Screening,Identification and Antagonism of Phosphate-Solubilizing Bacteria from the Compost Chinese Medicinal Herbal Residues

doi:10.13560/j.cnki.biotech.bull.1985.2021-0571

Abstract

Abstract:

To develop new,safe and efficient microbial fertilizer,Chinese medicine residues after two months composting was used as test material to screen phosphate-solubilizing and antagonistic bacteria. Organophosphorus bacteria medium was used to preliminarily screen 5 strains of phosphate-solubilizing bacteria. Phosphorus solubilizing ring method and phosphorus-molybdenum blue colorimetry were employed to determine phosphorus solubility of the strains. The strain with the best phosphorus solubility was YP5. Based on the colony morphology,physiological and biochemical characteristics,and 16S rDNA sequence analysis,YP5 was identified as Pseudomonas aeruginosa. The results of promoting and antagonist factors showed that YP5 secreted iron carrier and a large amount of extracellular protease. YP5 was highly sensitive to norfloxacin,chloramphenicol,gentamicin,enrofloxacin and amikacin. The results from the plate confrontation method and the virulent medium method revealed that YP5 fermentation broth and cell-free filtrate presented promising antagonism on 9 plant pathogens such as Fusarium solani,and the maximum inhibition rate was > 80%. The highest yield of phenazino-1-carboxylic acid of YP5 in KB medium at 28℃was 17.73 mg/L by high performance liquid chromatography.

Key words: phosphate-solubilizing bacteria, Pseudomonas aeruginosa, phosphate-solubilizing, antagonistic action

HU Shan, LIANG Wei-qu, HUANG Hao, XU Cong, LUO Hua-jian, HU Chu-wei, HUANG Xiao-yan, CHEN Shi-li. Screening,Identification and Antagonism of Phosphate-Solubilizing Bacteria from the Compost Chinese Medicinal Herbal Residues[J]. Biotechnology Bulletin, 2022, 38(3): 92-102.

Figures/Tables 11

References 37

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菌株 Strain	溶磷圈直径 Diameter of phosphate Solubilizing halo D/mm	菌落直径 Colony diameter d /mm	D/d
YP1	13.43±0.24d	4.77±0.22b	2.83±0.15c
YP2	18.53±0.33b	5.13±0.13b	3.62±0.15b
YP3	14.60±0.23c	6.47±0.24a	2.26±0.05d
YP4	18.17±0.17b	6.13±0.07a	2.96±0.03c
YP5	21.67±0.24a	5.10±0.25b	4.27±0.20a

菌株 Strain	溶磷圈直径 Diameter of phosphate Solubilizing halo D/mm	菌落直径 Colony diameter d /mm	D/d
YP1	13.43±0.24d	4.77±0.22b	2.83±0.15c
YP2	18.53±0.33b	5.13±0.13b	3.62±0.15b
YP3	14.60±0.23c	6.47±0.24a	2.26±0.05d
YP4	18.17±0.17b	6.13±0.07a	2.96±0.03c
YP5	21.67±0.24a	5.10±0.25b	4.27±0.20a

生理生化试验Physiological and biochemical test	检测结果Result	生理生化试验Physiological and biochemical tests	检测结果Result
硝酸盐还原 Nitrate reduction	-	色氨酸酶Tryptophanase	-
精氨酸双水解酶 Arginine dihydrolase	+	脲酶Urease	-
明胶液化 Gelatine liquefication	+	β-半乳糖苷酶β-galactosidase	-
阿拉伯糖Arabinose fermentation	-	甘露糖Mannose	-
麦芽糖Maltose fermentation	-	葡萄糖酸盐Gluconate	+
己二酸Adipic Acid	+	苹果酸Malic acid	+
苯乙酸Phenylacetic acid	-	氧化酶Oxidase	+
葡萄糖酸Gluconic acid	-	七叶灵水解 Hydrolysis of aescin	-
葡萄糖 Glucose fermentation	+	N-乙酰-葡萄糖胺n-acetyl-glucosamine	+
柠檬酸Citric acid	+	甘露醇Mannitol	+
癸酸 Capric acid	+

生理生化试验Physiological and biochemical test	检测结果Result	生理生化试验Physiological and biochemical tests	检测结果Result
硝酸盐还原 Nitrate reduction	-	色氨酸酶Tryptophanase	-
精氨酸双水解酶 Arginine dihydrolase	+	脲酶Urease	-
明胶液化 Gelatine liquefication	+	β-半乳糖苷酶β-galactosidase	-
阿拉伯糖Arabinose fermentation	-	甘露糖Mannose	-
麦芽糖Maltose fermentation	-	葡萄糖酸盐Gluconate	+
己二酸Adipic Acid	+	苹果酸Malic acid	+
苯乙酸Phenylacetic acid	-	氧化酶Oxidase	+
葡萄糖酸Gluconic acid	-	七叶灵水解 Hydrolysis of aescin	-
葡萄糖 Glucose fermentation	+	N-乙酰-葡萄糖胺n-acetyl-glucosamine	+
柠檬酸Citric acid	+	甘露醇Mannitol	+
癸酸 Capric acid	+

抗菌素Antibiotics	含药量Drug concentration/（μg·μL^-1）	抑菌圈直径Inhibitory zone diameter d/mm	敏感性Sensitivity
氨苄西林Ampicillin	0.5	0	R
诺氟沙星Norfloxacin	0.5	32.00±0.00	S
氯霉素Chloramphenicol	1.5	17.67±0.33	S
利福平 Rifampin	0.25	0	R
亚胺培南Imipenem	0.5	0	R
卡那霉素 Kanamycin	1.5	8.67±0.67	R
庆大霉素 Gentamicin	0.5	19.00±0.58	S
四环素Tetracycline	1.5	0	R
萘啶酸Nalidixic acid	1.5	13.33±0.33	I
链霉素Streptomycin	0.5	12.00±0.00	I
恩诺沙星Enrofloxacin	0.5	21.33±0.67	S
丁胺卡那霉素Amikacin	1.5	20.67±0.33	S
多西环素Doxycycline	1.5	8.67±0.33	R
万古霉素Vancomycin	1.5	0	R
阿莫西林Amoxicillin	1	0	R