高效解磷菌筛选优化及其对弱碱土小麦促生效应

doi:10.13560/j.cnki.biotech.bull.1985.2025-1452

生物技术通报 ›› 2026, Vol. 42 ›› Issue (8): 263-275.doi: 10.13560/j.cnki.biotech.bull.1985.2025-1452

• 研究报告 • 上一篇

高效解磷菌筛选优化及其对弱碱土小麦促生效应

吕红珍¹^,², 徐莉莉³^,⁴, 张琳², 高敏², 谭镇炜², 王亚雯², 张秀格⁵, 王硕⁶, 马爱军¹, 王承民²^,³^,⁴()

^1.五邑大学药学与食品工程学院，江门 529000
^2.广东省科学院动物研究所广东省动物保护与资源利用重点实验室广东省野生动物保护与利用公共实验室，广州 510260
^3.优宜邦生物科技（上海）有限公司，上海 201103
^4.南陵万泽微生物工程研究院有限公司，南陵 241300
^5.河北外国语学院，石家庄 050011
^6.武汉生物工程学院，武汉 430000

收稿日期:2025-12-30 出版日期:2026-08-26 发布日期:2026-08-17
通讯作者: 王承民wangchm@giz.gd.cn
基金资助:
优宜邦生物科技（上海）有限公司产业化项目（UYB20220001-01,UYB（研）20230001-02）

Screening and Optimization of High-efficiency Phosphate-solubilizing Bacteria and Wheat Growth Promotion in Slightly Alkaline Soil

LYU Hong-zhen¹^,², XU Li-li³^,⁴, ZHANG Lin², GAO Min², TAN Zhen-wei², WANG Ya-wen², ZHANG Xiu-ge⁵ WANG Shuo⁶ MA Ai-jun⁵, WANG Cheng-min⁶

^1.School of Pharmacy and Food Engineering, Wuyi University, Jiangmen 529000
^2.Guangdong Key Laboratory of Animal Conservation and Resource Utilization, Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Guangdong Institute of Zoology, Guangdong Academy of Sciences, Guangzhou 510260
^3.Uebang Biotechnology (Shanghai) Co. , Ltd. , Shanghai 201103
^4.Nanling Wanze Microbial Engineering Research Institute Co. , Ltd. , Nanling 241300
^5.Hebei International Studies University, Shijiazhuang 050011
^6.Wuhan University of Bioengineering, Wuhan 430000

Received:2025-12-30 Published:2026-08-26 Online:2026-08-17

摘要/Abstract

摘要：

目的针对弱碱性土壤磷素固化严重、植物利用率低的问题，筛选高效解磷菌株并优化培养条件，阐明其解磷机制，验证其土壤磷活化及小麦促生效应。方法以7个属26株菌株为材料，通过平板透明圈法初筛与液体培养法复筛获得高效解磷菌株；通过单因素试验与响应面法优化培养条件；采用HPLC分析有机酸组分，结合相关性分析揭示解磷机制；通过小麦盆栽试验评估菌株的土壤应用效果。结果筛选获得阿氏肠杆菌（Enterobacter asburiae）N4129-2AT，该菌株在15 ℃低温及pH 10高碱环境下仍保持稳定溶磷能力；经响应面优化（35 ℃、pH 6、接种量7.2%）后，72 h溶磷量达823.471 mg/L，较优化前提升23.7%，培养周期缩短96 h。该菌株可稳定分泌6种小分子有机酸，其解磷核心机制为有机酸介导的酸化作用，溶磷效能依赖强关联的有机酸而非含量占比，且总有机酸含量仅起辅助作用。小麦盆栽试验中，各处理组均显著提升土壤有效溶磷量以及小麦植株生长指标，其中优化后菌全液中剂量处理效果最优，土壤有效磷含量提升376.76%，小麦地下部干重提升157.4%，根冠比降低22.69%，土壤有效磷与小麦生长指标呈极显著正相关。结论阿氏肠杆菌N4129-2AT兼具耐低温、耐高碱及高效解磷能力，其基于强关联有机酸主导的酸谱定向优化解磷机制为同类菌株改良提供理论参考，该菌株及优化培养方案可为弱碱性土壤磷素活化与小麦高产提供技术支撑。

关键词: 解磷菌, 菌株筛选, 响应面法, HPLC, 磷活化, 小麦促生

Abstract:

Objective To address the severe phosphorus (P) immobilization and low plant availability in slightly alkaline soils, this study aimed to screen high-efficiency and stress-tolerant phosphate-solubilizing bacteria (PSB), optimize their culture conditions, clarify the P-solubilizing mechanism, and verify their effects on soil P activation and wheat growth promotion. Method A total of 26 candidate strains from 7 genera were used as test materials. High-efficiency PSB were isolated via primary screening by the plate transparent zone method (using inorganic P selective medium, with phosphorus-solubilizing index as the evaluation criterion) and secondary screening by the liquid culture method (soluble P content determined by molybdenum antimony anti-spectrophotometry). Single-factor experiments and response surface methodology (RSM) were conducted to optimize the cluture conditions. High-performance liquid chromatography (HPLC) was used to identify organic acid components, and Pearson correlation analysis was performed to reveal the relationship between organic acid profiles and P-solubilizing efficiency. Wheat pot experiments were carried out to evaluate soil application effects. Result An efficient PSB strain was identified as Enterobacter asburiae N4129-2AT. This strain maintained stable P-solubilizing capacity under low temperature (15 ℃) and high alkalinity (pH 10). After RSM optimization, the optimal culture conditions were confirmed to be 35 ℃, initial pH 6, and an inoculum size of 7.2%. Under these conditions, the soluble P content in fermentation broth reached 823.471 mg/L within 72 h, which was 23.7% higher than that before optimization, and the culture cycle was shortened by 96 h. E. asburiae N4129-2AT stably secreted six types of small-molecule organic acids. The core P-solubilizing mechanism was acidification mediated by organic acids, and the key factor improving P-solubilizing efficiency was organic acids strongly associated with P solubilization rather than their content proportion; total organic acid content only played an auxiliary role. In wheat pot experiments, all PSB treatments significantly increased soil available P content and wheat growth parameters compared with the control. The medium-dose optimized whole bacterial broth treatment showed the best performance: soil available P content increased by 376.76%, wheat the root dry weight increased by 157.4%, and root-shoot ratio decreased by 22.69%. A highly significant positive correlation was observed between soil available P and wheat growth parameters. Conclusion E.asburiae N4129-2AT possesses excellent stress tolerance (low temperature and high alkalinity) and high P-solubilizing efficiency. Its P-solubilizing mechanism, dominated by strongly associated organic acids through targeted optimization of organic acid profiles, breaks the traditional cognition that total organic acid content determines P-solubilizing capacity, providing a theoretical reference for the improvement of similar strains. This strain and its optimized culture conditions offer reliable technical support for P activation in slightly alkaline soils and high-yield wheat production.

Key words: phosphate-solubilizing bacteria, strain screening, response surface methodology, HPLC, phosphorus activation, wheat growth promotion

吕红珍, 徐莉莉, 张琳, 高敏, 谭镇炜, 王亚雯, 张秀格, 王硕, 马爱军, 王承民. 高效解磷菌筛选优化及其对弱碱土小麦促生效应[J]. 生物技术通报, 2026, 42(8): 263-275.

LYU Hong-zhen, XU Li-li, ZHANG Lin, GAO Min, TAN Zhen-wei, WANG Ya-wen, ZHANG Xiu-ge⁵ WANG Shuo⁶ MA Ai-jun, WANG Cheng-min. Screening and Optimization of High-efficiency Phosphate-solubilizing Bacteria and Wheat Growth Promotion in Slightly Alkaline Soil[J]. Biotechnology Bulletin, 2026, 42(8): 263-275.

图/表 15

表1 响应面分析因素与水平

Table 1 Factors and levels for response surface analysis

水平编码 Level code	因素 Factors
水平编码 Level code	A温度 A Temperature (℃)	B pH B pH	C接种量 C Inoculum size (%)
-1	25	6	5
0	30	8	7
1	35	10	9

表2 响应面实验设计方案及响应值

Table 2 Experimental design and response values for response surface methodology

编号 Number	A 温度 A Temperature （℃）	B pH	C 接种量 C Inoculum size （%）	可溶性磷含量 Soluble phosphorus content （mg/L）
1	25	6	7	641.026
2	35	6	7	813.609
3	25	10	7	433.925
4	35	10	7	510.355
5	25	8	5	599.112
6	35	8	5	608.974
7	25	8	9	493.097
8	35	8	9	692.801
9	30	6	5	714.990
10	30	10	5	436.390
11	30	6	9	653.353
12	30	10	9	483.235
13	30	8	7	641.026
14	30	8	7	643.491
15	30	8	7	663.215
16	30	8	7	658.284
17	30	8	7	663.215

图1 不同解磷菌株在培养过程中可溶性磷含量的动态变化

Fig. 1 Dynamic changes in soluble phosphate content during the cultivation of different phosphate-solubilizing strains

