马桑内酯对粘虫幼虫体内3种酶活性的影响

采用3种马桑内酯——羟基马桑毒素、马桑亭和马桑宁饲喂和注射粘虫幼虫,测定和分析了虫体组织中乙酰胆碱酯酶(AChE)、谷胱甘肽S-转移酶(GST)和多酚氧化酶(PPO)活性变化.结果表明,3种马桑内酯对试虫体内乙酰胆碱酯酶活性的影响存在差异,在4~48 h大多时间段具有激活特性;而谷胱甘肽S-转移酶的活性或被激活或被抑制,饲喂用药后4~8 h中肠组织中酶被激活,12 h酶活性被抑制,整个过程呈现S型;多酚氧化酶活性受3种毒素的影响不明显.     

PxCAD1与Cry1Ac蛋白混合物对小菜蛾生长发育及体内酶活性的影响

[目的]PxCAD1肽段可显著增强Cry1Ac的杀虫活性,探究其与Cry1Ac混合物对靶标昆虫生物学及体内酶活性的影响对明确其增效机制具有重要意义.[方法]采用叶片浸渍法,以PxCAD1肽段与Cry1Ac毒素的混合蛋白饲喂小菜蛾Plutella xylostella3龄幼虫,观察并统计小菜蛾死亡率、化蛹率和羽化率,并测定小菜蛾幼虫体体内羧酸酯酶、谷胱甘肽-S-转移酶、总蛋白酶和类胰蛋白酶的变化趋势.[结果]与对照相比,处理组幼虫的死亡率显著升高,化蛹率减少,羽化率显著降低;3龄幼虫在取食PxCAD1肽段与Cry1Ac毒素混合蛋白24 h和48 h后,羧酸酯酶活性显著升高,72 h显著降低,谷胱甘肽-S-转移酶活性在24 h显著升高,48 h显著降低,72 h则无显著差异,而总蛋白酶活性仅在72 h显著低于对照,类胰蛋白酶活性在各时间点无显著变化.[结论]PxCAD1不仅对Cry1Ac具有显著增效作用,同时对小菜蛾幼虫的生长发育及体内羧酸酯酶、谷胱甘肽-S-转移酶及总蛋白酶的活性产生不利影响.该结果将有助于进一步研究中肠受体蛋白对Cry1Ac的增效机制.     

苍耳甾醇物质对菜青虫取食、血淋巴和中肠酶活性及中肠组织的影响

甾醇是植物体内的重要次生物质,具有多种生物活性。为探明植物甾醇类物质对害虫的作用机理,采用叶碟饲喂法进行取食处理后研究了苍耳Xanthium sibiricum中分离纯化的甾醇类组分（甾醇A和甾醇B）对4龄菜青虫Pieris rapae的取食、酶活性以及中肠组织的影响。结果表明： 苍耳甾醇类组分甾醇A和甾醇B能明显抑制菜青虫的取食,拒食中浓度AFC50分别为0.0229 和0.0147 mg/mL;同时,显著降低菜青虫中肠蛋白酶、淀粉酶和羧酸酯酶活性,其中,甾醇B的作用效果较强,处理后24 h和36 h, 对蛋白酶活性抑制率分别为23.74%和58.59%,对中肠羧酸酯酶的活性抑制率分别为49.01%和83.03%;降低血淋巴蛋白质含量,诱导菜青虫血淋巴羧酸酯酶活性的提高;破坏昆虫中肠上皮组织,微杆模糊不清呈消融状,杯状细胞的杯腔基部微绒毛消失。这些结果说明苍耳甾醇类物质对菜青虫的取食抑制可能与对中肠消化酶活性的抑制以及对中肠上皮组织的破坏有关,植物甾醇组分的不同配比影响其对昆虫的作用效果。     

