草甘膦胁迫对茶树叶片中莽草酸含量的影响

刘洪霞; 刘颖颖; 陈红平; 柴云峰

doi:10.13305/j.cnki.jts.2023.05.005

茶叶科学 >

2023 , Vol. 43 >Issue 5: 657 - 666

DOI: https://doi.org/10.13305/j.cnki.jts.2023.05.005

研究报告

草甘膦胁迫对茶树叶片中莽草酸含量的影响

刘洪霞 ,
刘颖颖 ,
陈红平 ,
柴云峰

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1.中国农业科学院茶叶研究所,浙江杭州 310008;
2.中国农业科学院研究生院,北京 100081;
3.农业农村部茶叶产品质量安全风险评估实验室（杭州）,浙江杭州 310008

刘洪霞,女,硕士研究生,主要从事茶叶质量安全研究。

收稿日期: 2023-03-08

修回日期: 2023-07-09

网络出版日期: 2023-11-06

基金资助

国家自然科学基金（21775164）、现代农业产业技术体系建设专项资金（CARS-23）、三农九方项目（2022SNJF037）

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Glyphosate-stress Effects on Shikimic Acid in Tea Leaves

LIU Hongxia ,
LIU Yingying ,
CHEN Hongping ,
CHAI Yunfeng

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1. Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008, China;
2. Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, China;
3. Laboratory of Quality and Safety Risk Assessment for Tea (Hangzhou), Ministry of Agriculture and Rural Affairs, Hangzhou 310008, China

Received date: 2023-03-08

Revised date: 2023-07-09

Online published: 2023-11-06

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摘要

为探明草甘膦胁迫对茶树生长及莽草酸代谢的影响,通过水培试验考察草甘膦对茶苗的表观药害,采用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱对叶片中的非挥发性代谢物进行非靶向分析,并对叶片中的莽草酸和草甘膦进行定量测定。结果表明,高剂量草甘膦（200 mg·L^-1）处理组茶苗出现药害特征,而低剂量草甘膦（50 mg·L^-1）处理组和对照组茶苗未出现表观药害。质谱检测和统计学分析表明,发生药害的茶树叶片中莽草酸途径代谢物的含量发生显著变化,其中莽草酸是主要的差异代谢物之一。在试验期内（0~21 d）,茶树叶片中莽草酸的积累量与草甘膦的吸收量和作用时间高度正相关,当草甘膦吸收量达到28 mg·kg^-1以上时,茶树的莽草酸代谢受到明显抑制,导致叶片中莽草酸大量积累,与对照组相比,发生药害的茶树叶片中莽草酸的含量约高16倍。本研究表明莽草酸是茶树响应草甘膦胁迫的主要代谢物之一。

关键词： 茶树; 草甘膦; 莽草酸; 药害; 液质联用

本文引用格式

刘洪霞 , 刘颖颖 , 陈红平 , 柴云峰 . 草甘膦胁迫对茶树叶片中莽草酸含量的影响[J]. 茶叶科学, 2023 , 43(5) : 657 -666 . DOI: 10.13305/j.cnki.jts.2023.05.005

Abstract

To investigate the effect of glyphosate stress on the growth and shikimic acid metabolism of tea (Camellia sinensis L.) plants, tea seedlings were cultured in nutrient solution with different concentrations of glyphosate and the visual phytotoxicity on tea leaves was observed. The non-targeted analysis of non-volatile metabolites in the leaves and quantitative determination of shikimic acid and glyphosate in the leaves were carried out by ultra-high performance liquid chromatography-quadrupole orbitrap high-resolution mass spectrometry. The results show that the tea seedlings under the high dose of glyphosate (200 mg·L^-1) treatment exhibited characteristics of pesticide damage, while the tea seedlings under the low dose of glyphosate (50 mg·L^-1) treatment and control did not show apparent pesticide damage. Mass spectrometric and statistical analysis indicates that there were significant changes in the contents of shikimic acid pathway metabolites in the leaves of glyphosate-treated tea seedlings, with shikimic acid being one of the main differential metabolites. Within 21 d, the accumulation of shikimic acid in leaves was highly positively correlated with the absorption amount and action time of glyphosate. When the absorption amount of glyphosate was larger than 28 mg·kg^-1, the shikimic acid metabolism in tea plants was significantly inhibited, resulting in a large accumulation of shikimic acid in tea leaves. Compared with the control group, the content of shikimic acid in tea leaves affected by pesticides increased about 16-fold. This study shows that shikimic acid is one of the main metabolites of tea plants in response to glyphosate stress.

