Phosphate-solubilizing Properties and Optimization of Cultivation Conditions of Penicillium rubens: A Highly Efficient Phosphate Solubilizer

doi:10.13560/j.cnki.biotech.bull.1985.2023-0268

Abstract

Abstract:

In order to reduce the use of chemical fertilizers in agricultural production, a fungal strain JL-1 with high efficiency in degrading tricalcium phosphate was isolated from phosphorus-rich tea（Camellia sinensis L.）rhizosphere soil by using inorganic phosphorus screening medium, and identified as Penicillium rubens. The changes of pH value, phosphorus content and organic acid content of fermentation broth of strain JL-1 in the process of phosphorus solubilization showed that there was a significant negative correlation between pH value and gluconic acid content, a significant negative correlation between pH value and phosphorus content, and a significant positive correlation between gluconic acid and phosphorus content in the fermentation broth of strain JL-1. And the surface erosion marks of tricalcium phosphate particles were observed under the electron microscope. Further analysis uncovered that strain JL-1 eroded tricalcium phosphate and dissolved phosphorus by secreting gluconic acid. The culture conditions were optimized through a series of experiments including single factor experiment, Plackett-Burman design experiemnt, steepest climb experiment and central combination experiment. The results showed that glucose and ammonium sulfate were the optimal carbon and nitrogen sources for dissovling tricalcium phosphate by strain JL-1. Glucose content, tricalcium phosphate content and temperature were significant factors affecting the phosphorus solubility of strain JL-1. Under the conditions of 29.8 g/L glucose, 7.1 g/L tricalcium phosphate and 31.9℃, the phosphorus solubility of strain JL-1 in inorganic phosphorus medium, the solubilized phosphorus of the strain reached 1194.15 mg/L, which was nearly three times of the initial value. In phosphorus-deficient soil, this strain significantly promoted the growth of T. aestivum L., with its root length, plant height and total fresh weight increased by 57.9%, 36.4% and 42.9% respectively, indicating that the strain JL-1 had good functions of phosphate-dissolving and growth promotion. With the excellent phosphorus-dissolving characteristics and phosphate-dissolving ability, the strain JL-1 will provide a reference for the development and application in the field of biological fertilizer.

Key words: Penicillium rubens, rhizosphere of Camellia sinensis L., phosphorus-dissolving, tricalcium phosphate, gluconic acid, response surface optimization, growth promotion

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.

Figures/Tables 16

References 42

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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

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

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

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