生物技术通报 ›› 2023, Vol. 39 ›› Issue (7): 241-253.doi: 10.13560/j.cnki.biotech.bull.1985.2022-1450

• 研究报告 • 上一篇    下一篇

一株多功能菌株的发掘、鉴定及解磷条件优化

谢东(), 汪流伟, 李宁健, 李泽霖, 徐子航, 张庆华()   

  1. 江西农业大学生物科学与工程学院,南昌 330045
  • 收稿日期:2022-11-25 出版日期:2023-07-26 发布日期:2023-08-17
  • 通讯作者: 张庆华,男,博士,教授,研究方向:环境微生物;E-mail: zqh_net@163.com
  • 作者简介:谢东,男,硕士研究生,研究方向:环境微生物;E-mail: 2651553709@qq.com
  • 基金资助:
    江西省自然科学基金重点项目(20212ACB205003);江西省高层次高技能领军人才项目,江西省杰出青年人才资助计划项目(20171BCB23044)

Exploration, Identification and Phosphorus-solubilizing Condition Optimization of a Multifunctional Strain

XIE Dong(), WANG Liu-wei, LI Ning-jian, LI Ze-lin, XU Zi-hang, ZHANG Qing-hua()   

  1. School of Biological Science and Engineering, Jiangxi Agricultural University, Nanchang 330045
  • Received:2022-11-25 Published:2023-07-26 Online:2023-08-17

摘要:

从菌糠鸡粪堆肥中筛选高效的解磷菌并优化其解磷能力,发掘多功能微生物资源以实现农业废弃物的有效利用。利用无机磷选择培养基从堆肥中筛选高效解磷菌,结合形态学观察、生理生化试验、分子生物学鉴定,对菌株进行鉴定。利用单因素和响应面试验优化菌株的解磷培养条件;通过解磷、解钾和产吲哚乙酸能力的测定确定菌株的堆肥潜力。筛选得到一株高效解磷菌株Pb1,经鉴定发现菌株隶属于Bacillus smithii strain;培养条件为葡萄糖15.0 g/L、硫酸铵3 g/L、磷酸三钙7.0 g/L、无机盐0.48 g/L、温度55.0℃、摇床转速225 r/min、初始pH 7.0、装液量60.0 mL/250 mL、接种量3.0%(V/V)时,菌株Bacillus smithii Pb1的解磷能力最强;进一步通过响应面优化发现,当磷酸三钙6.4 g/L、摇床转速207 r/min、装液量65.0 mL/250 mL时,菌株Pb1培养3 d,发酵液中溶磷量可达534.68 mg/L,是优化前的1.58倍。同时,发现B. smithii Pb1解有机磷、解钾、产吲哚乙酸能力,分别为238.99 mg/L、81.06 mg/L、23.26 mg/L。菌株B. smithii Pb1在高温(55℃)条件下展现出了解无机磷、有机磷、解钾和产吲哚乙酸的能力,为菌株B. smithii Pb1开发成好氧堆肥菌剂奠定基础。

关键词: 多功能菌株, 筛选, 鉴定, 解磷特征, 响应面优化

Abstract:

In order to explore multifunctional microorganism resources to achieve the effective utilization of agricultural waste, some efficient phosphate-solubilizing bacteria were isolated from the compost of mushroom chaff and chicken manure, and their phosphate-solubilizing abilities were subsequently optimized. Efficient phosphorus-solubilizing bacteria were screened from the compost by inorganic phosphorus selective medium. The selected strain was identified through the combination of morphological observation, physiological and biochemical test and molecular biology identification. Single factor and response surface experiments were used to optimize the inorganic phosphorus-solubilizing conditions of this strain. In addition, the compost potential of this strain was evaluated by determining the ability of releasing organophosphorus and potassium, and indole acetic acid production. An efficient phosphate-solubilizing strain Pb1 was isolated and identified as belonging to Bacillus smithii strain. The highest phosphorus-solubilizing capacity of strain B. smithii Pb1 was achieved when the culture condition was kept as: glucose 15.0 g /L, ammonium sulfate 3 g/L, tricalcium phosphate 7.0 g/L, inorganic salt 0.48 g/L, temperature 55.0℃, rotating speed 225 r/min, initial pH 7.0, loaded liquid 60.0 mL/250 mL, and inoculum volume 3.0%(V/V). Further response surface optimization showed that under the conditions of tricalcium phosphate 6.4 g/L, rotating speed 207 r/min and loaded liquid 65.0 mL/250 mL, the solubilized phosphorus content in the fermentation broth of strain Pb1 cultured for 3 dreached 534.68 mg/L, which was 1.58 times higher than that before optimization. Moreover, B. smithii Pb1 was found to be capable of detoxifying organophosphates, detoxifying potassium and producing indole-3-acetic acid with the content of 238.99 mg/L, 81.06 mg/L and 23.26 mg/L, respectively. B. smithii Pb1 demonstrated the ability of releasing inorganic phosphorus, organophosphates and potassium, and producing indoleacetic acid at high temperature(55℃), laying the foundation for the development of B. smithii Pb1 as an aerobic composting bacterial agent.

Key words: multifunctional strain, screening, identification, characteristics of phosphorus solubilizing, response surface optimization