铅污染土壤中解磷菌对玉米根际土壤性质和微生物群落结构的影响

doi:10.13560/j.cnki.biotech.bull.1985.2024-0313

摘要/Abstract

摘要：

【目的】探究解磷菌及其发酵产物在铅污染土壤中对玉米（Zea mays L.）根际土壤性质和微生物群落组成和多样性的影响。【方法】在筛选具有耐铅解磷功能的巴氏克雷伯氏菌（Klebsiella pasteurii）的基础上，采用盆栽实验在铅污染土壤中培育种植玉米，在其根际施加LB培养基、上清液（菌剂分泌物）、菌液（只含菌体细胞）和发酵液（上清液+菌体细胞），并设置无菌水对照，探究解磷菌对根际土壤理化性质及微生物群落结构的影响。【结果】巴氏克雷伯氏菌的上清液、菌液和发酵液对玉米根际细菌群落多样性并无显著影响，而菌液显著增加了土壤真菌群落的Shannon指数和Chao指数；菌株的上清液、菌液、发酵液均增加了拟杆菌门（Bacteroidetes）与放线菌门（Actinobacteria）等耐重金属的微生物类群的相对丰度，同时上清液、发酵液增加了变形菌门（Proteobacteria）和被孢霉门（Mortierellomycota）的丰度，上清液提高了鞘氨醇单胞菌属（Sphingomonas）、芽球菌属（Blastococcus）、慢生根瘤菌属（Bradyrhizobium）、古根菌属（Archaeorhizomyces）的相对丰度。此外，对玉米根际土壤差异菌属进行皮尔逊相关性分析，有7组差异菌属之间为正相关性，揭示了不同微生物菌属之间更趋向于形成互利共生关系。菌株的上清液、菌液、发酵液均显著增加了土壤酸性磷酸酶（Acp）的活性，其中在发酵液处理组中玉米根际土壤Acp活性最高为574.44 mg/g，24^-1，施用上清液、菌液显著增加了根际土壤中碱解氮（AN）的含量，较对照组分别增加了47.4%、39.5%，三个处理组均显著降低了土壤pH值。通过冗余分析（RDA）发现土壤AN、Acp、pH值、可溶性磷（AP）是影响微生物群落结构的主要影响因素。【结论】外源施用解磷菌及其发酵产物有利于改善铅污染土壤肥力质量，影响土壤微生物群落的组成和结构，为铅污染农田接种解磷菌并提高土壤养分、改善土壤微生物群落结构提供了理论依据。

关键词: 重金属, 污染土壤, 解磷菌, 微生物群落结构, 土壤性质

Abstract:

【Objective】To explore the effects of phosphate-solubilizing bacteria（PSB）and their fermentation products on the properties of maize（Zea mays L.）rhizosphere soil and the composition and diversity of microbial community in lead-contaminated soil. 【Method】Based on the screening of Klebsiella pasteurii with lead-resistant and phosphate-solubilizing function, corn was cultivated in lead-contaminated soil by pot experiment. LB medium, supernatant（bacterial secretion）, bacterial solution（only bacterial cells）and fermentation broth（supernatant + bacterial cells）were applied to the rhizosphere, and sterile water control was set to explore the effects of phosphate-solubilizing bacteria on the physical and chemical properties of rhizosphere soil and microbial community structure.【Result】The supernatant, bacterial liquid and fermentation liquid of K.baumannii had no significant effect on the diversity of bacterial community in maize rhizosphere, while the bacterial liquid significantly increased the Shannon index and Chao index of soil fungal community. The supernatant, bacterial liquid and fermentation liquid of the strain increased the relative abundances of heavy metal-resistant microbial groups such as Bacteroidetes and Actinobacteria, while the supernatant and fermentation liquid increased the abundances of Proteobacteria and Mortierellomycota. The supernatant increased the relative abundances of Sphingomonas, Blastococcus, Bradyrhizobium and Archaeorhizomyces. In addition, Pearson correlation analysis of differential genera in the maize rhizosphere soil showed that there were positive correlations among 7 groups of differential genera, which revealed that different microbial genera tended to form mutually beneficial symbiotic relationships. The supernatant, bacterial liquid and fermentation liquid of the strain significantly increased the activities of soil acid phosphatase（Acp）. Among them, the Acp activity of maize rhizosphere soil in the fermentation liquid treatment group was the highest（574.44 mg/g, 24^-1）. The application of supernatant and bacterial liquid significantly increased the content of alkali-hydrolyzed nitrogen（AN）in the rhizosphere soil, which was 47.4% and 39.5% higher than that of the control, respectively. The three treatment groups significantly reduced the soil pH value. Through redundancy analysis（RDA）, it was found that soil AN, Acp, pH value and available phosphorus（AP）were the main factors affecting the microbial community structure.【Conclusion】This study reveales that exogenous application of PSB and their fermentation products was beneficial to improving the fertility of lead-contaminated soil, and affectes the composition and structure of soil microbial community, which provides a theoretical basis for inoculating PSB and improving soil nutrients and soil microbial community structure in lead-contaminated farmland.

