Effect of Bacillus thuringiensis on Habrobracon hebetor during combined biological control of Plodia interpunctella

Abstract  The suitability of combining microbial pesticides and an insect parasitoid for pest management of stored cereal in China was evaluated using laboratory assays. For this purpose, interactions between Bacillus thuringiensis ( Bt ), Bt -intoxicated host larvae and the parasitoid Habrobracon hebetor (Say) (Hymenoptera: Braconidae) were tested against Plodia interpunctella Hübner (Lepidoptera: Pyralidae) . Bt or H. hebetor alone caused 41.67% and 35.35% P. interpunctella larval mortality respectively. The Bt –parasitoid combined treatment significantly increased mortality of P. interpunctella (86%). Progeny development of H. hebetor was dependent upon its susceptibility to Bt . Fewer parasitoids emerged from Bt –parasitoid combined treatment than in non- Bt treatments. However, since Bt did not prevent parasitoid development, a combined treatment with Bt and parasitoid release could produce better protection against P. interpunctella than either treatments when used singly, because their lethal effects were additive to each other.     

Effectiveness of spinosad on four classes of wheat against five stored-product insects

Spinosad is a commercial reduced-risk pesticide that is naturally derived. Spinosad's performance was evaluated on four classes of wheat (hard red winter, hard red spring, soft red winter, and durum wheats) against adults of the lesser grain borer, Rhyzopertha dominica (F.); rice weevil, Sitophilus oryzae (L.); sawtoothed grain beetle, Oryzaephilus surinamensis (L.); red flour beetle, Tribolium castaneum (Herbst); and larvae of the Indianmeal moth, Plodia interpunctella (Hübner). Beetle adults (25) or P. interpunctella eggs (50) were exposed to untreated wheat and wheat treated with spinosad at 0.1 and 1 mg (AI)/kg of grain. On all untreated wheat classes, adult beetle mortality ranged from 0 to 6%, and P. interpunctella larval mortality ranged from 10 to 19%. The effects of spinosad on R. dominica and P. interpunctella were consistent across all wheat classes. Spinosad killed all exposed R. dominica adults and significantly suppressed progeny production (84-100%) and kernel damage (66-100%) at both rates compared with untreated wheat. Spinosad was extremely effective against P. interpunctella on all wheat classes at 1 mg/kg, based on larval mortality (97.6-99.6%), suppression of egg-to-adult emergence (93-100%), and kernel damage (95-100%), relative to similar effects on untreated wheats. The effects of spinosad on S. oryzae varied among wheat classes and between spinosad rates. Spinosad was effective against S. oryzae, O. surinamensis and T. castaneun only on durum wheat at 1 mg/kg. Our results suggest spinosad to be a potential grain protectant for R. dominica and P. interpunctella management in stored wheat.     

Efficacy of an esfenvalerate plus methoprene aerosol for the control of eggs and fifth instars of Plodia interpunctella (Lepidoptera: Pyralidae)

Abstract  Aerosol insecticides may provide an alternative to fumigants for control of the Indianmeal moth, Plodia interpunctella (Hübner), the Indianmeal moth, a major insect pest of stored processed food. In this study, eggs and larvae (5th instars) of P . interpunctella were exposed to aerosol applications of the pyrethroid esfenvalerate and insect growth regulator methoprene, alone and in combination, in open and obstructed positions inside small sheds. When larvae were exposed to methoprene alone, adult emergence from those exposed larvae was 7.1%± 1.5%. In contrast, adult emergence was 92.5%± 3.5% when larvae were exposed to esfenvalerate alone. When eggs were exposed to methoprene, adult emergence of those exposed eggs was approximately 75%; however, when eggs were exposed to esfenvalerate, adult emergence was approximately 35%. In the combination treatment of methoprene plus esfenvalerate at their respective label rates, adult emergence following larval exposure was 0.91%± 0.61% compared to 16.3%± 9.6% when eggs were exposed. Based on our results, methoprene alone is highly effective in reducing adult emergence after larval exposure. However, it is not as effective on eggs as esfenvalerate. A combination treatment of esfenvalerate plus methoprene could be used to control eggs and the wandering-phase larval stages of P . interpunctella . An economic risk analysis also supports a strategy of combining methoprene and esfenvalerate.     

