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1.
A Hillenweck D E Corpet J C Killeen M Bliss J P Cravedi 《Journal of agricultural and food chemistry》1999,47(7):2898-2903
The metabolic fate of chlorothalonil, a broad spectrum fungicide that is known to be metabolized via glutathione conjugation, was examined through the analysis of urine and bile metabolites. The role of digestive microflora in the metabolism of chlorothalonil was assessed by comparing the metabolic patterns in germ-free and conventional rats. Low urinary and biliary excretion of radioactivity was observed in both conventional and germ-free rats. However, the urinary excretion of radioactivity was higher in conventional than in germ-free rats. Radio-HPLC analysis of urine and bile showed a complex metabolic profile in both conventional and germ-free rats. Methylthio metabolites of chlorothalonil were determined in ethyl acetate extracts of urine and bile of conventional and germ-free rats. These metabolites were excreted in a higher amount in the urine of conventional rats than in the urine of germ-free rats. This study shows the complexity of chlorothalonil metabolism and the role of the digestive microflora in chlorothalonil metabolism. 相似文献
2.
采用未有百菌清用药史的土壤定向培养49d,分离出50株百菌清耐受菌株,经初筛、复筛后得到1株降解能力最强的细菌BJQ2,经形态学、生理生化试验、16SrDNA序列同源性比对及系统发育树的分析,初步确定为阴沟肠杆菌（EnterobactercloacaeB9）,BJQ2可以以百菌清为唯一碳源于基础培养基中生长。BJQ2降解百菌清的最适宜条件为：培养温度30~35℃,初始pH值为7.0~8.0,百菌清的初始浓度为20mg·L-1,选用菌体母液（OD600=0.35）,以10%的接种量接种,6d后提取百菌清,利用气相色谱进行检测分析,计算后发现百菌清的降解率可以达到79.2%。 相似文献
3.
Gene isolation, analysis of expression, and in vitro synthesis of glutathione S-transferase from orange fruit [Citrus sinensis L. (Osbeck) 总被引：1，自引：0，他引：1
Glutathione S-transferases (GSTs) (EC 2.5.1.18) are ubiquitous enzymes that have a defined role in xenobiotic detoxification, but a deeper knowledge of their function in endogenous metabolism is still lacking. In this work, we isolated the cDNAs as well as the genomic clones of orange GSTs. Having considered gene organization and homology data, we suggest that the isolated GST gene is probably involved in the vacuolar import of anthocyanins. We also found that the blood and blond orange GSTs shared the same nucleotide sequences, but as expected, the GST expression in the nonpigmented orange cultivar [Citrus sinensis L. (Osbeck)] (Navel and Ovale) was strongly reduced as compared to that of the pigmented orange (Tarocco). Interestingly, in the crude extracts of pigmented orange fruits, the GST activity was reproducibly detected by providing either 1-chloro-2,4 dinitrobenzene (CDNB) or cyanidin-3-O-glucoside (C-3-G) as substrates; moreover, we have shown that cyanidin-3-O-glucoside acted as a powerful competitive inhibitor of 1-chloro-2,4 dinitrobenzene conjugation to reduced glutathione (GSH) in the pigmented orange, confirming that this molecule might easily bind to the active site of the enzyme and functions as a putative substrate. In addition, we have reported here the successful in vitro expression of orange GST cDNAs leading to a GST enzyme that is active against cyanidin-3-O-glucoside, thus suggesting the probable involvement of the isolated gene in the tagging of anthocyanins for vacuolar import. This last result will help to study the kinetic and structural properties of orange fruit GST avoiding time-consuming protein purification procedures. 相似文献
4.
紫杆柽柳谷胱甘肽硫转移酶基因的克隆及功能鉴定 总被引：1，自引：0，他引：1
根据GenBank已公布的植物谷胱甘肽硫转移酶基因EST序列，设计引物利用3’和5’RACE方法，获得紫杆柽柳GST基因(TaGST)全长cDNA序列，全长为1175 bp，开放读码区672 bp, 编码224个氨基酸。其所编码的蛋白与大豆的GST10同源性最高为48%。利用pET-28a（+）构建了紫杆柽柳GST基因的原核表达载体，转入BL21大肠杆菌进行了原核表达。诱导表达蛋白的SDS-PAGE检测表明, 所表达的蛋白分子量约为26 kD，与预测的分子量大小一致。初步的酶学活性的检测表明所克隆的基因编码的肽段具有催化GST蛋白通用底物CDNB和GSH反应的活性。 相似文献
5.
