Regulation of Growth, Cell Shape, Cell Division, and Gene Expression by Second Messengers (p)ppGpp and Cyclic Di-GMP in Mycobacterium smegmatis

The alarmone (p)ppGpp regulates transcription, translation, replication, virulence, lipid synthesis, antibiotic sensitivity, biofilm formation, and other functions in bacteria. Signaling nucleotide cyclic di-GMP (c-di-GMP) regulates biofilm formation, motility, virulence, the cell cycle, and other functions. In Mycobacterium smegmatis, both (p) ppGpp and c-di-GMP are synthesized and degraded by bifunctional proteins Rel(Msm) and DcpA, encoded by rel(Msm) and dcpA genes, respectively. We have previously shown that the Delta rel(Msm) and Delta dcpA knockout strains are antibiotic resistant and defective in biofilm formation, show altered cell surface properties, and have reduced levels of glycopeptidolipids and polar lipids in their cell wall (K. R. Gupta, S. Kasetty, and D. Chatterji, Appl Environ Microbiol 81:2571-2578, 2015, http://dx.doi.org/10.1128/AEM.03999-14). In this work, we have explored the phenotypes that are affected by both (p) ppGpp and c-di-GMP in mycobacteria. We have shown that both (p) ppGpp and c-di-GMP are needed to maintain the proper growth rate under stress conditions such as carbon deprivation and cold shock. Scanning electron microscopy showed that low levels of these second messengers result in elongated cells, while high levels reduce the cell length and embed the cells in a biofilm-like matrix. Fluorescence microscopy revealed that the elongated Delta rel(Msm) and Delta dcpA cells are multinucleate, while transmission electron microscopy showed that the elongated cells are multiseptate. Gene expression analysis also showed that genes belonging to functional categories such as virulence, detoxification, lipid metabolism, and cell-wall-related processes were differentially expressed. Our results suggests that both (p) ppGpp and c-di-GMP affect some common phenotypes in M. smegmatis, thus raising a possibility of cross talk between these two second messengers in mycobacteria. IMPORTANCE Our work has expanded the horizon of (p) ppGpp and c-di-GMP signaling in Gram-positive bacteria. We have come across a novel observation that M. smegmatis needs (p) ppGpp and c-di-GMP for cold tolerance. We had previously shown that the Delta rel(Msm) and Delta dcpA strains are defective in biofilm formation. In this work, the overproduction of (p) ppGpp and c-di-GMP encased M. smegmatis in a biofilm-like matrix, which shows that both (p) ppGpp and c-di-GMP are needed for biofilm formation. The regulation of cell length and cell division by (p) ppGpp was known in mycobacteria, but our work shows that c-di-GMP also affects the cell size and cell division in mycobacteria. This is perhaps the first report of c-di-GMP regulating cell division in mycobacteria.

Metallothionein as an indicator of water quality: assessment of the bioavailability of cadmium, copper, mercury and zinc in aquatic animals at the cellular level

The study of metallothioneins (MTs) has greatly improved our understanding of body burdens, metal storage and detoxification in aquatic organisms subjected to contamination by the toxic heavy metals, Cd, Cu, Hg and Zn. These studies have shown that in certain organisms MT status can be used to assess impact of these metals at the cellular level and, whilst validation is currently limited to a few examples, this stress response may be linked to higher levels of organisation, thus indicating its potential for environmental quality assessment. Molluscs, such as Mytilus spp., and several commonly occurring teleost species, are the most promising of the indicator species tested. Natural variability of MT levels caused by the organism's size, condition, age, position in the sexual cycle, temperature and various stressors, can lead to difficulties in interpretation of field data as a definitive response-indicator of metal contamination unless a critical appraisal of these variables is available. From laboratory and field studies these data are almost complete for teleost fish. Whilst for molluscs much of this information is lacking, when suitable controls are utilised and MT measurements are combined with observations of metal partitioning, current studies indicate that they are nevertheless a powerful tool in the interpretation of impact, and may prove useful in water quality assessment.

Dendrorremediación: El uso de Betula alba para monitorizar y reducir el contenido de metales en suelos mineros.