CO_2浓度升高对粘虫幼虫体内酶活性的影响

【目的】阐明大气CO_2浓度升高对粘虫Mythimna separata(Walker)幼虫不同龄期体内酶活性的影响机制。【方法】测定分析了人工气候箱内不同CO_2浓度(800mL/L和400mL/L)条件下,粘虫幼虫体内4种解毒酶[羧酸酯酶(Car E)、乙酰胆碱酯酶(Ach E)、微粒体多功能氧化酶(MFO)和谷胱甘肽-S-转移酶(GST)]、3种保护酶[超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)]和4种蛋白酶(总蛋白酶、强碱性类胰蛋白酶、弱碱性类胰蛋白酶和类胰凝乳蛋白酶)活性的变化。【结果】(1)粘虫不同龄期幼虫体内4种解毒酶的活力均随着CO_2浓度升高而显著上升(P<0.05);(2)粘虫幼虫体内的保护酶中SOD酶活性在CO_2浓度为800mL/L条件下低于对照(CO_2浓度为400mL/L)条件下的酶活性,其中4至6龄幼虫的下降趋势较为显著(P<0.05),而POD、CAT酶活性均呈上升趋势,800mL/L浓度下的CAT活力均显著高于对照(P<0.05);(3)高CO_2浓度条件下粘虫总蛋白酶的酶活性随着龄期的增加呈现先上升后下降的趋势,强碱/弱碱性类胰蛋白和类胰凝乳蛋白3种酶的比活力均低于400mL/L CO_2浓度下的比活力,其中类胰凝乳蛋白酶比活力显著低于对照条件下的比活力(P<0.05)。【结论】在高CO_2浓度条件下,粘虫幼虫体内的大多数酶系活力呈现下降趋势,大部分解毒酶和蛋白酶活性受到抑制,SOD、CAT和POD 3种保护酶的动态平衡受到干扰而影响其正常的生理代谢,机体受损伤并随龄期的增加而加重。     

雷公藤总生物碱对粘虫幼虫神经系统酶活性及神经递质含量的影响

【目的】明确雷公藤Tripterygium wilfordii Hook. f.生物碱对粘虫Mythimna separata (Walker)神经系统的影响,为阐明其杀虫作用机制提供依据。【方法】采用载毒叶片法测定粘虫5龄幼虫经雷公藤总生物碱处理后体内乙酰胆碱酯酶（AChE）、Na⁺, K⁺-ATPase、Ca²⁺, Mg²⁺-ATPase、谷丙转氨酶（GPT）和谷氨酸脱羧酶（GAD）等重要神经系统酶活性及乙酰胆碱（ACh）、谷氨酸（Glu）和γ-氨基丁酸（GABA）等神经递质的含量。【结果】雷公藤总生物碱处理对粘虫5龄幼虫AChE无明显影响,麻醉期处理粘虫幼虫体内ACh相对含量与同期对照无显著差异。处理粘虫幼虫在轻度麻醉期、深度麻醉期和复苏期体内GABA和Glu含量显著升高,GABA含量分别升高了89.86%, 49.28%和20.29%,Glu含量分别升高了24.55%, 23.33%和8.13%。处理粘虫幼虫GPT活性明显受到抑制,而GAD活性无明显变化。处理明显抑制粘虫幼虫头部Na⁺, K⁺-ATPase和Ca²⁺, Mg²⁺-ATPase活性,但对中肠两种ATPase活性影响不大。【结论】研究结果有助于了解雷公藤生物碱对昆虫神经系统的影响,也为进一步阐明其作用靶标奠定了基础。     

百合鳞片细胞形态发生中生理生化特性研究

四倍体百合鳞片外植体脱分化形成愈伤组织过程其蛋白质、核酸含量与蛋白酶、淀粉酶、过氧化物酶ＰＯＤ、酸性磷酸酯酶活性先升高后降低,淀粉与ＡＴＰ含量呈降－升－降变化,ＳＯＤ活性先降低后升高,愈伤组织分化芽过程,除ＡＴＰ含量降低外,其余指标均不断升高．鳞片外植体直接分化发育不定芽过程中,仅见蛋白质、淀粉、ＡＴＰ含量与ＳＯＤ、酸性磷酸酯酶活性在５ｄ前降低,其它时间各项指标均升高,直到不定芽发育完成再下降．器官发生型比器官型经历了更为深刻的内部生理生化变化．     

