., Wang, F., Zhang, Z., Wu, D., & Bao, J. (2021). Characterization of gluten proteins in
351 different parts of wheat grain and their effects on the textural quality of steamed bread.
352 Journal of Cereal Science, 102, 103368.
353 Guttieri, M.J., Bowen, D., Gannon, D., O'Brien, K., & Souza, E. (2001). Solvent retention
354 capacities of irrigated soft white spring wheat flours. Crop Science, 41, 1054-1061.
355 Hallfrisch, J., & Behall, K.M. (2000). Mechanisms of the effects of grains on insulin and glucose
356 responses. Journal of the American College of Nutrition, 19, 320S-325S.

15
357 Hammed, A.M., Ozsisli, B., Ohm, J.B., & Simsek, S. (2015). Relationship between solvent
358 retention capacity and protein molecular weight distribution, quality characteristics, and
359 breadmaking functionality of hard red spring wheat flour. Cereal Chemistry, 92, 466-474.
360 Han, N., Fan, J.L., Chen, N., & Chen, H.Q. (2022). Effect of ball milling treatment on the
361 structural, physicochemical and digestive properties of wheat starch, A-and B-type starch
362 granules. Journal of Cereal Science, 104, 103439.
363 Issarny, C., Cao, W., Falk, D., Seetharaman, K., & Bock, J.E. (2017). Exploring functionality of
364 hard and soft wheat flour blends for improved end‐ use quality prediction. Cereal
365 Chemistry, 94, 723-732.
366 Jazaeri, S., Bock, J.E., Bagagli, M.P., Iametti, S., Bonomi, F., & Seetharaman, K. (2015).
367 Structural modifications of gluten proteins in strong and weak wheat dough during
368 mixing. Cereal Chemistry, 92, 105-113.
369 Katyal, M., Virdi, A.S., Kaur, A., Singh, N., Kaur, S., Ahlawat, A.K., & Singh, A.M. (2016).
370 Diversity in quality traits amongst Indian wheat varieties I: flour and protein
371 characteristics. Food Chemistry, 194, 337-344.
372 Katyal, M., Singh, N., Virdi, A.S., Kaur, A., Chopra, N., Ahlawat, A.K., & Singh, A.M. (2017).
373 Extraordinarily soft, medium-hard and hard Indian wheat varieties: composition, protein
374 profile, dough and baking properties. Food Research International, 100, 306-317.
375 Katyal, M., Virdi, A.S., Singh, N., Chopra, N., Kaur, A., Ahlawat, A.K., & Singh, A.M. (2018).
376 Fractionation and grain hardness effect on protein profiling, pasting and rheological
377 properties of flours from medium-hard and extraordinarily soft wheat varieties. Journal
378 of Food Science and Technology, 55, 4661-4674.
379 Katyal, M., Singh, N., Chopra, N., & Kaur, A. (2019). Hard, medium-hard and extraordinarily
380 soft wheat varieties: Comparison and relationship between various starch properties.
381 International Journal of Biological Macromolecules, 123, 1143-1149.
382 Kaur, A., Shevkani, K., Katyal, M., Singh, N., Ahlawat, A.K. & Singh, A.M. (2016).
383 Physicochemical and rheological properties of starch and flour from diff erent durum
384 wheat varieties and their relationships with noodle quality. Journal of Food Science and
385 Technology, 53, 2127-2138
386 Kaur, A., Singh, N., Ahlawat, A.K., Kaur, S., Singh, A.M., Chauhan, H., & Singh, G.P. (2013).
387 Diversity in grain, flour, dough and gluten properties amongst Indian wheat cultivars
388 varying in high molecular weight subunits (HMW-GS). Food Research International, 53,
389 63-72.
390 Kaur, A., Singh, N., Kaur, S., Ahlawat, A.K., & Singh, A.M. (2014). Relationships of flour
391 solvent retention capacity, secondary structure and rheological properties with the cookie
392 making characteristics of wheat cultivars. Food Chemistry, 158, 48-55.
16
393 Kim, K.H., Feiz, L., Martin, J.M., & Giroux, M.J. (2012). Puroindolines are associated with
394 decreased polar lipid breakdown during wheat seed development. Journal of Cereal
395 Science, 56, 142-146.
396 Ktenioudaki, A., Butler, F., & Gallagher, E. (2010). Rheological properties and baking quality of
397 wheat varieties from various geographical regions. Journal of Cereal Science, 51, 402-
398 408.
399 Kumar, R., Kumar, A., Sharma, N.K., Kaur, N., Chunduri, V., Chawla, M., Sharma, S., Singh,
400 K., & Garg, M. (2016). Soft and hard textured wheat differ in starch properties as
401 indicated by trimodal distribution, morphology, thermal and crystalline properties.
402 PloSone, 11, e0147622.
403 Kweon, M., Slade, L., & Levine, H. (2011). Solvent retention capacity (SRC) testing of wheat
404 flour: Principles and value in predicting flour functionality in different wheat‐ based food
405 processes and in wheat breeding-A review. Cereal Chemistry, 88, 537-552.
406 Labuschagne, M.T., Koen, E., & Dessalegn, T. (2004). Use of size exclusion high-performance
407 liquid chromatography for wheat quality prediction in Ethiopia. Cereal Chemistry, 81, 533-
408 537.
409 León, A.E., Barrera, G.N., Pérez, G.T., Ribotta, P.D., & Rosell, C.M. (2006). Effect of damaged
410 starch levels on flour-thermal behaviour and bread staling. European Food Research and
411 Technology, 224, 187-192.
412 Li, W., Shan, Y., Xiao, X., Luo, Q., Zheng, J., Ouyang, S., & Zhang, G. (2013). Physicochemical
413 properties of A-and B-starch granules isolated from hard red and soft red winter wheat.
414 Journal of Agricultural and Food Chemistry, 61, 6477-6484.
415 Lu, Z., & Seetharaman, K. (2014). Suitability of Ontario‐ grown hard and soft wheat flour
416 blends for noodle making. Cereal Chemistry, 91, 482-488.
417 Malalgoda, M., Ohm, J.B., Meinhardt, S., Chao, S., & Simsek, S. (2017). Cluster analysis of
418 historical and modern hard red spring wheat cultivars based on parentage and HPLC
419 analysis of gluten‐ forming proteins. Cereal Chemistry, 94, 560-567.
420 Marti, A., Augst, E., Cox, S., & Koehler, P. (2015). Correlations between gluten aggregation
421 properties defined by the GlutoPeak test and content of quality-related protein fractions
422 of winter wheat flour. Journal of Cereal Science, 66, 89-95.
423 Mohler, V., Schmolke, M., Paladey, E., Seling, S., & Hartl, L. (2012). Association analysis of
424 Puroindoline-D1 and Puroindoline b-2 loci with 13 quality traits in European winter
425 wheat (Triticum aestivum L.). Journal of Cereal Science, 56, 623-628.

