Hormonal studies of uridine utilization in an insect cell line CP-1268 derived from the codling mothLaspeyresia pomonella

Summary Ecdysterone decreased cellular growth and the incorporation of uridine into RNA following 4 days of hormone exposure. This hormone did not affect uridine incorporation following short-term exposure up to 25 hours. Juvenile hormone and farnesol both significantly decreased uridine uptake and incorporation into RNA; however, uridine uptake was inhibited to a greater extent than uridine incorporation. Cyclic AMP increased the incorporation of uridine into RNA but had no demonstrable effect on the uptake process. This stimulation was not the result of cAMP degradation products. Cyclic AMP and ecdysterone together produced a significant increase in urdine incorporation into RNA. These studies demonstrate the potential utilization of insect cell lines for studying the mode of action of insect developmental hormones.     

Utilization of uridine for RNA synthesis in the insect cell line CP-1268 derived from the codling moth,Laspeyresia pomonella

Summary The utilization of [³H]-5-uridine by CP-1268 cells was studied. Uridine was rapidly transported into these cells by a concentration dependent, saturable process. Exogenous uridine rapidly equilibrated with cellular nucleotide pools and virtually all of the uridine transported into the cells was phosphorylated. Uridine incorporation into RNA was studied by continuous and pulse-labeling techniques in the presence or absence of actinomycin D and cordycepin. These studies have shown that the pattern of unstable RNA precursor and relatively stable RNA product relationship known to exist in mammalian cells similarly exists in insect cells in vitro. This pattern varied markedly with pulse-labeling time and required the addition of RNA inhibitors to block reincorporation of intracellular labeled metabolites during the chase.     

Utilization of uridine for RNA synthesis in the insect cell line CP-1268 derived from the codling moth, Laspeyresia pomonella.

The utilization of [3H]-5-uridine by CP-1268 cells was studied. Uridine was rapidly transported into these cells by a concentration dependent, saturable process. Exogenous uridine rapidly equilibrated with cellular nucleotide pools and virtually all of the uridine transported into the cells was phosphorylated. Uridine incorporation into RNA was studied by continuous and pulse-labeling techniques in the prescence or absence of actinomycin D and cordycepin. These studies have shown that the pattern of unstable RNA precursor and relatively stable RNA product relationship known to exist in mammalian cells similarly exists in insect cells in vitro. This pattern varied markedly with pulse-labeling time and required the addition of RNA inhibitors to block reincorporation of intracellular labeled metabolites during the chase.     

Characterization of the respiratory activity of mitochondria isolated from an insect cell line CP-1268 Laspeyresia pomonella

Mitochondria have been isolated from the codling moth Laspeyresia pomonella, CP-1268 cell line. The mitochondrial fraction was isolated from pooled 4 d, exponential growth phase, cultures. The mitochondria were determined to be intact based on the demonstration of respiratory control, the effects of 2,4 dinitrophenol and oligomycin on respiration, the inability to oxidize NADH, and the inability of cytochrome c to enhance respiration. The isolated mitochondria were able to oxidize succinate, pyruvate, malate, alpha-ketoglutarate, and alpha-glycerophosphate efficiently. Of the substrates tested, the CP-1268 mitochondria oxidized succinate most efficiently. The respiratory control ratios ranged from a high of 4.6 for pyruvate to a low of 1.7 with alpha-glycerophosphate. These findings confirm that the mitochondria were tightly coupled. The data also confirm the presence of three sites of oxidative phosphorylation because NAD-linked substrates had ADP-to-O ratios approaching 3 and flavoprotein linked substrates had values approaching 2.     

Characterization of the respiratory activity of mitochondria isolated from an insect cell line CP-1268Laspeyresia pomonella

Summary Mitochondria have been isolated from the codling mothLaspeyresia pomonella, CP-1268 cell line. The mitochondrial fraction was isolated from pooled 4 d, exponential growth phase, cultures. The mitochondria were determined to be intact based on the demonstration of respiratory control, the effects of 2,4 dinitrophenol and oligomycin on respiration, the inability to oxidize NADH, and the inability of cytochromec to enhance respiration. The isolated mitochondria were able to oxidize succinate, pyruvate, malate, α-ketoglutarate, and α-glycerophosphate efficiently. Of the substrates tested, the CP-1268 mitochondria oxidized succinate most efficiently. The respiratory control ratios ranged from a high of 4.6 for pyruvate to a low of 1.7 with α-glycerophosphate. These findings confirm that the mitochondria were tightly coupled. The data also confirm the presence of three sites of oxidative phosphorylation because NAD-linked substrates had ADP-to-O ratios approaching 3 and flavoprotein linked substrates had values approaching 2.     