图2 菌株N4129-2AT的溶血性测定（A）与植物致病性观察（B）CK：空白对照；T：接种菌株N4129-2AT

Fig. 2 Hemolytic activity test (A) and phytopathogenicity assay (B) of strain N4129-2ATCK: blank control; T: inoculated with strain N4129-2AT

图3 不同单因素对N4129-2AT溶磷量的影响

Fig. 3 Effects of single factors on P-solubilizing capacity of N4129-2AT

表3 方差分析表

Table 3 Analysis of variance (ANOVA) table

来源 Source	平方和 Sum of squares	自由度 Degree of freedom	均方 Mean square	F值 F value	P值 P value
模型 Model	1.720E+05	9	19 113.08	117.05	<0.001
A-温度 Temperature	26 286.91	1	26 286.91	160.98	<0.001
B-pH	1.150E+05	1	1.150E+05	704.11	<0.001
C-接种量 Inoculum size	170.96	1	170.96	1.05	0.340 3
AB	2 311.37	1	2 311.37	14.15	0.007 1
AC	9 010.03	1	9 010.03	55.18	0.000 1
BC	2 942.05	1	2 942.05	18.02	0.003 8
A²	802.63	1	802.63	4.92	0.062 1
B²	6 841.92	1	6 841.92	41.90	0.000 3
C²	7 266.73	1	7 266.73	44.50	0.000 3
残差Residual	1 143.06	7	163.29
失拟项Lack of fit	676.23	3	225.41	1.93	0.266 2
纯误差Pure error	466.83	4	116.71
总和 Total	1.732E+05	16

图4 不同因素两两交互影响解磷菌解磷能力的曲面图和等高线图

Fig. 4 Response surface and contour plots for pairwise interactions of factors affecting P-solubilizing capacity of phosphate-solubilizing bacteria

图5 最优条件下N4129-2AT培养过程中可溶性磷含量与pH的动态变化

Fig. 5 Dynamic changes in soluble phosphate content and pH during incubation of N4129-2AT under optimal conditions

图6 N4129-2AT在不同条件下有机酸与pH、动态溶磷量的相关性热图Ini：初始条件；Str：逆境条件；Opt：最优条件；P：可溶性磷含量；TA：酒石酸；FA：甲酸；MA：苹果酸；LA：乳酸；SA：琥珀酸；AA：乙酸。*P<0.05，**P<0.01，***P<0.001，下同

Fig. 6 Correlation heatmap of organic acids, pH, and P-solubilization of N4129-2AT under different conditionsIni: Initial condition; Str: stress condition; Opt: optimal condition; P: soluble phosphorus content; TA: tartaric acid; FA: formic acid; MA: malic acid; LA: lactic acid; SA: succinic acid; AA: acetic acid. *P<0.05, **P<0.01, ***P<0.001, The same below

图7 不同培养条件下总有机酸与pH的动态变化及核心有机酸占比

Fig. 7 Dynamic changes of total organic acids and pH under different culture conditions as well as the proportion of core organic acids

图8 不同条件下7 d培养期内不同核心溶磷有机酸含量的动态变化

Fig. 8 Dynamic changes in content of core phosphate-solubilizing organic acids during the 7-day incubation period under different conditions

图9 不同处理对土壤有效磷含量的影响及土壤有效磷与pH的相关性

Fig. 9 Effects of different treatments on soil available P content and its correlation with pH

图10 不同处理对小麦幼苗生长指标的影响

Fig. 10 Effects of different treatments on growth indicators of wheat seedlings

表4 土壤有效磷与小麦幼苗生长指标的相关性

Table 4 Correlation between soil available phosphorus and wheat seedling growth indices

关联指标组合 Indicator combination	相关系数 Correlation coefficient	P-value
AP-株高 AP-Plant height	0.855^**	<0.001
AP-主根长 AP-Taproot length	0.752^**	<0.001
AP-地上部干重 AP-Shoot dry weight	0.965^**	<0.001
AP-地下部干重 AP-Root dry weight	0.665^**	<0.001
AP-根冠比 AP-Root-shoot ratio	-0.701^**	<0.001

图11 土壤有效磷与小麦地上部干重的相关性

Fig. 11 Correlation between soil available phosphorus and wheat shoot dry weight

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高效解磷菌筛选优化及其对弱碱土小麦促生效应

Screening and Optimization of High-efficiency Phosphate-solubilizing Bacteria and Wheat Growth Promotion in Slightly Alkaline Soil

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