17
426 Mondal, S., Hays, D.B., Alviola, N.J., Mason, R.E., Tilley, M., Waniska, R.D., Bean, S.R., &
427 Glover, K. D. (2009). Functionality of gliadin proteins in wheat flour tortillas. Journal of
428 Agricultural and Food Chemistry, 57, 1600-1605.
429 Morel, M.H., Pincemaille, J., Chaveau, E., Louhichi, A., Voilleau, F., Menut, P., Ramos, L., &
430 Banc, A. (2020). Insight into gluten structure in a mild chaotropic solvent by asymmetrical
431 flow field flow fractionation (AsF1FFF). Food Hydrocolloids,103, 105676
432 Morris, C.F., Luna, J., & Caffe‐ Treml, M. (2021). The vromindolines of cv. Hayden oat (Avena
433 sativa L.)–A review of the Poeae and Triticeae indolines and a suggested system for
434 harmonisation of nomenclature. Journal of Cereal Science, 97, 103135.
435 Nadolska-Orczyk, A., Gasparis, S., & Orczyk, W. (2009). The determinants of grain texture in
436 cereals. Journal of Applied Genetics, 50, 185-197.
437 Quayson, E.T., Marti, A., Bonomi, F., Atwell, W., & Seetharaman, K. (2016a). Structural
438 modification of gluten proteins in strong and weak wheat dough as affected by mixing
439 temperature. Cereal Chemistry, 93, 189-195.
440 Quayson, E.T., Atwell, W., Morris, C.F., & Marti, A. (2016b). Empirical rheology and pasting
441 properties of soft-textured durum wheat (Triticum turgidum ssp. durum) and hard-
442 textured common wheat (T. aestivum). Journal of Cereal Science, 69, 252-258.
443 Schopf, M., Wehrli, M.C., Becker, T., Jekle, M., & Scherf, K.A. (2021). Fundamental
444 characterization of wheat gluten. European Food Research and Technology, 247, 985-
445 997.
446 Shang, J., Li, L., Liu, C., Hong, J., Liu, M., Zhao, B., & Zheng, X. (2021). Relationships of flour
447 characteristics with isolated starch properties in different Chinese wheat varieties. Journal
448 of Cereal Science, 99, 103210.
449 Shang, J., Li, L., Zhao, B., Liu, M., & Zheng, X. (2020). Comparative studies on
450 physicochemical properties of total, A-and B-type starch from soft and hard wheat
451 varieties. International Journal of Biological Macromolecules, 154, 714-723.
452 Sharma, S., Katyal, M., Singh, N., Singh, A.M., & Ahlawat, A.K. (2022). Comparison of effect
453 of using hard and soft wheat on the high molecular weight-glutenin subunits profile and the
454 quality of produced cookie. Journal of Food Science and Technology, 59, 2545-2561.
455 Shevkani, K., Singh, N., Singh, S., Ahlawat, A.K. & Singh, A.M. (2011). Relationship between
456 physicochemical and rheological properties of starches from Indian wheat lines.
457 International Journal of Food Science and Technology, 46, 2584-2590.
458 Shevkani, K., Singh, N., Bajaj, R., & Kaur, A. (2017). Wheat starch production, structure,
459 functionality and applications-a review. International Journal of Food Science and
460 Technology, 52, 38-58.