Hormonal regulation of pupation in the codling moth, Laspeyresia pomonella

ABSTRACT. According to the different reactions to the juvenoid Altosid®, the last larval instar (L₅) of Laspeyresia pomonella (L.) (Tortricidae) reared under 'long day' conditions (constant light) was subdivided into three sensitive phases: an additional larval instar, a larval–pupal intermediate, or a pupa. Under short day conditions, the prothoracotropic effect of juvenile hormone (JH) in L₅, which have a continuous high titre of JH during the whole instar, indicated that it is not a particular titre of JH but a rise in the titre that can induce the production of moulting hormone. Neck-ligation experiments showed that JH acts not directly on the prothoracic glands but via the head, probably via the neurosecretory system. The meaning of the JH-peak in mature L₅ reared under long days was determined either by injections with the anti-JH, precocene II, in combination with applications of Altosid, or by forcing precocene-treated larvae to a precocious moult by injecting them with ecdysterone. Precocene delayed, and JH accelerated pupation if administered 4.5 days after the L₅ -moult. JH was also found to stimulate the growth and differentiation of the imaginal discs. Moulting hormone in long-days reared insects was detected one day after the larvae had spun their cocoon, with a maximum on the second day after spinning. The hormone was also present in freshly moulted pupae. Neck-ligation of mature larvae indicated that the delay between activation of the prothoracic glands and the production of an effective amount of moulting hormone is less than one day.     

Effects of predation on clutch size and egg dispersion in the codling moth Laspeyresia pomonella

ABSTRACT.

• 1 The effects of predation on clutch size and egg dispersion in the codling moth Laspeyresia pomonella (L.) were experimentally studied in an orchard by recording survival of eggs glued onto different substrates.
• 2 Egg mortality due to sucking insects was readily distinguishable from that due to chewing insects.
• 3 In both the field and laboratory, females preferred to deposit eggs singly on the uppersides of leaves.
• 4 Eggs on the uppersides of leaves and on fruit survived significantly better than did those on the undersides of leaves or on twigs.
• 5 Egg losses were highest during the early part of the flight season.
• 6 The smallest (one egg) and largest (nine eggs) clutches, associated with one egg-bearing leaf per twig, both suffered the lowest rates of predation. Factors selecting against oviposition on fruits and production of large clutch sizes are presented and discussed.

     

Dosage-mortality and time-mortality studies of a granulosis virus in a laboratory strain of the codling moth,Laspeyresia pomonella

Different doses of a granulosis virus were administered to first- and fifth-instar larvae of the codling moth Laspeyresia pomonella. Virus was very pathogenic for both larval instars. The LD₅₀ values for first- and fifth-instar larvae were 5 and 49 capsules/larva, respectively. However, fifth-instar larvae were much more variable in their response to virus than first-instar larvae. Using probit methods it was calculated that 1 capsule could cause death in about 25% of both larval instars but 1578 capsules were required to cause 70% mortality of fifth-instar larvae as compared to 12 capsules for first-instar larvae. This is the first report of a decided difference in variability of response to virus by two larval instars of the same species. A bimodal response by both larval instars was observed in time-mortality studies. Apparently, about 20% of the larvae were very resistant to virus infections.     

Apple orchard tests on the efficacy of the granulosis virus of the codling moth,Laspeyresia pomonella [Lep.: Olethreutidae]

The protection of apples against damage by the codling moth,Laspeyresia pomonella (L.), by applications of the granulosis virus ofL. pomonella was assessed in apple orchard tests at 4 locations in Canada in 1974–1978. Sprays containing 3×10⁹ to 4×10¹⁰ granules/litre, applied 2 or 3 times per generation of codling moth larvae, reduced injury to apples by 44 to 85% compared to reductions of 72 to 98% by applications of azinphos-methyl or phosmet. Applications of the virus did not reduce numbers of arthropods predaceous on pest insects and mites.

