微塑料对外生菌根真菌生长和抗氧化系统的影响

doi:10.3969/j.issn.1004-1524.2022.05.20

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (5): 1049-1060.DOI: 10.3969/j.issn.1004-1524.2022.05.20

微塑料对外生菌根真菌生长和抗氧化系统的影响

李玉婷¹(), 李莎¹, 曹杰¹, 李骄杨¹, 张亮¹^,^*(), 许晓风¹^,²

1.南京师范大学泰州学院化学与生物工程学院,江苏泰州 225300
2.南京师范大学生命科学学院,江苏南京 210023

收稿日期:2021-06-04 出版日期:2022-05-25 发布日期:2022-06-06
通讯作者: 张亮
作者简介:* 张亮,E-mail: liangzai0061@126.com
李玉婷(1999—),女,江苏南通人,本科生,研究方向为环境污染物降解与修复。E-mail: 16180320@nnutc.edu.cn
基金资助:
江苏省高等学校自然科学基金(20KJD180003);江苏省高校实验室研究会立项课题(GS2019YB06);2020年江苏省大学生创新创业训练计划(202013843017);2021年度泰州市“凤城英才”青年科技人才托举工程

Effects of microplastics on growth and antioxidant system of ectomycorrhizal fungi

LI Yuting¹(), LI Sha¹, CAO Jie¹, LI Jiaoyang¹, ZHANG Liang¹^,^*(), XU Xiaofeng¹^,²

1. School of Chemistry and Bioengineering, Nanjing Normal University Taizhou College, Taizhou 225300, Jiangsu, China
2. School of Life Sciences, Nanjing Normal University, Nanjing 210023, China

Received:2021-06-04 Online:2022-05-25 Published:2022-06-06
Contact: ZHANG Liang

摘要/Abstract

摘要：

土壤中的微塑料污染及其毒理学效应已逐渐引起广泛关注,但微塑料对外生菌根真菌的毒性研究仍不多见。为此,以彩色豆马勃(Pisolithus tinctorius,Pt)和松乳菇(Lactarius delicious,Ld)作为供试菌株,选用粒径80 nm和4 μm的单分散聚苯乙烯塑料微球(PS-MPs)作为试验材料,采用固体平板法和液体培养法研究不同粒径不同质量浓度(10~300 mg·L^-1)的PS-MPs对外生菌根真菌生长情况、丙二醛(MDA)含量、可溶性蛋白含量、抗氧化酶活性、组织电导率等指标的影响。结果显示,高浓度(200~300 mg·L^-1)的PS-MPs显著(P<0.05)抑制2株真菌的生长,且粒径4 μm的PS-MPs较粒径80 nm的对两株真菌的生物量表现出更强的抑制作用。随PS-MPs质量浓度的升高,Pt和Ld的超氧化物歧化酶活性始终显著(P<0.05)高于对照,且表现出先升高后降低的趋势。当暴露于2种粒径的PS-MPs之中时,Pt和Ld的过氧化氢酶活性均在300 mg·L^-1处理下最低,且显著(P<0.05)低于对照;而过氧化物酶活性在各处理下均显著(P<0.05)高于对照。与对照相比,PS-MPs处理下,Pt和Ld的MDA含量显著(P<0.05)升高(除10 mg·L^-1 4 μm PS-MPs处理下的Pt和200 mg·L^-1 4 μm PS-MPs处理下的Ld外),可溶性蛋白含量显著(P<0.05)降低(除10、20 mg·L^-1 80 nm PS-MPs处理下的Pt和10 mg·L^-1 80 nm PS-MPs处理下的Ld外),菌丝组织电导率显著(P<0.05)升高。据此推测,PS-MPs对外生菌根真菌的影响机制可能涉及氧化应激反应,且不同菌株对不同粒径PS-MPs的响应不同。研究结果可为揭示微塑料对土壤外生菌根真菌的急性毒性提供依据。