Key words： tea plant; glyphosate; shikimic acid; phytotoxicity; LC-MS

参考文献

[1] Duke S O, Powles S B.Glyphosate: a once-in-a-century herbicide[J]. Pest Management Science, 2008, 64(4): 319-325.
[2] Acquavella J F, Alexander B H, Mandel J S, et al.Glyphosate biomonitoring for farmers and their families: results from the farm family exposure study[J]. Environmental Health Perspectives, 2004, 112(3): 321-326.
[3] 陶波, 蒋凌雪, 沈晓峰, 等. 草甘膦对土壤微生物的影响[J]. 中国油料作物学报, 2011, 33(2): 162-168, 179.
Tao B, Jiang L X, Shen X F, et al.Effects of glyphosate on soil microorganisms[J]. Chinese Journal of Oil Crop Sciences, 2011, 33(2): 162-168, 179.
[4] Aparicio V C, De Gerónimo E, Marino D, et al.Environmental fate of glyphosate and aminomethylphosphonic acid in surface waters and soil of agricultural basins[J]. Chemosphere, 2013, 93(9): 1866-1873.
[5] 郭永春, 陈金发, 赵峰, 等. 草甘膦及其代谢物氨甲基膦酸在茶树体中的分布研究[J]. 茶叶科学, 2020, 40(4): 510-518.
Guo Y C, Chen J F, Zhao F, et al.Study on the distribution of glyphosate and its metabolite aminomethylphosphonic acid in Camellia sinensis[J]. Journal of Tea Science, 2020, 40(4): 510-518.
[6] Servaites J C, Tucci M A, Geiger D R.Glyphosate effects on carbon assimilation, ribulose bisphosphate carboxylase activity, and metabolite levels in sugar beet leaves[J]. Plant Physiology, 1987, 85(2): 370-374.
[7] Zobiole L H S, Kremer R J, De Oliveira Jr. R S, et al. Glyphosate effects on photosynthesis, nutrient accumulation, and nodulation in glyphosate-resistant soybean[J]. Journal of Plant Nutrition and Soil Science, 2012, 175(2): 319-330.
[8] Reddy K N, Bellaloui N, Zablotowicz R M.Glyphosate effect on shikimate, nitrate reductase activity, yield, and seed composition in corn[J]. Journal of Agricultural and Food Chemistry, 2010, 58(6): 3646-3650.
[9] Helander M, Pauna A, Saikkonen K, et al.Glyphosate residues in soil affect crop plant germination and growth[J]. Scientific Reports, 2019, 9: 19653. doi: 10.1038/s41598-019-56195-3.
[10] Malalgoda M, Ohm J B, Howatt K A, et al.Effects of pre-harvest glyphosate use on protein composition and shikimic acid accumulation in spring wheat[J]. Food Chemistry, 2020, 332: 127422. doi: 10.1016/j.foodchem.2020.127422.
[11] Anderson K A, Cobb W T, Loper B R.Analytical method for determination of shikimic acid: Shikimic acid proportional to glyphosate application rates[J]. Communications in Soil Science and Plant Analysis, 2001, 32(17/18): 2831-2840.
[12] 高万君, 张永志, 童蒙蒙, 等. 茶园常用除草剂田间药效试验与残留动态[J]. 茶叶科学, 2019, 39(5): 587-594.
Gao W J, Zhang Y Z, Tong M M, et al.Weeds control effect and residues of several herbicides in tea gardens[J]. Journal of Tea Science, 2019, 39(5): 587-594.
[13] Tong M M, Gao W J, Jiao W T, et al.Uptake, translocation, metabolism, and distribution of glyphosate in nontarget tea plant (Camellia sinensis L.)[J]. Journal of Agricultural and Food Chemistry, 2017, 65(35): 7638-7646.
[14] 郭永春, 王淑燕, 王鹏杰, 等. 草甘膦对茶树叶片主要生化成分的影响[J]. 天然产物研究与开发, 2021, 33(3): 394-401, 499.
