Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (9): 71-83.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0268
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ZHAO Guang-xu1(), YANG He-tong1, SHAO Xiao-bo1, CUI Zhi-hao1, LIU Hong-guang2, ZHANG Jie1()
Received:
2023-03-24
Online:
2023-09-26
Published:
2023-10-24
Contact:
ZHANG Jie
E-mail:guangxuzhao2022@163.com;zhangjie@qlu.edu.cn
ZHAO Guang-xu, YANG He-tong, SHAO Xiao-bo, CUI Zhi-hao, LIU Hong-guang, ZHANG Jie. Phosphate-solubilizing Properties and Optimization of Cultivation Conditions of Penicillium rubens: A Highly Efficient Phosphate Solubilizer[J]. Biotechnology Bulletin, 2023, 39(9): 71-83.
序号 No. | 葡萄糖 Glucose/(g·L-1) | 硫酸铵 Ammonium sulfate/(g·L-1) | 无机盐 Inorganic salt/(g·L-1) | 磷酸三钙 Tricalcium phosphate/(g·L-1) | 温度 Temperature/℃ | 转速 Rotate speed/(r·min-1) | 初始 pH Initial pH | 接种量 Inoculated amount/% |
---|---|---|---|---|---|---|---|---|
1 | 15 | 0.75 | 1.44 | 5.0 | 26 | 160 | 8 | 1 |
2 | 15 | 0.75 | 0.96 | 5.0 | 26 | 200 | 6 | 3 |
3 | 10 | 0.75 | 1.44 | 7.5 | 26 | 160 | 6 | 3 |
4 | 10 | 0.75 | 1.44 | 5.0 | 30 | 200 | 8 | 1 |
5 | 10 | 0.50 | 1.44 | 5.0 | 30 | 200 | 6 | 3 |
6 | 10 | 0.50 | 0.96 | 5.0 | 26 | 160 | 6 | 1 |
7 | 10 | 0.50 | 0.96 | 7.5 | 26 | 200 | 8 | 1 |
8 | 15 | 0.50 | 1.44 | 7.5 | 26 | 200 | 8 | 3 |
9 | 15 | 0.75 | 0.96 | 7.5 | 30 | 200 | 6 | 1 |
10 | 10 | 0.75 | 0.96 | 7.5 | 30 | 160 | 8 | 3 |
11 | 15 | 0.50 | 1.44 | 7.5 | 30 | 160 | 6 | 1 |
12 | 15 | 0.50 | 0.96 | 5.0 | 30 | 160 | 8 | 3 |
Table 1 Plackett-Burman design
序号 No. | 葡萄糖 Glucose/(g·L-1) | 硫酸铵 Ammonium sulfate/(g·L-1) | 无机盐 Inorganic salt/(g·L-1) | 磷酸三钙 Tricalcium phosphate/(g·L-1) | 温度 Temperature/℃ | 转速 Rotate speed/(r·min-1) | 初始 pH Initial pH | 接种量 Inoculated amount/% |
---|---|---|---|---|---|---|---|---|
1 | 15 | 0.75 | 1.44 | 5.0 | 26 | 160 | 8 | 1 |
2 | 15 | 0.75 | 0.96 | 5.0 | 26 | 200 | 6 | 3 |
3 | 10 | 0.75 | 1.44 | 7.5 | 26 | 160 | 6 | 3 |
4 | 10 | 0.75 | 1.44 | 5.0 | 30 | 200 | 8 | 1 |
5 | 10 | 0.50 | 1.44 | 5.0 | 30 | 200 | 6 | 3 |
6 | 10 | 0.50 | 0.96 | 5.0 | 26 | 160 | 6 | 1 |
7 | 10 | 0.50 | 0.96 | 7.5 | 26 | 200 | 8 | 1 |
8 | 15 | 0.50 | 1.44 | 7.5 | 26 | 200 | 8 | 3 |
9 | 15 | 0.75 | 0.96 | 7.5 | 30 | 200 | 6 | 1 |
10 | 10 | 0.75 | 0.96 | 7.5 | 30 | 160 | 8 | 3 |
11 | 15 | 0.50 | 1.44 | 7.5 | 30 | 160 | 6 | 1 |
12 | 15 | 0.50 | 0.96 | 5.0 | 30 | 160 | 8 | 3 |
Fig. 3 Phosphorous solubility of JL-1 to different insoluble phosphates a-d is inorganic phosphorus medium with aluminum phosphate, iron phosphate, zinc phosphate and tricalcium phosphate as phosphorus source respectively
