F1 Larvae Research Articles

Fitness Costs of Cry1Ab Resistance in a Field-Derived Strain of <I>Ostrinia nubilalis</I> (Lepidoptera: Crambidae)

The study of fitness costs associated with resistance to toxins from Bacillus thuringiensis Berliner (Bt) in Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) is important for understanding resistance evolution and for evaluating resistance management practices that prevent or mitigate resistance to transgenic corn, Zea mays L. Resistant individuals identified from a field collection in Kandiyohi, MN, were used to generate a Cry1Ab-resistant strain. We used susceptible and resistant strains with similar genetic background to establish crosses and estimate dominance of fitness costs by measuring fitness components and population parameters determined by fertility life tables. Spermatophore volume and mating frequency also were compared to identify potential effects of resistance on fertility. Inheritance of fitness costs in O. nubilalis varied from recessive to incompletely recessive among the parameters evaluated. Selection for resistance to Cry1Ab significantly reduced the fitness of O. nubilalis. Resistant insects exhibited reduced pupal weight and increased developmental time compared with susceptible and F1 larvae derived from reciprocal crosses of resistant and susceptible parents. In addition, it was observed that resistant insects exhibited a higher proportion of unsuccessful matings and lower fertility than the susceptible strain. Despite the differences observed in resistant insects, our results did not indicate strong evidence of fitness costs in the F1 progeny.

Ultraviolet radiation and handling medium osmolarity affect chimaerism success in zebrafish

The effects of a predefined ultraviolet radiation dose (0.529 mW/cm2 for 30s) together with two different micromanipulation medium osmolarities (30 mOsm/kg vs 300 mOsm/kg) were tested on embryo survival at different developmental stages and on the somatic (skin) and germ-line chimaerism rates. Somatic (13%, 6/47 adults) and germ-line chimaerism (50% pigmented F1 larvae) were detected only in the UV-treated recipient embryos micromanipulated in a 300 mOsm/kg medium. From the results obtained, we concluded that the conditions cited above were the most suitable to improve somatic and germ-line chimaerism rates in zebrafish.

Influence of bicarbonate and humic acid on effects of chronic waterborne lead exposure to the fathead minnow ( Pimephales promelas)

Historically, the USEPA has only considered water hardness when establishing acute and chronic water quality criteria (WQC) for lead (Pb) in freshwater. Yet, recent evidence suggests that hardness may not be protective during chronic Pb exposure and that other factors (e.g., dissolved organic carbon (DOC) and alkalinity) influence toxicity. In fact, we have recently shown that Ca 2+ (as CaSO 4) does not protect against Pb accumulation in fathead minnows ( Pimephales promelas) during chronic exposures whereas DOC as humic acid (HA) clearly does. To more clearly define the water chemistry parameters mediating chronic Pb toxicity we carried out 300 d exposures to study the influence of DOC and alkalinity on Pb accumulation and toxicity to fathead minnows at 2 different Pb concentrations (170 and 580 nM (35 and 120 μg/L)). Alkalinity was adjusted by addition of 500 μM NaHCO 3 and DOC by addition of 4 mg/L HA. Fish were collected at 4, 30, 150 and 300 d of exposure to measure growth and Pb accumulation. Breeding assays (21 d) were performed at the end of these exposures to assess reproductive and larval behavioral endpoints. To determine whether effects were acute or chronic, switched breeding exposures were performed in which control breeders were transferred to either high or low Pb conditions and Pb-exposed breeders transferred to tap water without Pb. Mortality and growth effects were observed primarily in the high Pb treatments and within the first 10 d of exposure. Strong protection against Pb accumulation was afforded by increased DOC at both Pb concentrations. Increased alkalinity also appeared to moderately reduce Pb accumulation although not to the level of statistical significance. Tissue distribution of Pb was analyzed at 300 d and was found to accumulate mostly in bone, gill, intestine and kidney. Unexpectedly, high Pb reduced total reproductive output and increased average egg mass in the HCO 3 − and DOC treatments but not in the control water (+Pb) treatments. No statistically significant differences in egg hatchability or egg Pb accumulation were observed. Results from switched exposures suggest that embryo Pb accumulation arose from acute exposure to embryos rather than parental transfer. Finally, prey capture assays revealed potential Pb-induced motor/behavioral impairment in 10-d-old F1 larvae exposed to high Pb in all water treatments.