18
461 Shevkani, K., Singh, N., Chen, Y., Kaur, A., & Yu, L. (2019). Pulse proteins: secondary
462 structure, functionality and applications. Journal of Food Science and Technology, 56,
463 2787-2798.
464 Shevkani, K., Singh, N., Isono, N., & Noda, T. (2021). Structural and functional properties of
465 amaranth starches from residue obtained during protein extraction. Journal of Food
466 Measurement and Characterization, 15, 5087-5096.
467 Shewry, P. (2023). Wheat grain proteins: Past, present, and future. Cereal chemistry, 100, 9-22.
468 Shewry, P.R., & Lafiandra, D. (2022). Wheat glutenin polymers 1. Structure, assembly and
469 properties. Journal of Cereal Science,106, 104386
470 Shewry, P.R., & Halford, N.G. (2002). Cereal seed storage proteins: structures, properties and
471 role in grain utilization. Journal of Experimental Botany, 53, 947-958.
472 Singh, M., & Khatkar, B.S. (2005). Structural and functional properties of wheat storage
473 proteins: a review. Journal of Food Science and Technology, 42, 455-471.
474 Singh, N., Gujral, H.S., Katyal, M., & Sharma, B. (2019). Relationship of Mixolab
475 characteristics with protein, pasting, dynamic and empirical rheological characteristics of
476 flours from Indian wheat varieties with diverse grain hardness. Journal of Food Science
477 and Technology, 56, 2679-2686.
478 Singh, N., Kaur, N., Katyal, M., Kaur, A., & Shevkani, K. (2017). Characteristics of starch
479 separated from coarse and fine flour fractions obtained from hard, medium‐ hard, and
480 soft Indian wheat cultivars. Starch‐ Stärke, 69, 1600012.
481 Singh, N., Shevkani, K., Kaur, A., Thakur, S., Parmar, N., & Singh Virdi, A. (2014).
482 Characteristics of starch obtained at different stages of purification during commercial wet
483 milling of maize. Starch‐ Stärke, 66, 668-677.
484 Singh, N., Virdi, A.S., Katyal, M., Kaur, A., Kaur, D., Ahlawat, A.K., Singh, A.M., & Sharma,
485 R.K. (2021). Evaluation of heat stress through delayed sowing on physicochemical and
486 functional characteristics of grains, whole meals and flours of India wheat. Food
487 Chemistry, 344, 128725.
488 Tatham, A.S., Hayes, L., Shewry, P.R., & Urry, D.W. (2001). Wheat proteins exhibit a complex
489 mechanism of protein elasticity. Biochemica et Biophysica Acta, 1548, 187–193.
490 Tao, H., Huang, J.S., Xie, Q.T., Zou, Y.M., Wang, H.L., Wu, X.Y., & Xu, X.M. (2018). Effect
491 of multiple freezing-thawing cycles on structural and functional properties of starch
492 granules isolated from soft and hard wheat. Food Chemistry, 265, 18-22.
493 Tester, R.F., & Karkalas, J. (2001). The effects of environmental conditions on the structural
494 features and physico‐ chemical properties of starches. Starch‐ Stärke, 53(10), 513-519.

