Phytotoxicity of Roundup Ultra 360 SL in aquatic ecosystems: Biochemical evaluation with duckweed (Lemna minor L.) as a model plant

Glyphosate-based herbicides (e.g. Roundup Ultra 360 SL) are extensively used in aquatic environment. Although glyphosate is more environmental favorable than many other herbicides, it may be exceptionally dangerous for aquatic ecosystems through high water solubility. Thus, the aim of the work was quantification of influence of Roundup Ultra 360 SL (containing isopropylamine salt of glyphosate as an active ingredient) on biomass and chlorophyll content within duckweed (Lemna minor L.). Moreover, changes in polyamine content and activity of such antioxidative enzymes as catalase (CAT) and ascorbate peroxidase (APX) were assayed in order to determine the biochemical mechanisms of L. minor response to the herbicide treatment. Obtained results showed that phytotoxicity of the herbicide was connected with decrease in chlorophyll-a, b and a+b content, and reduction of biomass growth. Roundup, similarly to some abiotic and biotic stressors, caused over-accumulation of putrescine, spermidine and total polyamines (PAs) within duckweed tissues. In addition an increase in CAT and APX activities suggested that stress generated by the herbicide treatment was at least partially connected with oxidative burst. Intensity of the duckweed responses to the herbicide was dependent on the applied herbicide level and/or duration of treatment.     

Elevated antioxidant response and induction of tau-class glutathione S-transferase after glyphosate treatment in Vigna radiata (L.) Wilczek

Glyphosate (N-(phosphonomethyl) glycine) is a broad-spectrum herbicide, acting on the shikimic acid pathway inhibiting 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), thus obstructing the synthesis of tryptophan, phenylalanine, tyrosine and other secondary products. It has also been reported to generate oxidative stress which influences the antioxidant response of target plants. The effect of glyphosate application on total protein, CAT, POD and GST activities was investigated and elevated expression of the oxidative stress enzymes was obtained after glyphosate treatment.Tau-class GSTs are plant-specific, and are chiefly involved in xenobiotics and oxidative stress metabolisms. Many herbicides and safeners have been known to selectively induce tau-class GSTs in different plant species. Here we also report the induction of tau-class GSTs after glyphosate treatment in the seedling roots of two Vigna radiata varieties (PDM11 and PDM54). GSH-agarose affinity chromatography and mass spectrometry revealed that the tau-class GSTs induced in the two varieties were different; the tau-class GSTs present in the untreated controls were also different in the two varieties. The present study highlights the elevated antioxidant response, the induction of tau-class GST and the genotypic variation in the type of tau-GST in control and glyphosate treated varieties of V. radiata.     

黄萎病对海/陆嫁接棉苗叶片中H2O2、抗病相关酶活性和GST、PAL基因表达的影响

为探讨棉花黄萎病抗性机制,以抗黄萎病的海岛棉材料Pima90(R)为砧木,以在湖北省感病的陆地棉品种鄂棉22(S)为接穗,获得海/陆嫁接棉苗(S/R),研究海岛棉Pima90(R)、陆地棉鄂棉22(S)和海/陆嫁接棉苗(S/R)接种后的抗病反应,测定叶片中的H_2O_2含量,比较三者POD、PAL酶活性水平和PAL、GST基因表达模式的差异。结果显示:以黄萎病菌大丽轮枝菌菌株V991接种棉苗根部,海/陆嫁接棉苗(S/R)的萎蔫程度介于砧木Pima90(R)和接穗鄂棉22(S)之间,表明利用抗病海岛棉砧木(R)对感病陆地棉(S)嫁接可提高棉苗对黄萎病的抗性;接种后,鄂棉22(S)、Pima90(R)和海/陆嫁接棉苗(S/R)三者叶片中的H_2O_2含量、POD和PAL活性均有明显升高;其中感病品种鄂棉22(S)中H_2O_2含量和POD、PAL活性上升相对较缓慢,抗病品种海岛棉Pima90(R)上升较快,而海/陆嫁接棉苗(S/R)上升幅度介于R和S二者之间,且与抗病砧木Pima90(R)表现出相似的趋势;接种后不同时段对GST和PAL基因的real-time PCR分析结果显示,GST基因表达在Pima90(R)、鄂棉22(S)和海/陆嫁接棉苗(S/R)均表现出先缓慢上升后下降的趋势;在感病品种鄂棉22(S)中高水平表达持续时间短(8～16h),而在抗病海岛棉品种Pima90(R)和海/陆嫁接棉苗(S/R)中,高水平表达持续时间较长(8～48h);PAL基因在Pima90(R)、鄂棉22(S)和嫁接棉苗(S/R)中具有相似的表达模式,即均表现为上升-下降-上升(-下降)的表达趋势,在8h和48h出现2个表达峰值;在感病品种鄂棉22(S)中PAL表达量较低,2个峰值较小;而在抗病海岛棉品种Pima90(R)中,PAL表达水平高,2个峰值较大;海/陆嫁接棉苗(S/R)中PAL表达量介于R和S二者之间。     