关键词: 微塑料, 外生菌根真菌, 抗氧化酶

Abstract:

Microplastics pollution in soil and its toxicological effects have gradually attracted wide attention, but the toxicity of microplastics to ectomycorrhizal fungi (ECMF) is not clear yet. Therefore, Pisolithus tinctorius(Pt) and Lactarius delicius(Ld) were used as test strains in the present study, and monodisperse polystyrene microspheres (PS-MPs) with particle sizes of 80 nm and 4 μm were used as experimental materials. The effects of PS-MPs with different particle sizes and different concentrations (10-300 mg·L^-1) on the growth, malondialdehyde (MDA) content, soluble protein content, antioxidant enzymes activities and tissue conductivity of ECMF were explored by solid plate method and liquid culture method. The results showed that 200-300 mg·L^-1 PS-MPs significantly (P<0.05) inhibited the growth of Pt and Ld, and the PS-MPs with particle size of 4 μm showed stronger inhibition effect than PS-MPs with particle size of 80 nm. With the elevated concentration of PS-MPs, the superoxide dismutase activities of Pt and Ld exhibited an increasing first and then decreasing trend, and were all significantly (P<0.05) higher than that of control. When exposed to PS-MPs with different particle sizes, the catalase activity reached the lowest point under 300 mg·L^-1 PS-MPs treatment for Pt and Ld, and was significantly (P<0.05) lower than that of the control, while peroxidase activity was significantly (P<0.05) higher than that of control under all treatments. Besides, compared with the control, MDA content of Pt and Ld was significantly (P<0.05) elevated under other treatments (excluding Pt under 10 mg·L^-1 4 μm PS-MPs and Ld under 200 mg·L^-1 4 μm PS-MPs), soluble protein content was significantly (P<0.05) decreased (excluding Pt under 10, 20 mg·L^-1 80 nm PS-MPs and Ld under 10 mg·L^-1 80 nm PS-MPs), and the electrical conductivity of mycelium tissue was significantly (P<0.05) increased. Thus, it was inferred that the influence mechanism of PS-MPs on ECMF may involve oxidative stress, and different strains have different responses to PS-MPs with different particle sizes. These results would provide basis for the study regarding acute toxicity of microplastics on soil ECMF.

Key words: microplastics, ectomycorrhizal fungi, antioxidant enzymes

中图分类号:

X172

李玉婷, 李莎, 曹杰, 李骄杨, 张亮, 许晓风. 微塑料对外生菌根真菌生长和抗氧化系统的影响[J]. 浙江农业学报, 2022, 34(5): 1049-1060.

LI Yuting, LI Sha, CAO Jie, LI Jiaoyang, ZHANG Liang, XU Xiaofeng. Effects of microplastics on growth and antioxidant system of ectomycorrhizal fungi[J]. Acta Agriculturae Zhejiangensis, 2022, 34(5): 1049-1060.

图/表 7

图1 聚苯乙烯塑料微球(PS-MPs)的形貌与粒径分布 A,80 nm PS-MPs的透射电子显微镜图;B,4 μm PS-MPs的扫描电子显微镜图;C,80 nm PS-MPs的粒径分布;D,4 μm PS-MPs的粒径分布。

Fig.1 Morphology and particle size distribution of polystyrene microspheres (PS-MPs) A,Transmission electron microscopy image of 80 nm PS-MPs; B,Scanning electron microscopy image of 4 μm PS-MPs; C,Particle size distribution of 80 nm PS-MPs; D, Particle size distribution of 4 μm PS-MPs.