19
495 Tian, J., Hu, R., Deng, Z., & Wang, Y. (2007). The variation and stability analysis of wheat
496 dough stability time. Agricultural Sciences in China, 6, 143-149.
497 Tsilo, T.J., Ohm, J.B., Hareland, G.A., & Anderson, J.A. (2010). Association of size‐ exclusion
498 HPLC of endosperm proteins with dough mixing and breadmaking characteristics in a
499 recombinant inbred population of hard red spring wheat. Cereal Chemistry, 87, 104-111.
500 Wang, P., Xu, L., Nikoo, M., Ocen, D., Wu, F., Yang, N., Jin, Z., & Xu, X. (2014). Effect of
501 frozen storage on the conformational, thermal and microscopic properties of gluten:
502 Comparative studies on gluten-, glutenin-and gliadin-rich fractions. Food Hydrocolloids,
503 35, 238-246.
504 Wrigley, C.W., Békés, F., Bushuk, W. (2006). Gluten: A balance of gliadin and glutenin. The
505 unique balance of wheat quality. C. Wrigley, F. Békés and W. Bushuk, eds. AACC
506 International, St. Paul, MN 2006, 3-34.
507 Wu, F., Li, J., Yang, N., Chen, Y., Jin, Y., & Xu, X. (2018). The roles of starch structures in the
508 pasting properties of wheat starch with different degrees of damage. Starch‐ Stärke, 70,
509 1700190.
510 Xiao, Z.S., Park, S.H., Chung, O.K., Caley, M.S., & Seib, P.A. (2006). Solvent retention
511 capacity values in relation to hard winter wheat and flour properties and straight‐ dough
512 breadmaking quality. Cereal Chemistry, 83, 465-471.
513
514
515
516

20
517 Legends to figures and tables
518
519 Figure 1. Size-exclusion chromatograms of hard and soft wheat (HW and SW, respectively)
520 flours (Source: Katyal et al., 2017)
521 Figure 2. Electrophoretic profile of reduced and alkylated glutenins of flours from soft
522 (QBP12-9, QBP12-10, QBP12-8 and QBP12-11), medium-hard (HD2967, HI977, UP2672 and
523 DBW39) and hard (K307, HD2733, DPW621-50 and HD2932) wheat varieties (Source: Katyal
524 et al., 2017)
525 Figure 3. Farinograms of flours from different soft and hard Indian wheat varieties (Source:
526 Katyal et al., 2017)
527 Table 1. Physicochemical, rheological (farinographic), chromatographic, solvent retention and
528 secondary structural properties of soft and hard wheat flours
529
530

21
531

532
533 Figure 1. Size-exclusion chromatograms of hard and soft wheat (HW and SW, respectively)
534 flours (Source: Katyal et al., 2017)

535

536
537 Figure 2. Electrophoretic profile of reduced and alkylated glutenins of flours from soft
538 (QBP12-9, QBP12-10, QBP12-8 and QBP12-11), medium-hard (HD2967, HI977, UP2672 and
539 DBW39) and hard (K307, HD2733, DPW621-50 and HD2932) wheat varieties; HMW: high
540 molecular weight; LMW: low molecular weight (Source: Katyal et al., 2017)

22
541

542

543

544 Soft wheat (GHI=29) Soft wheat (GHI=19)

545

546

547 Hard wheat (GHI=89) Hard wheat (GHI=73)

548

549 Figure 3. Farinograms of flours from different soft and hard Indian wheat varieties; GHI: grain
550 hardness index (Source: Katyal et al. 2017)

551

552
553
554
23
555

556 Table 1. Physicochemical, rheological (farinographic), chromatographic, solvent retention

557 and secondary structural properties of soft and hard wheat flours

Soft Hard
Parameter Source(s)
wheat wheat
Katyal et al. (2017); Katyal et al.,
Grain hardness index 17-35 71-95
(2018); Shang et al. (2020)
Katyal et al. (2017); Issarny et al.
Protein content (%) 7-10.5 10-13
(2017)
Damaged starch content (%) 2-3 7-10 Singh et al. (2019)
Farinographic analysis
Water absorption (%) 52-56 58-65 Cao et al. (2017); Hammed et al.
Dough development time (2015); Issarny et al. (2017); Lu
Up to 1.4 1.5-9
(min) and Seetharaman (2014);
Dough stability (min) Up to 1.3 7-17 Malalgoda et al. (2017)

Chromatographic characterization of proteins

Un-extractable polymeric
42 50-62
proteins (%)
Un-extractable monomeric
58 38-50
proteins (%)
Katyal et al. [2]; Gupta et al. [44]
Extractable polymeric proteins
27 33-46
(%)
Extractable monomeric
73 54-67
proteins (%)
Solvent retention capacity
Lactic acid (%) 87-102 105-178 Issarny et al. (2017); Cao et al.
Sucrose (%) 74-111 99-135 (2017); Hammed et al. (2015);
Sodium bicarbonate (%) 68-76 79-98 Xiao et al. (2006); Katyal et al.
(2018)
Water (%) 58-60 65-81
Protein secondary structure
α-helix (%) 3-9 3-10
Intermolecular β-sheets (%) 19 15 Cao et al. (2016); Katyal et al.
β-sheets (%) 32-71 37-71 (2017)
β-turns (%) 7-42 8-39
558 Values shown have been subjected to rounding-off.

24