[EN] The intense industrial activity that took place over the past century resulted in large contaminated áreas. This is an important risk to human health and environmental safety. Recent biotechnological techniques for bioremediation include phytoremediation, which uses plants to remove or stabilize contaminants in soils. In our study we choose birch (Betula alba) as the preferred species to remedy mining soils, due to it produces a large biomass and can accumulate high levels of toxic elements in its tissues. The aim of this study was (i) to determine the possibility of using this species in reforestation and/or remediation of mining soils (ii) to elucidate the potential of tocopherol levels as indicators of heavy metal pollution. Trees growing in mining soils with high concentrations of Zn, Cd and Pb were sampled and the metal content in various organs and in tree rings was analyzed. α-tocoferol levels were also analyzed as an indicator of stress. The results showed a different distribution of metals in plant tissues. Zn and Cd had a higher accumulation in leaves, whereas Pb was stored in the timber. In addition, the metal content in tree rings was higher in older rings, leading to a conclusion that older tissues present a detoxification strategy. Furthermore, we saw how the presence of α- tocoferol on branches can be an indicator of metal stress in plants and it can be also used as a monitoring factor.

Elementos-traço (mercúrio total, selênio e cobre) em penas de pinguins do gênero Pygoscelis da Ilha Rei George, Antártica

Este estudo teve como objetivo avaliar a magnitude da exposição de pinguins do gênero Pygoscelis da Antártica ao mercúrio total (HgT), selênio (Se) e cobre (Cu) e verificar a variabilidade destes elementos-traço de acordo com suas características biológicas, para tal foram utilizadas penas do peito de Pygoscelis papua (n=30), P. antarctica (n=21) e P. adeliae (n=48) que corresponderam àquelas coletadas em animais da Ilha Rei George, Antártica. Encontrou-se níveis aparentemente inferiores de HgT e Se em P. papua tendo também sido a espécie que mais se distinguiu das outras duas. Além disto, houve correlação entre os níveis de HgT e Se para as espécies P. antarctica e P. adeliae indicando que é possível que ocorra nos pinguins um processo de destoxificação do mercúrio orgânico por meio do selênio, porém isto não pôde foi confirmado. Ressalta-se que o atual estudo é o primeiro a relatar níveis de HgT em penas do peito da espécie P. antarctica, sendo esta informação relevante para o entendimento e conservação desta espécie de pinguim da Antártica. De modo geral o cobre (8346/32545 ng/g p.s. mín/máx) foi o elemento que apareceu em concentrações maiores se comparado ao selênio (618,7/7923,5 ng/g p.s. mín/máx) e mercúrio total (114,6/1180,2 ng/g p.s. mín/máx). Estas elevadas concentrações de cobre nas penas possivelmente se deve a incorporação via alimentação composta essencialmente de krill, que é um crustáceo de altas concentrações deste elemento essencial. Genericamente foi possível observar que todas as informações reportadas no atual trabalho, tais como as correlações entre os elementos-traço (HgT e Se) e características biológicas, mostram que as três espécies de pinguins analisadas seguem padrões distintos de distribuição de elementos-traço possivelmente porque apresentam dieta diferenciada, além disto, é provável que as características individuais influenciem nestes padrões, e por isto devem ser avaliadas para um melhor entendimento da distribuição dos elementos-traço nestes animais. Observa-se que as concentrações do elemento-traço não essencial HgT aparentemente não constituiu uma ameaça aos pinguins porque foi inferior ao nível considerado prejudicial a aves, enquanto o selênio é um elemento que deve ser constantemente monitorado pois exibiu concentrações próximas àquelas considerada danosa para aves quando incorporadas via dieta. Verificou-se que a presença dos elementos-traço não constitui uma ameaça a saúde dos Pygoscelis da Ilha Rei George, apesar disso, faz-se necessária a realização de um programa de monitoramento constante nesta região, que é uma área ainda considerada como não impactada pela ação antrópica.

Construção, clonagem e expressão do fragmento B da toxina diftérica de Corynebacterium diphtheriae (cepa PW- 8) em Mycobacterium bovis BCG sub-cepa Moreau

