Protein metabolism in Spodoptera litura (F.) is influenced by the botanical insecticide azadirachtin

Azadirachtin, as a botanical insecticide, affects a wide variety of biological processes, including reduction of feeding, suspension of molting, death of larvae and pupae, and sterility of emerged adults in a dose-dependent manner. However, the mode of action of this toxin remains obscure. By using proteomic techniques, we analyzed changes in protein metabolism of Spodoptera litura (F.) induced by azadirachtin. Following feeding 4th instar larvae of Spodoptera litura (F.) with an artificial diet containing 1 ppm azadirachtin until pupation, 48 h old pupae were collected and protein samples prepared. Total soluble protein content was measured and the results showed that azadirachtin significantly influenced protein level. Moreover, the proteins were separated by 2-DE (two-dimensional polyacrylamide gel electrophoresis) and 10 proteins were significantly affected by azadirachtin treatment when compared to an untreated control. Six of these proteins were identified with peptide mass fingerprinting using MALDI-TOF-MS after in-gel trypsin digestion. These proteins are involved in various cellular functions. One identified protein may function as an ecdysone receptor, which regulates insect development, and reproduction. It is suggested that the botanical insecticide azadirachtin affects protein expression and the azadirachtin-related proteins would be essential for a better understanding of the mechanisms by which neem toxins exert their effects on insects.     

Effect of fraxinellone on growth and digestive physiology of Asian corn borer, Ostrinia furnacalis Guenee

A limonoid, fraxinellone significantly reduced the relative growth rate, food consumption rate as well as the efficiency of conversion of ingested food of the Asian corn borer, Ostrinia furnacalis when incorporated into artificial diets at concentration of 10 ppm and above. After being fed with diets that contained fraxinellone for 24 h, the larval midguts of O. furnacalis possess significantly lower activities of α-amylase and non-specific proteases and higher activities of cytochrome P450s. In vitro, the compound did not inhibit the activities of α-amylase and non-specific proteases extracted from the larval midguts. The modes of action of fraxinellone against the insect were discussed.     

Sublethal effects of larval treatment with deltamethrin on moth sex pheromone communication system of the Asian corn borer, Ostrinia furnacalis

The first, third, and fifth instar larvae of Asian corn borer, Ostrinia furnacalis (Guenée) (Lepidoptera: Pyralidae), were treated with a lethal dose 30% of deltamethrin to determine its sublethal effects on chemical communication of the adult survivors. The capability of all males surviving deltamethrin to contact the pheromone source in a wind tunnel was significantly reduced, but those treated as the first and third instars also exhibited lower percentages of `orientation,' `up-wind flight,' and `approaching' behaviors than the control males. Female moths from the deltamethrin treatment exhibited similar calling periodicity and percentages to the control moths during 10-h scotophase. However, approximately 20% of the females that survived the pyrethroid in the first and third instars contained significantly higher amounts of sex pheromone and wider coefficients of variation of the ratio of E12-tetradecenyl acetate to its Z isomer in those female survivors. There was no significant difference in the pheromone titers and coefficients of variation in control females and the females treated in the fifth instars. The present study indicates that the Asian corn borer may possess a compensation system by which the males that survived treatment at larval stage present a lower response to the pheromone, while the female survivors produce and release more pheromone.     

丁布胁迫对亚洲玉米螟氧化还原系统的影响

为探明丁布胁迫对亚洲玉米螟氧化还原系统的影响,采用酶活力测定方法和实时荧光定量PCR技术,研究了取食丁布含量差异显著的3个玉米品种登海662、浚单20和隆玉602叶片及模拟其叶片丁布含量的人工饲料后,玉米螟3龄幼虫氧化还原系统主要酶活性和谷氧还蛋白(Grx)mRNA转录表达的变化。结果显示:取食3个玉米品种叶片24 h后,亚洲玉米螟幼虫过氧化物酶和超氧化物歧化酶活性均显著高于对照,且在3个品种处理间差异显著,由大到小依次为登海662(118.30 U/mg和182.19 U/mg)、浚单20(107.13 U/mg和90.02 U/mg)和隆玉602(77.54 U/mg和93.59 U/mg)。取食浚单20可显著促进幼虫脂肪体Grx的表达,取食登海662可显著促进其中肠Grx的表达;取食模拟登海662叶片丁布含量的人工饲料12 h后,幼虫脂肪体中Grx表达量高于对应玉米叶片饲喂处理,而中肠部位则低于对应玉米叶片饲喂处理;取食模拟浚单20的结果与模拟登海662相反。表明亚洲玉米螟氧化还原系统在防御丁布胁迫过程中发挥着重要作用。     

