安全剂R-28725保护玉米免受氯嘧磺隆药害的机理研究

研究了安全剂R-28725对玉米的保护作用及对氯嘧磺隆的解毒机理。当氯嘧磺隆的使用量为5、10、15 g/hm2时,加入R-28725能够明显提高玉米株高、株鲜重和产量,直接增加玉米体内谷胱甘肽(GSH)含量,增加氯嘧磺隆与谷胱甘肽的轭合,从而达到解毒的目的。     

芸苔素内酯保护玉米免受胺苯磺伤害的作用及其机理

采用土培法和田间小区试验法测定了芸苔素内酯(BR)对玉米的保护作用,并对其作用机理进行了初步研究.结果表明:BR的浸种浓度在0.01～0.04mg·L-1范围内,能不同程度地缓解16.0μg·kg-1胺苯磺隆对玉米的药害,BR浓度为0.02mg·L-1时,解毒效果最好.用0.02mg·L-1的BR浸种,能不同程度地缓解2.0～32.0μg·kg-1的胺苯磺隆对玉米生长的抑制,但当胺苯磺隆浓度增加到64.0μg·kg-1时,BR浸种无解毒作用;按15g ai/hm2的剂量施用胺苯磺隆对玉米的减产率达11.8%,用BR 0.02、0.04 mg·L-1浸种处理则减产率分别为0.2%和0.1%;BR提高胺苯磺隆处理的玉米幼苗中ALS活性可能是其保护玉米免受胺苯磺隆伤害的机理之一.     

大豆田土壤中氯嘧磺隆残留动态研究

采用高效液相色谱法对大豆田土壤中的氯嘧磺隆残留动态进行了研究。结果表明,氯嘧磺隆在大豆田土壤中的降解半衰期为7～10d,施药后60d其残留量为0．004mg／kg。     

安全剂AD-67减轻单嘧磺隆对高粱药害的作用

采用生物测定的方法对安全剂AD-67保护高粱免受单嘧磺隆药害进行研究,并对其保护作用的机理进行了初步探讨。结果表明:单嘧磺隆在浓度0.05～0.2mg/kg时,对高粱的生长有较为强烈的抑制作用;当AD-67的浓度在0.5～10mg/kg时,能不同程度地缓解单嘧磺隆对高粱的药害。在单嘧磺隆毒土浓度为0.05mg/kg时,使用5mg/kgAD-67浸种处理,高粱的主根长、株高和谷胱甘肽(GSH)含量分别是对照(CK)的118.45%、107.47%和108.33%。AD-67能够提高单嘧磺隆处理后的高粱幼苗中的GSH含量,促进单嘧磺隆与GSH的轭合作用,这是安全剂AD-67保护高粱免受单嘧磺隆药害的重要途径之一。     

安全剂AD-67保护玉米免受单嘧磺隆药害的作用及其机理

采用生物测定法研究了安全剂AD-67对除草剂单嘧磺隆造成玉米伤害的保护作用,并对其作用机理进行了初步探讨。结果表明:单嘧磺隆在浓度0.025～0.050mg/kg时,对玉米的生长有较为强烈的抑制作用;当AD-67的浓度在0.5～10mg/kg时,能不同程度地缓解单嘧磺隆对玉米的药害。当单嘧磺隆毒土浓度为0.025mg/kg,使用1mg/kgAD-67浸种处理,玉米的主根长和株高分别是对照的107.3%和100.82%;AD-67浸种处理的玉米,幼苗中谷胱甘肽(GSH)含量和乙酰乳酸合成酶(ALS)活性分别是对照的109.76%和113.33%。     

