绿盲蝽和中黑盲蝽解毒代谢酶活性的检测

害虫解毒代谢增强是对杀虫剂产生抗性的重要机制。为此,本研究建立了Bt棉田重要害虫绿盲蝽和中黑盲蝽成虫多功能氧化酶、酯酶和谷胱甘肽S转移酶3种解毒酶活性的测定方法。研究结果表明:以DCNB(1,2二氯4硝基苯)为底物,不能检测到棉盲蝽谷胱甘肽S转移酶的活性;棉盲蝽多功能氧化酶活性也不宜选用动力学法进行测定。同时2种棉盲蝽成虫的日龄对解毒酶活性影响不明显。因此,对由不同日龄成虫组成的棉盲蝽田间种群可以直接进行解毒酶活性的检测。     

植物信号分子介导抗病反应的研究进展

植物受到病原菌侵染时会发生一系列防卫反应：通过各种信号反应事件,如抗病信号的产生、传导及互作,最终诱导各种抗病防卫基因的表达和代谢的变化进而产生抗性。在防卫反应过程中,各类信号分子如激素类（水杨酸、茉莉酸、乙烯、脱落酸）,第二信使类(钙离子、活性氧、一氧化氮),低分子肽类(谷胱甘肽),脂类以及糖类发挥着重要作用。最近几年,这些信号分子介导植物抗病反应研究取得重要进展,就此进行了综述,并对今后的研究作出展望。     

GPX4失活通过JNK通路缓解LPS诱导巨噬细胞炎症反应

【目的】研究硒蛋白谷胱甘肽过氧化物酶4(glutathione peroxidases 4,GPX4)失活如何参与调控脂多糖(lipopolysaccharide, LPS)诱导的RAW264.7巨噬细胞炎症反应及其潜在的分子机制。【方法】体外培养RAW264.7巨噬细胞,以DMSO为对照,使用0.1～5.0μmol/L GPX4抑制剂FIN56处理,通过CCK-8法检测细胞活力和Western blotting检测GPX4蛋白表达水平,确定抑制剂最适浓度。将RAW264.7巨噬细胞分为4组：对照组,添加DMSO培养24 h; FIN56(GPX4抑制剂)组,添加0.5μmol/L FIN56培养24 h; DMSO-LPS组,DMSO培养24 h后使用LPS(100 ng/mL)刺激3 h; FIN56-LPS组,FIN56培养24 h后使用LPS刺激3 h。各组细胞经培养后,利用荧光探针2′,7′-二氯二氢荧光素二乙酸酯(2′,7′-dichlorodi-hydrofluorescein diacetate, H₂DCFDA)检测细胞内活性氧(reactive...     

氨氮胁迫对四鼻须鲤鱼肝脏GST酶活力的影响

实验采用生态毒理学方法,以NH4Cl为实验药物,设置0、50、150和250mg/L实验浓度。通过对野生和养殖群体的四鼻须鲤鱼氨氮对肝脏谷胱甘肽巯基转移酶（GST）活力的影响,比较两个群体抗氨氮能力。结果表明：在相同的氨氮浓度和胁迫时间下,四鼻须鲤鱼养殖群体GST酶活力的变化趋势与野生群体大致相同。除250mg/L浓度组在胁迫5d和10d时,野生群体酶活性略低于养殖群体外,在其余时间点的野生群体酶活性均高于养殖群体,而且,在高浓度氨氮胁迫下,野生群体肝脏的GST酶活性达到峰值的时间比养殖群体更短,显示出较强的抗氨氮能力。     

Oxidative stress drives divergent evolution of the glutathione peroxidase (GPX) gene family in mammals