Mortality of Indianmeal moth (Lepidoptera: Pyralidae) populations under fluctuating low temperatures: model development and validation

A model for predicting mortality of Indianmeal moth larvae [Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae)] under fluctuating low-temperature conditions was developed. The time and temperature combinations required to achieve 100% mortality of field-collected, cold-acclimated P. interpunctella larvae obtained from laboratory mortality experiments were used to develop the mortality model. Accumulation of mortality rate over time was called the cumulative lethality index (CLI). Complete mortality of insect populations would occur when CLI equals 1. Observed mortality of field-collected, cold-acclimated P. interpunctella larvae in five 76.2-T (3,000-bu) shelled corn bins located in Rosemount, MN, during the winters of 2003-2004 and 2004-2005 were used to validate the CLI model (i.e., mortality model). Excellent agreement between predicted and measured time to 100% larval mortality was observed. The CLI model would be useful for developing low-temperature aeration management strategies for controlling overwintering P. interpunctella in grain bins. In addition, this model will be useful when determining if additional control measures will be required as a result of above-seasonal ambient temperatures.     

Biology and development of some stored grain pests as affected by δ‐endotoxin and β‐exotoxin of Bacillus thuringiensis

Exposure of Plodia interpunctella and Sitotroga cerealella to sublethal concentrations of Dipel 2X® (Bacillus thuringiensis var.kurstaki‐HD‐1)with its spores and endotoxin crystals led to an increase in the sum of larval‐pupal duration. A decrease in moth emergence, egg production and fertility was observed with increase of Dipel 2X concentration, while the longevity of the moths was not affected by treatment. Sublethal concentrations of β‐exotoxin (ABG‐6162 A) had no significant effect on the sum of larval—pupal duration of either insect species. Percentage of moth emergence decreased with the increase of β‐exotoxin concentrations. The longevity of both male and female adults of P. interpunctella and of females of S. cerealella were shorter at high concentrations. Egg production in P. interpunctella was reduced by all concentrations of the β‐exotoxin, and hatching of eggs was reduced by concentrations of 15 ppm and above.     

Histological development of the digestive system of the Amazonian pimelodid catfish Pseudoplatystoma punctifer

The organogenesis of the digestive system was described in the Amazonian pimelodid catfish species Pseudoplatystoma punctifer from hatching (3.5 mm total length, TL) to 41 days post-fertilization (dpf) (58.1 mm TL) reared at 28°C. Newly hatched larvae showed a simple digestive tract, which appeared as a straight undifferentiated and unfolded tube lined by a single layer of columnar epithelial cells (future enterocytes). During the endogenous feeding period, comprised between 20 and 96 h post-fertilization (3.5 to 6.1 mm TL), the larval digestive system experienced a fast transformation with the almost complete development and differentiation of most of digestive organs (buccopahrynx, oesophagus, intestine, liver and exocrine pancreas). Yolk reserves were not completely depleted at the onset of exogenous feeding (4 dpf, 6.1 mm TL), and a period of mixed nutrition was observed up to 6 to 7 dpf (6.8 to 7.3 mm TL) when yolk was definitively exhausted. The stomach was the organ that latest achieved its complete differentiation, characterized by the development of abundant gastric glands in the fundic stomach between 10 and 15 dpf (10.9 to 15.8 mm TL) and the formation of the pyloric sphincter at the junction of the pyloric stomach and the anterior intestine at 15 dpf (15.8 mm TL). The above-mentioned morphological and histological features observed suggested the achievement of a digestive system characteristic of P. punctifer juveniles and adults. The ontogeny of the digestive system in P. punctifer followed the same general pattern as in most Siluriform species so far, although some species-specific differences in the timing of differentiation of several digestive structures were noted, which might be related to different reproductive guilds, egg and larval size or even different larval rearing practices. According to present findings on the histological development of the digestive system in P. punctifer, some recommendations regarding the rearing practices of this species are also provided in order to improve the actual larval rearing techniques of this fast-growing Neotropical catfish species.     

