磺酰脲类除草剂残留的微生物降解研究进展

磺酰脲类除草剂是一类高效、低毒和高选择性的除草剂, 此类除草剂能有效地防除阔叶杂草, 其中有些品种对禾本科杂草也有抑制作用。由于该类除草剂易残留药害及容易对地表水造成污染, 因而其在环境中的持久性和环境安全性备受人们关注。本文综述了磺酰脲类除草剂的应用概况及其作用机理、降解磺酰脲类除草剂的常见微生物种类及影响微生物降解效率的因素, 最后展望了微生物修复技术与抗除草剂的转基因作物是解决除草剂残留药害的最佳途径。     

二苯醚类除草剂的微生物降解研究进展

二苯醚类除草剂是一类广谱、高效、高选择性的除草剂,广泛应用于大豆、花生等农田一年生和多年生阔叶杂草的防除。由于该类除草剂不易降解,多年连续使用会导致其在土壤环境中的大量积累。本文概述了二苯醚类除草剂的基本结构及其对生物的影响,总结了降解二苯醚类除草剂的微生物种类、降解途径和降解过程中关键酶及其基因,分析了影响微生物降解二苯醚类除草剂的因素,对二苯醚类除草剂微生物降解未来的研究方向进行了展望,为深入研究二苯醚类除草剂的生物降解提供参考。     

除草剂的微生物降解研究进展

微生物降解是环境中农药消解的重要因素,分离筛选纯培养的农药降解微生物并阐述其降解机制为微生物修复环境的应用提供重要的菌株资源和理论依据。本文简述了广泛使用的8类除草剂(包括有机磷类、磺酰脲类、氯乙酰胺类、均三嗪类、芳氧基苯氧基丙酸酯类、苯氧乙酸类、二硝基苯胺类和硫代氨基甲酸酯类除草剂)的降解微生物资源及其降解途径和降解基因的研究进展,并分析了目前除草剂污染修复存在的问题及未来的发展方向。     

三酮类除草剂微生物降解的研究进展

三酮类除草剂是一类高效、广谱、高选择性的除草剂,能够有效地防除玉米地多种阔叶杂草和禾本科杂草。该类除草剂在土壤、水体中残留,易造成地表和地下水污染。有关研究表明,微生物降解有望成为解决该类除草剂残留的有效措施。本文分析了2种三酮类除草剂带来的生态效应,总结了已报道的微生物降解资源,简述了降解基因/酶的研究进展及可能的降解途径,为深入研究三酮类除草剂的生物降解提供一定的信息支撑。     

二硝基苯胺类除草剂微生物降解研究进展

二硝基苯胺类除草剂是一类广谱、高效且广泛使用的除草剂,微生物的降解代谢作用是其在环境中消解的最主要因素。分离筛选除草剂的高效降解菌株、分析其降解途径并阐明其微生物降解机制,可为除草剂残留污染的微生物降解修复提供理论依据和优良的降解菌株、降解基因和酶资源。本文简述了二硝基苯胺类除草剂的微生物降解菌株、降解代谢途径和降解基因/酶的研究进展,为进一步研究该类除草剂的微生物降解及其污染生物修复提供理论依据和资源。     

灭幼脲Ⅲ号在好气水环境中的降解代谢的初步研究

本文研究了新农药灭幼脲Ⅲ号在好气水环境中的降解与代谢。在避光条件下,观察了灭菌组与实验组中灭幼脲Ⅲ号及其主要代谢产物的消长过程,比较了它的化学水解与微生物降解的差异。在室内模拟好气系统中,研究了母体化合物在水体中的残留动态和生物降解半衰期,及其初期主要代谢途径的转化产物,同时分别用高效液相色谱法,紫外吸收光谱扫描,以及特征有机质谱图,对灭幼脲Ⅲ号的两种主要代谢产物进行了定性定量测定。结果表明:灭幼脲Ⅲ号在室内好气环境中较易水解,而且水中微生物的存在能加速它的降解,母体化合物在水体中初期代谢主要途径为分子中的苯甲酰碳与脲氮键首先开裂,生成邻氯苯甲酸(CBA)和对氯苯基脲素(CPU)。     

