小球藻(Chlorella vulgaris)对泰乐菌素的胁迫响应与耐受性

doi:10.3969/j.issn.1004-1524.20231269

浙江农业学报 ›› 2024, Vol. 36 ›› Issue (10): 2316-2327.DOI: 10.3969/j.issn.1004-1524.20231269

小球藻(Chlorella vulgaris)对泰乐菌素的胁迫响应与耐受性

聂红丽¹(), 成琪璐², 孙万春², 马进川², 林辉², 马军伟²^,^*()

1.浙江农林大学环境与资源学院,浙江杭州 311300
2.浙江省农业科学院省部共建农产品质量安全危害因子与风险防控国家重点实验室,环境资源与土壤肥料研究所,浙江杭州 310021

收稿日期:2023-11-10 出版日期:2024-10-25 发布日期:2024-10-30
作者简介:聂红丽(1993—),女,河南商丘人,硕士研究生,从事微藻生态毒理研究。E-mail:648600770@qq.com
通讯作者: ^*马军伟,E-mail:majw@zaas.ac.cn
基金资助:
国家自然科学基金(42207285);浙江省科技创新领军人才项目(2021R52045)

Stress response and tolerance of Chlorella vulgaris to tylosin

NIE Hongli¹(), CHENG Qilu², SUN Wanchun², MA Jinchuan², LIN Hui², MA Junwei²^,^*()

1. School of Environment and Resources, Zhejiang A&F University, Hangzhou 311300, China
2. State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Environment, Resources, Soil and Fertilizer, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2023-11-10 Online:2024-10-25 Published:2024-10-30

摘要/Abstract

摘要：

抗生素是一类典型的新兴污染物,对水生生物有较高的生态风险。微藻作为自然水体的初级生产者,极易受到环境变化尤其是污染物暴露的影响。但是目前关于微藻对泰乐菌素(tylosin, TYN)胁迫的响应研究还十分有限。本研究选用模式藻种小球藻(Chlorella vulgaris)为试验材料,从细胞生长、生理、代谢、基因表达等层面研究TYN处理下微藻的胁迫耐受性。结果表明,TYN对C. vulgaris的生长呈典型的hormesis效应:即低质量浓度(≤0.5 mg·L^-1)促进藻的生长;随着TYN处理浓度的升高,其对C. vulgaris生长的抑制作用越明显,并导致细胞形态改变、质壁分离、类囊体降解等不同程度的氧化损伤。转录组测序发现,高质量浓度TYN处理8 d后,C. vulgaris中光合作用、核糖体生物合成、抗氧化等相关的基因表达水平显著下调,进一步表明高质量浓度TYN对微藻细胞有抑制效应。值得注意的是,C. vulgaris可通过增加胞外多糖和蛋白质含量提高TYN与胞外聚合物中-OH、-COOH、-NH等官能团的结合水平,以应对高质量浓度TYN对细胞的胁迫效应。研究系统揭示了C. vulgaris对TYN的胁迫响应与耐受性,相关结果可以为抗生素的水体生态风险评估提供参考依据。

关键词: 新兴污染物, 微藻, 胁迫响应, 转录组学分析, 胞外聚合物, 逆境耐受

Abstract:

Antibiotics are typical emerging pollutants with high ecological risk to aquatic organisms. As the primary producers of natural water, microalgae are highly susceptible to environmental changes, especially exposure to pollutants. However, the studies on the response of microalgae to tylosin (TYN) stress are still limited. In this study, Chlorella vulgaris, a model algae species, was selected as the experimental material to investigate the stress tolerance of microalgae exposed to TYN from the aspects of microalgal growth, physiology, metabolism, and gene expression. The results showed that TYN had a typical hormesis effect on the growth of C. vulgaris. Microalgal growth was promoted by low concentration of TYN (≤ 0.5 mg·L^-1), with the increase of TYN exposure concentration, its inhibitory effect on the growth of C.vulgaris was more obvious, and it led to different degrees of oxidative damage such as cell morphological changes, plasmolysis, and thylakoid degradation. Transcriptome sequencing showed that the expression levels of genes related to photosynthesis, ribosome biosynthesis, and antioxidant in C. vulgaris were significantly down-regulated after 8 days of high-concentration TYN treatment, further indicating that high-concentration TYN had an inhibitory effect on C. vulgaris cells. It was worth noting that C. vulgaris could improve the binding level of TYN to functional groups such as —OH, —COOH and —NH in extracellular polymeric substance by increasing the content of extracellular polysaccharides and proteins, so as to cope with the stress effect of high concentration TYN on cells. This study systematically revealed the stress responses and tolerance of C. vulgaris to TYN, and the relevant results provided a reference for the ecological risk assessment of antibiotics in water.

Key words: emerging contaminant, microalgae, stress response, transcriptomic analysis, extracellular polymeric substance, stress tolerance

中图分类号:

S184

聂红丽, 成琪璐, 孙万春, 马进川, 林辉, 马军伟. 小球藻(Chlorella vulgaris)对泰乐菌素的胁迫响应与耐受性[J]. 浙江农业学报, 2024, 36(10): 2316-2327.

NIE Hongli, CHENG Qilu, SUN Wanchun, MA Jinchuan, LIN Hui, MA Junwei. Stress response and tolerance of Chlorella vulgaris to tylosin[J]. Acta Agriculturae Zhejiangensis, 2024, 36(10): 2316-2327.

图/表 7

表1 BG11培养基组分

Table 1 The component of BG11 cultural medium mg·L-1

成分 Ingredient	含量 Content	成分 Ingredient	含量 Content
硝酸钠NaNO₃	1 500	碳酸钠Na₂CO₃	20
三水磷酸氢二钾K₂HPO₄·3H₂O	40	硼酸H₃BO₃	2.860
七水硫酸镁MgSO₄·7H₂O	75	一水氯化锰MnCl₂·H₂O	1.810
二水氯化钙CaCl₂·2H₂O	36	七水硫酸锌ZnSO₄·7H₂O	0.222
柠檬酸C₆H₈O₇	6	五水硫酸铜CuSO₄·5H₂O	0.079
柠檬酸铁铵C₆H₁₁FeNO₇	6	二水钼酸钠Na₂MoO₄·2H₂O	0.390
乙二胺四乙酸EDTA	1	六水硝酸钻Co(NO₃)₂·6H₂O	0.049
A5(微量金属溶液)A5 (Trace mental solution)	1 000

图1 C. vulgaris在不同质量浓度TYN处理8 d内的生长情况

Fig.1 Growth of C. vulgaris during 8-day exposure to different mass concentrations of TYN

图2 不同质量浓度(0、0.50、5.00 mg·L-1)TYN处理4 d后C. vulgaris细胞的透射电镜图 CW、T、P、S分别代表细胞壁、类囊体、蛋白核和淀粉鞘。

Fig.2 Transmission electron micrographs of C. vulgaris cells after 4-day exposure to TYN at mass concentration of 0, 0.50, and 5.00 mg·L-1 CW, T, P, and S represent cell wall, thylakoid, pyrenoid and starch sheath, respectively.

图3 不同质量浓度TYN处理下C. vulgaris的EPS分泌情况 A、B、C分别为暴露2、4、8 d时C. vulgaris中EPS总量、胞外多糖含量、胞外蛋白含量变化,以105个细胞计;D为暴露8 d时C. vulgaris细胞表面的FTIR图。

Fig.3 EPS secretion of C.vulgaris under different mass concentrations of TYN treatment A, B and C, Changes in contents of total EPS, extracellular polysaccharides and extracellular proteins in 105 C. vulgaris cells after 2, 4, and 8 days of exposure, respectively; D, FTIR spectra of the C. vulgaris cell surface after 8 days of exposure.