Biological control of Indianmeal moth (Lepidoptera: Pyralidae) on finished stored products using egg and larval parasitoids

Biological control using hymenopteran parasitoids presents an attractive alternative to insecticides for reducing infestations and damage from the Indianmeal moth, Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae), in retail and warehouse environments. We examined the potential for using combinations of the egg parasitoid Trichogramma deion Riley (Hymenoptera: Trichogrammatidae), and the larval parasitoid Habrobracon hebetor (Say) (Hymenoptera: Braconidae) for preventing infestations of P. interpunctella in coarse-ground cornmeal as well as the influence of packaging on parasitoid effectiveness. Treatments included one or both parasitoids and either cornmeal infested with P. interpunctella eggs or eggs deposited on the surface of plastic bags containing cornmeal. H. hebetor had a significant impact on the number of live P. interpunctella, suppressing populations by approximately 71% in both unbagged and bagged cornmeal. In contrast, T. deion did not suppress P. interpunctella in unbagged cornmeal. However, when released on bagged cornmeal, T. deion significantly increased the level of pest suppression (87%) over bagging alone (15%). When H. hebetor was added to bagged cornmeal, there was a significant reduction of live P. interpunctella compared with the control (70.6%), with a further reduction observed when T. deion was added (96.7%). These findings suggest that, in most situations, a combined release of both T. deion and H. hebetor would have the greatest impact; because even though packaging may protect most of the stored products, there are usually areas in the storage landscape where poor sanitation is present.     

Relationship between supercooling point and mortality at low temperatures in Indianmeal moth (Lepidoptera: Pyralidae)

Indianmeal moth, Plodia interpunctella (Hübner), is classified as a freeze-intolerant organism and one of the most cold-tolerant stored-product pests. The objective of this study was to determine the relationship between mortality at low temperatures after minimum exposure and the supercooling point (SCP) for laboratory-reared P. interpunctella at different stages of development. This relationship also was analyzed for field-collected, cold-acclimated fifth instars. Mean SCP of laboratory-reared larvae (i.e., feeding stage) was consistently above approximately -16 degrees C. Mean SCP of laboratory-reared pupae and adults (i.e., nonfeeding stages) and field-collected, cold-acclimated fifth instars was consistently below approximately -21 degrees CP seemed to be the boundary between survival and death for larvae. However, it seemed that a 1-min exposure was not sufficient to cause larval mortality at the SCP. Alternatively, for both pupae and adults, the SCP seemed not to play an important role in their survival at low temperatures, with significant mortality observed at temperatures higher than the mean SCP. Adults were the most susceptible to low temperatures with no survival occurring at -20 degrees C, > 3 degrees C above its mean SCP. Results of this investigation demonstrate that P. interpunctella has a different response to low temperatures depending on stage of development and cold acclimation. Classifying P. interpunctella only as a freeze-intolerant organism disregards the occurrence of prefreeze mortality in this species. Therefore, a reclassification of this species (e.g., chill tolerant or chill susceptible) based on the extent of prefreeze mortality and the temperature and time of exposure at which it occurs is suggested.     