摘要：利用基因重组技术将鸡传染性支气管炎病毒S1基因分段（A段和B段）克隆到pGEX-6p-1载体中，并利用IPTG诱导表达。表达产物裂解后，经SDS-PAGE电泳和考马司亮蓝染色，有明显的融合表达蛋白条带。通过特异性抗体的Western-blot检测，两段表达产物均在相应位置出现了明显的条带，证明表达产物具有部分抗原性。用Glutathione Sepharose 4B纯化出GST融合蛋白。将纯化得到的融合蛋白免疫家兔，连续免疫3次，将得到的抗体与200TOCID50/0.1ml病毒工作液在37℃温育1h进行气管环试验，计算结果 A段抗体的50%血清中和终点为1：18，而B段抗体的50%血清中和终点为1：53，从而表明这两段的表达产物具有较好的免疫原性，且B段表达产物的免疫原性比A段高。 相似文献
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7.
甘蓝型油菜柱头特异启动子的克隆和序列分析 总被引：1，自引：0，他引：1
芸薹属自交不亲和相关基因SLR1只在柱头中特异表达，根据已知的SLR1基因启动子序列设计特异引物，用PCR法从几个甘蓝型油菜品种和品系中分离得到相应的启动子片断，进行序列比较分析后，并构建它们与GFP融合的植物表达载体，转化拟南芥，验证所分离到的启动子的时空特异性。研究表明，来自自交不亲和羽衣甘蓝和来自自交亲和的甘蓝型油菜的SLR1启动子具有相似的序列结构和相同的组织特异表达功能。该结果将为下一步研究S-locus基因在甘蓝型油菜中的表达和相互作用奠定基础。 相似文献
8.
Hafize Dilek Tepe 《Journal of plant nutrition》2017,40(9):1287-1299
Boron (B) is one of the essential nutrients for the growth of plants, but its high concentrations are toxic for plants. Thus, B toxicity is a big challenge in crop cultivation. Nitric oxide (NO) is a small signaling molecule that has cytoprotective roles in plants. We investigated whether exogenous sodium nitroprusside (SNP), which is a NO donor, may succeed to alleviate B-induced toxicity in wheat cultivars. Seedlings were grown for 10 days in a growth chamber at 25°C with 16 hr light–8 hr dark photo cycle. After high B application, the effects of SNP on growth parameters; electrolyte leakage (EL); changes in reactive oxygen species [contents of hydrogen peroxide (H2O2), malondialdehyde (MDA), and proline]; the activities of antioxidant enzymes [glutathione peroxidase (GSHPx), glutathione reductase (GR), and glutathione S-transferase (GST)] and nitrate reductase (NR); and low molecular weight organic acid (LMWOAs) contents and also chlorophyll and total carotenoid contents were investigated in both shoots and roots of two different wheat cultivars. All experiments were carried out in triplicate. 0.2 mM SNP application ameliorated the chlorophyll and total carotenoid contents, and growth parameters such as shoot length, root length, and fresh weight in both wheat cultivars exposed to B stress. SNP reduced the B-induced lipid peroxidation, EL, and proline and H2O2 content in wheat cultivars. SNP application also increased the activities of NR and antioxidant enzymes, including GSHPx, GR, and GST in wheat cultivars exposed to B toxicity. All of the tested LMWOAs including succinic, propionic, butyric, oxalic, formic, malic, malonic, and benzoic acids were increased by SNP treatment in the shoots and roots of both wheat cultivars exposed to B toxicity. In conclusion, results obtained from this study have demonstrated that interactive effects of SNP with B considerably reduced the toxic effects of B in wheat cultivars. 相似文献
9.
Thibon C Cluzet S Mérillon JM Darriet P Dubourdieu D 《Journal of agricultural and food chemistry》2011,59(4):1344-1351
Volatile thiols, compounds that contribute strongly to the varietal aroma, are present in much higher concentrations in sweet wines than in dry wines. This positive effect, due to the presence of Botrytis cinerea on the berries, in fact results from a strong enrichment of cysteine S-conjugate precursors in botrytized berries. In the present study, a convenient model was investigated to reproduce and therefore study this phenomenon. A Vitis vinifera cell culture was used as a simple model, and we focused on S-3-(hexan-1-ol)-l-cysteine (P-3SH), the cysteinylated precursor of 3-sulfanylhexanol. We demonstrated that grapevine cells were able to produce P-3SH and that the presence of B. cinerea considerably increased the precursor level (up to 1000-fold). This positive result was determined to be due to metabolites secreted by the fungus. These molecules were temperature sensitive, unstable over time, and their production was activated in the presence of grapevine cells. Moreover, part of the pathway leading to P-3SH was deciphered: it was directly derived from the cleavage of S-3-(hexan-1-ol)-l-glutathione, which itself was generated after a conjugation of glutathione on (E)-2-hexenal. 相似文献
10.