不同玉米品种对氯吡嘧磺隆的耐药性差异及其机制

为科学评价氯吡嘧磺隆对玉米的安全性,采用室内盆栽法和酶活力测定法探讨不同玉米品种对氯吡嘧磺隆的耐药性差异及其机制。不同玉米品种对氯吡嘧磺隆的耐药性有较大差异,其中甜玉米耐药性最差,IC10仅为94.87g/hm2(有效成分,余同)。花糯1号、鲁单981、五岳97-1等品种表现出中等耐药性,耐药性倍数为1.51～1.97;五岳21、郑单958、强盛16等耐药性较强,耐药性倍数为2.05～2.43。强盛16、鲁单981和甜玉米对氯吡嘧磺隆的耐药性差异显著,但其靶标酶乙酰乳酸合成酶(ALS)在离体条件下对该药剂的敏感性差异不显著;在活体条件下,三者ALS活力均可很快恢复,且耐药性品种恢复能力较强。与甜玉米相比,经氯吡嘧磺隆处理后强盛16的谷胱甘肽-S-转移酶(GSTs)对氯吡嘧磺隆的反应幅度较大和反应时间较短,第72h相对活力就达到4.2的峰值,而甜玉米变化幅度相对较小,第96 h才达到2.6的峰值。研究表明,不同玉米品种对氯吡嘧磺隆耐药性的差异源于GSTs对氯吡嘧磺隆的代谢差异,而与靶标酶ALS对氯吡嘧磺隆的敏感程度无关。     

高效液相色谱-串联质谱法检测稻田中吡嘧磺隆和苯噻酰草胺残留

采用高效液相色谱-串联质谱法(HPLC-MS/MS),建立了稻田水、土壤、水稻植株、稻秆、稻壳及糙米基质中吡嘧磺隆和苯噻酰草胺的残留分析方法。样品经20 m L V(乙腈)∶V(水)=70∶30的混合溶液提取,提取液用20 mg石墨化碳黑(GCB)与30 mg乙二氨基-N-丙基硅烷(PSA)净化,HPLC-M S/M S检测。吡嘧磺隆在上述各基质中的添加回收率在76%~107%之间,相对标准偏差(RSD)在1.5%~14%之间,定量限为0.004~0.01 mg/kg;苯噻酰草胺的添加回收率在77%~101%之间,RSD在2.4%~13%之间,定量限为0.001~0.01 mg/kg。实现了对两种除草剂同时简便、快速测定的要求。采用该方法测定了26%吡嘧磺隆·苯噻酰草胺水面扩展粒剂在稻田施用后,其有效成分吡嘧磺隆和苯噻酰草胺在实际样品中的残留量。结果表明,两种除草剂均属于易降解农药,在本试验条件下其在糙米中的残留量均低于我国最大残留限量(MRL)(吡嘧磺隆0.1 mg/kg;苯噻酰草胺0.05 mg/kg)。     

氯嘧磺隆高效降解细菌的分离与筛选

从北京市、河北省一些农田和农药厂排污口活性污泥中分离得到7株具有明显降解氯嘧磺隆作用的降解细菌,这些菌株能够以氯嘧磺隆为唯一氮源生长,对氯嘧磺隆的降解率为65．85％至94．78％不等。其中F-1和E-1两株菌为本试验的最高效率的降解菌,在40ml培养基、细菌起始浓度7.6×10^7cfu／ml、30℃恒温、180r／min条件下培养72h,F-1菌株对底物氯嘧磺隆50mg／L的降解效率为67．5％;E-1菌株对底物氯嘧磺隆10mg／L的降解率为94．78％。     

氯嘧磺隆高效降解菌株黑曲霉（TR-H）最佳降解条件的研究

试验采用高效液相色谱法测定不同条件下反应液中氯嘧磺隆除草剂的降解率,确定氯嘧磺隆高效降解菌株黑曲霉(TR-H)的最佳降解条件。结果表明:当氯嘧磺隆的初始浓度为10.0mg/L、接种量为5.0mL菌悬母液、反应液的温度为30.0℃、恒温振荡培养7d,真菌黑曲霉(TR-H)可以降解96.4%以上的氯嘧磺隆。     