阿维菌素对甘蔗绵蚜解毒酶系活性的影响

采用叶片浸渍法测定了阿维菌素对甘蔗绵蚜的毒力,并用分光光度计法检测了不同浓度药剂处理后活虫体内羧酸酯酶和乙酰胆碱酯酶活性的变化。结果表明,阿维菌素对甘蔗绵蚜的LC50为2.400×10-2mg·mL-1。甘蔗绵蚜体内β-NA羧酸酯酶活性高于α-NA羧酸酯酶,随着阿维菌素处理质量浓度的升高,α-NA羧酸酯酶活性逐渐升高,当质量浓度升高到4.500×10-2mg·mL-1时,活性达到最大,随后开始缓慢下降,而β-NA羧酸酯酶的活性总体呈上升趋势,说明甘蔗绵蚜体内β-NA羧酸酯酶的活性与阿维菌素的质量浓度可能存在一定的相关,β-NA羧酸酯酶活性的升高可能会使甘蔗绵蚜对阿维菌素产生抗性。甘蔗绵蚜体内乙酰胆碱酯酶的活性在阿维菌素处理质量浓度为2.250×10-2mg·mL-1时最高,而在处理质量浓度为4.500×10-2mg·mL-1时则降至最低,后又随阿维菌素处理质量浓度的加大而缓慢上升,乙酰胆碱酯酶的活性与阿维菌素的质量浓度不存在线性相关。     

He-Ne激光和增强UV-B辐射对小麦幼苗蛋白质代谢的影响

采用He-Ne激光(5 mW.mm-2)和增强UV-B(10.08 kJ.m-2.d-1)辐照‘晋麦8号’小麦幼苗,5 d后测定各处理组小麦叶片的蛋白酶、转氨酶活性以及可溶性蛋白质含量与组成的变化。结果显示,增强UV-B辐射使小麦叶片蛋白酶活性极显著升高,转氨酶活性降低,可溶性蛋白质含量极显著下降,蛋白质谱带增加;单独He-Ne激光处理使蛋白酶活性下降,转氨酶活性升高,可溶性蛋白质含量增加,对蛋白质条带影响不明显;与单独UV-B辐射相比,经He-Ne激光辐照和UV-B辐射复合处理后,蛋白酶的活性明显降低,转氨酶的活性增加,可溶性蛋白质含量增加,并且使UV-B辐射诱导出的新带减弱或消失。研究发现,增强UV-B辐射能减弱小麦幼苗蛋白代谢中正常基因的表达,但又激活了一些抗性基因的表达而诱导产生新的胁迫蛋白;一定剂量的He-Ne激光辐照可解除UV-B对小麦幼苗正常基因表达的抑制而使其蛋白质代谢加强。     

脱氧鬼臼毒素对粘虫几种代谢酶系的影响

脱氧鬼臼毒素是砂地柏Sabina vulgaris Ant.中的主要杀虫活性成分之一。为探讨其作用机理,以叶碟饲喂法处理粘虫Mythimna separata Walker 4龄幼虫,测定了饲喂处理12h、24 h、36 h、48 h和72 h后试虫的羧酸酯酶（CarE）、酸性磷酸酯酶(ACP)、碱性磷酸酯酶(AKP)、谷胱甘肽S-转移酶（GSTs）和多功能氧化酶(MFO)的活性。结果表明：脱氧鬼臼毒素对粘虫羧酸酯酶的活性无明显影响;对酸性磷酸酯酶具有明显的抑制作用,且随着处理时间的延长,抑制作用增强;对碱性磷酸酯酶的影响较为复杂,表现为先抑制,后有所恢复,再被抑制的变化过程;对谷胱甘肽S-转移酶处理12 h后活性变化不明显,24 h后被抑制,36 h后逐渐激活,与对照差异极显著;对细胞色素P450酶系的O-脱甲基酶活性具有明显的抑制作用。     

敌百虫对荔枝蝽几种代谢酶活性的影响

采用微量点滴法测定了荔枝蝽对敌百虫的敏感性,并对不同浓度敌百虫处理后荔枝蝽体内羧酸酯酶、酸性磷酸酯酶和碱性磷酸酯酶的活性进行了比较。结果表明,敌百虫对荔枝蝽的LC50为28.840 mg.mL-1;随着敌百虫处理浓度的升高,荔枝蝽体内羧酸酯酶活性显著升高,不同处理剂量间存在显著差异,酸性磷酸酯酶和碱性磷酸酯酶活性各药剂处理剂量间没有差异。表明羧酸酯酶活性的升高可能会导致荔枝蝽对敌百虫的敏感性下降。     

