基于转录组学揭示腐殖酸影响纳米三氧化二铬对小球藻的毒性机制

doi:10.16258/j.cnki.1674-5906.2024.08.013

生态环境学报 ›› 2024, Vol. 33 ›› Issue (8): 1289-1297.DOI: 10.16258/j.cnki.1674-5906.2024.08.013

• 研究论文【环境科学】 • 上一篇下一篇

基于转录组学揭示腐殖酸影响纳米三氧化二铬对小球藻的毒性机制

李玉彩¹^,²(), 杨垒¹^,²^,^*(), 梁贤³, 孟红艳¹^,², 刘焕焕¹^,², 石辉¹^,², 任勇翔¹^,²

1.西安建筑科技大学/陕西省环境工程重点实验室，陕西西安 710055
2.西安建筑科技大学/西北水资源与环境生态教育部重点实验室，陕西西安 710055
3.西安市燃气规划设计院有限公司，陕西西安 710055

收稿日期:2024-05-01 出版日期:2024-08-18 发布日期:2024-09-25
通讯作者: *杨垒。E-mail: yangleigps@xauat.edu.cn
作者简介:李玉彩（1997年生)，女，硕士研究生，研究方向为污水生态处理技术。E-mail: 3563274797@qq.com
基金资助:
国家自然科学基金项目(52300216);陕西省自然科学基础研究计划项目(2022JM-217);西安建筑科技大学前沿交叉领域培育专项(X20230076)

Transcriptomics Analysis Reveals the Impact of Humic Acid on the Toxicity of Nano-Cr₂O₃ to Chlorella sp.

LI Yucai¹^,²(), YANG Lei¹^,²^,^*(), LIANG Xian³, MENG Hongyan¹^,², LIU Huanhuan¹^,², SHI Hui¹^,², REN Yongxiang¹^,²

1. Shaanxi Key Laboratory of Environmental Engineering/Xi’an University of Architecture and Technology, Xi’an 710055, P. R. China
2. Key Laboratory of Northwest Water Resource, Environment and Ecology, Ministry of Education/Xi’an University of Architecture and Technology, Xi’an 710055, P. R. China
3. Xi’an Gas Planning and Design Institute Co., Ltd., Xi’an 710055, P. R. China

Received:2024-05-01 Online:2024-08-18 Published:2024-09-25

摘要/Abstract

摘要：

纳米材料（MNPs）暴露于天然水体中会与溶解性有机物（DOM）相互作用，影响其生物效应，但对于DOM作用下MNPs毒性机制的研究还不充分。分析了不同DOM对纳米三氧化二铬（nCr₂O₃）胁迫条件下小球藻生长生理影响，同时基于转录组学揭示了腐殖酸（HA）和nCr₂O₃共暴露条件下对小球藻的毒性作用机理。结果表明，加入HA后，小球藻的细胞密度和光合色素含量均显著提高，这在一定程度上缓解了nCr₂O₃对藻细胞的光合活性的影响。同时，海藻酸（AA）在长期作用下加重了nCr₂O₃对小球藻的毒性效应，而牛血清蛋白（BSA）对藻细胞密度和光合活性并未产生显著的影响。在1-10 mg·L⁻¹范围内，HA对nCr₂O₃的毒性缓解作用与HA浓度呈正相关，且随着培养时间的增加，HA对nCr₂O₃的毒性缓解作用更加明显。相应的，低浓度的HA就可显著减轻由nCr₂O₃所引起藻细胞内的氧化应激，减少活性氧（ROS）的累积。通过细胞结构观察，HA可以减轻由内化的nCr₂O₃对叶绿体造成的细胞器损伤，减少细胞空泡化，同时形成具有解毒功能的电子致密颗粒。转录组结果显示，加入HA使DNA复制相关基因表达上调，以及通过影响藻细胞碳水化物代谢和脂质代谢过程来缓解nCr₂O₃的毒性作用。因此，HA通过与nCr₂O₃相互作用，诱导小球藻内相关基因表达水平的变化来缓解nCr₂O₃的毒性效应。该研究从分子层面对进一步明确纳米颗粒在实际水体中的生物效应及控制策略提供理论依据。

关键词: 纳米颗粒, 溶解性有机物, 腐殖酸, 小球藻, 氧化应激, 转录组学

Abstract:

Nanomaterials (MNPs) can interact with dissolved organic matter (DOM) and affect its biological effects when exposed to natural water. However, the toxicity mechanism of MNPs under the action of DOM has not yet been fully studied. This study analyzed the effects of different DOM concentrations on the growth and physiology of Chlorella sp. under nCr₂O₃ stress conditions. Furthermore, transcriptomic analysis was performed to elucidate the toxic mechanisms of humic acid (HA) and nCr₂O₃ in Chlorella sp. The results showed that the cell density and photosynthetic pigment content of Chlorella sp. increased significantly after HA addition, which alleviated the effect of nCr₂O₃ on the photosynthetic activity of algal cells to a certain extent. Simultaneously, alginate (AA) aggravated the toxic effect of nCr₂O₃ on Chlorella sp. under long-term action, whereas the addition of bovine serum albumin (BSA) had no significant effect on algal cell density and photosynthetic activity. Within the concentration range of 1‒10 mg·L⁻¹, the mitigating effect of HA on nCr₂O₃ toxicity was positively correlated with the HA concentration, and the toxicity mitigation effect of HA on nCr₂O₃ was more obvious with increasing treatment time. Correspondingly, low concentrations of HA significantly reduced oxidative stress induced by nCr₂O₃ in algal cells and decreased reactive oxygen species (ROS) accumulation. Through observation of the cell structure, HA could alleviate organelle damage caused by internalized nCr₂O₃ in chloroplasts, reduce cell vacuolization, and form electron-dense particles with detoxifying functions. Transcriptomic analysis indicated that HA addition upregulated the expression of DNA replication-related genes and alleviated the toxic effects of nCr₂O₃ by affecting carbohydrate and lipid metabolism in algal cells. Therefore, HA mitigates the toxic effects of nCr₂O₃ through interactions that induce changes in the expression of the relevant genes in Chlorella sp. This study provides a theoretical basis for further clarifying the biological effects and control strategies of nanoparticles in water bodies at the molecular level.