降解菌2N3对被氯嘧磺隆污染土壤的生物修复

在实验室条件下,研究了高效降解菌2N3(克雷伯氏菌属,Klebsiella sp.)对被氯嘧磺隆污染土壤的修复作用及其影响因素。当土壤中氯嘧磺隆的添加浓度为20 mg/kg,每 1克土壤中2N3的接菌量为1×106个菌体时,第30 d时土壤中氯嘧磺隆的降解率为84.6%,对照仅为13.4%;相同2N3接菌浓度下,当土壤中氯嘧磺隆浓度为100 mg/kg时,其降解率为31.1%。以小麦、玉米、黄瓜为供试作物,在土壤中施加 20 mg/kg的氯嘧磺隆, 当每 1克土壤中2N3的接菌量为1×106个 菌体时,小麦、玉米、黄瓜的出苗率分别为85%,82%和79%,且处理组株高高于对照,表明降解菌2N3具有明显减轻氯嘧磺隆药害的作用。研究表明,人工接种降解菌2N3可提高土壤中氯嘧磺隆的降解率,有效降低其在土壤中的残留。     

化合物YF-206防除乙草胺药害研究

本文采用土培法对本课题组所合成的化合物5-二氯乙酰基-6-甲基-9-氧杂-1,5-二氮杂二环[4.3.0]壬烷(YF-206)进行了室内活性测定实验。当化合物YF-206浓度为25mg/kg时,玉米各项生长指标恢复率达到最大,其株高、株鲜重的恢复率分别为:55.39%和55.86%;玉米幼苗根部GSH含量增幅达到50.59%,叶片中增幅为26.02%,玉米根部GST比活力为2.694 nmol/s/mg。实验结果表明化合物YF-206对玉米有较好的安全活性,能在一定程度上解除乙草胺对玉米的药害。     

Glutathione transferase activities toward herbicides used selectively in soybean

Using extracts from suspension-cultured cells of soybean (Glycine max cv. Mandarin) as a source of active enzymes, the activities of glutathione transferases (GSTs) catalysing the conjugation of 1-chloro-2,4-dinitrobenzene (CDNB) and selective herbicides were determined to be in the order CDNB≫ fomesafen>metolachlor=acifluorfen>chlorimuron-ethyl. GST activities showed a thiol dependence in a substrate-specific manner. Thus, GST activities toward acifluorfen and fomesafen were greater when homoglutathione (hGSH), the endogenously occurring thiol in soybean, was used as the co-substrate rather than glutathione (GSH). Compared with GSH, hGSH addition either reduced or had no effect on GST activities toward other substrates. In the absence of enzyme, the rates of hGSH conjugation with acifluorfen, chlorimuron-ethyl and fomesafen were negligible, suggesting that rapid hGSH conjugation in soybean must be catalysed by GSTs. GST activities were subsequently determined in 14-day-old plants of soybean and a number of annual grass and broadleaf weeds. GST activities of the plants were then related to observed sensitivities to post-emergence applications of the four herbicides. When enzyme activity was expressed on a mg^-1 protein basis, all grass weeds and Abutilon theophrasticontained considerably higher GST activity toward CDNB than soybean. With fomesafen as the substrate, GST activities were determined to be in the order soybean≫Echinochloa crus-galli>Digitaria sanguinalis>Sorghum halepense=Setaria faberi with none of the broadleaf weeds showing any activity. This order related well to the observed selectivity of fomesafen, with the exception of A. theophrasti, which was partially tolerant to the herbicide. Using metolachlor as the substrate the order of the GST activities was soybean>A. theophrasti≫S. halepense>Amaranthus retroflexus>Ipomoea hederacea, with the remaining species showing no activity. GST activities toward metolachlor correlated well with the selectivity of the herbicide toward the broadleaf weeds but not toward the grass weeds. Acifluorfen and chlorimuron-ethyl were selectively active on these species, but GST activities toward these herbicides could not be detected in crude extracts from whole plants. © 1997 SCI     