环氧苍耳素Ⅰ对菜青虫中肠消化酶和羧酸酯酶活性的影响

【目的】探明苍耳Xanthium sibiricum活性物质--环氧苍耳素Ⅰ对菜青虫的作用机理。【方法】采用饲喂法处理4龄菜青虫Pieris rapae, 测试从苍耳中分离提纯的环氧苍耳素Ⅰ（为倍半萜内酯类物质）对菜青虫中肠蛋白酶、 淀粉酶和羧酸酯酶活性的影响。【结果】环氧苍耳素Ⅰ对蛋白酶活性的抑制最强, 处理后12, 24和48 h, 菜青虫中肠蛋白酶抑制率分别为20.95%,29.38%和50.06%; 其次为淀粉酶, 抑制率分别为11.89%,39.01%和31.92%。同时,中肠羧酸酯酶活性在处理后12 h与对照之间无显著变化, 24 h 时活性被显著抑制, 而48 h时活性却明显高于对照。【结论】环氧苍耳素Ⅰ对昆虫中肠消化酶活性的抑制, 可能是引起昆虫表现取食抑制和生长发育不良的重要原因之一。     

蛋白酶抑制剂对绿豆象幼虫中肠蛋白酶活性的影响

为明确蛋白酶抑制剂对绿豆象幼虫中肠蛋白酶活性的影响,采用室内人工接虫和生化测定的方法,研究了在离体条件和饲喂条件下4种蛋白酶抑制剂对绿豆象幼虫中肠蛋白酶的抑制作用,并测定了绿豆象幼虫取食不同含量的绿豆胰蛋白酶抑制剂(MBTI)的人工绿豆后,其中肠内总蛋白酶、类胰蛋白酶和类胰凝乳蛋白酶活性的变化.结果表明:在离体条件下,供试4种蛋白酶抑制剂对绿豆象幼虫总蛋白酶、类胰蛋白酶和类胰凝乳蛋白酶活性均有明显的抑制作用,且浓度越大,抑制效果越显著,其中以20μg·mL^-1的MBTI对3种酶活性的抑制效果最强,3种酶活性分别比对照降低了62.5%、41.2%和38.7%,而卵粘蛋白抑制剂(OI)抑制效果最弱.绿豆象幼虫取食含不同抑制剂的人工绿豆后,中肠内3种酶活性也均受到一定的抑制作用,取食后随龄期的延长,3种酶活性有所升高但仍显著低于对照,且以MBTI的抑制作用最强.当绿豆象幼虫取食不同含量MBTI的人工绿豆后,随MBTI含量的增加,对总蛋白酶活性和类胰蛋白酶活性的抑制作用均逐渐增强,但对类胰凝乳蛋白酶活性的抑制作用并不显著,只有当MBTI含量达20%时,对类胰凝乳蛋白酶活性才表现出明显的抑制作用.     

Varroa treatment with bromfenvinphos markedly suppresses honeybee biochemical defence levels

We examined the influence of bromfenvinphos, a commonly used acaricide, on activities of many metabolic enzymes affecting the biochemical defences/physiology of the western honeybee, Apis mellifera L. (Hymenoptera: Apidae), as well as on some metabolic compound concentrations, percentage of global DNA methylation, and Nosema spp. infection levels. Bromfenvinphos‐treated workers had decreased haemolymph volumes and higher protein concentrations on their cuticle but lower protein concentrations in the haemolymph. They had higher global DNA methylation levels independent of the age‐related variants. Bromfenvinphos decreased the activities of antioxidant enzymes (SOD, GPx, CAT, GST), acidic, neutral, and alkaline protease inhibitors and enzymatic physiological markers (AST, ALT, ALP), and concentrations of urea, uric acid, creatinine, cholesterol, glucose, Mg²⁺, and Ca²⁺ in worker haemolymph, depending on the age of the bees. Protease activities were higher only in the haemolymph of young bromfenvinphos‐treated bees in comparison with untreated bees. This compound decreased the activities of alkaline proteases and neutral protease inhibitors on the cuticle. Unexpectedly, in the treated bees, the activities of acidic and neutral proteases, and acidic and alkaline protease inhibitors, were higher in the young bees and lower in the older workers in comparison to the untreated group. The bromfenvinphos‐treated workers were more heavily infested with Nosema spp. Thus, bromfenvinphos not only supressed many levels of biochemical defences, and therefore stress‐resistance‐related biochemical pathways but also visibly increased the Nosema spp. infection levels.     