Systemic effects of a neem insecticide onLiriomyza huidobrensis larvae

In an effort to expand the spectrum of larvicides effective against the pea leafminer,Liriomyza huidobrensis (Blanchard), we studied the effects of a neem-based insecticide (Neemix-45) on the development of the leafminer under laboratory conditions. Bean plants were treated with a soil drench of 1, 5, 10 or 25 ppm azadirachtin or by dipping leaves in 1 or 15 ppm azadirachtin at various times before or during the development of the leafminer. Treating the plants with the neem insecticide before exposure to egg-laying adults had a greater effect on inhibiting the development of pupae and adult eclosion than treatment at the 1st-instar larval stage. The systemic effects from a soil drench had a greater adverse effect on pupation and adult eclosion than leaf dipping. Drenching plants with 1 ppm azadirachtin 24 h before exposure to adults had a greater effect (0% adult eclosion) than leaf dipping at the same time period and concentration (15. 6% adult eclosion). Similar results were obtained when drenching plants infested with lst-instar larvae with 1 ppm azadirachtin (11. 7% eclosion)vs dipping leaves at the same time period and concentration (44. 7% eclosion).     

硒对亚洲玉米螟生长发育和生殖行为的影响

为探讨硒(selenium)对亚洲玉米螟Ostrinia furnacalis(Guenée)的营养作用及其生长发育、生殖行为的影响,选用添加不同浓度硒的人工饲料饲喂3龄幼虫,观察了蛹、成虫的生长发育及成虫生殖行为情况。结果表明,不同浓度的硒对蛹和成虫的发育影响差异较大,其中硒浓度为0.75 mg/kg时生长发育最好,化蛹率、雄蛹体重和雄蛾寿命显著高于对照,分别提高了7.4%、12.7%和19.1%;硒浓度为5.00 mg/kg时生长发育最差,化蛹率和羽化率最低,雌、雄蛹重最小,成虫寿命最短。当硒浓度为0.75 mg/kg时,雌蛾求偶百分率和求偶持续时间均有所降低,且求偶高峰期推迟1~2 h。以正常雌蛾为诱源时,硒雄蛾的定向能力显著低于对照,而以硒雌蛾为诱源时,硒雄蛾的定向能力显著高于对照雄蛾;硒雌蛾的产卵量和卵孵化率显著提高,硒雌蛾×硒雄蛾组合提高最多,分别为28.9粒和9.3%。研究表明,适量硒可以促进亚洲玉米螟生长发育,并能显著影响成虫的生殖行为。     

灭幼脲类对玉米螟幼虫的生物活性及防治效果

本文报道灭幼脲类——TH_(6040)对玉米螟幼虫具有很高的生物活性及显著的防治效果。室内测定证明:玉米螟低龄幼虫在含有3ppm浓度TH_(6040)的人工饲料中取食后,存活率降低88～100％。田间试验证明:在春玉米心叶末期施用一次0.045％TH_(6040)颗粒剂,第一代幼虫存活数量降低80％。由此看出,具有改变昆虫生理功能作用的TH_(6040)可应用于玉米螟的综合防治。     

The toxicity and physiological effect of neem limonoids on Cnaphalocrocis medinalis (Guenée) the rice leaffolder

The effect of neem (Azadirachta indica) limonoids azadirachtin, salannin, deacetylgedunin, gedunin, 17-hydroxyazadiradione, and deaceytlnimbin on gut enzyme activity of the rice leaffolder larvae was investigated. When fed a diet of rice leaves treated with limonoids in bioassays, gut tissue enzymes—acid phosphatases (ACP), alkaline phosphatases (ALP), and adenosine triphosphatases (ATPase) activities of rice leaffolder (Cnaphalocrocis medinalis) larvae are affected. Azadirachtin was most potent in all experiments. Larvae that were chronically exposed to limonoids showed a reduction in weight (59-89%) and exhibited a significant reduction in ACP, ALP, and ATPase activities. These results indicate neem limonoids affects gut enzyme activities. These effects are most pronounced in early instars.     