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Résumé L'efficacité de la protection des pommes contre le carpocapse,Laspeyresia pomonella (L.), par des applications du virus de la granulose deL. pomonella a été évaluée par des assais dans 4 vergers du Canada entre 1974 et 1978. Des pulvérisations contenant de 3×10⁹ à 4×10¹⁰ granules/litre, appliquées 2 ou 3 fois par génération de larves du carpocapse des pommes, ont réduit les dommages aux pommes de 44 à 85% comparé à des réductions de 72 à 98% obtenues avec des pulvérisations d'azinphos-méthyl ou de phosmet. Les applications de virus n'ont pas réduit les populations d'arthropodes prédateurs d'insectes nuisibles et d'acariens.

     

Pleiotropy of insecticide resistance in the codling moth, Cydia pomonella

The codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), has developed resistance to various insecticides. Relative fitness of one susceptible strain (Sv) and two strains selected for resistance to diflubenzuron (Rt) and deltamethrin (Rv), respectively, was measured in the absence of insecticide selection pressure. Mating rate, fecundity, fertility, developmental time, fifth instar weight, and adult longevity were compared. Both resistant strains were less fecund and fertile, developed more slowly, weighed less, and had shorter life-spans than the susceptible strain. These results indicate that biological constraints are associated with insecticide resistance in the codling moth. We also found that fitness estimates of the Rv strain did not differ statistically from those of the Rt strain. Enhanced mixed-function oxidase and glutathione-S-transferase activities have been shown to be involved in insecticide resistance in both Rt and Rv strains. This suggests that the fitness cost described in both resistant strains was mainly associated to metabolic resistance. The impact of such deleterious pleiotropy of insecticide resistance in C. pomonella in terms of resistance management in the field is discussed.     

Acetylcholinesterase mutation in an insecticide-resistant population of the codling moth Cydia pomonella (L.)

Two strains of Cydia pomonella (L.) (Lepidoptera: Tortricidae) were selected in the lab by exposure to increasing concentrations of diflubenzuron (Rdfb strain) or azinphos-methyl (Raz strain). Insecticide bioassays showed that the adults of the Rdfb strain exhibited a 2.6-fold and a 7.7-fold resistance ratio to azinphos-methyl and carbaryl, respectively compared to a susceptible strain (S) whereas the adults of the Raz strain exhibited a 6.7-fold resistance ratio to azinphos-methyl and a 130-fold resistance ratio to carbaryl. In the Raz strain, a target site resistance mechanism was suggested by the inhibition of acetylcholinesterase (AChE) activity. In fact the ki values did not discriminate the S and Rdfb strains, while the Raz strain exhibited a 1.7-fold and a 14-fold increase in ki value compared to the S strain for azinphos-methyl oxon and carbaryl, respectively. To verify this hypothesis, two cloned AChE cDNAs sequences (named cydpom-ace2 e cydpom-ace1) were compared between the susceptible and the resistant strains. No difference in the deduced amino acid sequence was found in cydpom-ace2 (orthologous to the Drosophila melanogaster AChE). In the putative cydpom-ace1 (paralogous to the Drosophila AChE), a single amino acid substitution F399V was exclusively present in the Raz strain. The F399 lined the active site of the enzyme and the F399V substitution likely could influence the accessibility of different types of inhibitors to the catalytic site of the insensitive cydpom-ace1.     

Mapping the disjunct distribution of introduced codling moth Cydia pomonella in China

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Ascorbic acid influence on larval blood cell number and susceptibility to bacterial and fungal infection in the codling moth, Laspeyresia pomonella (Lepidoptera: Tortricidae)

The optimum concentration of ascorbic acid in a semisynthetic diet for codling moth larvae was 0.6–0.8%. The number of circulating hemocytes and the percentage of prohemocytes and phagocytes decreased in larvae reared on the diet with suboptimal and supraoptimal ascorbic acid levels. These larvae were significantly more susceptible to Bacilus thuringiensis and Beauveria bassiana.     