Midgut amylase,lysozyme, aminopeptidase,and trehalase from larvae and adults of Musca domestica

Based on polyacrylamide gel electrophoresis, density-gradient ultracentrifugation and thermal inactivation, there is only one major molecular species of each of the following larval enzymes (soluble in water or solubilized in Triton X-100): membrane-bound aminopeptidase (pH optimum 8.5; K_m 0.21 mM L-leucine p-nitroanilide; M_r 322,000), amylase (pH optimum 6.5; K_m 0.14% starch; M_r 66,000), lysozyme (pH optimum 3.5; K_m 0.3 mg/ml; Mr 24,000); and membrane-bound trehalase (pH optimum 5.0; K_m 1.09 mM trehalose; M_r 94,000). Except for lysozyme, the properties of adult digestive enzymes are different from those described for larval enzymes. Larval aminopeptidase and trehalase were purified by electrophoresis and larval lysozyme (contaminated with amylase) by density-gradient ultracentrifugation, and were used to raise antibodies in a rabbit. Antibodies raised against larval aminopeptidase, trehalase, and amylase did not recognize the imaginal enzymes, whereas those against larval lysozyme recognize imaginal lysozyme. The data suggest that the genes coding for digestive enzymes (except for lysozyme) are different in larvae and imagoes.     

Phenoloxidase in larvae of Plodia interpunctella (Lepidoptera: Pyralidae): molecular cloning of the proenzyme cDNA and enzyme activity in larvae paralyzed and parasitized by Habrobracon hebetor (Hymenoptera: Braconidae)

Phenoloxidase (PO) is a major component of the insect immune system. The enzyme is involved in encapsulation and melanization processes as well as wound healing and cuticle sclerotization. PO is present as an inactive proenzyme, prophenoloxidase (PPO), which is activated via a protease cascade. In this study, we have cloned a full-length PPO1 cDNA and a partial PPO2 cDNA from the Indianmeal moth, Plodia interpunctella (Hubner) (Lepidoptera: Pyralidae) and documented changes in PO activity in larvae paralyzed and parasitized by the ectoparasitoid Habrobracon hebetor (Say) (Hymenoptera: Braconidae). The cDNA for PPO1 is 2,748 bp and encodes a protein of 681 amino acids with a calculated molecular weight of 78,328 and pI of 6.41 containing a conserved proteolytic cleavage site found in other PPOs. P. interpunctella PPO1 ranges from 71-78% identical to other known lepidopteran PPO-1 sequences. Percent identity decreases as comparisons are made to PPO-1 of more divergent species in the orders Diptera (Aa-48; As-49; and Sb-60%) and Coleoptera (Tm-58; Hd-50%). Paralyzation of host larvae of P. interpunctella by the idiobiont H. hebetor results in an increase in phenoloxidase activity in host hemolymph, a process that may protect the host from microbial infection during self-provisioning by this wasp. Subsequent parasitization by H. hebetor larvae causes a decrease in hemolymph PO activity, which suggests that the larval parasitoid may be secreting an immunosuppressant into the host larva during feeding.     

The solubility of inclusion proteins from Bacillus thuringiensis is dependent upon protoxin composition and is a factor in toxicity to insects.

Bacillus thuringiensis subsp. aizawai HD133 is one of several strains particularly effective against Plodia interpunctella selected for resistance to B. thuringiensis subsp. kurstaki HD1 (Dipel). B. thuringiensis subsp. aizawai HD133 produces inclusions containing three protoxins, CryIA(b), CryIC, and CryID, and the CryIC protoxin has been shown to be active on resistant P. interpunctella as well as on Spodoptera larvae. The CryIA(b) protoxin is very similar to the major one in B. thuringiensis subsp. kurstaki HD1, and as expected, this protoxin was inactive on resistant P. interpunctella. A derivative of B. thuringiensis subsp. aizawai HD133 which had been cured of a 68-kb plasmid containing the cryIA(b) gene produced inclusions comprising only the CryIC and CryID protoxins. Surprisingly, these inclusions were much less toxic for resistant P. interpunctella and two other Lepidoptera than those produced by the parental strain, whereas the soluble protoxins from these strains were equally effective. In contrast, inclusions from the two strains were about as active as soluble protoxins for Spodoptera frugiperda larvae, so toxicity differences between inclusions may be due to the solubilizing conditions within particular larval guts. Consistent with this hypothesis, it was found that a higher pH was required to solubilize protoxins from inclusions from the plasmid-cured strain than from B. thuringiensis subsp. aizawai HD133, a difference which is probably attributable to the absence of the CryIA(b) protoxin in the former. The interactions of structurally related protoxins within an inclusion are probably important for solubility and are thus another factor in the effectiveness of B. thuringiensis isolates for particular insect larvae.     