图4 0.05、5.00 mg·L-1 TYN暴露2、8 d后的差异表达基因数及其在GO功能注释中显著富集的前3种类别

Fig.4 The number of differentially expressed genes, and the top 3 categories of genes significantly enriched in the GO functional annotation after 0.05 and 5.00 mg·L-1 TYN exposure for 2 and 8 days

图5 5.00 mg·L-1 TYN暴露8 d后C. vulgaris细胞内DEG显著富集的KEGG通路、关键代谢途径 T5代表5.00 mg·L-1 TYN处理。

Fig.5 The significantly enriched KEGG pathways of DEGs and the key metabolic pathways in C. vulgaris cells after 8 days of exposure to 5.00 mg·L-1 TYN T5 represents 5.00 mg·L-1 TYN treatment.

图6 C. vulgaris对TYN的去除情况 A,1.00、5.00 mg·L-1 TYN暴露2、4、8 d时培养体系中TYN的去除率;B,C. vulgaris EPS的FTIR图;C和D,培养8 d后,对照组与5.00 mg·L-1 TYN处理中C. vulgaris EPS的3D-EEM图;EM是发射波长,EX是激发波长。

Fig.6 The removal of TYN by C.vulgaris A, Removal rate of 1.00 and 5.00 mg·L-1 TYN in the culture system on 2, 4, and 8 days; B, The fourier transform infrared spectrum of C. vulgaris EPS; C and D, 3D-EEM plots of C. vulgaris EPS in the control and 5.00 mg·L-1 TYN treatment on day 8; EM is the emission wavelength and EX is the excitation wavelength.