The effects of azadirachtin and nucleopolyhedrovirus on midgut enzymatic profile of Spodoptera litura Fab. (Lepidoptera: Noctuidae)

The effects of azadirachtin (AZA) and nucleopolyhedrovirus (NPV) on midgut enzyme activity in Spodoptera litura Fabricius (Lepidoptera: Noctuidae) (Tobacco cutworm) were evaluated. Gut enzyme activities were decreased by AZA and NPV individually and in combination. When S. litura larvae were fed a diet of castor leaves treated with AZA and NPV in bioassays, gut enzyme—acid phosphatases, alkaline phosphatases, adenosine triphosphatases, and lactate dehydrogenase—activities were decreased. There were statistically significant differences (P ? 0.05) in enzyme activities between combined and individual treatment. A synergistic effect of botanical insecticides and virus was found when combined in low doses. These effects are most pronounced in early instars. Maximum weight loss (59-72%) occurred, when AZA and NPV were combined.     

亚洲玉米螟的寄主植物种类

为探明亚洲玉米螟Ostrinia furnacalis(Guenée)的寄主植物种类,对13科38种植物进行了田间笼罩接虫试验和室内生物测定,并通过组建实验种群生命表分析了不同寄主植物对亚洲玉米螟生长发育和繁殖的影响。田间试验结果表明,亚洲玉米螟寄主植物有8科20种,酸模叶蓼是最适宜寄主植物,其次为玉米、谷子、薏苡等;在酸模叶蓼、玉米等11种植物上还发现了第2代幼虫。室内试验结果表明,在室内饲养条件下,亚洲玉米螟能在9科25种寄主植物上完成生活史,其中以向日葵为寄主植物的亚洲玉米螟种群增长最快,其次为玉米。研究表明,亚洲玉米螟的寄主植物共有9科27种,较适宜寄主植物有酸模叶蓼、玉米、谷子、苍耳和葎草,其中在玉米上的生长发育和繁殖最佳。     

Induction of prolonged larval feeding stage by juvenile hormone analogues inTribolium Castaneum

Dietary ZR-512 and ZR-619 at concentrations of 10 — 1000 ppm induced prolongation of the larval feeding period up to tenfold, increasing larval weight up to double that of untreated larvae. A comparison study of four juvenoids, using 200 ppm of ZR-512, ZR-515, ZR-619 or ZR-777, showed that ZR-515 elicits the highest larval weight (6.2 mg) and ZR-777 the lowest (3.6 mg). In all cases a pronounced enhancement of larval weight — of 50 — 250% relative to untreated larvae (2.4 mg) — was obtained.Tr. castaneum larvae reared up to their 3rd instars on a diet containing 100 ppm of ZR-512, ZR-515, ZR-619 or ZR-777 and then transferred to a juvenile hormone-free diet, were not affected. The period between 4th instar larva and pupation should therefore be considered as critical for juvenile hormone effect. The induced prolongation of the larval stage after juvenile hormone treatment was followed by a pronounced enhancement of cuticle phenoloxidase activity, indicating an alteration of the larval biochemical processes. Although juvenile hormone treatment inhibitsTr. castaneum pupation and emergence, it markedly prolongs larval feeding stage and weight and thus accelerates damage.     

Effect of diflubenzuron on growth and carbohydrate hydrolases ofTribolium castaneum

The potency of diflubenzuron is much greater in inhibiting growth and development of 1st instar larvae ofTribolium castaneum than of 4th instar larvae, as expressed by death at the apolytic stage and retardation of larval development. A dose-dependent decrease in the activity of trehalase, invertase and amylase was obtainedin vivo with the increase in diflubenzuron concentration. At 5 ppm dietary concentration, a reduction of 37 and 27% in invertase and trehalase activity, respectively, was obtained in 4th instar larvae fed for 3 days on treated diet. The amylase activity was affected to a lesser extent. The observed disturbances of trehalase activity might hamper the supply of glucose needed for chitin build-up and those of invertase and amylase activity might affect feeding. Diflubenzuron does not inhibit these enzymesin vitro; hence, thein vivo effect seems to result from general disturbances in carbohydrate metabolism.     

Host plant induction of microsomal monooxygenase activity in relation to diazinon metabolism and toxicity in larvae of the tobacco budworm Heliothis virescens (F.)

