6种植物次生物质对斜纹夜蛾解毒酶活性的影响

草食性昆虫取食植物时遇到宿主植物中大量次生物质的化学防御,研究昆虫适应植物毒素的反防御策略具有重要的科学意义。分别添加0.01%肉桂酸、0.01%水杨酸、0.01%花椒毒素、0.02%槲皮素、0.05%黄酮和0.1%香豆素等6种植物次生物质的人工饲料饲养斜纹夜蛾(Spodoptera litura)五龄幼虫48 h后,测定斜纹夜蛾幼虫中肠和脂肪体中谷胱甘肽S-转移酶(GSTs)、羧酸酯酶(CarE)、P450的酶含量及头部乙酰胆碱酯酶(AChE)的活性,利用半定量RT-PCR检测中肠和脂肪体中CYP4M14和CYP4S9的基因表达水平。结果表明:取食肉桂酸和香豆素后,斜纹夜蛾中肠中CarE的酶活性分别提高了1.67和1.37倍,取食6种次生物质均能显著提高斜纹夜蛾脂肪体中GSTs酶活性。取食肉桂酸和香豆素48 h后,脂肪体中P450酶含量比对照增加2.93和14.50倍。取食肉桂酸、花椒毒素、槲皮素和香豆素后,斜纹夜蛾头部AchE酶活性与对照相比提高了1.53、1.80、2.36和1.56倍。6种次生物质均可诱导脂肪体中CYP4M14基因表达,槲皮素、肉桂酸和香豆素强烈诱导CYP4S9在脂肪体中表达。表明,斜纹夜蛾具有利用植物次生物质诱导其解毒酶的能力,进而提高其对毒素的抗性。     

绿原酸对美国白蛾幼虫生长发育和解毒相关蛋白活性的影响

【目的】明确植物次生代谢物质绿原酸对美国白蛾Hyphantria cunea幼虫生长发育和解毒相关蛋白活性的影响。【方法】将含不同浓度绿原酸(0, 0.125%, 0.250%, 0.500%, 1.000%和2.000%)的人工饲料饲养美国白蛾幼虫,测定各组中5龄幼虫6 d内的死亡率, 各组中5龄幼虫在取食48 h时的营养效应指标,0.500%绿原酸处理组中幼虫(3龄幼虫开始)的生长发育,以及在取食36 h时各组中5龄幼虫肠道中的解毒相关蛋白活性。【结果】绿原酸处理组中美国白蛾5龄幼虫6 d内的死亡率随着人工饲料中绿原酸浓度的增大而逐渐升高,且显著高于对照组(取食含10% DMSO人工饲料的)。绿原酸对美国白蛾5龄幼虫的营养效应指标有显著影响,各处理组幼虫的相对取食量和近似消化率显著高于对照组,食物转化率显著低于对照组,除0.125%绿原酸处理组外,其余处理组幼虫的食物利用率和相对生长速率也显著低于对照组。0.500%绿原酸处理使美国白蛾3-5龄幼虫发育历期比对照组显著延长28.24%(3.7 d),老熟幼虫(6-7龄幼虫)的发育历期缩短8.97%(0.7 d),且幼虫总成活率、化蛹率、羽化率、雌雄性比及产卵量等各项指标均显著低于对照组。不同浓度绿原酸处理对美国白蛾幼虫肠道中的细胞色素P450酶(CYP450)、谷胱甘肽S-转移酶(GSTs)、尿苷二磷酸葡萄糖醛酸转移酶(UGT)和ABC转运蛋白(ABC transporters)的活性具有显著影响,但对羧酸酯酶(CarE)活性未见显著影响。【结论】绿原酸可影响美国白蛾幼虫的存活率、食物利用效率、生长发育和繁殖,而美国白蛾幼虫可能通过调节食物利用率和诱导其解毒相关蛋白活性从而对食物中绿原酸产生适应性。     