Diapause induction in the codling moth, Cydia pomonella: effect of prediapause temperatures

The effect of four prediapause temperatures (18, 22, 26 and 30°C) on the photoperiodic response of the codling moth, Cydia pomonella (L.), was studied under controlled conditions. The highest rates of diapause were recorded, for all day-lengths, at temperatures of 22 and 26°C while relatively lower rates of diapause were elicited at 18 and 30°C. The same trend was demonstrated by projecting the values of the critical photoperiod which induces 50% diapause (=CPhP₅₀) over the prediapause temperature. The change in diapause incidence as a function of photoperiod, at all prediapause temperatures, exhibited a response characteristic of long-day insects, i.e. high rates of diapause at short days (12–13.5 h) and a decrease in diapause incidence at long days (14–15 h). The results for temperatures 22, 26 and 30°C support the view that lower prediapause temperatures enhance diapause induction, at a give photoperiod, while higher temperatures tend to avert or diminish the process. On the other hand, the low rates of diapause obtained at 18°C contradict this view. Nevertheless, high correlation was found between the laboratory evidence and field data, indicating the adaptability of the Israeli codling moth to subtropical climate.     

苹果蠹蛾在中国的扩散与危害

【背景】苹果蠹蛾原产于欧亚大陆,是世界著名的严重危害苹果生产的入侵害虫,也是中国的重要对外检疫性有害生物。【方法】通过收集整理历史文献、资料,并于2005~2013年使用标准性信息素诱芯对全国各苹果、梨主产区的苹果蠹蛾发生情况进行系统监测,明确该虫在我国的分布现状及扩散历史;此外,使用特定公式对苹果蠹蛾在发生区造成的当前经济损失和扩散完成后的潜在经济损失进行了系统评估,以明确该虫在我国的现有和潜在危害。【结果】苹果蠹蛾于1953年首次在中国新疆被发现,目前在中国分布于新疆、甘肃、内蒙古、宁夏、黑龙江、辽宁和吉林7省的144个县(市、区、旗),主要集中分布在东经74.56°~106.8°以及东经120.02°~132.95°,发生面积约49410 hm~2;经济损失评估结果显示,苹果蠹蛾导致中国苹果和梨的潜在减产数量为185.01万和81.33万t。【结论与意义】该虫已在中国形成东西2个分布区,对占据中国苹果产量80%的西北黄土高原(陕西为主)和渤海湾(山东、河北、辽宁为主)两大苹果主产区构成了严重威胁,须引起高度重视。     

Pheromone-trap catch in relation to the phenology of codling moth (Cydia pomonella)

Relationships between a pheromone-trap catch, adult emergence and penetration of fruit by first-instar larvae of Cydia pomonella were investigated from 1975 to 1977 in an orchard in South West England. For the first generation the times of moth emergence and catch in the pheromone trap were not significantly different; nor were male and female emergence times. The catch of moths of the first generation in the pheromone trap anticipated the appearance of their larvae in the fruit by 140–169 day-degrees > 10 °C. Eggs hatched after 94 day-degrees in the laboratory but in the orchard, wind and sunshine modified the microclimate so that the number of day-degrees required for egg development, as measured by standard meteorological instruments, was affected by wind and sunshine but development lasted on average about 90 day-degrees. This indicated a lag of 50–80 day-degrees between the curves for trap catch and oviposition: the pre-oviposition period in the orchard was shorter than expected from laboratory studies. In 1975 and 1976, some larvae developed to produce a second generation of moths which gave rise to a second generation of eggs and larvae, after an interval of 161 day-degrees in 1975, but only 41 day-degrees in 1976, indicating that in 1976 some eggs were laid before moths of the second generation were trapped. These results indicate that the first insecticidal spray against first-generation larvae should be applied about 140 day-degrees after the start of the reference week in which five moths of the spring brood are caught per trap. A second spray, if required to maintain insecticidal cover, should be applied about 100 day-degrees later. A spray should be applied against second-generation larvae immediately after the reference week for second-generation moths.