Physical grain characteristics, composition, and malting quality of CDC Copeland containing a high percentage of an undesignated DNA profile

Objective

The aim of this project was to determine if the presence of an undesignated DNA profile in CDC Copeland barley affected physical grain characteristics, composition, malting performance, and malting quality.

Materials and methods

Four samples of CDC Copeland barley, grown in 2022 in Saskatchewan, which had a relatively high number of kernels with an undesignated DNA profile were collected for this project (Table 1). Two other sets of CDC Copeland samples were selected from the Grain Research Laboratory’s 2022 Harvest Survey of Malting Barley Program and from the Cargo Monitoring Program (Table 1). DNA was extracted from 144 randomly selected kernels from each CDC Copeland sample and the genetic makeup was determined by comparing the amplified simple sequence repeat (SSR) markers with those in the Grain Research Laboratory’s reference catalogue for barley varieties registered in Canada. An undesignated DNA profile was found and quantified in several CDC Copeland barley samples (Table 1).

Samples were tested for moisture and protein by near-infrared technology (Infratec 1241, Foss). Germination energy was measured following the ASBC method for barley germination. Test weight was determined according to the method outlined in the Canadian Grain Commission’s Official Grain Grading Guide using a 0.125-litre cup due to limited sample size. Samples were scanned using a Cgrain Value™ seed analyzer (Cgrain AB, Uppsala, Sweden) to measure several physical grain characteristics including kernel weight and kernel dimensions (length, width). Kernel dimensions were measured in pixels and transformed into millimetres by the instrument software. Other physical parameters were derived through calculations by the Cgrain software. Barley was malted in an automated Phoenix Micromalting System and malt quality analysis was conducted according to the ASBC Official Methods of Analysis.

Table 1  CDC Copeland malting barley samples used in this study.
Sample type Province of origin Kernels with undesignated DNA profile (%)
Samples with a high % of the undesignated DNA profile
Sample 1 SK 16
Sample 2 SK 22
Sample 3 SK 17
Sample 4 SK 20
2022 Harvest Survey of Malting Barley Program
Harvest 1 AB 3
Harvest 2 SK 4
Harvest 3 SK 3
Harvest 4 AB/SK 3
Harvest 5 SK 1
Harvest 6 AB 1
Harvest 7 SK 5
Harvest 8 MB/SK 4
2022 Cargo Monitoring Program
Cargo 1 unknown origin 3
Cargo 2 unknown origin 4
Cargo 3 unknown origin 0
Cargo 4 unknown origin 5
Cargo 5 unknown origin 3
Cargo 6 unknown origin 1
Cargo 7 unknown origin 3
Cargo 8 unknown origin 1

Results

Physical characteristics and composition of CDC Copeland samples

  1. Four CDC Copeland samples, grown in Saskatchewan in 2022 and selected for this project, contained a significantly higher percentage (16 to 22%) of the undesignated DNA profile compared to CDC Copeland samples collected from the 2022 Harvest Survey of Malting Barley Program (1 to 5% of the undesignated DNA) and from the Cargo Monitoring Program (0 to 5% of the undesignated DNA) (Table 1).
  2. The commonly measured physical grain characteristics, including thousand kernel weight, plumpness, and test weight, of CDC Copeland samples with a high percentage of the undesignated DNA profile did not differ from the Harvest Survey of Malting Barley Program and Cargo Monitoring Program samples of CDC Copeland as shown in Figure 1. One exception was Sample 3, which was characterized by a slightly lower kernel weight compared to the other samples.
  3. The protein content of CDC Copeland samples with a high percentage of the undesignated DNA profile ranged from 11.6 to 12.8% and fell within the range of protein content observed for the Harvest Survey of Malting Barley Program and Cargo Monitoring Program samples of CDC Copeland (Figure 2).
  4. All four CDC Copeland samples with a high percentage of the undesignated DNA profile exhibited the required germination energy of ≥ 95%. Their germination rates, determined by the germination index, was also within the range observed for the Harvest Survey of Malting Barley Program and Cargo Monitoring Program samples of CDC Copeland (Figure 2).
  5. Supplementary physical grain characteristics, including kernel weight, dimensions, and colour, were obtained with an imagine analyzer (Cgrain ValueTM). Several other parameters, such as kernel density, volume, and roundness, were derived from the measured physical characteristics (Table 2). None of the four CDC Copeland samples with a high percentage of the undesignated DNA profile exhibited unusual or unique physical parameters that would distinguish their kernels from those of the Harvest Survey of Malting Barley Program or Cargo Monitoring Program samples of CDC Copeland. The average kernel weight of Sample 3 was slightly lower compared to the other samples, but the difference was not statistically significant given a high degree of variability within each sample as indicated by relatively high standard deviation values (Table 2).