Effects of cotton condensed tannin on feeding and digestion in the cotton pest, Heliothis zea

Condensed tannin of cotton incorporated into an artificial diet caused growth inhibition in H. zea larvae whose susceptibility was dependent upon the concentration of the dietary tannin, and also upon the size of the feeding larvae.Larvae feeding on tannin-treated diet exhibited both decreased protease and invertase activities in the midgut caecal wall, as well as lowered total protein and sugar levels in the haemolymph as compared to controls. These differences, however, had no effect on assilimation and efficiency of conversion of digested matter into animal biomass and therefore may be secondary effects. Ultimately, inhibition of growth is attributed to a reduction in food consumption.     

Feeding behaviour, digestive physiology and lipid content in macropterous females of Pyrrhocoris apterus (L.) (Heteroptera: Pyrrhocoridae)

Abstract. Macropterous females of Pyrrhocoris apterus (L.) reared under short-day conditions (LD 12:12 h) were analysed for temporal patterns of feeding and drinking behaviour, activities of digestive enzymes in the gut, and lipid and glycogen content in the haemolymph and fat body. Peaks of drinking activity were recorded at days 3, 7 and 10 during the first 14 days after imaginal ecdysis. Feeding activity peaked on the third day, ceasing completely after the fourth day of adult life. Esterase, protease, amylase and aminopeptidase activities exhibited the highest overall activity in the first days after imaginal emergence; then enzyme activities decreased. In the fat body, the content of lipids was highest on day 5, then a decrease of about 40% was observed at day 14; the amount of glycogen was highest on day 1 at 11 μg of glucose equivalents/mg of fat body, then decreased to 2 μg at day 14 after the imaginal moult. In the haemolymph, the lipid content rose until day 8 when it reached almost 0.3 μmol/μl; at day 14 the value was slightly lower. The association of fasting with reproductive arrest in macropterous females of P. apterus, accompanied by a decrease in digestive enzyme activities and a mobilization of lipid reserves from the fat body, was demonstrated.     

The control of the proventriculus in the honeybee (Apis mellifera carnica L.) II. Feedback mechanisms

The mechanisms underlying the control of solution transport rates through the proventriculus in foraging honeybees were investigated in individuals trained to collect defined amounts of sugar solutions. Following feeding, bees were injected either with metabolisable (glucose, fructose, trehalose), or non-metabolisable (sorbose) sugars, in order to distinguish between haemolymph osmolarity and haemolymph sugar levels as factors controlling the solution transport rates through the proventriculus. After a fixed period, workers were dissected in order to measure crop content and haemolymph sugar titers. Between feeding and dissection, the metabolic rate of every investigated forager was measured using open-flow respirometry. Bees injected with metabolisable sugars 15 min after feeding were observed to reduce their solution transport rates through the proventriculus, but injection of non-metabolisable sugars had no influence on them. This suggests that the solution transport rate through the proventriculus is controlled by the concentration of metabolisable compounds in the haemolymph, and not by the haemolymph osmolarity. A period of 10 min after injection of metabolisable sugars was enough to observe reduced solution transport rates. However, if bees were injected only 5 min after feeding, no reduced solution transport rates were observed 10 min after injection.     