A vacina anti-diftérica de uso corrente no Brasil (DTP), embora de alta eficácia na prevenção da difteria, está associada com episódios de toxicidade e reatogenicidade no recipiente vacinal, resultantes de proteínas residuais derivadas do processo de produção ou detoxificação. Estratégias para o desenvolvimento de vacinas menos reatogênicas e ao mesmo tempo mais eficazes e economicamente viáveis contra a difteria têm sido alvo de intensa investigação. A alternativa proposta por nosso grupo é a utilização da vacina contra a tuberculose (Mycobacterium bovis BCG sub-cepa Moreau), como vetor do gene que codifica o fragmento B da toxina diftérica (dtb) de 58,3 kDa. Neste trabalho o dtb foi clonado no vetor micobacteriano bifuncional (pUS977) de expressão citoplasmática e os clones recombinantes (pUS977dtbPW8), após a transformação do BCG, foram testados com relação a expressão do DTB em BCG e quanto a antigenicidade frente a anticorpos policlonais anti-toxóide diftérico por Immunobloting. A integridade do gene dtb e a identidade das sequências de DNA da construção plasmidial pUS977dtbPW8 foram confirmadas por sequenciamento de DNA e análise de similaridade. A imunogenicidade do BCGr pUS977dtbPW8 expressando o DTB foi investigada em camundongos BALB/c, os resultados obtidos revelaram uma soroconversão específica (IgG). A infectividade e atividade microbicida do BCGr pUS977dtbPW8 no ambiente intracelular foi avaliada através da infecção de linhagens de células de monócitos humano (THP-1), os dados obtidos indicaram que houve sobrevivência intracelular em até 12 dias. Nesse contexto, esplenócitos dos camundongos imunizados com 30 e 60 dias foram extraídos, mostrando que o BCGr pUS977dtbPW8 persistiu até 60 dias na ausência de pressão seletiva e a viabilidade celular não sofreu alteração significativa durante o período testado. Por outro lado, o BCGr pUS977dtbPW8, quando submetido a seis sub-cultivos consecutivos in vitro não apresentou diferença significativa na capacidade de expressar o DTB, demonstrando portanto a persistência da estabilidade funcional da linhagem recombinante. A estabilidade estrutural da construção pUS977dtbPW8 também foi avaliada por PCR confirmando a presença do gene dtb em colônias do BCGr pUS977dtbPW8 . Adicionalmente, foi possível avaliar preliminarmente in vitro a capacidade soroneutralizante dos soros de camundongos imunizados com BCGr pUS977dtbPW8 após 30 e 60 dias em células VERO. A ação citotóxica da toxina diftérica entre as diluições de 1/4 e 1/16 foram neutralizadas com o pool de soros imunes com 60 dias. Finalmente, em nosso estudo foi possível avaliar o potencial da vacina BCG como vetor de expressão de um antígeno de Corynebacterium diphtheriae in vitro e in vivo.

砷超富集植物蜈蚣草(Pteris vittata L.)中砷解毒机制的研究

砷是一种具有致癌、致畸、致突变的有毒元素，在地表的含量本来很低。然而，随着现代社会的发展和工业活动的增加导致砷污染日趋严重。土壤和水体中的砷污染可以通过食物链进入人体，对人类的健康造成极大的危害。植物修复是一种利用植物对污染物的超富集能力来清除或减低污染的新型环境生物技术。植物修复的实际应用依赖于超富集植物的发现和超富集机制的阐明，特别是砷解毒过程(砷的吸收、还原和区域化)的研究及相关基因的克隆。砷超富集植物蜈蚣草(Pteris vittata L.)中砷解毒机制的阐明将为砷污染的植物修复及新型工程植物的研发提供理论基础。 本论文以蜈蚣草为试材，针对蜈蚣草的砷解毒机制取得了如下研究进展： 1.以砷超富集植物蜈蚣草为材料，建立了一个适于研究蜈蚣草砷吸收和解毒机制的新系统—愈伤组织悬浮培养体系。首次证明蜈蚣草愈伤组织与其孢子体及配子体一样具有对砷的抗性和砷超富集的能力。 2.以蜈蚣草愈伤组织为材料，通过比较亚砷酸盐、砷酸盐和二甲基胂酸盐对蜈蚣草和拟南芥植物毒性的差异，表明砷的还原可能是蜈蚣草对砷解毒的重要机制之一而砷的甲基化对蜈蚣草的砷解毒作用甚微。 3.以蜈蚣草愈伤组织为材料，通过对砷在蜈蚣草愈伤组织细胞中的亚细胞定位，首次直接证明植物液泡对砷具有非常明显的区隔化作用。暗示区隔化作用在蜈蚣草对砷的解毒过程中发挥着重要的作用。 4.通过测定蜈蚣草愈伤组织对不同化学态的砷处理下抗氧化物质的变化发现酸溶性巯基在蜈蚣草砷解毒中也发挥着重要作用。 5.以蜈蚣草愈伤组织为材料，发现磷和砷的吸收在高浓度范围下（﹥0.2 mM）存在明显的协同效应。对蜈蚣草高亲和磷酸盐转运蛋白基因－PvPHT基因功能的初步分析则表明PvPHT参与了蜈蚣草对磷和砷的吸收过程。