取食含不同植物源成分的饲料对香樟齿喙象幼虫生长发育及体内解毒酶活性的影响

【目的】明确取食含不同植物源成分的饲料对香樟齿喙象Pagiophloeus tsushimanus幼虫生长发育以及体内解毒酶活性的影响,以探究寄主植物对香樟齿喙象食性与抗性的作用。【方法】以3种樟科植物(香樟Cinnamomum camphora和浙江桂Cinnamomum chekiangensis和浙江楠Phoebe chekiangensis)枝条粉末为植物源成分,配制成半人工饲料。以半人工饲料进行室内饲养后,分别测定取食半人工饲料后香樟齿喙象幼虫的发育历期、体重、死亡率和化蛹率,以及饥饿12 h后取食含不同植物源成分的半人工饲料的4龄幼虫体内解毒酶(谷胱甘肽-S-转移酶、羧酸酯酶和乙酰胆碱酯酶)活性时序变化。【结果】香樟齿喙象幼虫的发育历期在取食含不同植物源成分的半人工饲料之间存在显著差异。取食含香樟成分的半人工饲料的幼虫发育历期最短(100.13 d),取食含浙江楠成分的半人工饲料的幼虫发育历期最长 (123.33 d);整个幼虫期的死亡率以取食含浙江桂成分的半人工饲料的幼虫最高(41.67%),以取食含香樟成分的半人工饲料的幼虫最低(10.00%);化蛹率以取食含香樟成分的半人工饲料的最高(87.32%);取食含香樟成分的半人工饲料的幼虫体重增长速率也明显高于取食含浙江桂或浙江楠成分的半人工饲料的幼虫。饥饿12 h后取食含不同植物源成分的半人工饲料的香樟齿喙象4龄幼虫体内GST和 AChE活性并无显著差异(P＞0.05);而取食含香樟成分的半人工饲料的幼虫CarE活性在60 h时大幅度上升。【结论】说明香樟齿喙象对天然寄主香樟具有较强的生理适应性,这可能与其专食性密切相关;同时,也表明香樟齿喙象可以通过调节自身生长发育和解毒代谢功能来适应不同寄主植物。     

寄主植物对甜菜夜蛾感染核型多角体病毒后黑化反应关键酶活性的影响

【目的】明确寄主植物对感染核型多角体病毒的甜菜夜蛾Spodostera exigua (Hübner)幼虫体内参与黑化反应的关键酶的影响。【方法】借助紫外分光光度计或酶标仪,测定了取食不同食物(蕹菜、甘蓝、黄豆和人工饲料)的感毒与未感毒甜菜夜蛾幼虫血淋巴中多酚氧化酶、酪氨酸羟化酶和多巴脱羧酶含量。【结果】两因素分析显示,感毒和食物两个因子及其交互作用显著影响幼虫血淋巴中这3种酶的活性;进一步比较发现,取食蕹菜的感毒幼虫多酚氧化酶活性最高,取食甘蓝的次之,而以黄豆和人工饲料为食物的多酚氧化酶活性最低,感毒幼虫血淋巴中其他两种酶活性也表现相同的趋势。【结论】寄主植物能够调控感毒甜菜夜蛾幼虫体内参与黑化反应的关键酶活性,而这些关键酶活性的变化可能与寄主植物调控甜菜夜蛾对核型多角体病毒的敏感性有关。     