Malting and malt quality

  1. All barley samples were malted using a Phoenix Micromalting System under identical conditions of steeping, germination and kilning. The parameters collected during malting included grain moisture after steeping and germination, malt yield, and roots yield. The results presented in Figure 3 do not indicate any unusual processing performance of the four CDC Copeland samples with a high percentage of the undesignated DNA profile.
  2. The concentration of proteins in malt and the levels of starch degrading enzymes, as measured by diastatic power and α-amylase levels, for the four CDC Copeland samples with a high percentage of the undesignated DNA profile were generally within the range of values observed for the Harvest Survey of Malting Barley Program and Cargo Monitoring Program samples of CDC Copeland (Figure 4). One exception was Sample 3, which exhibited slightly lower level of α- amylase compared to other samples.
  3. The level of malt extract and the concentration of β-glucans in the wort of the CDC Copeland samples with a high percentage of the undesignated DNA profile were within the range of values observed for the Harvest Survey of Malting Barley Program and Cargo Monitoring Program samples of CDC Copeland (Figure 5).
  4. Sample 3 exhibited somewhat higher concentrations of soluble proteins and free amino nitrogen compounds in the wort compared to Samples 1, 2, and 4, and compared to the Harvest Survey of Malting Barley Program and Cargo Monitoring Program samples of CDC Copeland (Figure 6). These results point to a higher activity of proteolytic enzymes in the malt of Sample 3. A higher wort viscosity (Figure 5) and a darker colour of wort (Figure 6) of Sample 3 were likely associated with the higher concentration of proteins and free amino nitrogen content in the wort of Sample 3. The origin of a relatively higher proteolytic activity of malt of Sample 3 cannot be fully explained at this point. One plausible explanation could be related to a different and/or higher microbial load on Sample 3 barley. The presence of the undesignated DNA profile in Sample 3 is not likely to be associated with the higher activity of proteolytic enzymes in its malt since this sample did not contain the highest percentage of this DNA material.

Conclusion

The results of this study indicated that physical grain characteristics, composition, and malting quality of four samples of CDC Copeland barley containing a relatively high percentage of the undesignated DNA profile (16 to 22%) did not differ from the properties exhibited by CDC Copeland barley with much lower percentages (0 to 5%) of this DNA material.

Figure 1  Physical grain characteristics of Samples 1 to 4 (with a high percentage of the undesignated DNA profile) in comparison to the Harvest Survey of Malting Barley Program and Cargo Monitoring Program samples of CDC Copeland.
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Thousand kernel weight (g)
Samples Harvest Cargos
45.4 44.0 43.8
45.4 46.9 46.2
42.2 44.4 44.3
46.8 45.3 43.1
N/AFootnote 1 45.4 44.4
N/A 45.6 46.7
N/A 43.9 44.2
N/A 43.7 42.5
Plump, >6/64 sieve (%)
Samples Harvest Cargos
92.9 94.1 90.6
92.3 96.0 93.4
90.9 95.7 92.3
95.2 95.0 92.1
N/A 96.6 90.7
N/A 95.3 93.4
N/A 93.7 90.7
N/A 94.5 89.8
Test weight (kg/hL)
Samples Harvest Cargos
67.8 67.7 71.2
67.6 68.3 72.0
66.1 66.5 70.9
69.5 66.0 71.4
N/A 66.9 71.3
N/A 68.7 71.9
N/A 67.1 70.8
N/A 68.2 71.0
Figure 2  Protein content, germination energy, and germination index of barley grain of Samples 1 to 4 (with a high percentage of the undesignated DNA profile) in comparison to the Harvest Survey of Malting Barley Program and Cargo Monitoring Program samples of CDC Copeland.
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Protein (%, db)
Samples Harvest Cargos
12.3 11.9 12.9
12.8 11.3 12.1
11.6 11.5 12.1
11.8 11.7 12.5
N/A 12.1 12.3
N/A 11.7 12.0
N/A 11.9 12.6
N/A 11.6 12.1
Germination engergy, 4ml (%)
Samples Harvest Cargos
98 100 98
99 100 99
97 100 99
98 98 97
N/A 99 99
N/A 100 96
N/A 99 98
N/A 99 99
Germination index, 4ml
Samples Harvest Cargos
8.1 7.1 8.2
8.2 6.7 7.9
7.7 7.4 7.7
7.5 7.4 8.0
N/A 7.2 8.7
N/A 7.0 8.0
N/A 7.3 8.4
N/A 7.1 8.0
Table 2  Germination energy, RVA viscosity, and supplementary physical grain characteristics of CDC Copeland barley samples.
Sample type Province of origin Kernels with undesignated DNA profile (%) Germination energy 4 mL (%) Final viscosity (RVU) Supplementary grain characteristics
Kernel weight (mg) Kernel density (g/cm)
 