方差来源Source | 平方和Sum of squares | 自由度df | 均方Mean square | F 值F value | P值P value |
---|---|---|---|---|---|
Model | 1.603E+05 | 8 | 20 033.75 | 46.95 | 0.004 6 |
A-Glucose | 1.369E+05 | 1 | 1.369E+05 | 320.83 | 0.000 4 |
B-Ammonium sulfate | 3.17 | 1 | 3.17 | 0.007 4 | 0.936 7 |
C-Inorganic salt | 2.23 | 1 | 2.23 | 0.005 2 | 0.946 9 |
D-Tricalcium phosphate | 12 973.79 | 1 | 12 973.79 | 30.41 | 0.011 7 |
E-Temperature | 7 137.98 | 1 | 7 137.98 | 16.73 | 0.026 4 |
F-Rotational speed | 1 879.75 | 1 | 1 879.75 | 4.41 | 0.126 7 |
G-Initial pH | 1 119.21 | 1 | 1 119.21 | 2.62 | 0.203 8 |
H-Inoculated quantity | 264.05 | 1 | 264.05 | 0.618 9 | 0.488 9 |
Residual | 1 280.01 | 3 | 426.67 | ||
Cor Total | 1.616E+05 | 11 |
Table 2 Plackett-Burman design analysis of variance
方差来源Source | 平方和Sum of squares | 自由度df | 均方Mean square | F 值F value | P值P value |
---|---|---|---|---|---|
Model | 1.603E+05 | 8 | 20 033.75 | 46.95 | 0.004 6 |
A-Glucose | 1.369E+05 | 1 | 1.369E+05 | 320.83 | 0.000 4 |
B-Ammonium sulfate | 3.17 | 1 | 3.17 | 0.007 4 | 0.936 7 |
C-Inorganic salt | 2.23 | 1 | 2.23 | 0.005 2 | 0.946 9 |
D-Tricalcium phosphate | 12 973.79 | 1 | 12 973.79 | 30.41 | 0.011 7 |
E-Temperature | 7 137.98 | 1 | 7 137.98 | 16.73 | 0.026 4 |
F-Rotational speed | 1 879.75 | 1 | 1 879.75 | 4.41 | 0.126 7 |
G-Initial pH | 1 119.21 | 1 | 1 119.21 | 2.62 | 0.203 8 |
H-Inoculated quantity | 264.05 | 1 | 264.05 | 0.618 9 | 0.488 9 |
Residual | 1 280.01 | 3 | 426.67 | ||
Cor Total | 1.616E+05 | 11 |
显著影响因素 Significant influencing factor | 水平 Level | 溶磷量Amount of dissolved phosphorus/(mg·L-1) |
---|---|---|
A-Glucose/(g·L-1) | 25 | 809.63 |
30 | 822.28 | |
35 | 805.27 | |
B-Tricalcium phosphate/(g·L-1) | 6 | 419.92 |
7 | 437.79 | |
8 | 426.46 | |
C-Temperature/℃ | 30 | 456.92 |
32 | 521.49 | |
34 | 486.18 |
Table 3 Experiment results of the steepest climb
显著影响因素 Significant influencing factor | 水平 Level | 溶磷量Amount of dissolved phosphorus/(mg·L-1) |
---|---|---|
A-Glucose/(g·L-1) | 25 | 809.63 |
30 | 822.28 | |
35 | 805.27 | |
B-Tricalcium phosphate/(g·L-1) | 6 | 419.92 |
7 | 437.79 | |
8 | 426.46 | |
C-Temperature/℃ | 30 | 456.92 |
32 | 521.49 | |
34 | 486.18 |
Run | 葡萄糖 Glucose/ (g·L-1) | 磷酸三钙 Tricalcium phosphate/ (g·L-1) | 温度 Tempe- rature/℃ | 溶磷量 Amount of dissolved phosphorus/(mg·L-1) |
---|---|---|---|---|
1 | 35 | 7 | 30 | 991 |
2 | 30 | 7 | 32 | 1 123 |
3 | 25 | 6 | 32 | 791 |
4 | 35 | 8 | 32 | 731 |
5 | 30 | 7 | 32 | 1 135 |