Protein Profiles in Zebrafish (Danio rerio) Embryos Exposed to Perfluorooctane Sulfonate

Perfluorooctane sulfonate (PFOS) is widely distributed and persistent in the environment and in wildlife, and it has the potential for developmental toxicity. However, the molecular mechanisms that lead to these toxic effects are not well known. In the present study, proteomic analysis has been performed to investigate the proteins that are differentially expressed in zebrafish embryos exposed to 0.5 mg/l PFOS until 192 h postfertilization. Two-dimensional electrophoresis coupled with mass spectrometry was employed to detect and identify the protein profiles. The analysis revealed that 69 proteins showed altered expression in the treatment group compared to the control group with either increase or decrease in expression levels (more than twofold difference). Of the 69 spots corresponding to the proteins with altered expression, 38 were selected and subjected to matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (TOF/TOF) analysis; 18 proteins were identified in this analysis. These proteins can be categorized into diverse functional classes such as detoxification, energy metabolism, lipid transport/steroid metabolic process, cell structure, signal transduction, and apoptosis. Overall, proteomic analysis using zebrafish embryos serves as an in vivo model in environmental risk assessment and provides insight into the molecular events in PFOS-induced developmental toxicity.

Genome-wide identification and characterization of cytochrome P450 monooxygenase genes in the ciliate Tetrahymena thermophila

Background: Cytochrome P450 monooxygenases play key roles in the metabolism of a wide variety of substrates and they are closely associated with endocellular physiological processes or detoxification metabolism under environmental exposure. To date, however, none has been systematically characterized in the phylum Ciliophora. T. thermophila possess many advantages as a eukaryotic model organism and it exhibits rapid and sensitive responses to xenobiotics, making it an ideal model system to study the evolutionary and functional diversity of the P450 monooxygenase gene family. Results: A total of 44 putative functional cytochrome P450 genes were identified and could be classified into 13 families and 21 sub-families according to standard nomenclature. The characteristics of both the conserved intron-exon organization and scaffold localization of tandem repeats within each P450 family clade suggested that the enlargement of T. thermophila P450 families probably resulted from recent separate small duplication events. Gene expression patterns of all T. thermophila P450s during three important cell physiological stages (vegetative growth, starvation and conjugation) were analyzed based on EST and microarray data, and three main categories of expression patterns were postulated. Evolutionary analysis including codon usage preference, sit-especific selection and gene-expression evolution patterns were investigated and the results indicated remarkable divergences among the T. thermophila P450 genes. Conclusion: The characterization, expression and evolutionary analysis of T. thermophila P450 monooxygenase genes in the current study provides useful information for understanding the characteristics and diversities of the P450 genes in the Ciliophora, and provides the baseline for functional analyses of individual P450 isoforms in this model ciliate species.

The effect of cyanobacterial crude extract on the transcription of GST mu, GST kappa and GST rho in different organs of goldfish (Carassius auratus)

The glutathione S-transferases play important roles in the detoxification of microcystin. Core-sequences of three classes of GST (mu, kappa and rho) were cloned from goldfish (Carassius auratus L) i.p. injected with cyanobacterial crude extract at two doses (50 and 200 mu g MC-LReq kg(-1) BW). The relative changes of the mRNA abundance in liver, kidney and intestine were analyzed by real-time PCR. The transcription of GST mu was inhibited in intestine at both doses and the transcription of GST kappa was inhibited from 12 to 48 h in kidney at both doses. The decreased transcription of GST rho was detected in all three organs at the high dose. It is suggested that transcription inhibition of GST rho might be significant in MCs toxicity at higher toxin concentration in omnivorous freshwater fish. Alteration in transcription of GSTs stimulated by MCs implicates an increased health risk to fish. (C) 2008 Published by Elsevier B.V.