甜菜夜蛾体内酶活性对三种棉酚含量棉花的响应

在室内用3种不同棉酚含量的棉花饲养甜菜夜蛾幼虫,并在不同取食时间测定了甜菜夜蛾幼虫的营养物质、消化酶、保护酶和解毒酶活性的变化.实验结果表明,甜菜夜蛾幼虫取食高棉酚含量的棉花M9101品种6h后,体内的游离脂肪酸较取食其他两种低酚含量棉花出现了显著性的降低.甜菜夜蛾幼虫取食高棉酚含量的棉花M9101品种24h后,体内的胰蛋白酶较取食1,4,6h后出现了显著性的增加.甜菜夜蛾幼虫取食高棉酚含量的棉花M9101品种1,4,6,24h后,体内的过氧化氢酶和总超氧化物歧化酶显著高于取食低酚棉花品种ZMS13和HZ401,但体内显著较低的羧酸酯酶和乙酰胆碱酯酶酶活性显著低于取食低酚棉花品种ZMS13和HZ401.多因素方差分析表明,不同棉酚含量棉花品种除对甜菜夜蛾幼虫体内淀粉酶活性无影响外,对其他消化酶、保护酶和解毒酶活性均有显著性的影响.棉花品种和甜菜夜蛾不同为害时间的交互作用可显著影响甜菜夜蛾幼虫体内的羧酸酯酶和过氧化氢酶活性.测定对甜菜夜蛾不同为害时间体内酶活性对不同棉酚含量的棉花的响应,可有效评估植食性昆虫在植物次生代谢物质压力下的生理反应和抗性变化.     

不同寄主植物继代饲养后草地贪夜蛾幼虫生理酶活性差异

为了明确继代饲喂不同寄主植物对草地贪夜蛾幼虫体内生理酶活性的影响,在室内分别用3种寄主植物叶片饲养草地贪夜蛾多代,测定其F3代5龄幼虫体内保护酶（SOD,POD,CAT）、消化酶（脂肪酶、胃蛋白酶和α-淀粉酶）和解毒酶（GSTs,CarEs,CYP450s）活性差异,并分析不同寄主植物次生代谢物含量与幼虫体内酶活及其生长发育的相关性。结果表明,取食马铃薯叶片的F3代幼虫总发育历期最长,取食小麦的最短,两者间差异显著;小麦叶片中的单宁、总酚、类黄酮和生物碱含量均显著低于玉米和马铃薯;草地贪夜蛾幼虫体内保护酶活性均为取食小麦叶片的最低,取食马铃薯叶片的最高,其中取食马铃薯叶片的草地贪夜蛾体内POD和SOD酶活性达取食小麦的1.34和1.26倍;以3种寄主为食的F3代幼虫脂肪酶和α-淀粉酶活性变化不大,但持续饲喂马铃薯叶片3个世代可导致草地贪夜蛾幼虫体内胃蛋白酶活性显著升高,达10.502 U/mg. prot,为持续饲喂玉米叶片幼虫（1.508 U/mg. prot）的6.96倍以上。连续饲喂3种寄主叶片后的幼虫GSTs和CarEs活性无显著性差异,但取食玉米叶片的幼虫体内细胞色素P450s（CYP450s）活性显著高于取食小麦和马铃薯的幼虫。可见,连续多代取食次生物质含量不同的寄主植物会使草地贪夜蛾体内部分酶活性发生变化,并对幼虫的生长发育产生影响。     