联苯肼酯与炔螨特混配对朱砂叶螨的联合毒力作用

为了科学合理使用2008年在中国获得登记的新型酰基乙腈联苯肼类杀螨剂联苯肼酯,延长其使用寿命,本研究采用生物测定法和紫外分光光度法针对性地开展了联苯肼酯与炔螨特混配对朱砂叶螨的联合毒力作用评价。结果表明,43%联苯肼酯悬浮剂和73%炔螨特乳油按有效成分215∶73混配对朱砂叶螨的共毒系数为163.0,共毒系数大于120,表现出良好的增效作用,经其20.00、14.29、11.11、9.09和7.69 mg/L的浓度处理24 h后,朱砂叶螨的谷胱甘肽S-转移酶(GST)活性显著降低,分别为3.371、3.645、3.932、4.045和4.089 mmol/(mg Pro·min),显著低于相同浓度联苯肼酯单剂[7.250、6.818、6.714、6.441、5.811 mmol/(mg Pro·min)]和炔螨特单剂[10.130、9.789、9.142、8.932、8.441 mmol/(mg Pro·min)],说明联苯肼酯与炔螨特混配对朱砂叶螨GST具有显著的联合抑制作用;经联苯肼酯与炔螨特混配液处理24 h后,朱砂叶螨的多功能氧化酶(MFO)、羧酸酯酶(Car E)和乙酰胆碱酯酶(Ach E)活性与联苯肼酯和炔螨特单剂处理的结果无显著差异。本研究结果为合理混配使用联苯肼酯与炔螨特防治朱砂叶螨的发生与危害提供了理论依据。     

大花红景天谷胱甘肽 S-转移酶基因的分离与鉴定

以西藏大花红景天为试材,利用生物信息学和实时荧光定量PCR技术对谷胱甘肽S-转移酶(GST)基因家族中的11个成员进行分析,以期为红景天属GST家族基因的功能研究提供参考。结果表明:基于蛋白质序列比对可以将11个大花红景天RcGSTs基因分为6个亚组,分别为tau类(RcGSTU1、RcGSTU2、RcGSTU3);phi类(RcGSTF1、RcGSTF2、RcGSTF3、RcGSTF4);lambd类RcGSTL;微粒体类RcGSTM;theta类RcGST1;zeta类RcGSTZ。在线网站分析结果显示仅RcGSTM具有跨膜结构域,并定位于细胞膜,其它RcGSTs均无跨膜结构域并定位于细胞质。组织特异性与胁迫诱导表达模式结果表明,RcGSTs基因具有不同的组织表达特异性;低温胁迫下4个基因表达水平上调,7个基因下调;且脱落酸(ABA)可以诱导RcGSTs基因表达。     

Acute effects of cadmium on liver phase I and phase II enzymes and metallothionein accumulation on sea bream Sparus aurata

This research was designed to study Sparus aurata (sea bream) biotransformation and detoxification responses to acute exposure to cadmium (Cd). Sexually immature gilthead sea bream were treated by intraperitoneal injection of Cd chloride (200 μg kg⁻¹) for 6, 12, 24 and 48 h. Cd accumulation was quantified in sea bream liver by graphite furnace atomic absorption spectroscopy after the various exposure periods. The following biological responses were measured: (1) ethoxyresorufin-O-deethylase (EROD) activity as phase I biotransformation parameter, (2) liver glutathione-S-transferase (GST) activity as a phase II conjugation enzyme and metallothionein (MT) content as specific response to Cd contamination. Cd bioaccumulation in the liver resulted in an increasing uptake up to 10.3 μg g⁻¹ wet weight after 48 h of exposure. EROD showed a significant activation only after 6 h exposure and a return to control levels after 12 h. GST revealed significant activation starting from 12 h exposure. MT accumulation in liver showed the same behavior as GST activation.     

北京地区家蝇抗药性与酶活性的关系

试验结果表明，不同抗性水平种群的羧酸酯酶活性与其对拟除虫菊酯的抗性程度明显相关．北京崇文区、宣武区和密云等地区的家蝇种群以及用残杀威选育的种群和用残杀威、７５０４、溴氰菊酯选育后合并的种群，多功能氧化酶（ＭＦＯ）活性明显高于农大敏感品系（ＳＡ），最多相差近１０倍，但是用溴氰菊酯选有的种群和用残杀威选育后加敏感品系稀释的种群，ＭＦＯ活性没有明显的提高。说明ＭＦＯ活性的提高与家蝇对残杀威的抗性有密切关系，而与对溴氰菊酯的抗性关系次之。抗性家蝇种群的乙酰胆碱酯酶（ＡｃｈＥ）对敌百虫的敏感度明显低于ＳＡ品系的ＡｃｈＥ，说明ＡｃｈＥ的敏感度降低是家蝇对有机磷杀虫剂的抗性机制之一。酸性磷酸酯酶、碱性磷酸酯酶以及谷胱甘肽－Ｓ－转移酶（ＧＳＴ）活性在不同家蝇种群中没有明显差异，说明这三个酶系可能与家蝇的抗药性无关。     

转CpTI基因杨树对美国白蛾幼虫中肠解毒酶及乙酰胆碱酯酶的影响

以转CpTI基因毛白杨回交杂种 [(Populustomentosa×Populusbolleana)×Populustomentosa]叶片饲喂 4～ 5龄美国白蛾 (HyphantriacuneaDrury)幼虫 ,对其中肠解毒酶和乙酰胆碱酯酶活性进行了测定。结果表明 ,酯酶、羧酸酯酶的活力受到明显抑制 ,且随时间的延长抑制强度增加 ,饲喂 48h后 ,上述两种酶的活力分别比对照降低 76 .42 %和 73.91% ;多功能氧化酶在饲喂的前 12h ,表现为受到抑制 ,最大抑制率为 35 .78% ,2 4h后酶活力反而高于对照 ;谷胱甘肽S -转移酶和乙酰胆碱酯酶活力受到抑制 ,而且两者表现极为相似 ,均在饲喂后 4h抑制作用最强 ,酶活力分别比对照降低 5 1.40 %和 40 .5 7% ,但在整个试验过程中 ,均没有表现出随时间加强的趋势。