Characterization of a novel insect digestive DNase with a highly alkaline pH optimum.

A novel DNase from the digestive tract of the spruce budworm (Choristoneura fumiferana) has been isolated and characterized. This DNase has two features that distinguish it from other known DNases: (1) it has a pH optimum of 10.5 to 11; (2) it plays an important role in the conversion of the insecticidal crystal protein from Bacillus thuringiensis to the active DNA-free toxin in the larval gut. Only one digestive DNase with an apparent molecular mass of 23 kDa was found and no associated carbohydrate was detected. It has some similarities to pancreatic DNase I in that divalent alkaline metal ion is required for activity and it is inhibited by monovalent cations. In particular, Mg(2+) and Ca(2+) were the most effective activators. Transition metal ions also activated the enzyme but were less effective. The enzyme is an endonuclease that hydrolyzes single and double stranded DNA but shows a higher specificity for single stranded DNA. The purified enzyme acted synergistically with proteases on crystals from Bacillus thuringiensis to yield the DNA-free toxin. To our knowledge, this is the first characterization of DNase activity in insect larvae and provides strong evidence that a DNase is an integral component of the larval digestive system.     

明亮熊蜂繁育室内印度谷斑螟的形态特征与生物学特性

明亮熊蜂Bombus lucorum L.是一种重要的温室果菜传粉昆虫,印度谷斑螟Plodia interpunctella(Hbner)是危害明亮熊蜂繁育的主要害虫之一。2年的研究结果表明,印度谷斑螟的形态特征与危害蜜蜂巢房的大蜡螟Gallerie mellonellaL.和小蜡螟Achroia grisella Fabricius明显不同。印度谷斑螟以饲喂熊蜂的花粉为载体传播进熊蜂繁育室,在熊蜂繁育室内可以连续繁殖,1年发生6～8代,且世代重叠。印度谷斑螟幼虫主要以蜂群内的剩花粉为食,当花粉不足时,就开始取食巢房和蜂蛹,对熊蜂的规模化繁育危害较大,每年的5～8月、11～2月为危害高峰期。印度谷斑螟的发育受环境和食物的影响很大,在温度为28℃、相对湿度为60%熊蜂繁育室内,印度谷斑螟的卵期为4d,幼虫期为19～23d,蛹期为11d,成虫期为4～20d。成虫羽化后3～4h即可进行交配,雌蛾交配后当天就开始产卵,产卵期4～10d不等,平均产卵量107.8粒。在熊蜂繁育室消毒期间,食物短缺,印度谷斑螟幼虫进入休眠状态,各龄幼虫均可发生休眠现象。印度谷斑螟不危害蜜蜂,在蜜蜂巢房中不能存活。     

中国印度谷螟性信息素次级组份的研究

印度谷螟是我国危害性较严重的仓储害虫之一,食性很广,几乎危害每一种植物性仓储物.印度谷螟雌虫性信息素主要成份顺9,反12-十四碳二烯醇醋酸酯(TDA)早已被鉴定,并利用TDA单一组份诱捕印度谷螟雄蛾也较为有效,但该虫的完整信息素系统尚不清楚.本研究采用单个雌蛾性信息素腺体抽提技术,利用毛细管气相色谱GC保留时间定性方法和气质联用GC-MS分析鉴定技术,从印度谷螟雌虫腺体提取物中鉴定了顺9,反12-十四碳二烯醇醋酸酯(A),顺9,反12-十四碳二烯醇(B),顺9,反12-十四碳二烯醛(C)和顺9-十四碳醋酸醋(D)4种主要信息素成份,其比例为A∶B∶C∶D=100∶22∶12∶9.风洞试验结果表明,以该4种主要成分按A∶B∶C∶D=8∶2∶1∶0.8比例配制成的诱芯,其引诱雄蛾的活性接近于雌蛾腺体提取物.     