Kernel area (mm2) Kernel length (mm) Kernel width (mm) Kernel thickness (mm) Kernel volume (mm3) Kernel roundness Kernel lightnessFootnote 2
Mean SDFootnote 3 Mean SD Mean SD Mean SD Mean SD Mean SD Mean SD
Samples with a high % of the undesignated DNA profile
Sample 1 SK 16 92 160 46.4 9.6 1.186 18.9 3.4 8.41 0.84 3.61 0.32 2.80 0.31 39.1 8.1 0.5579 0.0461 0.543
Sample 2 SK 22 89 159 44.6 9.6 1.166 18.9 3.4 8.37 0.82 3.59 0.34 2.76 0.33 38.2 8.2 0.5547 0.0503 0.542
Sample 3 SK 17 85 152 42.0 9.3 1.153 18.3 3.3 8.31 0.78 3.51 0.34 2.68 0.33 36.4 8.0 0.5490 0.0487 0.535
Sample 4 SK 20 90 151 46.2 10.1 1.197 18.7 3.6 8.24 0.95 3.64 0.34 2.79 0.31 38.6 8.4 0.5644 0.0558 0.544
2022 Harvest Survey of Malting Barley Program
Harvest 1 AB 3 100 158 44.8 8.9 1.166 19.1 3.2 8.73 0.84 3.53 0.29 2.76 0.28 38.3 7.5 0.5469 0.0432 0.673
Harvest 2 SK 4 94 155 46.0 9.4 1.168 19.3 3.5 8.60 0.93 3.65 0.33 2.80 0.31 39.4 7.9 0.5526 0.0536 0.577
Harvest 3 SK 3 90 153 45.1 8.0 1.145 19.2 3.2 8.68 0.80 3.65 0.30 2.79 0.27 39.3 7.0 0.5474 0.0431 0.560
Harvest 4 AB/SK 3 90 99 45.8 9.0 1.145 19.6 3.5 8.81 0.92 3.63 0.29 2.81 0.28 40.0 7.8 0.5469 0.0446 0.570
Harvest 5 SK 1 96 146 45.4 8.7 1.152 19.6 3.4 8.73 1.00 3.57 0.26 2.80 0.25 39.4 7.5 0.5509 0.0492 0.596
Harvest 6 AB 1 98 121 46.5 9.4 1.180 19.3 3.5 8.73 0.97 3.61 0.29 2.80 0.28 39.4 8.0 0.5491 0.0443 0.628
Harvest 7 SK 5 93 98 45.2 8.6 1.155 19.3 3.4 8.65 0.78 3.61 0.31 2.78 0.29 39.1 7.4 0.5481 0.0423 0.535
Harvest 8 MB/SK 4 90 156 43.7 8.8 1.170 18.4 3.4 8.36 0.90 3.60 0.31 2.77 0.29 37.4 7.5 0.5581 0.0484 0.568
2022 Cargo Monitoring Program
Cargo 1 unkFootnote 4 3 88 93 41.9 10.0 1.173 17.9 3.4 8.12 0.74 3.47 0.37 2.66 0.34 35.5 8.3 0.5542 0.0450 0.583
Cargo 2 unk. 4 92 121 46.7 9.9 1.217 18.7 3.4 8.27 0.69 3.60 0.34 2.75 0.32 38.4 8.1 0.5572 0.0423 0.596
Cargo 3 unk. 0 91 86 45.5 10.0 1.210 18.6 3.3 8.22 0.72 3.56 0.34 2.73 0.32 37.6 8.2 0.5574 0.0452 0.579
Cargo 4 unk. 5 87 45 43.8 9.7 1.207 17.9 3.4 8.09 0.66 3.52 0.37 2.68 0.33 36.3 8.0 0.5566 0.0448 0.576
Cargo 5 unk. 3 90 125 43.4 10.3 1.202 17.9 3.4 8.08 0.70 3.51 0.37 2.67 0.35 36.1 8.4 0.5553 0.0461 0.605
Cargo 6 unk. 1 88 97 44.6 11.0 1.204 18.3 3.4 8.12 0.70 3.54 0.37 2.70 0.36 37.1 9.1 0.5567 0.0478 0.577
Cargo 7 unk. 3 92 115 43.7 9.8 1.195 18.0 3.3 8.10 0.73 3.52 0.37 2.71 0.34 36.6 8.2 0.5605 0.0489 0.580
Cargo 8 unk. 1 70 89 43.9 10.0 1.203 18.0 3.4 8.04 0.70 3.52 0.36 2.71 0.34 36.5 8.3 0.5610 0.0470 0.576
Figure 3  Malting performance parameters of Samples 1 to 4 (with a high percentage of the undesignated DNA profile) in comparison to the Harvest Survey of Malting Barley Program and Cargo Monitoring Program samples of CDC Copeland.
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Steep-out moisture (%)
Samples Harvest Cargos
45.1 43.6 46.0
45.5 44.6 45.7
46.2 45.0 46.2
45.4 46.5 45.4
N/A 45.7 45.3
N/A 43.9 46.5
N/A 46.5 46.8
N/A 44.4 45.0
Germ-out moisture (%)
Samples Harvest Cargos