百菌清是一种广谱非内源性农药，在土壤中难以降解，已经成为农业环境污染的主要污染源之一。因此，其对环境的污染和被污染土壤的修复技术越来越受到关注。土壤环境中原位降解细菌的多样性对于评价环境毒理学、生物降解性、自净化能力和污染物的修复潜力具有重要价值。该研究从长期被百菌清污染的土壤中收集大量样本，分离到了14种能够明显降解百菌清的细菌。根据菌株形态和rDNA同源性分析，将它们分为假单胞菌属（Pseudomonas sp.）、无色杆菌属（Achromobacter sp.）、苍白杆菌属（Ochrobactrum sp.）、青枯菌属（Ralstonia sp.）和溶杆菌属（Lysobacter sp.）。其中溶杆菌属是该研究中新发现的具有百菌清降解能力的功能菌属，该发现扩大了已知的百菌清降解菌的菌属范围。通过进一步鉴定及生理生化分析，确定了该降解菌的分类地位及理化特征。此外，该研究通过基因文库法克隆到了发挥关键降解作用的水解酶基因，并发现该基因与转座子原件IS-Olup相连，二者组成代谢转座子，具有水平转移的分子基础。通过揭示降解基因在细菌间的漂移机制，为修复百菌清污染土壤的生物技术奠定了理论基础。 相似文献
11.
Glutathione (GSH) can be found in plants in considerable amounts. It is very important for numerous metabolic functions and for the detoxification of xenobiotics. The present study was carried out to show the inducibility of glutathione S-transferase (GST) isozymes in spruce by various substrates commonly used to assay GST activity, Inducers were: chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene (DCNB), para-nitrobenzoylchloride (pNBC), fluorodifen and n-ethylmaleinimide (NEM). Conjugation of CDNB was enhanced by the xenobiotics ranking in the order: CDNB > fluorodifen > pNBC > NEM > DCNB. 相似文献
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13.
Responses of soil microorganisms and enzymes to repeated applications of chlorothalonil 总被引：5，自引：0，他引：5
Yu YL Shan M Fang H Wang X Chu XQ 《Journal of agricultural and food chemistry》2006,54(26):10070-10075
Introduction of anthropogenic chemicals into soil may have lasting effects on soil microbial activities and thus soil health. This study was conducted with chlorothalonil to evaluate its effects on soil bacterial, fungal, and actinomycete populations and soil enzymes (acid phosphatase, alkaline phosphatase, urease, catalase, and invertase) after repeated applications. After the first addition of chlorothalonil, the soil bacterial and actinomycete populations were significantly reduced, whereas the population of soil fungi was unchanged. The most marked inhibition on soil microorganisms was observed after the second pesticide addition. However, after initial variations, soil bacteria, fungi, and actinomycetes adapted gradually to chlorothalonil, and the negative effects became transient and weaker following the third and fourth treatments. The inhibitory effect of repeated chlorothalonil applications on soil enzymes followed a similar trend to that on soil microorganisms. Repeated chlorothalonil applications did not result in significant changes in its persistence. Three bacterial strains capable of utilizing chlorothalonil as a sole source of carbon and energy for growth were isolated 21 days after the fourth treatment with chlorothalonil, which indicated that the capability of soil microorganisms for degrading chlorothalonil was formed during the experiment. 相似文献
14.
the degradation rate of chlorothalonil (2,4,5,6-tetrachloroisophthalonitrile) was significantly decreased after repeated application in field experiments. However, the degradation rate completely recovered after further application of chlorothalonil. Chlorothalonil was also degraded in a laboratory study, forming a stoichiometric amount of chloride anion. 4-Hydroxy-2,5,6-trichloroisophthalonitrile (TPN-OH) was also detected as a metabolite in the study. Bacterial which degrade chlorothalonil rapidly were isolated from the soil. The cell suspension of the bacteria transformed chlorothalonil to TPN-OH and chloride anion, but did not utilize chlorothalonil (=cometabolism). It seems that microbial acclimation has occurred during repeated application because the bacteria could not be detected in soil which was not treated with chlorothalonil.Dedicated to Professor J. C. G. Ottow on the occasion of his 60th birthday 相似文献
15.