Development of the larval ovary in the moth, Plodia interpunctella

The morphogenesis of ovaries and the organization of germ cells within them were visualized during the larval stages of the moth, Plodia interpunctella. The germ cells were observed by utilizing confocal microscopy coupled with immuno-fluorescent staining for the alpha-crystallin protein 25 (alphaCP25). The alphaCP25 was previously shown to be specific to germ cells of pupae and adults, and this study shows that alphaCP25 is present in larval germ cells as well. A cluster of 28 germ cells that stain for alphaCP25 was found in the gonads of newly hatched first instar larvae. The founding germ cells became segregated into four clusters, most likely by somatic cell intrusion, around the beginning of the second instar. Division of the primary germ cells began by the end of the second instar and the formation of all cystoblasts appeared to be completed within the four ovarioles by the end of the third instar. Within the ovarioles of third instar larvae, the germ cells were organized with a distal cap of seven germ cells which was segregated from the majority of the germ cells. The main body of germ cells was arranged around a central germ cell-free core as a spiral. Divisions of the cystoblasts to form cystocyte clusters were nearly completed during the fourth (last) larval instar. These features suggest that the strategy to produce follicles in moths is fundamentally different from the fruitfly, Drosophila. It appears that during the initial stages of ovary development in P. interpunctella, the primary germ cells undergo stage-complete divisions that are completed prior to the onset of the next set of divisions, which results in a complete complement of follicles available by the time of adult eclosion, while in Drosophila the primary germ cell divisions are initiated in the adult stage, and follicles are produced individually as resources are available.     

Attractancy and toxicity of an attracticide for Indianmeal moth, Plodia interpunctella (Lepidoptera: Pyralidae)

Plodia interpunctella (Hübner) is a serious and widespread postharvest pest on cereal products, dried fruits, candy, and pet food. Due to the strong positive anemotactic flight response of P. interpunctella males to the main component of the female-produced pheromone [(Z,E)-9,12-tetradecadienyl acetate, herein referred to as ZETA], we evaluated the potential of an attracticide for this pest, in which ZETA as attractant was combined with permethrin as the killing agent. Two concentrations of ZETA [0.16 and 0.32% (wt:wt)] and five concentrations of permethrin [0, 3, 6, 12, and 18% (wt:wt)] were incorporated into Last Call gel (10 different permethrin:ZETA combinations). All attracticide gels were evaluated in a toxicity test, in which either the tip of a leg or an antenna of a virgin P. interpunctella male was touched < 3 s into a dot of attracticide gel. These males were subsequently transferred to jars with virgin females. The toxicity test showed that a brief and gentle contact of P. interpunctella males with attracticide gel containing 3-18% permethrin caused a significant reduction in mating and killed males moths within 24 h. A wind tunnel test was conducted to evaluate the flight responses of P. interpunctella males to the same 10 attracticide gels. Male moths displayed significantly higher levels of positive anemotactic flight and more males made contact with the attracticide gel when the ZETA concentration was 0.16% compared with 0.32%. P. interpunctella males showed no signs of repellency to permethrin concentrations within a range of 0-18% in the attracticide gel. Three densities of P. interpunctella pairs were released into small warehouse rooms, and we found that the attracticide gel suppressed oviposition when the moth density was at a low level, but it was ineffective when the moth density exceeded one male-female pair per 11.3 m3.     