6 | 30 | 7 | 32 | 1 141 |
7 | 25 | 7 | 34 | 1 009 |
8 | 25 | 7 | 30 | 712 |
9 | 30 | 7 | 32 | 1 139 |
10 | 30 | 7 | 32 | 1 132 |
11 | 35 | 6 | 32 | 991 |
12 | 30 | 8 | 30 | 1 057 |
13 | 30 | 6 | 30 | 723 |
14 | 30 | 6 | 34 | 1 034 |
15 | 30 | 8 | 34 | 732 |
16 | 35 | 7 | 34 | 657 |
17 | 25 | 8 | 32 | 1 082 |
Table 4 Box-Behnken experiment results
Run | 葡萄糖 Glucose/ (g·L-1) | 磷酸三钙 Tricalcium phosphate/ (g·L-1) | 温度 Tempe- rature/℃ | 溶磷量 Amount of dissolved phosphorus/(mg·L-1) |
---|---|---|---|---|
1 | 35 | 7 | 30 | 991 |
2 | 30 | 7 | 32 | 1 123 |
3 | 25 | 6 | 32 | 791 |
4 | 35 | 8 | 32 | 731 |
5 | 30 | 7 | 32 | 1 135 |
6 | 30 | 7 | 32 | 1 141 |
7 | 25 | 7 | 34 | 1 009 |
8 | 25 | 7 | 30 | 712 |
9 | 30 | 7 | 32 | 1 139 |
10 | 30 | 7 | 32 | 1 132 |
11 | 35 | 6 | 32 | 991 |
12 | 30 | 8 | 30 | 1 057 |
13 | 30 | 6 | 30 | 723 |
14 | 30 | 6 | 34 | 1 034 |
15 | 30 | 8 | 34 | 732 |
16 | 35 | 7 | 34 | 657 |
17 | 25 | 8 | 32 | 1 082 |
来源Source | 平方和Sum of squares | 自由度df | 均方Mean square | F 值F value | P 值P value | 显著性Significance |
---|---|---|---|---|---|---|
Model | 5.26E+05 | 9 | 58 440.03 | 398.42 | < 0.000 1 | Significant |
A-Glucose | 6 272.00 | 1 | 6 272.00 | 42.76 | 0.000 3 | |
B-Tricalcium phosphate | 496.13 | 1 | 496.13 | 3.38 | 0.108 5 | |
C-Temperature | 325.13 | 1 | 325.13 | 2.22 | 0.180 1 | |
AB | 75 900.25 | 1 | 75 900.25 | 517.46 | < 0.000 1 | |
AC | 99 540.25 | 1 | 99 540.25 | 678.63 | < 0.000 1 | |
BC | 1.01E+05 | 1 | 1.01E+05 | 689.43 | < 0.000 1 | |
A² | 82 231.84 | 1 | 82 231.84 | 560.63 | < 0.000 1 | |
B² | 38 401.05 | 1 | 38 401.05 | 261.80 | < 0.000 1 | |
C² | 97 280.00 | 1 | 97 280.00 | 663.22 | < 0.000 1 | |
Residual | 1 026.75 | 7 | 146.68 | |||
Lack of fit | 826.75 | 3 | 275.58 | 5.51 | 0.066 3 | Not significant |
Pure error | 200.00 | 4 | 50.00 | |||
Cor total | 5.27E+05 | 16 |
Table 5 Box-Behnken design variance analysis
来源Source | 平方和Sum of squares | 自由度df | 均方Mean square | F 值F value | P 值P value | 显著性Significance |
---|---|---|---|---|---|---|
Model | 5.26E+05 | 9 | 58 440.03 | 398.42 | < 0.000 1 | Significant |
A-Glucose | 6 272.00 | 1 | 6 272.00 | 42.76 | 0.000 3 | |
B-Tricalcium phosphate | 496.13 | 1 | 496.13 | 3.38 | 0.108 5 | |
C-Temperature | 325.13 | 1 | 325.13 | 2.22 | 0.180 1 | |
AB | 75 900.25 | 1 | 75 900.25 | 517.46 | < 0.000 1 | |
AC | 99 540.25 | 1 | 99 540.25 | 678.63 | < 0.000 1 | |
BC | 1.01E+05 | 1 | 1.01E+05 | 689.43 | < 0.000 1 | |
A² | 82 231.84 | 1 | 82 231.84 | 560.63 | < 0.000 1 | |
B² | 38 401.05 | 1 | 38 401.05 | 261.80 | < 0.000 1 | |