To investigate whether the regulation of garlic allyl sulfides on biotransformation enzyme expression is tissue-specific, the expression of cytochrome P450 2B1 (CYP 2B1) and the placental form of glutathione S-transferase (PGST) in liver, lung, and intestine, which are the three major organs responsible for drug metabolism, was examined. Rats were orally administrated 0.5 or 2 mmol/kg BW diallyl sulfide (DAS) or 0.5 mmol/kg BW diallyl disulfide (DADS) or diallyl trisulfide (DATS) three times per week for 6 weeks. The final body weights and the body weight ratio of liver and lung were not changed by any of these three allyl sulfide treatments as compared to the control rats. An 11- and 12-fold increase of 7-pentoxyresorufin O-dealkylase (PROD) activities was noted in rats treated with 0.5 or 2 mmol/mg BW DAS, respectively, as compared with the controls (P < 0.05). In contrast, DADS and DATS significantly increased hepatic PGST activity toward ethacrynic acid by 30 and 40%, respectively, as compared with the control rats (P < 0.05). An increase in PGST activity was only noted at 2 mmol/kg BW DAS group (P < 0.05). In addition, similar increases in PGST activity due to DADS and DATS were also noted in lung and jejunum tissue (P < 0.05). Immunoblot assay shows that the changes in CYP 2B1 and PGST proteins due to the three garlic allyl sulfide treatments on liver, lung, and jejunum were consistent with those observed for PROD and PGST activities. Northern blot further revealed that the DADS and DATS increased PGST mRNA levels in both liver (2.9- and 3.0-fold, respectively) and lung (4.1- and 2.6-fold, respectively) and DAS dose-dependently increased CYP 2B1 mRNA levels in the liver. Garlic allyl sulfides differentially induced CYP 2B1 and PGST expression, and this up-regulation of these two biotransformation enzymes is tissue-specific. 相似文献
16.
Capone DL Pardon KH Cordente AG Jeffery DW 《Journal of agricultural and food chemistry》2011,59(20):11204-11210
Precursors to varietal wine thiols are a key area of grape and wine research. Several such precursors, in the form of odorless conjugates, have been closely studied in recent years. A new conjugate has now been identified as 3-S-cysteinylglycinehexan-1-ol (Cysgly-3-MH), being the dipeptide intermediate between cysteine and glutathione precursors of tropical thiol 3-mercaptohexan-1-ol (3-MH). Authentic Cysgly-3-MH was produced via enzymatic transformation of the glutathione conjugate and used to verify the presence of both diastereomers of Cysgly-3-MH in Sauvignon blanc juice extracts. Cysgly-3-MH was added into our HPLC-MS/MS precursor method, and the validated method was used to quantify this new analyte in a selection of Sauvignon blanc juice extracts. Cysgly-3-MH was found in the highest concentrations (10-28.5 μg/L combined diastereomer total) in extracts from berries that had been machine-harvested and transported for 800 km in 12 h. This dipeptide conjugate was much less abundant than the glutathione and cysteine conjugates in the samples studied. On the basis of the results, the new cysteinylglycine conjugate of 3-MH seemingly has a short existence as an intermediate precursor, which may explain why it has not been identified as a natural juice component until now. 相似文献
17.
在室内半静水的实验条件下,将翡翠贻贝（Pernaviridis）分别暴露于0．005、0．010、0．050、0．100mg·L-。的0#柴油水溶液（WSF）中,在污染后l、3、7、15d取样,于15d后转入清洁海水中进行7d污染释放试验,在18、22d采样。测定内脏团和外套膜丙二醛（MDA）、还原型谷胱甘肽（GSH）含量、超氧化物歧化酶（SOD）和谷胱甘肽-S-转移酶（GST）活性。结果表明,高剂量组（0．100、0．050mg·L^-1）暴污的翡翠贻贝内脏团和外套膜SOD活性和MDA含量变化比低剂量组（0．005mg·L^-1）显著（P〈0．01）,整个暴露过程中呈现先抑制后诱导再抑制的波动变化趋势,无明显的剂量-时间-效应关系。WSF暴露初期,内脏团组织中GSH含量和GST活性反应灵敏且受到抑制,此后受到明显诱导而升高,在暴露第7d达到最大值并显著高于空白组（P〈O．01）。外套膜各剂量组GSH含量和GST活性存在此消彼长的趋势,剂量-时间效应明显。污染解除后,4种生理指标均缓慢恢复至空白对照组水平。 相似文献
18.