菜青虫感染玫烟色拟青霉后血淋巴蛋白质含量及几种保护酶活力的变化

对玫烟色拟青霉Paecilomyces fumosoroseus侵染3、4龄菜青虫Pieris rapae后,其血淋巴中蛋白质含量、体内保护酶及谷胱甘肽S-转移酶活力进行了研究。结果表明,感病的3、4龄菜青虫血淋巴中蛋白质含量明显低于同期未感染的幼虫;感病虫体内超氧化物歧化酶、过氧化氢酶和过氧化物酶的活力发生了不同程度的改变,其中对3龄菜青虫体内酶活力的影响比4龄幼虫大。此外被侵染的3、4龄菜青虫体内谷胱甘肽S-转移酶活力在感病前期显著高于同期未感染的菜青虫,而在感病后期明显低于同期未感染的菜青虫。     

Brain-midgut short neuropeptide F mechanism that inhibits digestive activity of the American cockroach, Periplaneta americana upon starvation

Immunohistochemical reactivity against short neuropeptide F (sNPF) was observed in the brain-corpus cardiacum and midgut paraneurons of the American cockroach, Periplaneta americana. Four weeks of starvation increased the number of sNPF-ir cells in the midgut epithelium but the refeeding decreased the number in 3h. Dramatic rises in sNPF contents in the midgut epithelium and hemolymph of roaches starved for 4 weeks were confirmed by ELISA. Starvation for 4 weeks reduced α-amylase, protease and lipase activities in the midgut of P. americana but refeeding restored these to high levels. Co-incubation of dissected midgut with sNPF at physiological concentrations inhibited α-amylase, protease and lipase activities. sNPF injection into the hemocoel led to a decrease in α-amylase, protease and lipase activities, whereas PBS injection had no effects. The injection of d-(+)-trehalose and l-proline into the hemocoel of decapitated adult male cockroaches that had been starved for 4 weeks had no effect on these digestive enzymes. However, injection into the hemocoel of head-intact starved cockroaches stimulated digestive activity. Injection of d-(+)-trehalose and l-proline into the lumen of decapitated cockroaches that had been starved for 4 weeks increased enzymes activities and suppressed sNPF in the midgut. Our data indicate that sNPF from the midgut paraneurons suppresses α-amylase, protease and lipase activities during starvation. Injection of d-(+)-trehalose/l-proline into the hemocoel of head-intact starved cockroach decreased the hemolymph sNPF content, which suggests that sNPF could be one of the brain factors, demonstrating brain-midgut interplay in the regulation of digestive activities and possibly nutrition-associated behavioral modifications.     

嗜水气单胞菌感染对背角无齿蚌5种免疫相关酶活力的影响

本研究以背角无齿蚌为材料,利用嗜水气单胞菌为诱导刺激物,对背角无齿蚌进行注射感染,在注射后3、6、12 、24、48h分别取血淋巴、肝胰腺,测定其超氧化物歧化酶（SOD）、酚氧化酶 (PO)、酸性磷酸酶 (ACP)、碱性磷酸酶 (AKP) 及过氧化氢酶 (CAT) 的活力,并研究各项酶活力的变化规律。结果表明：除3h肝胰腺外,实验组的SOD活力均不同程度地高于对照组;血清实验组的PO活力开始显著高于对照组,然后降低,而肝胰腺实验组的PO活力持续高于对照组;另外,与对照组相比,实验组中血淋巴与肝胰腺的ACP、AKP和CAT活力在不同的时间段虽然有所增强,但两者之间无显著差异。因此,认为SOD、PO活性可以作为背角无齿蚌免疫抗病功能指标参数,而ACP、AKP及CAT活性能否作为该参数还有待于进一步研究。     

Interrelationships between haemolymph lipid and carbohydrate during starvation in Locusta

During starvation in adult female Locusta, the haemolymph total lipid concentration increases markedly while that of the total carbohydrate decreases. The majority of the increased haemolymph lipid is diglyceride and 75% of this is associated with a high molecular weight lipoprotein (A⁺) which disappears rapidly after feeding when the total lipid concentration is restored to the normal resting value. The effect of feeding can be mimicked by injecting or feeding starved locusts with sugars but not with protein. The lowering of the haemolymph total diglyceride concentration in starved locusts by injection of carbohydrates is dose-related and, at doses in excess of 4 mg per locust, almost normal values (for fed locusts) are obtained within 6 hr. It is suggested that in the haemolymph there is an inverse relationship between the concentration of diglyceride and that of trehalose.