The molecular basis for adaptations to extreme environments can now be understood by interrogating the ever-increasing number of sequenced genomes. Mammals such as cetaceans, bats, and highland species can protect themselves from oxidative stress, a disruption in the balance of reactive oxygen species, which results in oxidative injury and cell damage. Here, we consider the evolution of the glutathione peroxidase (GPX) family of antioxidant enzymes by interrogating publicly available genome data from 70 mammalian species from all major clades. We identified 8 GPX subclasses ubiquitous to all mammalian groups. Mammalian GPX gene families resolved into the GPX4/7/8 and GPX1/2/3/5/6 groups and are characterized by several instances of gene duplication and loss, indicating a dynamic process of gene birth and death in mammals. Seven of the eight GPX subfamilies (all but GPX7) were under positive selection, with the residues under selection located at or close to active sites or at the dimer interface. We also reveal evidence of a correlation between ecological niches (e.g. high oxidative stress) and the divergent selection and gene copy number of GPX subclasses. Notably, a convergent expansion of GPX1 was observed in several independent lineages of mammals under oxidative stress and may be important for avoiding oxidative damage. Collectively, this study suggests that the GPX gene family has shaped the adaption of mammals to stressful environments.     

还原型谷胱甘肽对罗非鱼生长和抗氧化性能的影响

将体长、体质量相近的奥尼罗非鱼Oreochromis niloticus×O.aureus 360尾,随机分为4组,每组3个重复,每个重复30尾.在Ⅰ、Ⅱ、Ⅲ、Ⅳ组的日粮中分别添加0、100、200、400 mg/kg剂量的还原型谷胱甘肽(GSH),试验期 60 d,检测了与生长性能、血清中类胰岛素样生长因子-I(IGF-I)水平及肌肉组织抗氧化能力相关的各项指标.结果表明:1)在试验期间,添加GSH的Ⅱ组和Ⅳ组的相对体质量生长率(RSGR)显著高于对照组Ⅰ组(P《0.05),而且第20 d时,Ⅱ组和Ⅳ组鱼体的平均体质量显著高于Ⅰ组(P《0.05);2)第40 d和第60 d,Ⅳ组鱼体的血中IGF-I水平均显著高于Ⅰ组(P《0.05);3)第20 d,各处理组鱼体肌肉组织的总抗氧化能力(T-AOC)显著高于Ⅰ组(P《0.05).第40 d,Ⅳ组的T-AOC和总超氧化物歧化酶(T-SOD)活力均显著高于Ⅰ组(P 《0.05).第60 d,Ⅱ组的 T-AOC 和T-SOD活力显著高于Ⅰ组(P 《0.05).以上结果显示,在饲粮中添加适宜剂量的GSH能改善鱼体肌肉组织的抗氧化能力,并使IGF-I水平在正常生理范围内有所升高,从而对鱼体的生长起促进作用.     

微囊藻毒素MC-LR对罗非鱼（Oreochromis niloticus）肝脏谷胱甘肽含量及其相关酶活性的影响

采用腹腔注射的方式研究了微囊藻毒素MC-LR（注射剂量为500μg.kg-1 BW）胁迫下罗非鱼（Oreochromis niloticus）肝脏谷胱甘肽（GSH）含量及其相关酶γ-谷氨酰半胱氨酸合成酶（γ-GCS）、谷胱甘肽硫转移酶（GST）、谷胱甘肽还原酶（GR）和谷胱甘肽过氧化物酶（GPx）活性的动态变化。结果表明,MC-LR能够对罗非鱼肝脏GSH含量及其相关酶活性产生明显影响。在MC-LR胁迫下,与对照组相比,GSH含量呈现先下降后上升趋势,总体上被诱导;罗非鱼肝脏γ-GCS和GST在试验过程中出现两次显著升高现象,在GST作用下,GSH与MC-LR结合会造成GSH的消耗,γ-GCS和GR的活性增强能够使GSH含量升高,从而使罗非鱼肝脏GSH能够维持一定水平;GR和GPx的活性均表现为先上升后下降,它们能有效调节罗非鱼肝脏GSH-GSSG缓冲系统,从而在减轻或消除由MC-LR侵入而造成的肝细胞氧化胁迫中发挥重要作用。     

小麦品种的耐盐性变化：甜菜碱和乙烯的作用

Four wheat (Triticum aestivum L.) cultivars 711, PBW343, 3765 and WH542 were screened for studying variations in glycinebetaine (GB) content and plant dry mass under 100 mmol L-1 NaCl stress. A tolerance index was calculated using plant dry mass data to select salt-tolerant and salt-sensitive types and find association between tolerance index and GB content. Tolerance index has been used as a good criterion to select the tolerant types under high salinity stress. Further, physiological differences in salt-tolerant cultivar 711 and salt-sensitive cultivar WH542 were examined. The salt-tolerant cultivar exhibited greater GB content, which was found correlative with ethylene. The cultivar also showed higher nitrogen (N) content and nitrate reductase activity, reduced glutathione and higher redox state resulting in maximal protection of plant dry mass than the salt-sensitive type. Thus, the content of GB may be considered as important physiological criteria for selecting salt-tolerant wheat types.     