Antioxidant enzyme activities of Microcystis aeruginosa in response to nonylphenols and degradation of nonylphenols by M-aeruginosa

The aim of this study was to examine the effects of chemical nonylphenols (NPs) on the antioxidant system of Microcystis aeruginosa strains. The degradation and sorption of NPs by M. aeruginosa were also evaluated. High concentrations of NPs (1 and 2 mg/l) were found to cause increases in superoxidase dismutase (SOD) and glutathione-S-transferase (GST) activities and in glutathione (GSH) levels. These results suggest that toxic stress manifested by elevated SOD and GST levels and GSH contents may be responsible for the toxicity of NPs to M. aeruginosa and that the algal cells could improve their antioxidant and detoxification ability through the enhancement of enzymatic and nonenzymatic prevention substances. The observed elevations in GSH levels and GST activities were relatively higher than those in SOD activities, indicating that GSH and GST contributed more in eliminating toxic effects than SOD. Low concentrations of NPs (0.05-0.2 mg/l) enhanced cell growth and decreased GST activity in algal cells of M. aeruginosa, suggesting that NPs may have acted as a protecting factor, such as an antioxidant. The larger portion of the NPs (> 60%) disappeared after 12 days of incubation, indicating the strong ability of M. aeruginosa to degrade the moderate persistent NP compounds. The sorption ratio of M. aeruginosa after a 12-day exposure to low nominal concentrations of NPs (0.02-0.5 mg/l) was relatively high (> 30%). The fact that M. aeruginosa effectively resisted the toxic effects of NPs and strongly degraded these pollutants indicate that M. aeruginosa cells have a strong ability to adapt to variations in environmental conditions and that low and moderate concentrations of organic compounds may favor its survival. Further studies are needed to provide detailed information on the fate of persistent organic pollutants and the survival of algae and to determine the possible role of organic pollutants in the occurrence of water blooms in eutrophic lakes.

Sequential ultrastructural and biochemical changes induced in vivo by the hepatotoxic microcystins in liver of the phytoplanktivorous silver carp Hypophthalmichthys molitrix

Up to now, in vivo studies on the toxic effects of microcystins (MCs) on the ultrastructures of fish liver have been very limited. The phytoplanktivorous silver carp was injected i.p. with extracted hepatotoxic microcystins (mainly MC-RR and -LR) at a dose of 1000 mu g MC-LReq. kg(-1) body weight, showing a time-dependent ultrastructural change in liver as well as significant increases in enzyme activity of plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH). We observed for the first time the occurrence of a large amount of activated secondary lysosomes, which might be an adaptive mechanism to eliminate or lessen cell damage caused by MCs through lysosome activation. Quantitative and qualitative determinations of MCs in the liver were conducted by HPLC and LC-MS2, respectively. MCs concentration in the liver reached the maximum (114.20 mu g g(-1) dry weight) after 3 h post-injection, and then rapidly dropped to 7.57 mu g g(-1) dry weight at 48 h, indicating a deputation of 99% accumulated MC-LReq. On the other hand, a decrease trend in glutathione (GSH) concentration was observed in the liver of silver carp while the activity of glutathione S-transferase (GST) increased significantly after injection. The high tolerance of silver carp to MCs might be due to the high basic GSH level in their liver, and/or an increased GSH synthesis. (C) 2007 Elsevier Inc. All rights reserved.

The role of NH4+ toxicity in the decline of the submersed macrophyte Vallisneria natans in lakes of the Yangtze River basin, China

Experimental and field studies were conducted to evaluate the effects of NH4+ enrichment on growth and distribution of the submersed macrophyte, Vallisneria natans L, in lakes of the Yangtze River in China, based on the balance between free amino acids (FAA) and soluble carbohydrates (SC) in the plant tissue. Increase of NH4+ rather than NO3- concentrations in the water column caused FAA accumulation and SC depletion of the plant. The plant showed a unimodal pattern of biomass distribution along both FAA/SC ratios and external NH4+ concentrations, indicating that a moderate NH4-N concentration (< 0.3 mg L-1) benefited the plant, whereas the high NH4-N concentration (> 0.56 mg L-1) eliminated the plant completely. Therefore, 0.56 mg NH4-N mg L-1 in the water column was taken as the upper limit for V. natans in lakes of the Yangtze River basin. The mesocosm experiment showed that at a high external NH4-N (0.81 mg L-1), V. natans failed to propagate with a loss of half SC content (5 mg g(-1) DW) in the rhizomes, indicating that the consumption of carbohydrates for detoxification of excess NH4+ into non-toxic FAA significantly diminished carbohydrate supply to the rhizomes. This might consequently inhibit the vegetative reproduction of the plant, and also might be an important cause for the decline and disappearance of the plant with eutrophication. The present study for the first time reports substantial ecophysiological evidences for NH4+ stress to submersed macrophytes, and indicates that NH4+ toxicity arising from eutrophication probably plays a key role in the deterioration of submersed macrophytes like V. natans.

