Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (2): 239-247.doi: 10.3864/j.issn.0578-1752.2021.02.001

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Improvement and Application of the Method for Determining Yellow Pigment Content in Wheat Grain

ZHAI ShengNan(),LIU AiFeng,LI FaJi,LIU Cheng,GUO Jun,HAN Ran,ZI Yan,WANG XiaoLu,LÜ YingYing,LIU JianJun()   

  1. Crop Research Institute, Shandong Academy of Agricultural Sciences, Jinan 250100
  • Received:2020-04-27 Accepted:2020-07-06 Online:2021-01-16 Published:2021-02-03
  • Contact: JianJun LIU E-mail:zsn19870322@163.com;ljjsaas@163.com

Abstract:

【Objective】The yellow pigment content (YPC) in wheat grain is an important factor affecting the color and nutritional quality of flour end-use products. To quickly, accurately and simply detect YPC in wheat grain, the traditional method for YPC determination, AACC 14-50, was improved, providing a technical support for genetic improvement of the color and nutritional quality of flour end-use products.【Method】In this study, improvements have been made to the AACC 14-50 method using a Multiskan Spectrum microplate reader. The accuracy and stability of the modified method were analyzed, and the influence of extraction time and preservation time on flour YPC was dissected. The YPC of 283 domestic and foreign wheat varieties were determined by the improved method to further verify its effectiveness and explore excellent resources for genetic improvement of the color and nutritional quality of flour end-use products. 【Result】 The results showed that YPC determined by the modified method was significantly and positively correlated with those by the traditional AACC 14-50 method, with a correlation coefficient of 0.994 (P<0.001), and analysis of variance showed that YPC measured by the two methods was significant among varieties. The YPC of 144 wheat varieties were measured repeatedly using the modified method, and the correlation coefficients of YPC among three replicates were 0.997, 0.998 and 0.998, respectively. The flour YPC of Gaoyou 5218, Jishi 02-2 and Zhengmai 366 reached the maximum value (0.93, 1.24 and 1.53 μg·g-1) at the extraction time of 30 min, and remained stable during 30-120 min (0.94-0.97 μg·g-1, 1.30-1.33 μg·g-1 and 1.59-1.62 μg·g-1). No significant changes were observed in the flour YPC of Gaoyou 5218, Jishi 02-2 and Zhengmai 366 stored at room temperature for 0-20 days (0.93-0.97 μg·g-1, 1.29-1.33 μg·g-1 and 1.60-1.64 μg·g-1; P>0.05). A wide range of YPC variation was identified in 283 domestic and foreign wheat varieties, with correlation coefficients ranging from 0.73 to 0.98 among different environments. Briefly, the mean YPC of varieties in the Huang-Huai wheat region was 1.15 μg·g-1, ranging between 0.51-2.42 μg·g-1. The average value of YPC in varieties from the Northern winter wheat region was 1.57 μg·g-1, ranging from 0.90 to 2.52 μg·g-1. The average value of YPC in the Middle-Lower reaches of the Yangtze River wheat region was 1.07 μg·g-1, and the variation ranged from 0.56 to 2.54 μg·g-1. The average YPC of foreign varieties was 1.61 μg·g-1, ranging from 0.94 to 2.48 μg·g-1. Twenty-four varieties with lower YPC and 26 with higher YPC were identified, which can be used as parents in wheat breeding programs to improve the color and nutritional quality of flour end-use products. Compared with the traditional AACC 14-50 method, the modified one reduces the workload, decreases the detection time, and increases the detection efficiency by about 12-15 times, and the consumption of samples and reagents is only 1/16 of those in the traditional method, which greatly reduces the cost. 【Conclusion】 The modified method for determining YPC in wheat grain established in this study is accurate, reliable, economical, efficient and simple, which can replace the traditional AACC 14-50 method for a large-scale determination of YPC.

Key words: Triticum aestivum, color of flour end-used products, nutritional quality, multiskan spectrum microplate reader, genetic improvement

Fig. 1

The correlation of yellow pigment content measured by the two methods"

Table 1

Analyses of variance of yellow pigment content measured by the two methods"

变异来源
Source
自由度
DF
传统AACC 14-50法 AACC 14-50 method 改良方法 Modified method
平方和 SS FF value 平方和 SS FF value
重复 Replication 2 0.174 1.54 0.007 1.31
品种 Cultivars 11 29.480 385.44** 4.370 1425.50**

Table 2

Comparison of two methods for detecting yellow pigment content"