天然芸苔素内酯缓解胺苯磺隆对后茬水稻药害的作用机理初探

通过测定天然芸苔素内酯对水稻幼苗谷胱甘肽(GSH)含量、谷胱甘肽-S-转移酶(GST)活性以及在离体和活体条件下对水稻幼苗乙酰乳酸合成酶(ALS)活力的影响,研究天然芸苔素内酯减轻胺苯磺隆对水稻药害的作用机理。结果表明,其作用机理与水稻幼苗GSH含量、GST活性变化没有太大相关性,可能与其能间接激活水稻幼苗ALS活力有较大相关。     

Protective effect of vitamin C against chlorpyrifos oxidative stress in male mice

Pesticides may induce oxidative stress leading to generate free radicals and alternate antioxidant or oxygen free radical scavenging enzyme system. This study was conducted to investigate the acute toxicity of chlorpyrifos toward male mice and the oxidative stress of the sub-lethal dose (1/10 LD₅₀) on the lipid peroxidation level (LPO), reduced glutathione content (GSH) and antioxidant enzymes; catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glucose-6-phosphate dehydrogenase (G6PD), and glutathione-S-transferase (GST) activities. Also, the protective effects of vitamin C (200 mg/kg body weight, bw) 30 min before or after administration of chlorpyrifos were investigated. The results demonstrated that the LD₅₀ value of chlorpyrifos was 134.95 mg/kg bw. The oral administration of 13.495 mg/kg chlorpyrifos significantly caused elevation in LPO level and the activities of antioxidant enzymes including CAT, SOD and GST. However, GPx activity remained unchanged, while the level of GSH and G6PD activity were decreased. Vitamin C treatment to chlorpyrifos intoxicated mice decreased LPO level and GST activity, normalized CAT, SOD and G6PD activities, while GSH content was increased. We conclude that vitamin C significantly reduces chlorpyrifos-induced oxidative stress in mice liver and the protective effect of the pre-treatment with vitamin C is better than the post-treatment.     

Effects of indoleacetic acid and kinetin on lipid peroxidation and antioxidant defense in various tissues of rats

This study aims to investigate the effects of indoleacetic acid (IAA) and kinetin (Kn), which are plant growth regulators (PGRs), on antioxidant defense systems [reduced glutathione (GSH), glutathione-S-transferase (GST), catalase (CAT)], and lipid peroxidation level (malondialdehyde, MDA) various tissues of rats. Rats (Sprague-Dawley albino) were exposed to 100 ppm IAA and Kn. One hundred parts per million of PGRs was administered orally to rats ad libitum for 21 days continuously. The PGRs treatments caused different effects on the content of MDA and antioxidant defense system in comparison to those of control rats. According to the results, the subchronic treatments of IAA caused significant decrease in the GSH concentration and CAT activity in erythrocyte. Kn decreased GSH concentration in erythrocyte too. While the MDA concentration in brain was increased significantly by IAA and Kn, Kn decreased significantly brain CAT and GST activity. The liver GST activity was decreased by IAA and Kn. But, liver CAT activity was increased by IAA. On the other hand, while IAA treatment caused a significant decrease kidney GST activity, Kn caused a significant decrease both kidney GST and CAT activity. Also, while heart CAT activity was decreased by IAA, heart GST activity was decreased by both IAA and Kn. Moreover, MDA concentration in heart was increased by Kn treatment. It was concluded that IAA might effect MDA and antioxidant defense on the animals at subchronic treatment.     

Effect of chlorimuron-ethyl on Bradyrhizobium japonicum and its symbiosis with soybean