The acute effects of microcystin LR on the transcription of nine glutathione S-transferase genes in common carp Cyprinus carpio L.

The glutathione S-transferases play important roles in the detoxification of microcystin. In this experiment, nine glutathione S-transferase genes including cytosolic GSTs (rho, mu, theta, alpha and pi), mitochondrial GST (kappa) and microsomal GSTs (mGST1, mGST2 and mGST3) were cloned from common carp Cyprinus carpio. The mRNA abundance of each carp GST isoform in liver was analyzed by real time PCR. The relative changes after stimulation with microcystin LR were also analyzed: increased levels of transcription of GST alpha, rho and mGST3 isoforms were detected at 6 h post stimulation; the transcription of mu, theta and mGST2 isoforms were relatively stable; and all the GST isoforms except GST kappa and rho recovered to original levels compared with controls at 72 h. It is suggested that MC-LR showed different effects on the transcription of nine carp GST isoforms. (c) 2006 Elsevier B.V. All rights reserved.

Identification of differentially expressed genes in Tetrahmena thermophila in response to dichlorodiphenyltrichloroethane (DDT) by suppression subtractive hybridization

The insecticide dichlorodiphenyltrichloroethane (DDT) is persistent in the environment, and continues to cause health problems. Tetrahymena has potential as a model organism for assaying low levels of DDT and for analysing the mechanisms of its toxicity. We constructed the suppression subtractive hybridization library of T thermophila exposed to DDT, and screened out 90 Expressed Sequence Tags whose expressions were significantly up- or downregulated with DDT treatment. From this, a series of important genes related to the DDT metabolism and detoxification were discovered, such as P450 gene, glutathione S-transferase gene and sterol carrier protein 2 gene. Furthermore, their expressions under different concentrations of DDT treatment were detected by real-time fluorescent quantitative PCR. The results show that Tetrahymena is a relevant and useful model organism for detecting DDT in the environment and for discovering biomarkers that can be used to develop specific bio-reporters at the molecular and genomic levels.

铜-农药污染的土壤生态化学脱毒行为研究

通过开展Cu与农药甲胺磷、草甘嶙吸附／解吸试验、Cu和甲胺磷的大豆根际行为试验，以及农药脱毒生态化学指示（土壤酶、有效P）试验，对东北地区典型土壤—黑土与棕壤污染物的化学脱毒行为进行研究，为东北地区农业清洁生产提供理论参考。污染物的吸附／解吸行为是土壤物理化学脱毒的体现。试验结果表明，Cu2＋的吸附／解吸等温线与Freundiich方程有较好的拟合性。黑土对cu2＋的吸附脱毒能力高于棕壤，且解吸率更低。Cu2＋的解吸量与吸附量之间呈二次幂函数关系。描述Cu2＋吸附／解吸动力学过程的最优模型为双常数方程。两种农药在土壤原胶体及去有机质土壤上的吸附仍符合Freundllch方程，但有机质去除后吸附量大为降低。无机矿物是甲胺磷吸附的主要载体，而有机质对草甘麟的吸附脱毒则更为重要。在土壤一农药-Cu2＋共存体系中，Cu2+的吸附量降低，解吸量增加。由于较强的络合能力，草甘麟对Cu2+吸附确军吸行为的影响大于甲胺磷。根际和土壤酶通过生物化学机制对污染物进行脱毒。大豆根系通过将Cu2＋转化为铁锰氧化物结合态和有机质结合态而减少吸收。同时，甲胺磷在大豆根际环境中的降解速率显著加快。土壤脱氢酶对农药反应敏感，多数情况表现出抑制作用。酸性磷酸酶在甲胺磷加入初期被刺激，随后被部分抑制；随着草甘磷浓度的升高，酸性磷酸酶的抑制作用增强。农药的施入增加了土壤有效P含量，但对有效P的贡献率仍较低。