微生物降解磺酰脲类除草剂的研究进展

磺酰脲类除草剂是一种高效、广谱、高选择性的除草剂,但其长期广泛使用对生态环境造成了严重破坏,因此对于如何科学合理使用磺酰脲类除草剂、有效防治作物药害和降低对人类的危害等问题成为近年来的研究热点。磺酰脲除草剂在土壤中以化学降解和生物降解方式为主,生物降解是自然界本身具有的一种降解污染物的方式,是一种可行性高、副作用小的方法。近年来,很多学者已经开始研究并利用真菌、细菌等微生物来降解磺酰脲类除草剂,取得了许多重要结果。本文总结了磺酰脲类除草剂的性质、结构以及降解机理、可降解该类除草剂的微生物种类和影响微生物降解效率的因素;最后指出了现阶段存在的问题并对磺酰脲类除草剂的未来发展趋势进行展望。     

环境中雌激素的微生物降解

环境中的雌激素是一类重要的环境内分泌干扰物,微生物降解是去除环境雌激素的主要途径。通过归纳已报道的雌激素降解细菌、总结其降解雌激素的机制、分析雌激素降解途径以及其他真核微生物的雌激素降解作用4个方面,概括阐述了雌激素的微生物降解作用,并对未来的研究方向提出展望。     

厌氧丁草胺降解菌BAD-20的分离鉴定及降解特性研究

【目的】分离并鉴定能够降解除草剂丁草胺的厌氧微生物菌株,研究其厌氧降解丁草胺的特性和代谢途径,为深入研究丁草胺厌氧降解机制提供依据。【方法】以丁草胺为碳源作为选择压力从水稻田土壤中富集驯化丁草胺降解菌,利用16S rRNA基因系统发育分析结合菌株培养特征对降解菌株进行初步鉴定,利用液相色谱-时间飞行质谱(LC-TOF-MS)检测菌株降解丁草胺的代谢产物。【结果】筛选到一株降解丁草胺的厌氧细菌,命名为BAD-20,初步鉴定为嗜蛋白质菌属(Proteiniphilum),菌株BAD-20降解丁草胺的最适条件为温度30–35℃、pH 7.5–8.0和0–0.5%NaCl,在有氧条件下该菌不能降解丁草胺。最适条件下,菌株BAD-20在10d降解90%的20mg/L丁草胺。菌株BAD-20还能降解甲草胺、乙草胺、丙草胺,降解效率从高到低依次为甲草胺乙草胺丙草胺丁草胺,对这些氯乙酰胺除草剂的降解动力学符合一级动力学方程。鉴定到2个丁草胺降解代谢产物,分别是N-(2,6-二乙基苯基)-N-(丁氧甲基)乙酰胺(DEPBMA)和N-(2,6-二乙基苯基)乙酰胺(DEPA),表明菌株BAD-20降解丁草胺的起始步骤为脱氯,随后脱去N-丁氧甲基。【结论】本研究富集分离到一株降解丁草胺的厌氧细菌嗜蛋白质菌属(Proteiniphilum) BAD-20,为深入研究丁草胺厌氧降解机制及研发含丁草胺废水厌氧生物处理技术提供依据。     

