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一种ABC转运蛋白介导的农药苯达松在水稻(Oryza sativa L.)中的转运与代谢。

An ABC transporter-mediated transport and metabolism of the pesticide bentazone in rice (Oryza sativa L.).

作者信息

Qiao Yuxin, Wang Yujue, Gu Yucheng, Zhang Nan, Yang Hong, Liu Jintong

机构信息

Jiangsu Key Laboratory of Pesticide Science, College of Sciences, Nanjing Agricultural University, Nanjing 210095, China; Nanjing Institute of Environmental Science, Ministry of Ecology and Environment of China, Nanjing 210042, China.

Syngenta Crop Protection AG, Rosentalstrasse 67, CH-4002 Basel, Switzerland.

出版信息

J Adv Res. 2024 Aug 20. doi: 10.1016/j.jare.2024.08.020.

Abstract

INTRODUCTION

Bentazon (BNTZ) is a selective contact herbicide widely used to control field weeds for crop production. Excessive use of BNTZ leads to its accumulation in soils and crops, becoming an environmental contaminant. Therefore, investigation of the mechanisms for BNTZ detoxification and degradation in crops is fundamentally important to reduce crop contamination and ensure food safety.

OBJECTIVES

This study aims to elucidate the mechanism of detoxification and degradation pathways of the BNTZ complex in rice by creating transgenic lines expressing a rice ATP-binding cassette (OsABC) transporter gene through genetic engineering techniques combined with chemical analytical techniques and metabolomics approaches.

METHODS

We established the rice transgenic lines overexpressing (OE) a rice OsABC transporter and its knockout lines by CRISPR-Cas9 to characterize the gene function and measured the accumulation of BNTZ residues in rice. The metabolites of BNTZ were characterized by LC/Q-TOF-HRMS/MS (Liquid chromatography/time of flight-high resolution mass spectrometry).

RESULTS

Overexpression of OsABC significantly conferred rice resistance to BNTZ toxicity by increasing plant elongation, dry weight, and chlorophyll content, and significantly reducing cell membrane damage and BNTZ accumulation in rice tissues. Six different metabolites and ten conjugates were well defined in chemical structures. The reduced BNTZ levels and degradation products in the grains of the OE lines supported the robust activity of the OsABC gene function. Using UPLC-Q-TOF/MS, we further identified accumulated basic metabolites of various carbohydrates, amino acids, hormones, and flavonoids, and found that these metabolites involved in BNTZ degradation were increased more in OE lines than in wild-type (WT) rice.

CONCLUSIONS

Our work demonstrates that the OsABC transporter plays a critical role in regulating the mobility and degradative metabolism of BNTZ in rice, thus revealing a regulatory mechanism underlying rice resistance to BNTZ toxicity and adaptation to the environmental stress.

摘要

引言

苯达松(BNTZ)是一种选择性触杀型除草剂,广泛用于控制作物生产中的田间杂草。过度使用BNTZ会导致其在土壤和作物中积累,成为一种环境污染物。因此,研究作物中BNTZ解毒和降解的机制对于减少作物污染和确保食品安全至关重要。

目的

本研究旨在通过基因工程技术结合化学分析技术和代谢组学方法,创建表达水稻ATP结合盒(OsABC)转运蛋白基因的转基因系,以阐明水稻中BNTZ复合物的解毒机制和降解途径。

方法

我们通过CRISPR-Cas9技术建立了过表达(OE)水稻OsABC转运蛋白的转基因系及其敲除系,以表征该基因的功能,并测量水稻中BNTZ残留的积累。通过液相色谱/飞行时间-高分辨率质谱(LC/Q-TOF-HRMS/MS)对BNTZ的代谢产物进行表征。

结果

OsABC的过表达通过增加植株伸长、干重和叶绿素含量,显著赋予水稻对BNTZ毒性的抗性,并显著降低细胞膜损伤和水稻组织中BNTZ的积累。六种不同的代谢产物和十种共轭物的化学结构得到了明确界定。OE系籽粒中BNTZ水平和降解产物的降低支持了OsABC基因功能的强大活性。使用超高效液相色谱-飞行时间/质谱(UPLC-Q-TOF/MS),我们进一步鉴定了各种碳水化合物、氨基酸、激素和黄酮类化合物的积累的碱性代谢产物,并发现参与BNTZ降解的这些代谢产物在OE系中比在野生型(WT)水稻中增加得更多。

结论

我们的工作表明,OsABC转运蛋白在调节水稻中BNTZ的流动性和降解代谢中起关键作用,从而揭示了水稻对BNTZ毒性抗性和对环境胁迫适应性的调控机制。

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