中药渣堆肥中解磷细菌的筛选、鉴定及其拮抗作用

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

为开发新型、安全及高效的微生物菌肥,以堆肥2个月后的中药渣为试材,筛选解磷拮抗细菌。用有机磷细菌培养基初步筛选5株解磷细菌;采用溶磷圈法和磷钼蓝比色法测定初筛菌株的解磷作用,解磷作用最优的菌株是YP5;通过形态学特征观察、生理生化试验和16S rDNA序列分析鉴定,确定菌株YP5为铜绿假单胞菌(Pseudomonas aeruginosa)。促生拮抗因子测定结果表明菌株YP5能分泌铁载体和大量胞外蛋白酶;药敏实验结果显示菌株YP5对诺氟沙星、氯霉素、庆大霉素、恩诺沙星、丁胺卡那霉素高度敏感;分别采用平板对峙法和带毒培养基法测定结果表明菌株YP5发酵液和无菌滤液均对冬瓜根腐病等9种植物病原菌有良好的拮抗作用,抑菌率最高达80%以上。高效液相色谱测定菌株YP5在28℃、KB培养基中抗菌代谢物申嗪霉素的产量最高,为17.73 mg/L。

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.