多种农药与次氯酸钠、高锰酸钾和高铁酸钾的反应活性

为探究4类,10种广泛使用的农药(苯氧羧酸类,芳香酸类,取代脲类和烟碱类)与3种氧化剂(次氯酸钠,高锰酸钾和高铁酸钾)的反应活性,本研究在温度(25±2)℃、pH值为8的条件下,分析10种农药分别与3种氧化剂NaClO、KMnO4、K2Fe O4在不同浓度下的反应活性,采用HPLC检测法,对比降解效能,探究氧化剂性质与有机物结构导致的反应活性的差异。实验结果表明,不同种类农药的结构性质对反应活性有重要影响,3种氧化剂的氧化降解能力有明显差异。苯氧羧酸类和芳香酸类农药结构较简单,并含有稳定的苯环或吡啶环结构,氧化降解较困难。取代脲类和烟碱类农药结构较复杂,氧化剂可攻击其不饱和官能团,反应活性较高。NaClO对取代脲类农药的降解率明显优于其他2种氧化剂,3种氧化剂对烟碱类农药的氧化降解效果依次为NaClO>KMn O4>K2Fe O4。研究多种农药与次氯酸钠、高锰酸钾和高铁酸钾的反应活性对降解去除水体中的农药残留对水环境的治理具有重要意义。     

Degradation of Chlorbromuron and Related Compounds by the Fungus Rhizoctonia solani

The ability of the soil fungus Rhizoctonia solani to degrade phenyl-substituted urea herbicides was investigated. The fungus was able to transform chlorbromuron [3-(3-chloro-4-bromophenyl)-1-methyl-1-methoxyurea] to the demethylated product [3-(3-chloro-4-bromophenyl)-1-methoxyurea], which was isolated and identified. Evidence was obtained that further degradation of chlorbromuron occurred. Several other phenylurea compounds (chloroxuron, diuron, fenuron, fluometuron, linuron, metobromuron, neburon, and siduron) were also metabolized by the fungus, indicating that R. solani may possess a generalized ability to attack this group of herbicides.     

Phenylurea herbicides induce cytogenetic effects in Chinese hamster cell lines

The intensive use of herbicides over the last few decades has caused a general increase of environmental pollution. It is thus very important to evaluate the possible genotoxic properties of these chemical compounds as well as identifying their mode of action. Phenylurea herbicides are selective agents widely used for the control of infestant plants. Of these herbicides, which are widely used in agriculture, we analysed four of the less intensively studied molecules. More precisely, we investigated the genotoxic effects of fenuron, chlorotoluron, diuron, and difenoxuron by analyses of chromosomal aberrations (CAs) and sister chromatid exchange (SCE) in exposed mammalian cells. We used the Chinese hamster ovary (CHO) and epithelial liver (CHEL) cell lines, endowed with the absence or the presence, respectively, of an enzymatic system to activate pro-mutagenic compounds. Our results show that all herbicides tested induce, at high concentrations, an increasing number of CAs in non-metabolising CHO cells. Instead, in the exposed CHEL cell line, the four herbicides induced CAs also at the lowest dose-level. In the CHEL cells, a statistically significant increase of SCE was also observed. The phenylurea herbicides showed direct genotoxic activity, but the cytogenetic effects were greatly enhanced after metabolic conversion. These data, together with other information on phenylurea herbicides, are of great interest from the environmental point of view, and for human health. In fact, intensive use of herbicides contaminates soil, surface water, groundwater and agricultural products, and thus should be taken in particular consideration not only for those initiatives to specifically protect exposed workers, but also to safeguard the health of consumers of agricultural products.     

Isolation from agricultural soil and characterization of a Sphingomonas sp. able to mineralize the phenylurea herbicide isoproturon.

A soil bacterium (designated strain SRS2) able to metabolize the phenylurea herbicide isoproturon, 3-(4-isopropylphenyl)-1,1-dimethylurea (IPU), was isolated from a previously IPU-treated agricultural soil. Based on a partial analysis of the 16S rRNA gene and the cellular fatty acids, the strain was identified as a Sphingomonas sp. within the alpha-subdivision of the proteobacteria. Strain SRS2 was able to mineralize IPU when provided as a source of carbon, nitrogen, and energy. Supplementing the medium with a mixture of amino acids considerably enhanced IPU mineralization. Mineralization of IPU was accompanied by transient accumulation of the metabolites 3-(4-isopropylphenyl)-1-methylurea, 3-(4-isopropylphenyl)-urea, and 4-isopropyl-aniline identified by high-performance liquid chromatography analysis, thus indicating a metabolic pathway initiated by two successive N-demethylations, followed by cleavage of the urea side chain and finally by mineralization of the phenyl structure. Strain SRS2 also transformed the dimethylurea-substituted herbicides diuron and chlorotoluron, giving rise to as-yet-unidentified products. In addition, no degradation of the methoxy-methylurea-substituted herbicide linuron was observed. This report is the first characterization of a pure bacterial culture able to mineralize IPU.     

