铅污染矿区中耐铅解磷菌对玉米的促生及根际铅的固化效应

doi:10.16258/j.cnki.1674-5906.2024.02.013

生态环境学报 ›› 2024, Vol. 33 ›› Issue (2): 291-300.DOI: 10.16258/j.cnki.1674-5906.2024.02.013

• 研究论文【环境科学】 • 上一篇下一篇

铅污染矿区中耐铅解磷菌对玉米的促生及根际铅的固化效应

江润海¹(), 温绍福¹, 朱城强¹, 张梅¹, 杨润玲¹, 王春雪², 侯秀丽¹^,^*()

1.昆明学院农学与生命科学学院/高原湖泊生态与环境健康云南省高校协同创新中心，云南昆明 650214
2.云南磷化集团有限公司/国家磷资源开发利用工程技术研究中心，云南昆明 650600

收稿日期:2023-10-18 出版日期:2024-02-18 发布日期:2024-04-03
通讯作者: *侯秀丽。E-mail: hxlyn@aliyun.com
作者简介:江润海（1997年生），男，硕士研究生，研究方向为土壤重金属污染修复。E-mail: m15519322764@163.com
基金资助:
国家自然科学基金项目(42167009);国家自然科学基金项目(U1902202);云南省教育厅科学研究基金项目(2022Y712);云南省教育厅科学研究基金项目(2023Y0874);云南省高校联合基金项目(2018FH001-004);云南省教育厅云南高原湖泊-北美五大湖国际联合科技创新团队

Research on the Promotion of Maize Growth and Immobilization of Pb in the Rhizosphere by Pb-tolerant Phosphate Solubilizing Bacteria in Pb-contaminated Mining Areas

JIANG Runhai¹(), WEN Shaofu¹, ZHU Chengqiang¹, ZHANG Mei¹, YANG Runling¹, WANG Chunxue², HOU Xiuli¹^,^*()

1. Yunnan Collaborative Innovation Center for Plateau Lake Ecology and Environmental Health/College of Agronomy and Life Sciences, Kunming University, Kunming 650214, P. R. China
2. Yunnan Phosphorus Group Co., Ltd./National Engineering Research Center for Phosphorus Resource Development and Utilization, Kunming 650600, P. R. China

Received:2023-10-18 Online:2024-02-18 Published:2024-04-03

摘要/Abstract

摘要：

矿产资源的开采、冶炼活动造成了一定的生态环境问题，土壤中可溶性重金属随着地表径流和地下渗透造成矿区和周边农田重金属污染。而矿区中土壤微生物对重金属具有一定耐性，研究微生物对植物根际微生态环境的改善作用具有重要意义。在矿区废弃地土壤中筛选耐铅（Pb）解磷菌的基础上，将含有菌株分泌物的上清液、菌液、发酵液（上清液+菌株）分别施用到玉米（Zea mays L.）根际土壤中，对比三者对玉米的促生效果及根际土壤铅的形态变化，探究解磷菌对玉米的促生机制及其对土壤铅的固化作用。所筛菌株被鉴定为巴氏克雷伯菌（Klebsiella pasteurii），其通过分泌乙酸、乳酸、酒石酸和草酸对Ca₃(PO₄)₂的溶磷率为26.5%，并能分泌生长素（IAA）。在玉米根际土壤中施用菌株的上清液、菌液和发酵液后，较对照组玉米株高、茎直径、地上与地下生物量均显著增加，其中施用发酵液组增幅最高，较对照组分别增加了128%，216%、266%、147%。同时，3个处理组中玉米地上生物量中铅含量分别降低68.6%、58.1%、70.1%，地下部铅含量分别降低119%、36.7%、39.5%。施用菌株上清液、菌液和发酵液后均使玉米根际土壤中可溶态的铅向稳定态铅转化，这可能是巴氏克雷伯菌在解离土壤磷素的过程中，其解离的磷可能与铅形成稳定态的磷铅化合物，进而降低植物对铅的吸收。由此可以得出，在铅污染土壤中巴氏克雷伯菌能够显著固化重金属铅并抑制玉米对铅的吸收，对玉米具有促生作用，因此，巴氏克雷伯菌在土壤重金属修复和保障农作物食品安全方面具有重要价值。

关键词: 铅的固化, 铅污染矿区, 解磷菌, 促生, 铅的形态, 重金属, 土壤污染

Abstract:

The mining and smelting of mineral resources can cause environment issues not only contaminating mining areas but also surrounding farmland with heavy metals through surface runoff and underground infiltration. In the mining areas, soil microorganisms have developed resistance mechanisms to heavy metals, so it is important to investigate the role of these microbes in changing the microecological environment of the plant's rhizosphere. In this study, we isolated lead (Pb)-tolerant phosphate solubilizing bacteria (PSB) from the soil in a mining area. Clear supernatants, bacteria solution, and fermentation liquid (supernatant+bacteria) were applied in rhizosphere soil of maize (Zea mays L.) to compare their effects on promoting maize growth and changes of soil Pb speciation. The screened bacteria strain is Klebsiella pasteurii, which can dissolve Ca₃(PO₄)₂ with the solubilization capacity of 26.5% by secreting acetic acid, lactic acid, tartaric acid, and oxalic acid and secrete indole-3-acetic acid (IAA). After application of the clear supernatants, bacteria solution, and fermentation liquid of K. pasteurii to the rhizosphere soil of maize, the maize's height, stem thickness, and biomass, both aboveground and belowground, were significantly improved when compared to the control group. Especially in the fermentation liquid group, maize's height, stem thickness, and biomass, both aboveground and belowground, were significantly increased by 128%, 216%, 266%, and 147%, respectively, compared to the control group. Meanwhile, the Pb accumulation in aboveground biomass of three treatment groups decreased by 68.6%, 58.1%, and 70.1%, respectively. And the Pb accumulation in roots of maize decreased by 119%, 36.7%, and 39.5%, respectively. This study indicated that the application of clear supernatants, bacteria solution, and fermentation liquid resulted in the transformation of soluble Pb into stable Pb in the rhizosphere soil. It might be because that the phosphorus released by K. pasteurii chemically reacts with soluble Pb to form stable Pb-phosphate compounds, decreasing maize absorption of Pb. We conclude that in Pb-contaminated soil, K. pasteurii can significantly promote Pb immobilization and plant's growth, and inhibit maize's absorption of Pb. Therefore, K. pasteurii plays a significant role in the remediation of Pb-contaminated soil and ensuring the food safety of crops.

Key words: Pb immobilization, Pb-contaminated mining areas, phosphate solubilizing bacteria, promoting growth, form of Pb, heavy metals, soil pollution

中图分类号:

江润海, 温绍福, 朱城强, 张梅, 杨润玲, 王春雪, 侯秀丽. 铅污染矿区中耐铅解磷菌对玉米的促生及根际铅的固化效应[J]. 生态环境学报, 2024, 33(2): 291-300.

JIANG Runhai, WEN Shaofu, ZHU Chengqiang, ZHANG Mei, YANG Runling, WANG Chunxue, HOU Xiuli. Research on the Promotion of Maize Growth and Immobilization of Pb in the Rhizosphere by Pb-tolerant Phosphate Solubilizing Bacteria in Pb-contaminated Mining Areas[J]. Ecology and Environment, 2024, 33(2): 291-300.

图/表 10

表1 菌株的生理生化特征

Table 1 Physiological and biochemical identification of strains

生理生化实验指标	结果
接触酶	+
氧化酶	−
丙二酸	+
柠檬酸盐	+
葡萄	+
D-山梨醇	+
甲基红	+
V-P	−
吲哚	+
铁载体	+
革兰氏染色	−

图1 菌株基于16S rDNA基因序列的系统发育树

Figure 1 Phylogenetic tree of strain based on 16S rDNA gene sequences

图2 分离出的解磷菌株

Figure 2 Isolated phosphate-removing bacterial strain

图3 菌株溶磷量和培养液pH变化

Figure 3 Changes in phosphorus solubilization and culture medium pH of bacterial strain

图4 菌株对铅的耐受能力

Figure 4 The capacity of strain to tolerate lead

表2 菌株分泌物质量浓度

Table 2 The concentration of secretions from the strain

生长因子	质量浓度/(mg·L⁻¹)
吲哚-3-乙酸	49.5±0.81
乙酸	1 364±147
乳酸	25.6±0.66
酒石酸	40.1±4.03
草酸	50.2±3.47

图5 菌株对土壤铅胁迫下玉米生长的影响不同小写字母代表不同处理间差异显著（P<0.05），n=6

Figure 5 The impact of strain on the growth of maize under soil lead stress

图6 不同处理玉米根系扫描

Figure 6 Root systems of maize under different treatments

图7 玉米对铅的吸收数据进行多重比较分析，图中*代表不同处理间差异显著（*P<0.05，** P<0.01，*** P<0.001），n=3

Figure 7 The uptake of lead by maize

表3 菌株对土壤铅形态变化影响

Table 3 The impact of strain on soil lead forms mg·kg?1

土壤铅形态	处理
土壤铅形态	CK	T1	T2	T3	T4
水溶态	3.32±0.01c	3.34±0.00b	3.39±0.02a	3.34±0.00b	3.34±0.00b
可交换态	168±0.05b	175±3.61b	152±1.05d	155±0.98cd	157±0.95c
碳酸盐结合态	431±1.88b	459±1.64a	455±10.91a	407±7.00c	410±7.76c
铁锰氧化物结合态	1 046±46.48c	1 138±13.41ab	1 183±33.44a	1 129±26.10ab	1 089±17.32bc
强有机结合态	240±2.57d	308±2.12c	382±29.90a	335±12.91bc	341±2.91b
腐殖酸结合态	7.44±0.09b	7.35±0.13b	8.17±0.01a	8.14±0.01a	8.19±0.23a
残渣态	70.2±3.09d	69.0±3.23d	94.1±2.51b	86.2±2.06c	99.3±1.00a

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铅污染矿区中耐铅解磷菌对玉米的促生及根际铅的固化效应

Research on the Promotion of Maize Growth and Immobilization of Pb in the Rhizosphere by Pb-tolerant Phosphate Solubilizing Bacteria in Pb-contaminated Mining Areas

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