Key words: nanoparticles, dissolved organic matter, humic acid, Chlorella sp., oxidative stress, transcriptomics

中图分类号:

X171
X703

李玉彩, 杨垒, 梁贤, 孟红艳, 刘焕焕, 石辉, 任勇翔. 基于转录组学揭示腐殖酸影响纳米三氧化二铬对小球藻的毒性机制[J]. 生态环境学报, 2024, 33(8): 1289-1297.

LI Yucai, YANG Lei, LIANG Xian, MENG Hongyan, LIU Huanhuan, SHI Hui, REN Yongxiang. Transcriptomics Analysis Reveals the Impact of Humic Acid on the Toxicity of Nano-Cr₂O₃ to Chlorella sp.[J]. Ecology and Environment, 2024, 33(8): 1289-1297.

图/表 7

图1 外源DOM作用下nCr2O3（20 mg·L?1）对小球藻细胞密度和光合色素的影响不同小写字母代表各处理间具有显著性差异（p<0.05），n=3。下同

Figure 1 Effects of nCr2O3 (20 mg·L?1) on cell density and photosynthetic pigments of Chlorella sp. under the action of exogenous DOM

图2 不同质量浓度HA条件下nCr2O3（20 mg·L?1）对小球藻细胞密度和光合色素含量的影响

Figure 2 Effects of nCr2O3 (20 mg·L?1) on cell density and photosynthetic pigment content of Chlorella sp. under different concentrations of HA

图3 不同浓度HA影响下nCr2O3（20 mg·L?1）对小球藻氧化应激水平的影响

Figure 3 The effect of nCr2O3 (20 mg·L?1) on the oxidative stress level of Chlorella sp. under different concentrations of HA

图4 HA作用下小球藻受nCr2O3（20 mg·L?1）胁迫下的TEM图像条形图代表500 nm。白色箭头表示质壁分解和膜分解，红色箭头表LD示nCr2O3的细胞内化，黄色箭头表示电子密集颗粒。缩写：P，淀粉核；SG，淀粉粒；LD，脂滴；V，空泡化；CW，细胞壁；chl，叶绿体

Figure 4 TEM images of Chlorella sp. under the stress of nCr2O3 (20 mg·L?1) with HA

图5 10 mg·L?1 HA影响下nCr2O3（20 mg·L?1）对藻细胞内差异表达基因分析

Figure 5 Analysis of differentially expressed genes in algal cells under the influence of nCr2O3 (20 mg·L?1) under the influence of 10 mg·L?1 HA

图6 10 mg·L?1 HA影响下nCr2O3（mg·L?1）对藻细胞内DEGs的GO富集分析

Figure 6 GO enrichment analysis of DEGs in algal cells by nCr2O3 (20 mg·L?1) under the influence of 10 mg·L?1 HA

表1 DEGs的KEEG富集分析

Table 1 KEEG enrichment analysis of DEGs

组	通路ID	通路名称	水平2	上调基因	下调基因	p值	p值校正值
A Vs C₁	Ko03010	核糖体	翻译	RPL4, RPL35, RPL5，RPS4, RPL7A, RPLP0, RPL10A, RPL13, RPL15, RPL27, RPS6, RPS30		0.0012	0.0516
	Ko00910	氮代谢	能量代谢		GLUD, GLUL	0.0070	0.1229
	Ko00250	丙氨酸、天冬氨酸和谷氨酸代谢	氨基酸代谢		GLUD, GFPT, (glms), GLUL	0.0287	0.2805
	Ko00220	精氨酸生物合成	氨基酸代谢		GLUL, (glnA), gdha	0.0319	0.2805
	Ko00730	辅助因子和维生素代谢	硫胺素代谢		thiC, THI4, THI1, NAD+	0.0437	0.3203
C₁ Vs C₂	Ko03030	DNA复制	复制与修复	RPA, RPA1, RPA3，PCNA, POLA1, PRI1，POLD2, POLD3, Mcm4, Mcm5, Mcm6, PCNA, RFC2/4		1.06158E-08	2.68934E-0
C₁ Vs C₂	Ko00053	抗坏血酸与醛酸代谢	碳水化合物代谢	ALDH, L-ascorbate oxidase, oxidoreductase		0.047654392	0.235571983

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基于转录组学揭示腐殖酸影响纳米三氧化二铬对小球藻的毒性机制

Transcriptomics Analysis Reveals the Impact of Humic Acid on the Toxicity of Nano-Cr₂O₃ to Chlorella sp.

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基于转录组学揭示腐殖酸影响纳米三氧化二铬对小球藻的毒性机制

Transcriptomics Analysis Reveals the Impact of Humic Acid on the Toxicity of Nano-Cr2O3 to Chlorella sp.

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Transcriptomics Analysis Reveals the Impact of Humic Acid on the Toxicity of Nano-Cr₂O₃ to Chlorella sp.