参考文献 33

[1]	LILIANA S. Antimicrobials and antibiotic-resistant bacteria: a risk to the environment and to public health[J]. Water, 2020, 12(12): 3313.
[2]	ARSÈNE M M J, DAVARES A K L, ANDREEVNA S L, et al. The use of probiotics in animal feeding for safe production and as potential alternatives to antibiotics[J]. Veterinary World, 2021, 14(2): 319-328.
[3]	RIAZ L, MAHMOOD T, KHALID A, et al. Fluoroquinolones (FQs) in the environment: a review on their abundance, sorption and toxicity in soil[J]. Chemosphere, 2018, 191: 704-720.
[4]	ARSIC B, BARBER J, ČIKOŠ A, et al. 16-membered macrolide antibiotics: a review[J]. International Journal of Antimicrobial Agents, 2018, 51(3): 283-298.
[5]	LEKAGUL A, TANGCHAROENSATHIEN V, YEUNG S. Patterns of antibiotic use in global pig production: a systematic review[J]. Veterinary and Animal Science, 2019, 7: 100058.
[6]	BOXALL A, TIEDE K, BRYNING G, et al. Desk-based study of current knowledge on veterinary medicines in drinking water and estimation of potential levels[EB/OL]. [2023-11-09]. https://www.academia.edu/22907779/DESK_BASED_STUDY_OF_CURRENT_KNOWLEDGE_ON_VETERINARY_MEDICINES_IN_DRINKING_WATER_AND_ESTIMATION_OF_POTENTIAL_LEVELS.
[7]	SANDOZ M A, WOOTEN K J, CLENDENING S L, et al. Transport mechanisms for veterinary pharmaceuticals from beef cattle feedyards to wetlands: is aerial deposition a contributing source?[J]. Agriculture, Ecosystems & Environment, 2018, 252: 14-21.
[8]	MAIA C, SOUSA C A, SOUSA H, et al. Parabens removal from wastewaters by microalgae: ecotoxicity, metabolism and pathways[J]. Chemical Engineering Journal, 2023, 453: 139631.
[9]	LE V V, TRAN Q G, KO S R, et al. How do freshwater microalgae and cyanobacteria respond to antibiotics?[J]. Critical Reviews in Biotechnology, 2023, 43(2): 191-211.
[10]	BABIAK W, KRZEMIŃSKA I. Extracellular polymeric substances (EPS) as microalgal bioproducts: a review of factors affecting EPS synthesis and application in flocculation processes[J]. Energies, 2021, 14(13): 4007.
[11]	LI J R, WANG Y J, FAN Z Q, et al. Toxicity of tetracycline and metronidazole in Chlorella pyrenoidosa[J]. International Journal of Environmental Research and Public Health, 2023, 20(4): 3623.
[12]	CHEN S, WANG L Q, FENG W B, et al. Sulfonamides-induced oxidative stress in freshwater microalga Chlorella vulgaris: evaluation of growth, photosynthesis, antioxidants, ultrastructure, and nucleic acids[J]. Scientific Reports, 2020, 10(1): 8243.
[13]	LU D L, MA Z H, PENG J L, et al. Integrated comparison of growth and oxidative stress induced by tylosin in two freshwater algae Chlorella vulgaris and Raphidocelis subcapitata[J]. Ecotoxicology, 2022, 31(3): 376-384.
[14]	CHENG Q L, DU L N, XU L G, et al. Toxicity alleviation and metabolism enhancement of nonylphenol in green algae Dictyosphaerium sp. by NaHCO₃[J]. Science of the Total Environment, 2022, 848: 157698.
[15]	ZHOU G J, PENG F Q, ZHANG L J, et al. Biosorption of zinc and copper from aqueous solutions by two freshwater green microalgae Chlorella pyrenoidosa and Scenedesmus obliquus[J]. Environmental Science and Pollution Research International, 2011, 19(7): 2918-2929.
[16]	QU F S, LIANG H, HE J G, et al. Characterization of dissolved extracellular organic matter (dEOM) and bound extracellular organic matter (bEOM) of Microcystis aeruginosa and their impacts on UF membrane fouling[J]. Water Research, 2012, 46(9): 2881-2890.
[17]	XIAO M, LI M, DUAN P F, et al. Insights into the relationship between colony formation and extracellular polymeric substances (EPS) composition of the cyanobacterium Microcystis spp[J]. Harmful Algae, 2019, 83: 34-41.
[18]	ZHU W J, MA W, LI C X, et al. Well-designed multihollow magnetic imprinted microspheres based on cellulose nanocrystals (CNCs) stabilized Pickering double emulsion polymerization for selective adsorption of bifenthrin[J]. Chemical Engineering Journal, 2015, 276: 249-260.
[19]	VIDYADHARANI G, DHANDAPANI R. Fourier transform infrared (FTIR) spectroscopy for the analysis of lipid from Chlorella vulgaris[J]. Elixir Applied Biology, 2013, 61: 16753-16756.
[20]	MOHAN N H, CHOUDHURY M, AMMAYAPPAN L, et al. Characterization of secondary structure of pig hair fiber using fourier-transform infrared spectroscopy[J]. Journal of Natural Fibers, 2022, 19(11): 4223-4235.
[21]	CARPENTER J, SAHARAN V K. Ultrasonic assisted formation and stability of mustard oil in water nanoemulsion: effect of process parameters and their optimization[J]. Ultrasonics Sonochemistry, 2017, 35: 422-430.
[22]	LIU W H, MING Y, HUANG Z W, et al. Impacts of florfenicol on marine diatom Skeletonema costatum through photosynthesis inhibition and oxidative damages[J]. Plant Physiology and Biochemistry, 2012, 60: 165-170.
[23]	DONG X T, SUN S H, JIA R B, et al. Effects of sulfamethoxazole exposure on the growth, antioxidant system of Chlorella vulgaris and Microcystis aeruginosa[J]. Bulletin of Environmental Contamination and Toxicology, 2020, 105(3): 358-365.
[24]	MAO Y F, YU Y, MA Z X, et al. Azithromycin induces dual effects on microalgae: roles of photosynthetic damage and oxidative stress[J]. Ecotoxicology and Environmental Safety, 2021, 222: 112496.
[25]	VILLACIS R A R, FILHO J S, PIÑA B, et al. Integrated assessment of toxic effects of maghemite (γ-Fe₂O₃) nanoparticles in zebrafish[J]. Aquatic Toxicology, 2017, 191: 219-225.
[26]	CHENG Q L, LIU Y Z, XU L G, et al. Regulation and role of extracellular polymeric substances in the defensive responses of Dictyosphaerium sp. to enrofloxacin stress[J]. Science of the Total Environment, 2023, 896: 165302.
[27]	XIE Q T, LIU N, LIN D H, et al. The complexation with proteins in extracellular polymeric substances alleviates the toxicity of Cd (II) to Chlorella vulgaris[J]. Environmental Pollution, 2020, 263: 114102.
[28]	LENG L J, WEI L, XIONG Q, et al. Use of microalgae based technology for the removal of antibiotics from wastewater: a review[J]. Chemosphere, 2020, 238: 124680.
[29]	宋晓梅, 胡桂林, 江钰, 等. Fe³⁺对水溶液中泰乐菌素光解的影响[J]. 安全与环境学报, 2018, 18(4): 1569-1572.
	SONG X M, HU G L, JIANG Y, et al. Influence of Fe³⁺ on the tylosin photolysis in aqueous solution[J]. Journal of Safety and Environment, 2018, 18(4): 1569-1572. (in Chinese with English abstract)
[30]	YAHIAT S, FOURCADE F, BROSILLON S, et al. Removal of antibiotics by an integrated process coupling photocatalysis and biological treatment: case of tetracycline and tylosin[J]. International Biodeterioration & Biodegradation, 2011, 65(7): 997-1003.
[31]	CHEN Q H, ZHANG L, HAN Y H, et al. Degradation and metabolic pathways of sulfamethazine and enrofloxacin in Chlorella vulgaris and Scenedesmus obliquus treatment systems[J]. Environmental Science and Pollution Research International, 2020, 27(22): 28198-28208.
[32]	JIANG R X, WEI Y R, SUN J Y, et al. Degradation of cefradine in alga-containing water environment: a mechanism and kinetic study[J]. Environmental Science and Pollution Research International, 2019, 26(9): 9184-9192.
[33]	LU Z K, XU Y F, PENG L, et al. A two-stage degradation coupling photocatalysis to microalgae enhances the mineralization of enrofloxacin[J]. Chemosphere, 2022, 293: 133523.