Key words: heavy metals, contaminated soil, phosphate-solubilizing bacteria, microbial community structure, soil properties

温绍福, 江润海, 朱城强, 张梅, 余小琴, 杨杰惠, 杨小容, 侯秀丽. 铅污染土壤中解磷菌对玉米根际土壤性质和微生物群落结构的影响[J]. 生物技术通报, 2024, 40(9): 225-237.

WEN Shao-fu, JIANG Run-hai, ZHU Cheng-qiang, ZHANG Mei, YU Xiao-qin, YANG Jie-hui, YANG Xiao-rong, HOU Xiu-li. Effects of Phosphate-solubilizing Bacteria on the Rhizosphere Soil Properties and Microbial Community Structure of Maize in Lead-contaminated Soil[J]. Biotechnology Bulletin, 2024, 40(9): 225-237.

图/表 8

图1 细菌（A）和真菌（B）在OTUs水平上差异丰度在不同处理中的分布 CK：无菌水对照；T1: LB培养基；T2：上清液（菌剂分泌物）；T3：菌液（只含菌体细胞）；T4：发酵液（上清液+菌体细胞），下同

Fig. 1 Distribution of different abundances of bacteria（A）and fungi（B）at the OTUs level in different treatments CK: Sterile water control; T1: luria-bertani medium; T2: supernatant（bacterial secretion）; T3: bacterial liquid（only containing bacterial cells）; T4: fermentation broth（supernatant + bacterial cells）. The same below

图2 玉米根际土壤细菌和真菌群落Alpha多样性指数变化

Fig. 2 Changes of Alpha diversity index of bacterial and fungal communities in maize rhizosphere soil ***，P < 0.001；**，P < 0.01；*，P < 0.05. The same below

图3 玉米根际土壤细菌群落和真菌群落相对丰度

Fig. 3 Relative abundances of bacterial and fungal community in maize rhizosphere soil

图4 玉米根际土壤细菌属水平相对丰度在不同处理中的差异变化

Fig. 4 Differences in the relative abundances of bacterial genera in the rhizosphere soils of maize under different treatments

图5 玉米根际土壤真菌属水平相对丰度在不同处理中的差异变化

Fig. 5 Differences in relative abundances of fungi in the rhizosphere soils of maize under different treatments

图6 玉米根际土壤差异菌属的相关性分析

Fig. 6 Correlation analysis of differential bacterial genera in maize rhizosphere soil

表1 玉米根际土壤性质

Table 1 Properties of maize rhizosphere soil

处理 Treatment	有效磷 AP /（mg·kg^-1）	碱解氮 AN /（mg·kg^-1）	脲酶 Urease/（mg·kg^-1, 24^-1）	酸性磷酸酶 Acid phosphatase /（mg·kg^-1, 24^-1）	pH
CK	16.77 ±1.68a	133.00 ±9.90b	21783.70 ±3288.05a	270.00 ±33.00c	7.12 ±0.05a
T1	17.48 ±1.48a	122.50 ±4.95b	2901.60 ±274.99c	505.00 ±16.50ab	7.11 ±0.04a
T2	14.41 ±0.38b	196.00 ±19.80a	14216.60 ±1860.19b	486.67 ±103.71ab	6.70 ±0.08b
T3	13.88 ±0.13b	185.50 ±34.65a	4473.30 ±1357.42c	451.67 ±40.07b	6.57 ±0.03c
T4	12.36 ±0.54b	157.50 ±4.95ab	19522.25±46.03a	574.44 ±9.62a	6.71 ±0.05b

图7 差异细菌属（A）、真菌属（B）相对丰度与土壤性质的冗余分析

Fig. 7 Redundancy analysis of the relative abundances of differential bacteria（A）and fungi（B）and soil properties

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