图2 聚苯乙烯塑料微球(PS-MPs)对外生菌根真菌彩色豆马勃(Pisolithus tinctorius)(A)和松乳菇(Lactarius delicious)(B)生长情况的影响

Fig.2 Effect of polystyrene microspheres (PS-MPs) on growth of ectomycorrhizal fungi Pisolithus tinctorius (A) and Lactarius delicious (B)

图3 聚苯乙烯塑料微球(PS-MPs)对外生菌根真菌彩色豆马勃(Pisolithus tinctorius)(A)和松乳菇(Lactarius delicious)(B)生物量的影响柱上无相同字母的表示处理间差异显著(P<0.05),下同。

Fig.3 Effect of polystyrene microspheres (PS-MPs) on biomass of ectomycorrhizal fungi Pisolithus tinctorius (A) and Lactarius delicious (B) Bars marked without the same letters indicated significant difference at P<0.05. The same as below.

表1 微塑料对外生菌根真菌SOD、CAT和POD活性的影响

Table 1 Effects of microplastics on SOD, CAT and POD activities of extomycorrhizal fangi

菌株 Strain	PS-MPs质量浓度 PS-MPs concentration/(mg·L^-1)	4 μm PS-MPs			80nm PS-MPs
菌株 Strain	PS-MPs质量浓度 PS-MPs concentration/(mg·L^-1)	SOD/ (U·g^-1)	CAT/ (U·g^-1)	POD/ (U·g^-1)	SOD/ (U·g^-1)	CAT/ (U·g^-1)	POD/ (U·g^-1)
Pt	0	14.23 i	118.65 a	39.15 d	14.23 i	118.65 b	39.15 h
	10	35.32 h	96.69 b	39.67 d	40.82 h	50.80 i	116.20 e
	20	45.21 f	79.90 c	62.59 c	39.63 h	81.25 f	142.15 c
	50	76.34 b	41.48 g	90.67 a	58.54 f	94.35 e	278.18 a
	100	67.86 c	37.67 g	91.81 a	87.42 c	48.21 i	132.82 cd
	200	62.12 d	55.81 e	11.22 f	70.17 d	36.77 j	101.43 f
	300	45.21 f	9.19 j	10.60 f	44.25 gh	20.02 k	122.68 de
Ld	0	17.56 i	59.26 e	22.45 e	17.56 i	59.26 h	22.45 i
	10	68.52 c	49.62 f	19.01 e	117.26 b	98.34 d	186.99 b
	20	84.22 ab	68.03 d	41.93 d	120.43 ab	109.64 c	130.68 d
	50	89.32 a	54.68 e	39.24 d	126.67 a	129.65 a	136.24 c
	100	69.82 c	46.71 f	82.76 b	91.46 c	127.04 a	138.22 c
	200	53.47 e	15.38 h	44.29 d	62.18 e	75.38 g	126.90 d
	300	40.36 g	5.36 i	13.30 f	48.91 g	14.01 k	67.55 g

图4 聚苯乙烯塑料微球(PS-MPs)对外生菌根真菌彩色豆马勃(Pisolithus tinctorius)(A)和松乳菇(Lactarius delicious)(B)丙二醛(MDA)含量的影响

Fig.4 Effect of polystyrene microspheres (PS-MPs) on malondialdehyde (MDA) content of ectomycorrhizal fungi Pisolithus tinctorius (A) and Lactarius delicious (B)

图5 聚苯乙烯塑料微球(PS-MPs)对外生菌根真菌彩色豆马勃(Pisolithus tinctorius)(A)和松乳菇(Lactarius delicious)(B)可溶性蛋白含量的影响

Fig.5 Effect of polystyrene microspheres (PS-MPs) on soluble protein content of ectomycorrhizal fungi Pisolithus tinctorius (A) and Lactarius delicious (B)

图6 聚苯乙烯塑料微球(PS-MPs)对外生菌根真菌彩色豆马勃(Pisolithus tinctorius)(A)和松乳菇(Lactarius delicious)(B)菌丝电导率的影响

Fig.6 Effect of polystyrene microspheres (PS-MPs) on electrical conductivity of mycelium tissue of ectomycorrhizal fungi Pisolithus tinctorius (A) and Lactarius delicious (B)

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微塑料对外生菌根真菌生长和抗氧化系统的影响

Effects of microplastics on growth and antioxidant system of ectomycorrhizal fungi

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