苹果蠹蛾细胞色素P450基因CYP332A19和CYP337B19的克隆及表达分析

【目的】本研究旨在通过克隆苹果蠹蛾Cydia pomonella细胞色素P450基因CYP332A19和CYP337B19,并对其进行序列和表达分析,以更好地了解这两个P450基因在植物次生物质解毒方面的作用,为进一步的功能研究提供依据。【方法】采用本地BLAST搜索苹果蠹蛾转录组数据库获得细胞色素P450基因cDNA序列,采用RT-PCR技术克隆目的基因的编码区。利用生物信息学软件分析目的基因的序列特征及与其他近缘物种的P450基因的系统进化关系。采用RT-qPCR技术测定目的基因在苹果蠹蛾不同发育阶段(卵、1-5龄幼虫、蛹和成虫)、4龄幼虫不同组织(头部、表皮、脂肪体、中肠和马氏管)以及4龄幼虫分别取食添加0.1%香豆素和0.5%槲皮素的人工饲料2 d后的表达水平。【结果】克隆获得苹果蠹蛾细胞色素P450基因CYP332A19(GenBank登录号: MF574708)和CYP337B19(GenBank登录号: MF574697)的全长cDNA序列,开放阅读框(ORF)分别长1 518和1 491 bp,分别编码505和496个氨基酸,其蛋白质分子量分别为58.586和57.734 kD,理论等电点分别为8.99和7.61。结构域分析显示,CYP332A19和CYP337B19中均包含包括血色素结合区在内的5个保守的细胞色素P450结构域。系统发育树显示,苹果蠹蛾CYP332A19与苹淡褐卷蛾Epighyas postvittana CYP332A9等CYP332A基因聚在一枝,而CYP337B19与稻纵卷叶螟Cnaphalocrocis medinalis CYP337B12和六星灯蛾Zygaena filipendulae CYP337B11等CYP337B基因聚在另一枝。RT-qPCR分析结果表明,CYP332A19和CYP337B19在苹果蠹蛾幼虫期的表达水平高于卵期的,分别在4龄幼虫脂肪体和中肠中的表达量最高。取食分别含0.1%香豆素和0.5%槲皮素的人工饲料2 d后,4龄幼虫体内的CYP332A19和CYP337B19相对表达量显著高于对照组(取食含2%DMSO的人工饲料)。【结论】CYP332A19和CYP332B19分别在苹果蠹蛾幼虫脂肪体和中肠中高表达,且在取食含香豆素和槲皮素的人工饲料的苹果蠹蛾幼虫体内表达量升高,说明这两个基因可能参与苹果蠹蛾对外源物质的解毒代谢过程。本研究的结果有助于我们了解苹果蠹蛾对寄主次生物质解毒代谢机理,为苹果蠹蛾防治提供新思路。     

不同方式诱导植物抗性对甜菜夜蛾幼虫及其寄生蜂斑痣悬茧蜂发育表现的影响

【目的】明确诱导植物抗性对植食性昆虫甜菜夜蛾 Spodoptera exigua (Hübner) 及内寄生蜂斑痣悬茧蜂 Meteorus pulchricornis (Wesmael) 生长发育的影响。【方法】采用取食损伤、机械损伤及外源喷施3种浓度 (0.01, 0.1和1 mmol/L) 茉莉酸甲酯 (MeJA) 3种方式来诱导菜薹抗性,观察甜菜夜蛾初孵幼虫及其寄生蜂斑痣悬茧蜂的发育表现。【结果】取食取食损伤和茉莉酸甲酯处理的菜薹后,甜菜夜蛾幼虫存活率显著降低,茉莉酸甲酯浓度与幼虫存活呈负相关; 取食0.1 mmol/L MeJA处理的菜薹后,甜菜夜蛾幼虫体重显著低于清水对照。甜菜夜蛾取食取食损伤和0.1 mmol/L MeJA处理的菜薹,其寄生蜂斑痣悬茧蜂幼虫的死亡率显著增加,同时结茧率显著降低;0.1 mmol/L MeJA处理的菜薹使成蜂体重显著低于对照。不同处理间斑痣悬茧蜂的卵巢发育无明显差异,卵巢由8~10根卵巢管组成,羽化24 h后成熟卵数量为7~10个,成熟卵的长度和宽度分别为350和78 μm。【结论】结果说明,取食损伤和茉莉酸甲酯处理诱导的菜薹反应会影响甜菜夜蛾的存活和大小,进而抑制斑痣悬茧蜂生长。     