小球藻(Chlorella vulgaris)对泰乐菌素的胁迫响应与耐受性

Stress response and tolerance of Chlorella vulgaris to tylosin

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 7

参考文献 33

相关文章 5

编辑推荐

Metrics

本文评价

[1]	邱文怡, 王诗雨, 李晓芳, 徐恒, 张华, 朱英, 王良超. MYB转录因子参与植物非生物胁迫响应与植物激素应答的研究进展[J]. 浙江农业学报, 2020, 32(7): 1317-1328.
[2]	罗龙皂, 林小爱, 杨佳, 刘烨, 田光明. 微藻净化畜禽养殖废水影响因素研究进展[J]. 浙江农业学报, 2020, 32(3): 552-558.
[3]	边建文, 崔岩, 杨宋琪, 罗光宏, 孟宪刚. 衣藻和固氮鱼腥藻对盐胁迫下小麦幼苗生长的影响[J]. 浙江农业学报, 2020, 32(10): 1748-1756.
[4]	罗龙皂, 林小爱, 朱峰, 曾凡健, 张邦喜, 刘烨, 田光明. 曝二氧化碳气体对近具刺链带藻在高氨氮废水中生长的影响[J]. 浙江农业学报, 2019, 31(9): 1541-1548.
[5]	程鹏飞, 王艳, 杨期勇, 刘德富, 刘天中. 微藻贴壁培养对沼液废水的处理效果[J]. 浙江农业学报, 2017, 29(9): 1564-1569.