36.8 35.6 37.3
36.9 36.3 37.3
38.1 35.3 36.6
37.0 38.0 36.8
N/A 37.3 36.6
N/A 34.4 37.4
N/A 37.5 38.0
N/A 37.1 36.1
Malt yield (%)
Samples Harvest Cargos
90.7 91.3 90.4
90.1 91.0 90.1
89.4 90.6 91.2
89.6 89.1 90.1
N/A 89.6 89.2
N/A 92.1 89.2
N/A 90.2 88.7
N/A 89.6 91.6
Roots (%)
Samples Harvest Cargos
3.5 3.6 4.1
3.7 3.3 3.8
3.8 3.4 3.1
3.6 4.2 3.5
N/A 4.2 3.8
N/A 2.3 4.5
N/A 3.7 4.5
N/A 3.6 2.6
Figure 4  Malt protein content and enzyme levels in Samples 1 to 4 (with a high percentage of the undesignated DNA profile) in comparison to the Harvest Survey and Cargo Monitoring samples of CDC Copeland.
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Malt protein (%, db)
Samples Harvest Cargos
12.2 12.2 12.6
12.5 11.2 12.2
11.7 11.4 12.1
11.8 12.0 12.1
N/A 12.2 11.8
N/A 11.8 12.5
N/A 11.7 12.0
N/A 11.7 12.0
Diastatic power (°, db)
Samples Harvest Cargos
133.0 139.8 146.9
140.7 129.2 139.6
134.1 134.4 143.6
114.3 143.7 147.7
N/A 143.3 137.7
N/A 131.8 150.0
N/A 148.6 145.9
N/A 140.2 138.0
Alpha-amylase (DU, db)
Samples Harvest Cargos
54.3 56.6 58.5
56.4 58.3 55.7
51.8 56.2 55.1
57.6 57.8 59.0
N/A 60.0 54.1
N/A 57.5 56.7
N/A 62.0 59.7
N/A 60.4 59.4
Figure 5  Malt extract, concentration of β-glucans in wort, and wort viscosity of the Samples 1 to 4 (with high percentage of the undesignated DNA profile) in comparison to the Harvest Survey and Cargo Monitoring samples of CDC Copeland.
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Fine extract (%, db)
Samples Harvest Cargos
80.4 79.8 79.8
79.9 81.0 80.2
80.4 80.9 80.5
80.7 80.0 80.3
N/A 79.5 80.7
N/A 80.2 80.2
N/A 79.9 80.1
N/A 80.7 80.7
β-Glucan (mg/L)
Samples Harvest Cargos
92.6 75.9 67.4
89.6 83.3 52.5
47.2 58.8 62.1
73.8 43.6 64.7
N/A 45.7 46.9
N/A 114.4 47.8
N/A 45.8 44.4
N/A 57.5 111.0
Viscosity (cP)
Samples Harvest Cargos
1.46 1.45 1.45
1.45 1.46 1.44
1.57 1.43 1.46
1.47 1.42 1.45
N/A 1.43 1.46
N/A 1.45 1.45
N/A 1.42 1.46
N/A 1.44 1.46
Figure 6  Concentration of soluble proteins and free amino nitrogen compounds (FAN) in wort and wort color Samples 1 to 4 (with a high percentage of the undesignated DNA profile) in comparison to the Harvest Survey and Cargo Monitoring samples of CDC Copeland.
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Soluble protein (%, db)
Samples Harvest Cargos
4.99 4.08 4.59
4.82 4.30 4.86
5.42 4.90 4.60
4.69 4.58 4.52
N/A 4.15 4.70
N/A 4.02 4.63
N/A 4.37 4.85
N/A 4.80 4.28
FAN (mg/L)
Samples Harvest Cargos
188.6 136.1 165.2
180.6 155.8 179.3
227.9 197.3 169.6
180.6 176.8 165.1
N/A 153.2 179.6
N/A 136.3 169.8
N/A 160.9 186.5
N/A 193.7 158.0
Wort colour (°)
Samples Harvest Cargos
3.3 1.5 2.1
2.9 1.9 2.8
4.6 3.4 2.2
2.8 2.6 2.0
N/A 1.7 2.5
N/A 1.5 2.2
N/A 2.2 2.7
N/A 3.5 1.9

Acknowledgments

The DNA analysis was performed by staff of the Grain Biotechnology program and the Microbiology and Grain Genomics program at the Grain Research Laboratory under the supervision of Dr. Tigst Demeke and Dr. Sean Walkowiak.