Guo Y C, Wang S Y, Wang P J, et al.Effect of glyphosate on main biochemical components of tea leaves[J]. Natural Product Research and Development, 2021, 33(3): 394-401, 499.
[15] 郭永春, 王鹏杰, 金珊, 等. 基于WGCNA鉴定茶树响应草甘膦相关的基因共表达模块[J]. 中国农业科学, 2022, 55(1): 152-166.
Guo Y C, Wang P J, Jin S, et al.Identification of co-expression gene related to tea plant response to glyphosate based on WGCNA[J]. Scientia Agricultura Sinica, 2022, 55(1): 152-166.
[16] Holländer-Czytko H, Amrhein N.Subcellular compartment of shikimic acid and phenylalanine in buckwheat cell suspension cultures grown in the presence of shikimate pathway inhibitors[J]. Plant Science Letters, 1983, 29(1): 89-96.
[17] Pline W A, Wilcut J W, Duke S O, et al.Tolerance and accumulation of shikimic acid in response to glyphosate applications in glyphosate-resistant and nonglyphosate-resistant cotton (Gossypium hirsutum L.)[J]. Journal of Agricultural and Food Chemistry, 2002, 50(3): 506-512.
[18] 诸力, 陈红平, 周苏娟, 等. 超高效液相色谱-串联质谱法测定不同茶叶中草甘膦、氨甲基膦酸及草铵膦的残留[J]. 分析化学, 2015, 43(2): 271-276.
Zhu L, Chen H P, Zhou S J, et al.Determination of glyphosate, aminomethyl phosphonic acid and glufosinate in different teas by ultra performance liquid chromatography-tandem mass spectrometry[J]. Chinese Journal of Analytical Chemistry, 2015, 43(2): 271-276.
[19] Nilsson G.Effects of glyphosate on the amino acid content in spring wheat plants[J]. Swedish Journal of Agricultural Research, 1977, 7(3): 153-157.
[20] Wang C Y.Effect of glyphosate on aromatic amino acid metabolism in purple nutsedge (Cyperus rotundus)[J]. Weed Technology, 2001, 15(4): 628-635.
[21] 罗威, 高冬丽. 莽草酸含量测定方法综述[J]. 现代食品, 2020(10): 59-63.
Luo W, Gao D L.A review of the quantitative methods of shikimic acid content[J]. Modern Food, 2020(10): 59-63.
[22] Gachumi G, Purves R W, Hopf C, et al.Fast quantification without conventional chromatography, the growing power of mass spectrometry[J]. Analytical Chemistry, 2020, 92(13): 8628-8637.
[23] Campmajó G, Saurina J, Núñez O.FIA-HRMS fingerprinting subjected to chemometrics as a valuable tool to address food classification and authentication: application to red wine, paprika, and vegetable oil samples[J]. Food Chemistry, 2022, 373: 131491. doi: 10.1016/j.foodchem. 2021.
131491.
[24] Harring T, Streibig J C, Husted S.Accumulation of shikimic acid: a technique for screening glyphosate efficacy[J]. Journal of Agricultural and Food Chemistry, 1998, 46(10): 4406-4412.
[25] Schrübbers L C, Valverde B E, Sørensen J C, et al.Glyphosate spray drift in Coffea arabica-Sensitivity of coffee plants and possible use of shikimic acid as a biomarker for glyphosate exposure[J]. Pesticide Biochemistry and Physiology, 2014, 115: 15-22.
[26] 巩元勇, 郭书巧, 束红梅, 等. 1株抗草甘膦棉花突变体草甘膦抗性的初步鉴定[J]. 棉花学报, 2014, 26(1): 18-24.
Gong Y Y, Guo S Q, Shu H M, et al.Preliminary identification of glyphosate resistance of a new cotton mutant[J]. Cotton Science, 2014, 26(1): 18-24.

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