项目Item 传统AACC 14-50法 AACC 14-50 method 改良方法 Modified method
面粉用量 Flour consumption (g) 8.0 0.5
水饱和正丁醇用量
Amount of n-butanol saturated with water (mL)
40
2.5
检测方法 Detection method 分光光度计 Spectrophotometer 酶标仪 Multiskan spectrum microplate reader
上样容器 Container 比色皿 Cuvette 96孔酶标板 96-well microtiter plate
上样量 Loading quantity 1 mL 200 μL
检测效率
Detection efficiency
一次只能检测单个样本,1个样品/分钟
Only one sample can be tested at a time, one sample/min
同时检测多个样品,12—15个样品/分钟
Multiple samples can be tested simultaneously, 12-15 samples/min

Fig. 2

The correlation of yellow pigment content between three replications measured by the modified method"

Fig. 3

Effect of extraction time on yellow pigment content of flour"

Fig. 4

Effect of preservation time on yellow pigment content of flour"

Table 3

Information of yellow pigment content in 283 domestic and foreign wheat varieties"

麦区 Wheat region 平均值 Mean 标准差 SD 最小值 Minimum 最大值 Maximum 变异系数 CV
黄淮麦区Huang-Huai wheat region
2013-AY 1.17 0.44 0.51 2.42 0.38
2013-SX 1.13 0.40 0.53 2.24 0.36
2014-AY 1.26 0.43 0.61 2.38 0.35
2014-SX 1.07 0.38 0.52 2.16 0.36
平均值Mean 1.15 0.41 0.57 2.21 0.35
北部冬麦区Northern winter wheat region
2013-SJZ 1.58 0.37 1.02 2.42 0.23
2013-BJ 1.66 0.38 1.08 2.52 0.23
2014-SJZ 1.43 0.36 0.90 2.32 0.25
2014-BJ 1.61 0.36 1.07 2.40 0.23
平均值Mean 1.57 0.36 1.02 2.33 0.23
长江中下游麦区Middle-Lower reaches of the Yangtze River wheat region
2013-CD 0.99 0.27 0.56 1.77 0.28
2013-SX 0.98 0.26 0.64 1.71 0.27
2014-CD 1.30 0.38 0.81 2.54 0.29
2014-SX 1.03 0.27 0.66 1.86 0.26
平均值Mean 1.07 0.25 0.73 1.82 0.23
国外品种Foreign wheat varieties
2013-SJZ 1.72 0.35 1.22 2.48 0.20
2013-BJ 1.64 0.35 1.06 2.31 0.22
2014-SJZ 1.54 0.36 0.94 2.36 0.23
2014-BJ 1.54 0.28 1.07 2.23 0.18
平均值Mean 1.61 0.31 1.11 2.20 0.19

Table 4

The wheat varieties with lower and higher yellow pigment content in grain"

麦区
Wheat region
低黄色素含量小麦品种
Varieties with lower yellow pigment content
高黄色素含量小麦品种
Varieties with higher yellow pigment content
黄淮麦区
Huang-Huai wheat region
丰产3号、临旱2号、陕农981、小偃54、鲁麦23、西农979-005
Fengchan 3 hao, Linhan 2 hao, Shaannong 981, Xiaoyan 54, Lumai 23, Xinong 979-005
鲁麦14、淄选2号、陕麦509、豫麦7号、中麦895、鲁麦11、烟农18
Lumai 14, Zixuan 2 hao, Shaanmai 509, Yumai 7 hao, Zhongmai 895, Lumai 11, Yannong 18
北部冬麦区
Northern winter wheat region
中优9507、CA1055、秦农151、CA0548、CA1133
Zhongyou 9507, CA1055, Qinnong 151, CA0548, CA1133
京冬22、轮选987、科衡6654、京411、中麦415、北京841、新麦37
Jingdong 22, Lunxuan 987, Keheng 6654, Jing 411, Zhongmai 415, Beijing 841, Xinmai 37
长江中下游麦区
Middle-lower reaches of the Yangtze river wheat region
鄂麦14、川麦107、扬麦10号、川麦42、鄂麦23、鄂麦21
Emai 14, Chunmai 107, Yangmai 10 hao, Chunmai 42, Emai 23, Emai 21
徐州25、宁麦9号、中国春、鄂恩5号、蓉麦4号
Xuzhou 25, Ningmai 9 hao, Chinese Spring, Een 5 hao, Rongmai 4 hao
国外品种
Foreign wheat varieties
Jagger、Salmone、WGRC10、Mesofold、F98047G14-2INC、RE714、Fr03711 YANA、Lovrin10、Fr03732、Lovrin13、E、SELYANKA、C39
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