The effect of the wild tomato, Lycopersican hirsutam F. glabratam (accession PI 134417), on susceptibility to and metabolism of diazinon, 0,0-diethyl-0-(2-isopropyl-4-methyl-6-pyrimidinyl) phosphorothioate, in larvae of the tobacco budworm, Heliothis virescans F., was studied. The larvae were over 4-fold more tolerant to diazinon toxicity when fed on leaves of wild tomato than when fed on an artificial diet. Diazinon injected into fifth instar larvae is converted into two chloroform-soluble and five water-soluble metabolites. Degradation of diazinon was faster in larvae fed tomato leaves (88.1%) than in larvae fed on the artificial diet (68.4%). Some oxon (13.0%) was detected in the latter case but none in larvae fed tomato leaves. The major metabolite was hydroxypyrimidine and its formation was higher (73.2%) in larvae fed tomato leaves than in larvae fed the artificial diet (49.2%). In vitro studies revealed that both diazinon and its oxon were metabolized primarily by the microsomal cytochrome P-450-dependent monooxygenase system which was induced 2.5- to 3.7-fold by feeding on tomato leaves. It was concluded that diazinon was degraded in H. virescens larvae through desulfuration, hydroxylation of the ring-alkyl side chain, and oxidative dearylation reactions, all of which were increased by varying amounts after feeding on tomato leaves. Treatment of the larvae with 2-tridecanone, a naturally occurring toxin in tomato leaves, resulted in increased tolerance to diazinon and increased in vitro degradation of diazinon and its oxon, the induction being dependent on the magnitude of 2-tridecanone treatment. The microsomes of tomato fed larvae had a 1.5- to 2.1-fold higher concentration of cytochrome P-450, accompanied by a 1–2 nm shift in the λmax of the cytochrome P-450 carbon monoxide complex.     

苹果蠹蛾不同防治方法的控害效应比较

为了及时地在苹果蠹蛾Cydia pomonella(L.)入侵地实施有效的防控策略,通过评价活雌蛾诱捕器诱捕雄蛾数量、在树干上化蛹或越冬的幼虫数量以及对苹果果实的为害程度,比较性信息素诱杀技术、迷向技术和化学防治技术对苹果蠹蛾的控害效应。结果显示,采用迷向法控制苹果蠹蛾的防治效果与化学防治相当,其中在第1代幼虫为害盛末期,蛀果率可控制在1%以下,在第2代幼虫为害盛末期,可控制在2%左右;而诱捕区与对照区(无处理)的蛀果率相当,几乎无控制效应。     

The effect of some benzoyl phenylureas on the larvae of earias insulana

Diflubenzuron, PH 60-38, PH 6043, penfluron (PH 60-44), PH 6045, triflumuron, chlorfuazuron (IKI-7899), teflubenzuron (CME 134), XRD473 and Dowco 439 were tested for their efficacy against the larvae of the spiny bollworm,Earias insulana (Boisd.), in laboratory experiments. The compounds were incorporated at different concentrations in an artificial diet and 5-day-old larvae were introduced and grown on the treated diets until pupation and adult emergence. Teflubenzuron was active at 0.1 ppm, chlorfuazuron at 0.75 ppm and PH 60-38 at 10 ppm; triflumuron and diflubenzuron were active only at 50 ppm; all the rest of the compounds were even less active. When cotton bolls were dipped in teflubenzuron and offered to 6-day-old larvae in the laboratory, only 4% and 10% of the larvae penetrated inside the bolls treated with 50 and 25 ppm a.i., respectively, whereas 68% penetrated inside untreated bolls.     

转cry1Ab/cry2Aj玉米对亚洲玉米螟的抗性评价

为探讨转cry1Ab/cry2Aj玉米对亚洲玉米螟Ostrinia furnacalis(Guenée)的抗性,采用室内生测和田间人工接虫鉴定方法评价了6个转cry1Ab/cry2Aj玉米品系对亚洲玉米螟的杀虫效果,并利用酶联免疫吸附法(ELISA)测定了Cry1Ab蛋白在各品系主要组织的表达水平.结果显示,亚洲玉米螟幼虫取食转基因玉米各品系雄穗、苞叶、花丝和雌穗48 h后死亡率在90％左右,取食心叶的死亡率在70％左右,96h后大部分幼虫死亡,而取食非转基因玉米的死亡率为3.3％～8.7％;春播和夏播田转基因玉米叶片、茎秆、雌穗基本没有被害,而对照玉米叶片、茎秆、雌穗被害严重,春播和夏播玉米平均每株分别有2.9、2.3头幼虫和蛹,2.8、3.4个蛀孔,隧道长度为7.5、14.2 cm,雌穗被害级别为5.3和5.1;6个转基因玉米品系中Cry1Ab蛋白在心叶、雄穗、花丝、苞叶和雌穗中稳定表达.综合评价认为,转基因玉米各品系在整个生育期内对亚洲玉米螟有很好的抗性,其中N50品系抗虫效果最佳,可以作为抗虫转多基因玉米育种的备选材料.     