Isolation from Agricultural Soil and Characterization of a Sphingomonas sp. Able To Mineralize the Phenylurea Herbicide Isoproturon

A soil bacterium (designated strain SRS2) able to metabolize the phenylurea herbicide isoproturon, 3-(4-isopropylphenyl)-1,1-dimethylurea (IPU), was isolated from a previously IPU-treated agricultural soil. Based on a partial analysis of the 16S rRNA gene and the cellular fatty acids, the strain was identified as a Sphingomonas sp. within the α-subdivision of the proteobacteria. Strain SRS2 was able to mineralize IPU when provided as a source of carbon, nitrogen, and energy. Supplementing the medium with a mixture of amino acids considerably enhanced IPU mineralization. Mineralization of IPU was accompanied by transient accumulation of the metabolites 3-(4-isopropylphenyl)-1-methylurea, 3-(4-isopropylphenyl)-urea, and 4-isopropyl-aniline identified by high-performance liquid chromatography analysis, thus indicating a metabolic pathway initiated by two successive N-demethylations, followed by cleavage of the urea side chain and finally by mineralization of the phenyl structure. Strain SRS2 also transformed the dimethylurea-substituted herbicides diuron and chlorotoluron, giving rise to as-yet-unidentified products. In addition, no degradation of the methoxy-methylurea-substituted herbicide linuron was observed. This report is the first characterization of a pure bacterial culture able to mineralize IPU.     

Differential impact of environmental stresses on the pea mitochondrial proteome

Exposure to adverse environmental conditions causes oxidative stress in many organisms, leading either to disease and debilitation or to response and tolerance. Mitochondria are a key site of oxidative stress and of cellular response and play important roles in cell survival. We analyzed the response of mitochondria in pea (Pisum sativum) plants to the common stresses associated with drought, cold, and herbicides. These treatments all altered photosynthetic and respiratory rates of pea leaves to various extents, but only herbicides significantly increased lipid peroxidation product accumulation. Mitochondria isolated from the stressed pea plants maintained their electron transport chain activity, but changes were evident in the abundance of uncoupling proteins, non-phosphorylating respiratory pathways, and oxidative modification of lipoic acid moieties on mitochondrial proteins. These data suggest that herbicide treatment placed a severe oxidative stress on mitochondria, whereas chilling and particularly drought were milder stresses. Detailed analysis of the soluble proteome of mitochondria by gel electrophoresis and mass spectrometry revealed differential degradation of key matrix enzymes during treatments with chilling being significantly more damaging than drought. Differential induction of heat shock proteins and specific losses of other proteins illustrated the diversity of response to these stresses at the protein level. Cross-species matching was required for mass spectrometry identification of nine proteins because only a limited number of pea cDNAs have been sequenced, and the full pea genome is not available. Blue-native separation of intact respiratory chain complexes revealed little if any change in response to environmental stresses. Together these data suggest that although many of the molecular events identified by chemical stresses of mitochondria from a range of model eukaryotes are also apparent during environmental stress of plants, their extent and significance can vary substantially.     

Effects of tail-like substituents on the binding of competitive inhibitors to the Q(B) site of photosystem II