Possible side-effects of the acetolactate synthase (ALS)-inhibiting herbicide chlorimuron-ethyl on Bradyrhizobium japonicum (Kirchner & Jordan) in pure culture and on inoculated soybean plants growing under controlled conditions were investigated. Growth of B japonicum strain E109 was not affected by this herbicide even when exposed to concentrations 150 times higher than recommended field doses. However, nodulation of soybean plants treated 5 days after emergence with chlorimuron-ethyl at standard application rates was impaired: a 38% decrease in the number of nodules per plant was observed four weeks after treatment. Despite nodule number decrease, no changes in shoot nitrogen content could be detected. Total fresh biomass was diminished by 25% in herbicide-treated plants. Leghemoglobin content in nodules did not vary; nevertheless total nodule protein was diminished by 40% in the herbicide-treated group. ALS activity in different soybean tissues and their relative sensitivity to chlorimuron-ethyl were also investigated. Roots and bacteroids had the greatest specific ALS activities. On a fresh weight basis, the bacteroid fraction displayed the highest ALS activity and was also the most tolerant to in vitro chlorimuron addition: 72% of its activity was retained after including 10 microM chlorimuron-ethyl in the reaction mixture. These results indicate that standard application rates of chlorimuron-ethyl will have limited incidence on B japonicum survival, and effects on nodulation may have little long-term consequences on soybean nitrogen fixation potential. The differences found among soybean tissues not only in intrinsic ALS activity but also in their relative sensitivity to this herbicide suggests that, in leguminous plants living in symbiosis with rhizobia, nodules may contribute to an enhanced tolerance to ALS inhibitors.     

Dégradation du chlorimuron-éthyle déposé sur divers minéraux et sur acides humiques

Degradation of chlorimuron-ethyl The chemical degradation of chlorimuron-ethyl and chlorimuron on various dry minerals (kaolinite, silica, bentonite H⁺, montmorillonite K10) and on solid humic acid was investigated at 30°C. Hydrolysis of sulfonylurea function was therefore always observed, and followed the same pathway as previously described in acidic aqueous solution. The formed phenyl-sulfonamide was transformed into saccharin on only weakly acidic minerals (kaolinite, silica). A relative affinity between pyrimidamine (hydrolysis product) and strong acidic minerals (bentonite H⁺, montmorillonite K10) or humic materials was observed.     

毛细管电泳定量测定稻田土壤中痕量磺酰脲类除草剂残留

研究了提取浓缩和毛细管电泳相结合测定稻田土壤中低剂量多个磺酰脲类除草剂混合残留的分析方法。结果表明: 毛细管电泳可有效分离和定量测定稻田土壤中甲磺隆、氯磺隆和氯嘧磺隆混合残留。通过定量补偿甲磺隆、氯磺隆和氯嘧磺隆可使残留检测限达到ng/kg级,回收率>87%。表明毛细管电泳具有定量测定土壤中ng/kg级磺酰脲类除草剂残留的能力。     

Effects of carbaryl and azinphos methyl on juvenile rainbow trout (Oncorhynchus mykiss) detoxifying enzymes

In this study, the effects of sublethal exposures to the anticholinesterase insecticides azinphos methyl (AzMe) and carbaryl on the detoxifying responses of juvenile rainbow trout Oncorhynchus mykiss were investigated. Juvenile specimen were exposed to sublethal concentrations of AzMe (2.5 and 5 μg/L) and carbaryl (1 and 3 mg/L) for 24, 48 and 96 h. Carboxylesterase (CbE), catalase (CAT) and glutathione S-transferase (GST) activities as well as reduced glutathione (GSH) and cytochrome P450-1A (CYP1A) levels were monitored in liver and/or kidney. In all exposed groups liver CbE was significantly inhibited. Liver and kidney GSH level was reduced after sublethal exposure to both compounds. Carbaryl induced CAT activity during the first 48 h of exposure, followed by a significant decrease, whereas AzMe continuously decreased CAT activity. GST activity and CYP1A were transiently induced at 24 h by carbaryl exposure (3 mg/L) but sublethal exposure to AzMe did not affect GST activity or CYP1A. Our results show that the O. mykiss detoxifying system are a target for carbaryl and AzMe action, probably affecting redox balance. Although the responses showed similar trends in both organs, they were more important in liver than in kidney. The early inhibitory effect in CAT activity and GSH content produced by AzMe may be associated with a high degree of oxidative stress. Early induction of CYP1A, GST and CAT by carbaryl followed by enzyme inhibition suggests a milder or delayed oxidative stress, revealing differences between both pesticides metabolization. CbE inhibition is a good biomarker for AzMe and carbaryl exposure.