Crops ›› 2020, Vol. 36 ›› Issue (1): 61-66.doi: 10.16035/j.issn.1001-7283.2020.01.011

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Study on the Method of Determining Digestible Protein Content and Screening of Rice Resources

Chen Tingmu,Sun Zhiguang,Xing Yungao,Fang Zhaowei,Wang Baoxiang,Liu Yan,Xu Dayong()   

  1. Lianyungang Academy of Agricultural Sciences/Jiangsu Collaborative Innovation Center for Modern Crop Production, Lianyungang 222006, Jiangsu, China
  • Received:2019-06-05 Revised:2019-08-02 Online:2020-02-15 Published:2020-02-23
  • Contact: Dayong Xu E-mail:xudayong3030@sina.com

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Abstract:

Rice is the main food crop and an important source of protein. The proteins of rice seed include non-digestible gliadin and digestible glutenin, albumin and globulin. A feasible method for determination of digestible protein content in rice was studied in 804 genotypes imported from International Rice Research Institute. Satisfactory results were obtained by adding 0.4% NaOH in 60% ethanol as the extraction solvent and using 300W ultrasound to extract digestible protein three times, simultaneously the average digestible protein of resources was measured. The average digestible protein content of 804 materials was 7.80%. It provided an effective technical means for the determination of digestible protein content in rice and screened a number of rice breeding materials with high and low digestible protein content.

Key words: Rice protein, Digestible protein, Comassie brilliant blue G250, Determination method, Rice resource

Fig.1

Statistics of geographical sources of imported rice resources of International Rice Research Institute"

Table 1

Horizontal of factors in orthogonal test"

碱浓度(%)
Alkali
concentration		 乙醇浓度(%)
Ethanol
concentration		 固液比
Solid liquid
ratio		 超声时间(min)
Ultrasonic time
0.25 30 1g:10mL 30
0.30 40 1g:15mL 40
0.35 50 1g:20mL 50
0.40 60 1g:25mL 60

Table 2

Orthogonal test scheme"

处理
Treatment		 碱浓度(%)
Alkali concentration		 乙醇浓度(%)
Ethanol concentration		 固液比
Solid liquid ratio		 超声时间(min)
Ultrasonic time
T1 0.25 30 1g:10mL 30
T2 0.25 40 1g:15mL 40
T3 0.25 50 1g:20mL 50
T4 0.25 60 1g:25mL 60
T5 0.30 30 1g:20mL 60
T6 0.30 40 1g:25mL 50
T7 0.30 50 1g:10mL 40
T8 0.30 60 1g:15mL 30
T9 0.35 30 1g:25mL 40
T10 0.35 40 1g:20mL 30
T11 0.35 50 1g:15mL 60
T12 0.35 60 1g:10mL 50
T13 0.40 30 1g:15mL 50
T14 0.40 40 1g:10mL 60
T15 0.40 50 1g:25mL 30
T16 0.40 60 1g:20mL 40
T17 (CK) 0.30 70 1g:15mL 30

Fig.2

BSA G250 dyeing working curve"

Table 3

Orthogonal experiments results of protein extraction %"

处理
Treatment		 重复Repeat 平均
Average
Ⅰ Ⅱ Ⅲ
T1 2.7381 2.9181 2.6652 2.7738g
T2 3.8575 4.1175 4.1009 4.0253cd
T3 4.5201 4.0480 4.5995 4.3892abc
T4 4.1211 4.0053 4.3273 4.1512bcd
T5 1.9880 1.9904 1.7808 1.9197h
T6 3.8264 3.8296 4.1371 3.931de
T7 4.3076 4.0519 4.6434 4.3343bcd
T8 3.2721 3.7520 3.7740 3.5994ef
T9 1.7408 1.5948 1.8809 1.7388h
T10 3.2347 3.2947 3.6159 3.3818f
T11 3.9750 4.6421 4.8588 4.492ab
T12 4.3465 4.4786 4.5657 4.4636ab
T13 1.9913 1.9704 1.8209 1.9275h
T14 4.6533 4.0477 4.0604 4.2538bcd
T15 4.5498 4.1563 4.1454 4.2838bcd
T16 4.7898 4.6145 4.9990 4.8011a
T17(CK) 2.9284 2.7422 2.5562 2.7423g

Table 4

Variance analysis of orthogonal experiments for protein extraction"

变异来源
Source of variation		 变异平方和
Sum of squares of variation		 变异自由度
Variation degree of freedom		 变异均方
Variance mean square		 F P
碱浓度Alkali concentration 1.4469 3 0.4823 4.2284 0.0119
乙醇浓度Ethanol concentration 40.6212 3 13.5404 118.7114 0.0000
固液比Solid liquid ratio 1.5499 3 0.5166 4.5294 0.0087
超声时间Ultrasonic time 0.3472 3 0.1157 1.0147 0.3979
误差Error 3.9921 35 0.1141

Table 5

Multiple comparisons of orthogonal experiments for protein extraction"

碱浓度(%)
Alkali concentration		 蛋白质含量(%)
Protein content		 乙醇浓度(%)
Ethanol concentration		 蛋白质含量(%)
Protein content		 固液比
Solid liquid ratio		 蛋白质含量(%)
Protein content
0.25 3.8349aA 50 4.3748aA 1g:15mL 3.9563aA
0.40 3.8166aA 60 4.2538aA 1g:15mL 3.6229abA
0.35 3.5191abA 40 3.8980bA 1g:15mL 3.5262bA
0.30 3.4461bA 30 2.0900cB 1g:15mL 3.5110bA

Table 6

Re-optimization of orthogonal experiments for protein extraction"

试验号
Test number		 碱浓度(%)
Alkali concentration		 乙醇浓度(%)
Ethanol concentration		 超声时间(min)
Ultrasonic time		 固液比
Solid liquid ratio		 蛋白质含量(%)
Protein content
T20 0.40 60 40 1g:15mL 4.5953a
T16 0.40 60 40 1g:20mL 4.5260ab
T18 0.40 55 40 1g:15mL 4.4167ab
T19 0.40 50 40 1g:15mL 4.3001b

Fig.3

Digestible protein content and frequency of rice groups with different digestible protein contents Different capital and lowercase letters indicate extremely significant and significant difference at the 0.01 and 0.05 level, respectively"

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