The QB quinone-binding site of photosystem II is an important target for herbicides. Two major classes of herbicides are based on s-triazine and phenylurea moieties. A small library of triazine and phenylurea compounds has been synthesized which have tail-like substituents in order to test the effects of charge, hydrophobicity and size of the tail on binding properties. It is found that a tail can be attached to one of the alkylamino groups of triazine-type herbicides or to the para position of phenylurea-type herbicides without loss of binding, provided that the tail is hydrophobic. This indicates that the herbicides must be oriented in the QB site such that these positions point toward the natural isoprenyl tail-binding pocket that extends out of the Q(B) site. In turn, the requirement that the tail must extend out of the QB site constrains the size of the other herbicide substituents in the pocket. This is in agreement with the presumed orientation and fit of ligands in the QB site. When longer hydrophobic tails are used, the binding penalty that occurs upon adding a charged substituent at the distal end is reduced. This allows the use of a series of tail substituents possessing a distal charge as an approximate molecular ruler to measure the distance from the QB site to the aqueous phase. Even a 10-carbon alkyl chain still shows a 4-fold effect from the presence or absence of a distal charge. Such a chain does not appear to be long enough to extend from the bulk aqueous phase to the QB site because binding is completely lost when a large hydrophilic domain (PEG(4000)) is attached to the distal end. Longer tails are effective only if they are sufficiently hydrophobic. An effort was made to use tailed herbicides for affinity binding of photosystem II. It was found that hydrophobic linkers promote nonspecific binding, but careful choice of solvent conditions, such as the use of excess nonionic detergent well above its critical micelle concentration, might obviate this problem during affinity-binding applications.     

Degradation of Linuron and Some Other Herbicides and Fungicides by a Linuron-Inducible Enzyme Obtained from Bacillus sphaericus

Linuron [3-(3,4-dichlorophenyl)-1-methoxy-1-methylurea] induces the formation of an enzyme (acylamidase) responsible for the degradation of a large variety of different herbicides and fungicides of the acylanilide and phenylurea type. The former type is degraded at a rate at least 10 times higher than the latter.     

Inhibition of atrazine degradation by cyanazine and exogenous nitrogen in bacterial isolate M91-3

A variety of s-triazine herbicides and nitrogen fertilizers frequently occur as co-contaminants at pesticide manufacturing and distribution facilities. The degradation of atrazine and cyanazine by the bacterial isolate M91-3 was investigated in washed-cell suspensions and crude cellular extracts. Cyanazine competitively inhibited atrazine degradation. The maximum atrazine degradation rate (V _max) was 41 times higher and the half-saturation constant for the inhibitor (K _i) was 1.3 times higher in the crude cellular extract than in the washed-cell suspension, suggesting that cellular uptake influenced degradation of the s-triazines. Cultures that had received prior exposure to atrazine and simazine exhibited comparable atrazine degradation rates, while cells exposed to cyanazine, propazine, ametryne, cyanuric acid, 2-hydroxyatrazine, biuret, and urea exhibited a lack of atrazine-degradative activity. Growth in the presence of exogenous inorganic nitrogen inhibited subsequent atrazine-degradative activity in washed-cell suspensions, suggesting that regulation of s-triazine and nitrogen metabolism are linked in this bacterial isolate. These findings have significant implications for the environmental fate of s-triazines in agricultural settings since these herbicides are frequently applied to soils receiving N fertilizers. Furthermore, these results suggest that bioremediation of s-triazine-contaminated sites (common at pesticide distribution facilities in the cornbelt) may be inhibited by the presence of N fertilizers that occur as co-contaminants. Received: 3 March 1998 / Received revision: 24 September 1998 / Accepted: 11 October 1998     

Inhibition of Phenylamide Hydrolysis by Bacillus sphaericus with Methylcarbamate and Organophosphorus Insecticides

The degradation of the phenylamide herbicides monolinuron, linuron, and solan by cultures of Bacillus sphaericus ATCC 12123 was inhibited by the methylcarbamate insecticides metmercapturon, aldicarb, propoxur, and carbaryl and by the organophosphorus insecticides fenthion and parathion. The extent of inhibition was largest with metmercapturon and smallest with parathion inhibition of hydrolysis of the two phenylurea herbicides was greater than of the acylanilide compound. Tests with crude enzyme preparations of aryl acylamidase derived from B. sphaericus showed that the inhibition of the hydrolysis of linuron with methylcarbamates is a competitive one. The insecticides tested did induce the enzyme, nor could they serve as its substrate.