发芽绿豆苗的生理指标和抗氧化特性对邻苯二甲酸酯的响应特征

Single phytotoxicity of two representative phthalate esters （PAEs）, di-n-butyl phthalate （DnBP） and bis（2-ethylhexyl） phthalate （DEHP）, was tested in mung bean （Vigna radiata） seedlings germinated for 72 h in soils spiked with varying concentrations （0-500 mg kg-1 soil） of DnBP or DEHP. PAEs added at up to 500 mg kg-1 soil exerted no significant effect on germination but both pollutants significantly inhibited root elongation （P 〈 0.01）; DEHP inhibited shoot elongation （P 〈 0.01） and DnBP depressed biomass on a fresh weight basis （P 〈 0.05）. Seedling shoot and root malondialdehyde （MDA） Contents tended to be stimulated by DnBP but inhibited by DEHP. However, increases in superoxide dismutase, peroxidase, ascorbate peroxidase and polyphenol oxidase activities, as well as glutathione （GSH） content, were induced at higher concentrations （e.g., 20 mg kg-1） of both compounds. Accumulation of proline in both roots and shoots and the storage compounds, such as free amino acids and total soluble sugars, in whole plant was induced under the stress exerted by both PAEs. The general responses of mung bean seedlings indicated higher toxicity of DnBP than DEHP on primary growth, during which root elongation was a more responsive index. MDA and GSH were more sensitive parameters in the roots than in the shoots and they might be recommended as physiologically sensitive parameters to assess the toxicity of PAE compounds in soils in future long-term studies.     

玉米逆境响应基因ZmGST23克隆和表达分析

谷胱甘肽-S-转移酶(glutathione S-transferases,GSTs)是植物抗氧化系统的重要成员,在初生代谢、次生代谢、细胞信号转导及逆境响应等生物学过程中发挥着重要作用.本研究以GenBank中公布的玉米谷胱甘肽-S-转移酶23基因(ZmGST23)mRNA序列(GenBank登录号:NM_001111524)为依据,采用RT-PCR方法从玉米(Zea mays)自交系F83中克隆得到ZmGST23基因669 bp的完整开放阅读框(open reading frame,ORF),编码222个氨基酸,蛋白分子量为24.84kD,理论等电点为5.68.保守结构域分析表明,该基因具有GSTs所特有的N端及C端结构域,并含有典型的G位点及H位点,属于Tau类谷胱甘肽硫转移酶.系统进化分析表明,ZmGST23编码蛋白与高粱(Sorghum bieolor)GST蛋白亲缘关系最近,且在不同物种间存在明显的种属特性.启动子序列分析结果显示,ZmGST23基因启动子区含有多个响应逆境及植物激素的作用元件,包括脱落酸(abscisic acid,ABA)响应元件、生长素(auxin,IAA)响应元件、赤霉素(gibberellin,GA)响应元件、厌氧响应元件、防御及逆境响应元件以及干旱诱导的MYB转录因子(v-myb avian myeloblastosis viral oncogene homolog,MYB)结合位点.胁迫诱导表达分析表明,ZmGST23的表达显著受干旱脱水、水涝、盐、ABA、IAA、GA、低温及高温等非生物胁迫的诱导.组织特异性表达分析表明,ZmGST23基因在幼芽和成熟叶片中表达量较高,在幼根、花丝、苞叶、雌穗及雄穗中表达量较低,存在明显的组织特异性.将ZmGST23基因片段连接至原核表达载体pEASY-E1中,转化大肠杆菌(Escherichia coli)BL21,用0.5 mmol/L的异丙基硫代半乳糖苷(isopropylβ3-D-1-thiogalactopyranoside,IPTG)诱导表达1～4 h后获得30 kD的诱导蛋白.本研究结果为进一步利用该基因进行玉米抗性改良提供了一定的理论依据.