A hydroponic experiment was conducted to investigate the dynamic variations of cadmium (Cd) uptake and transport, non‐protein thiols (NPT) and glutathione (GSH) concentrations, glutathione S‐transferase (GST) activity and lipid peroxidation under Cd stress in order to clarify the role of NPT and GST in reducing Cd toxicity and translocation in rice (Oryza sativa L.). Cadmium accumulation was initially fast and then slowed down with increasing time of Cd exposure. However, the rice growth inhibition and lipid peroxidation were not intense until 5d after Cd treatment, even though Cd kept accumulating in root and shoot, suggesting that Cd may be effectively detoxified. The concentrations of NPT in root increased gradually until 5d after Cd stress, whereas those in shoot showed no significant changes. The concentration of shoot GSH was progressively enhanced upon Cd treatment, while it gradually declined in root after an initial increase. The GST activity varied similarly in root and shoot, reaching the maximum level on 3rd day, followed by a significant decrease 5 d after Cd application. Significant increases of lipid peroxidation and root‐to‐shoot translocation on 7th day indicate that the equilibrium in Cd‐thiol interaction in rice might be disturbed upon the prolonged Cd exposure. In summary, our results suggest that Cd may be retained and detoxified in rice root through chelation with thiol compounds and subsequent sequestration. 相似文献
19.
Transformation of the fungicide chlorothalonil by Fenton reagent 总被引：1，自引：0，他引：1
A modified Fenton reagent (Fe(3+)/H(2)O(2)) transformed the fungicide chlorothalonil within 60 min in aqueous solution at unadjusted pH. Transformation varied with ferric salt. Transformation was greatest with ferric nitrate and least when ferric sulfate was used. UV irradiation enhanced the transformation of chlorothalonil. The transformation of chlorothalonil was enhanced, which increased with ferric ion or hydrogen peroxide concentration. Maximum transformation was achieved at 2 mM ferric ion and 100 mM hydrogen peroxide. Additionally, chlorothalonil was more dechlorinated in the UV irradiation condition. The proposed reaction pathway includes reduction of chlorothalonil to trichloroisophthalonitrile, dichloroisophthalonitrile, and monochloroisophthalonitrile; oxidation of trichloroisophthalonitrile to trichloro-3-cyanobenzoic acid and 3-carbamyltrichlorobenzoic acid; and oxidation of hydroxychlorothalonil to trichloro-3-cyanohydroxybenzoic acid and trichlorocyanophenol. 相似文献
20.
《Soil biology & biochemistry》2001,33(7-8):1123-1130
The aim of this study was to investigate the effects of carbaryl, at different concentrations and exposure times, on the biochemical responses of the earthworm, Eisenia fetida andrei (E. andrei) to: (1) elucidate the mechanisms of action of carbamate compounds; and (2) explore the potential for using these responses as biomarkers to monitor carbamate-contaminated soils or for use in sublethal assays to test chemicals in the laboratory. Thus, earthworms were exposed to increasing concentrations of carbaryl (CA) (12, 25 and 50 mg kg−1 artificial soil) for different periods of time (2, 7 and 14 days) using the standard soil test method. The activities of the following enzymes were measured: catalase (CAT), acetylcholinesterase (AChE), glutathione reductase (GR), glutathione-S-transferase (GST), methoxyresorufin-O-deethylase (MROD) and NADH (NADH Red) and NADPH (NADPH Red) cytochrome reductases activities. Levels of lipid peroxides (LP), peroxidizable lipids (LPI), total glutathione (total GSH) and the percentage of oxidized glutathione (%GSSG) were also determined. Our results showed that cholinesterases are the main target of carbaryl in E. andrei. Carbaryl also inhibited biotransformation enzyme activities but did not induce oxidative stress. Changes were detected in phase I and acetylcholinesterase activities upon exposure to the lowest dose of carbaryl (12 mg kg−1). This demonstrated the sensitivity of these parameters in E. andrei. In addition, discrimination between doses and exposure times was clearer when all the responses were considered rather than a selective choice of biomarkers. This confirms our previous finding that a suite of biochemical responses could be used as a sublethal assay for testing chemicals in the laboratory or for soil contamination surveys. 相似文献