Insect juvenile hormones: Chemistry and action: Edited by Julius J. Menn and Morton Beroza. Academic Press,New York, 1972, $11.00

Heptachlor metabolism was determined in young male rats fed for 10 days diets containing 18% protein supplied either by gluten or casein. Rats on the casein diet were either pair-fed on an individual basis to the gluten-fed animals or they were fed ad lib. Saline, sodium phenobarbital (80 mg/kg) or diethylaminoethyl 2,2-diphenylvalerate HCl (SKF 525-A, 50 mg/kg) was administered ip to the animals before sacrifice. Weight gain, liver microsomal protein, and heptachlor metabolism (nmole of product/g liver/10 min), assayed in 9000g rat liver supernatants, were found to be significantly reduced in the gluten-fed animals. Animals pair-fed the casein diet had higher heptachlor epoxidase activity than those fed ad lib. Phenobarbital pretreatment significantly enhanced the metabolism of heptachlor in animals in all groups to varying magnitudes. Heptachlor epoxidase activity (units/g liver) was increased 11-fold in the gluten-fed animals, 6-fold in the pair-fed animals on casein diet, and 7-fold in the animals fed casein ad lib. Heptachlor metabolism was less inhibited by SKF 525-A in the rats fed gluten compared to the rats on casein diet, either pair-fed or ad lib. The degree of induction or inhibition of enzyme activity was not influenced by restriction of food intake (pair-fed vs ad lib). The results of these studies suggest an interaction of protein inadequacy with drug metabolism and its induction or inhibition.     

Host plant stimulation of detoxifying enzymes in a phytophagous insect

The midgut microsomal aldrin epoxidase of variegated cutworm larvae (Peridroma saucia, Hübner) fed bean or peppermint leaves was up to 10 and 45 times more active, respectively, than that of larvae fed a basic control diet. Large increases in oxidase activity and cytochrome P-450 levels also occurred in larvae fed mint plant constituents such as menthol menthone, α-pinene, and β-pinene. Mint-fed larvae were more tolerant of the insecticide, carbaryl, than bean-fed larvae.     

北方春玉米区亚洲玉米螟对Bt玉米的抗性风险分析

为了解我国北方春玉米区种植Bt玉米条件下主要靶标害虫亚洲玉米螟Ostrinia furnacalis的Bt抗性风险,探索合理的Bt玉米种植比例,根据亚洲玉米螟的特点构建种群动态与遗传模型,对不同的Bt玉米种植比例进行分析评估。结果显示,当以种子混合种植75%的Bt玉米+25%的非Bt玉米庇护所时,亚洲玉米螟的种群控制时间,即亚洲玉米螟种群密度下降到每百株玉米植株30头幼虫以下的时间,为1～2年,而害虫的抗性时间,即亚洲玉米螟种群抗性基因频率达到50%的时间,在1代区为40年、在2代区为20年。当以种子混合种植20%、15%、10%、5%的非Bt玉米庇护所时,亚洲玉米螟的种群控制时间仍为1～2年,而害虫的抗性时间缩短,在1代区分别是29、21、15、9年,在2代区分别是14、10、7、4年。表明在研究区域内以种子混合种植20%以上的非Bt玉米庇护所可维持对亚洲玉米螟的Bt抗性管理至少14年。     

Cry1Ac和Cry2Ab蛋白对大草蛉生长发育及酶活力的影响

为明确转Cry1Ac和Cry2Ab基因棉花对大草蛉的影响,运用Bt蛋白与人工饲料混合的方法,以大于转基因棉花叶片中蛋白含量20倍的剂量饲喂大草蛉初孵幼虫,初步研究了Cry1Ac和Cry2Ab对大草蛉生物学参数和消化酶、解毒酶活性的影响。结果表明：取食含Bt蛋白饲料的大草蛉幼虫的发育历期、体重、蛹重、成虫体重、羽化率等生物学参数与对照相比均没有显著差异;在大草蛉幼虫体内可以检测到含量较高的Cry1Ac和Cry2Ab蛋白,分别为974.92~1 282.39 ng/g鲜重和5 592.62~6 082.92 ng/g鲜重,而在大草蛉成虫体内检测到的Cry1Ac和Cry2Ab蛋白含量非常低,分别为0.29~0.39 ng/g鲜重和50.34~56.71 ng/g鲜重;取食含Bt蛋白的饲料对大草蛉幼虫的类胰蛋白酶、类胰凝乳蛋白酶、氨肽酶和谷胱甘肽-S-转移酶活性有一定的影响,但对大草蛉成虫影响与对照差异不显著。表明大草蛉取食含Bt蛋白的人工饲料后,虽然体内可以检测到一定含量的Bt蛋白,但对大草蛉的生长发育并没有显著的直接不利影响。