Activity of Plodia interpunctella (Lepidoptera: Pyralidae) in and around flour mills

Studies were conducted at two flour mills where male Indian meal moths, Plodia interpunctella (Hübner), were captured using pheromone-baited traps. Objectives were to determine the distribution of male P. interpunctella at different locations in and around the mills throughout the season, and to monitor moth activity before and after one of the mills was fumigated with methyl bromide to assess efficacy of treatment. Commercially available sticky traps baited with the P. interpunctella sex pheromone were placed at various locations outside and within the larger of the two mills (mill 1). Moths were captured inside mill 1 after methyl bromide fumigations. The highest numbers of P. interpunctella were caught outside the facility and at ground floor locations near outside openings. Additional traps placed in the rooms above the concrete stored-wheat silos at mill 1 during the second year captured more moths than did traps within the mill's production and warehouse areas. In another study, moths were trapped at various distances from a smaller flour mill (mill 2) to determine the distribution of moths outdoors relative to the mill. There was a negative correlation between moth capture and distance from the facility, which suggested that moth activity was concentrated at or near the flour mill. The effectiveness of the methyl bromide fumigations in suppressing moth populations could not be assessed with certainty because moths captured after fumigation may have immigrated from outside through opened loading bay warehouse doors. This study documents high levels of P. interpunctella outdoors relative to those recorded inside a food processing facility. Potential for immigration of P. interpunctella into flour mills and other stored product facilities from other sources may be greater than previously recognized. Moth entry into a food processing facility after fumigation is a problem that should be addressed by pest managers.     

Effects of transgenic Bacillus thuringiensis maize grain on B. thuringiensis-susceptible Plodia interpunctella (Lepidoptera: Pyralidae)

Percentage survivorship, developmental time, adult body length, and sex ratio of Plodia interpunctella (Hübner) reared on field-produced grain from sixteen cultivars of maize, Zea mays L., including several transgenic Bacillus thuringiensis (Bt) Berliner hybrids and selected non-Bt isolines, were evaluated under laboratory conditions. Compared with isolines, development was delayed and survivorship reduced for P. interpunctella reared on grain from transgenic hybrids with the CaMV/35s promoter that express Cry1Ab protein. Similarly, compared with non-Bt hybrids, a transgenic hybrid with the CaMV/35s promoter that expresses Cry9C protein delayed development, decreased survivorship, and caused reductions in adult body length of P. interpunctella. In contrast, no significant differences in P. interpunctella developmental times or survivorship were observed between transgenic hybrids with the PEPC promoter expressing Cry1Ab and their isolines. Additionally, developmental time, survivorship, and adult body length were similar between P. interpunctella reared on a transgenic hybrid with the CaMV/35s promoter expressing Cry1Ac and non-Bt hybrids. Our data demonstrate that transgenic Bt maize grain, especially grain from hybrids with the CaMV/35s promoter expressing Cry1Ab or Cry9C, can significantly affect B. thuringiensis-susceptible P. interpunctella populations up to 4 or 5 mo after harvest.     

Hormonal control of growth and differentiation of insect tissues cultured in vitro.

This paper reviews the effects of insect hormones on lepidopteran imaginal discs cultured in vitro. Beta-ecdysone showed that RNA and protein synthesis was required for evagination and cuticle deposition. In particular, studies with actinomycin D and cycloheximide (at nontoxic levels) showed that RNA and protein synthesis during the ecdysone-dependent period was essential for subsequent development. These findings support the hypothesis that stimulation of macromolecular synthesis is fundamental to the action of ecdysone on imaginal discs. The influence of beta-ecdysone on chitin synthesis was also examined. Beta-ecdysone stimulated uptake and incorporation of tritiated-glucosamine by cultured P.interpunctella wing discs. Addition of hexosamines to the culture medium had no influence on ecdysone-induced cuticle deposition, but inhibition of glucose-uptake by cytochalasin B prevented the formation of cuticle. The action of ecdysone on particular enzymes in the chitin pathway remains to be elucidated.timulated both evatination and cuticle deposition of wing discs of Plodia interpunctella(Hubner). However, evagination required a shorter exposure to ecdysone than did cuticle deposition. Cuticle deposition was obtained under the following conditions: (a) a 24-hour pulse of beta-ecdysone (0.5-5.0 mug/ml); (b) continuous treatment with 0.2 mug/ml beta-ecdysone; or (c) continuous treatment with 0.5 to 50.0 mug/ml beta-ecdysone in medium conditioned with larval fat body...