茉莉酸甲酯处理对棉铃虫生长和解毒能力的影响

【目的】为明确茉莉酸甲酯（methyl jasmonate, MeJA）的不同处理方式对昆虫生长和解毒代谢能力的影响,以及MeJA熏蒸处理昆虫对植物防御和昆虫解毒代谢之间关系的影响。【方法】本文比较了取食MeJA和MeJA熏蒸等两种处理对棉铃虫Helicoverpa armigera 3龄末幼虫生长和解毒代谢酶活性的影响,并系统研究了经MeJA熏蒸处理的棉铃虫3龄末幼虫,再取食经MeJA喷雾处理的棉叶,其中肠解毒酶活性的变化。【结果】取食MeJA和MeJA熏蒸两种不同处理,对棉铃虫幼虫的生长及解毒能力的影响有所不同。取食含2.9 μg/g MeJA的人工饲料显著抑制了棉铃虫的生长,同时诱导棉铃虫幼虫P450活性增加了1.92倍,而MeJA熏蒸虽然没有影响棉铃虫幼虫的生长,但MeJA熏蒸处理使棉铃虫幼虫P450活性和羧酸酯酶活性分别增加了2.94倍和1.16倍。经MeJA熏蒸处理后的棉铃虫幼虫,再取食正常的或经MeJA喷雾处理的棉叶,其幼虫P450活性显著增加,其中经MeJA熏蒸处理的棉铃虫,再取食经MeJA喷雾处理的棉叶,其P450酶显示了最高活性。【结论】MeJA熏蒸处理棉铃虫,不仅使棉铃虫中与P450酶相关的解毒能力显著增加,而且增加了棉铃虫对于MeJA所诱导的棉花防御反应的解毒能力。     

取食不同食物的甜菜夜蛾幼虫感染核型多角体病毒后中肠组织的变化

【目的】观察取食人工饲料和3种寄主植物的甜菜夜蛾Spodoptera exigua 3龄幼虫感染核型多角体病毒后,其中肠组织的病理变化。【方法】核型多角体病毒感染甜菜夜蛾后,剖取围食膜观察在不同时间点围食膜的变化,以及采用组织切片法探究取食人工饲料和不同寄主植物的甜菜夜蛾感染病毒前后其中肠组织的病理变化。【结果】幼虫感毒后的围食膜首先由无色透明、具有弹性的圆桶状结构,变成乳白色、弹性降低的单片状结构,随着感毒后时间的推移,围食膜破坏严重,变成无弹性的碎片状。取食不同寄主植物和人工饲料的甜菜夜蛾在未感染病毒时其中肠组织都没有明显的变化,而感染病毒后甜菜夜蛾中肠组织围食膜被破坏或消失;中肠柱状细胞伸长、变形,细胞间隙变大,排列混乱,细胞脱落,肠壁变厚。此外,核型多角体病毒对取食人工饲料和不同寄主植物的甜菜夜蛾中肠组织的影响也存在差异,对取食人工饲料和黄豆的影响类似且最大、甘蓝次之,蕹菜最小。【结论】核型多角体病毒对甜菜夜蛾中肠组织具有显著的影响,且取食不同寄主植物时其影响存在明显差异。     

Uniform hypothesis of cell behavior--movement, contact inhibition of movement, adhesion, chemotaxis, phagocytosis, pinocytosis, division, contact inhibition of division, fusion.

The cell has been represented as a charged liquid drop. Contrary to the DLVO-theory, the effect of the surface potential upon the value of the interfacial tension of the cell membrane has also been taken into consideration. The cell membrane has visco-elastic properties and its constituents may move against each other. Cell movement is caused by the appearance of a small number of the electrically charged constituents of the cell membrane on the leading edge of the cell. This produces a local decrease in the surface tension and the cell membrane expansion. At the moment of contact between two cells proton transfers occur between the strongly negatively charged microvilli of one cell and the body of the other, analogous to a condenser breakdown. This, through the effect on the surface tension, causes contact inhibition of movement. The distribution of the proton dissociable groups modifies the interaction between the cells (differentiation) and between the cell and the substratum (adhesion). Adsorption of the charged compounds at the surface of the cell membrane, decreasing the surface potential and increasing the surface tension, causes the phenomena of chemotaxis, phagocytosis and pinocytosis. Cell division, considered in the terms of the surface energy, requires an adequate supply of considerable quantities of energy inversely proportional to the surface potential value. In case of a reduction of the distance between the cells, their surface potential and the energetic barrier of the cell division processes increases, and causes contact inhibition of cell division. Due to their high charge, division of neoplastic cells is inhibited much later than division of normal cells, or is completely ininhibited due to geometric conditions. Fusion of the cell membrane in the intra-cellular and intercellular processes is a reverse process in relation to the cell division.     