C² | 97 280.00 | 1 | 97 280.00 | 663.22 | < 0.000 1 | |
Residual | 1 026.75 | 7 | 146.68 | |||
Lack of fit | 826.75 | 3 | 275.58 | 5.51 | 0.066 3 | Not significant |
Pure error | 200.00 | 4 | 50.00 | |||
Cor total | 5.27E+05 | 16 |
根长 Root length /cm | 株高 Plant height /cm | 总鲜重 Total fresh weight/g | 上部鲜重 Upper fresh weight/g | 上部干重 Upper dry weight/g | |
---|---|---|---|---|---|
CK | 12.1±1.36 | 15.37±0.95 | 0.136 4±0.003 2 | 0.100 4±0.004 1 | 0.011 2±0.008 0 |
P1 | 19.1±1.91* | 20.97±1.07 * | 0.194 9±0.006 8 * | 0.141 4±0.009 0 * | 0.015 0±0.001 5 |
P2 | 18.8±2.30 | 27.57±0.68 *** | 0.280 2±0.027 4 * | 0.199 3±0.028 6 * | 0.019 9±0.002 3 * |
Table 6 Growth promoting effect of JL-1 on wheat
根长 Root length /cm | 株高 Plant height /cm | 总鲜重 Total fresh weight/g | 上部鲜重 Upper fresh weight/g | 上部干重 Upper dry weight/g | |
---|---|---|---|---|---|
CK | 12.1±1.36 | 15.37±0.95 | 0.136 4±0.003 2 | 0.100 4±0.004 1 | 0.011 2±0.008 0 |
P1 | 19.1±1.91* | 20.97±1.07 * | 0.194 9±0.006 8 * | 0.141 4±0.009 0 * | 0.015 0±0.001 5 |
P2 | 18.8±2.30 | 27.57±0.68 *** | 0.280 2±0.027 4 * | 0.199 3±0.028 6 * | 0.019 9±0.002 3 * |
菌种 Strain | 最大溶磷量 Maximum phosphorus solubilization/(mg·L-1) | 来源 Source |
---|---|---|
产红青霉Penicillium rubens | 1 194.15 | JL-1(本研究 This study) |
黑曲霉Aspergillus niger | 1 741.59 | YJC191[ |
嗜松青霉Penicillium pinophilum | 1 390.00 | JP-NJ4[ |
绳状青霉Penicillium funicuiosum | 1 022.77 | P1[ |
烟曲霉菌Aspergillus fumigatiaffinis | 967.40 | JL-7[ |
青霉菌Penicillium sp. | 773.36 | PSF-FJ1[ |
黑曲霉Aspergillus niger | 616.81 | Xj-2[ |
阿达青霉Penicillium adametzii | 587.32 | FG54[ |
恶臭假单胞菌Pseudomonas putida | 424.14 | 623-3[ |
枯草芽孢杆菌Bacillus subtilis | 386.25 | JT-1[ |
产红青霉Penicillium rubens | 328.79 | R3[ |
欧文氏杆菌Erwinia carotovor | 248.00 | P9[ |
Table 7 Comparison of phosphorus solubilizing capacity of phosphorus solubilizing bacteria
菌种 Strain | 最大溶磷量 Maximum phosphorus solubilization/(mg·L-1) | 来源 Source |
---|---|---|
产红青霉Penicillium rubens | 1 194.15 | JL-1(本研究 This study) |
黑曲霉Aspergillus niger | 1 741.59 | YJC191[ |
嗜松青霉Penicillium pinophilum | 1 390.00 | JP-NJ4[ |
绳状青霉Penicillium funicuiosum | 1 022.77 | P1[ |
烟曲霉菌Aspergillus fumigatiaffinis | 967.40 | JL-7[ |
青霉菌Penicillium sp. | 773.36 | PSF-FJ1[ |
黑曲霉Aspergillus niger | 616.81 | Xj-2[ |
阿达青霉Penicillium adametzii | 587.32 | FG54[ |
恶臭假单胞菌Pseudomonas putida | 424.14 | 623-3[ |
枯草芽孢杆菌Bacillus subtilis | 386.25 | JT-1[ |
产红青霉Penicillium rubens | 328.79 | R3[ |
欧文氏杆菌Erwinia carotovor | 248.00 | P9[ |
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