The Effect of Age and Gender on Al, B, Ba, Ca, Cu, Fe, K, Li, Mg, Mn, Na, P, S, Sr, V, and Zn Contents in Rib Bone of Healthy Humans

The effect of age and gender on major, minor, and trace element contents in the intact rib bone of 80 relatively healthy 15–55-year-old women and men was investigated. Contents or upper limit of contents of 16 chemical elements in the rib bone were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES). Mean values (M?±?SΕΜ) for the mass fraction of Ba, Ca, Cu, Fe, K, Li, Mg, Na, P, S, Sr, and Zn (milligram per kilogram of dry bone) were as follows: 2.54?±?0.16, 171,400?±?4,050, 1.35?±?0.22, 140?±?11, 1,874?±?71, 0.049?±?0.011, 2,139?±?38, 5,378?±?88, 75,140?±?1,660, 1,881?±?51, 291?±?20, and 92.8?±?1.5, respectively. The upper limits of contents of Al, B, Mn, and V were <7.20, <0.65, <0.36, and <0.03, respectively. Statistically significant tendency for the Ca, Mg, and P content to decrease with age was found in the human rib bone, regardless of gender. The mass fraction of Fe in the male rib bone increases with age. It was shown that higher Ca, Mg, Na, P, and Sr mass fractions as well as lower Fe content were typical of female ribs as compared to those in male ribs.     

Distribution of N,P, K,Ca, Mg,S, Zn,Fe, Mn and Cu in groundnut

Summary Concentration of N, P, K, Ca, Mg and S in summer groundnut crop was higher than in kharif while Zn, Fe, Mn and Cu contents were higher in summer crop. Kernel's N, P and Zn; Leaflet's Ca and Mn; Stem's K and Fe; Root's S and Cu and Petiole's Mg contents were highest. Shell's N, P, K, Mg, S, Zn and Cu; Kernel's Ca, Fe and Mn contents were the least. N, P, K, S, Zn and Cu concentrations decreased linearly as the crop grew. Ca, Mg, Fe and Mn concentrations did not display any distinct pattern. Ca concentration was positively correlated with pod yield in both the seasons.     

The effect of malathion,diazinon, and various concentrations of zinc,copper, nickel,lead, iron,and mercury on fish

Acute and chronic toxicity tests for malathion, diazinon, copper (Cu), mercury (Hg), lead (Pb), zinc (Zn), nickel (Ni), and iron (Fe) were conducted. Mortalities ofBarilius vagra andCyprinus carpio (common carp) were variable but LC50-96 hr were similar for pesticides. AdultB. vagra seem to be more sensitive to malathion than juvenile carp. Both juvenile carp and adultB. vagra were extremely sensitive to diazinon. Long-term exposure to pesticides modified morphology and behavior. The LC50-96 values for Cu, Hg, and Pb were 0.3, 0.16, and 0.44, respectively, for smaller fish and 1.0, 0.77, and 1.33, respectively, for larger fish. Replicate LC50 values for Zn, Ni, and Fe were somewhat variable, and for these metals, the size of the fish seemed to affect response because LC50 values increased as fish size increased. Cooper, Pb, Zn, and Fe residues following exposure to sublethal concentrations of these metals for 15 d were significantly greater in whole juvenile common carp than in controls.     

Synthesis,tautomerism, and antimicrobial,anti-HCV,anti-SSPE,antioxidant, and antitumor activities of arylazobenzosuberones

2-Dimethylaminomethylene-1-benzosuberone 1 was coupled with diazotized aniline derivatives to afford a series of the hitherto unreported 2-arylazo-1-benzosuberones 3a–i. The tautomeric structure and the effect of substituents on the tautomeric form (s) of the products 3a–i were discussed. Similar coupling of the enaminone 1 with diazonium salts of heterocyclic amines gave the respective fused azolotriazino-benzosuberones. Some of the newly synthesized compounds showed potent antimicrobial, anti-HCV, antioxidant, antitumor (as topoisomerase I inhibitors), and antimicrobial activities.