Comparative data on the insecticide resistance of Anopheles albimanus in relation to agricultural practices in northern Belize, CA

Anopheles albimanus Wiedemann is one of the primary vectors of malaria in northern Belize. The primary breeding sites for this species in Central America are fresh water wetlands that are found in close proximity to agriculture and, in particular, sugarcane fields. The use of insecticides and herbicides on these crops leads to the introduction of chemicals into the surrounding wetlands. The potential for the development of insecticide resistance is, therefore, quite high. A study was undertaken to obtain comparative data on the resistance status of An. albimanus in northern Belize in relation to agriculture practices. Larval surveys were conducted at four collection points in marshes juxtaposed to different crop types: Dubloon (DB)—control site located 300 m from any agriculture; Little Belize (LB)—mixed crop agricultural with some human settlements; and Chan Chen (CC) and Buena Vista (BV)—year round sugarcane production. Time mortality curves were obtained by exposing F1 larvae to DDT 1 μg/ml, malathion 1 μg/ml, and permethrin 0.05 μg/ml. Significant differences in mortality rates from the four sites were observed only to malathion. The DB sites contained larvae that were the most susceptible overall, in congruence with its distance from crops. Larvae from LB exhibited a medium level of tolerance. Finally, the two populations collected at the edge of sugarcane fields (CC and BV) exhibited the highest tolerance to malathion. These results represent first data on the impact of agriculture on the resistance status of malaria vectors in northern Belize. The effect of malathion on An. albimanus insecticide resistance reinforces the concerns of the potential impact of agriculture usage on the efficacy of vector control program.

Transovarial Transmission ofFrancisella-Like Endosymbionts andAnaplasma phagocytophilumVariants inDermacentor albipictus(Acari: Ixodidae)

Dermacentor albipictus (Packard) is a North American tick that feeds on cervids and livestock. It is a suspected vector of anaplasmosis in cattle, but its microbial flora and vector potential remain underevaluated. We screened D. albipictus ticks collected from Minnesota white-tailed deer (Odocoileus virginianus) for bacteria of the genera Anaplasma, Ehrlichia, Francisella, and Rickettsia using polymerase chain reaction (PCR) gene amplification and sequence analyses. We detected Anaplasma phagocytophilum and Francisella-like endosymbionts (FLEs) in nymphal and adult ticks of both sexes at 45 and 94% prevalences, respectively. The A. phagocytophilum and FLEs were transovarially transmitted to F1 larvae by individual ticks at efficiencies of 10-40 and 95-100%, respectively. The FLEs were transovarially transmitted to F2 larvae obtained as progeny of adults from F1 larval ticks reared to maturity on a calf, but A. phagocytophilum were not. Based on PCR and tissue culture inoculation assays, A. phagocytophilum and FLEs were not transmitted to the calf. The amplified FLE 16S rRNA gene sequences were identical to that of an FLE detected in a D. albipictus from Texas, whereas those of the A. phagocytophilum were nearly identical to those of probable human-nonpathogenic A. phagocytophilum WI-1 and WI-2 variants detected in white-tailed deer from central Wisconsin. However, the D. albipictus A. phagocytophilum sequences differed from that of the nonpathogenic A. phagocytophilum variant-1 associated with Ixodes scapularis ticks and white-tailed deer as well as that of the human-pathogenic A. phagocytophilum ha variant associated with I. scapularis and the white-footed mouse, Peromyscus leucopus. The transovarial transmission of A. phagocytophilum variants in Dermacentor ticks suggests that maintenance of A. phagocytophilum in nature may not be solely dependent on horizontal transmission.

O.472 Ultrasound activated pin flxation in pediatric maxillo-facial traumas

Maternal thyroid hormones (THs) play an essential role in the embryonic and larval development of fish. Previous studies in fish have reported that parental exposure to thyroid disrupting chemicals (TDCs) changed maternal TH levels in the offspring; however, whether this transgenerational thyroid endocrine disruption can further disturb the early development of the offspring still remains largely unknown. Bisphenol S (BPS), a substitute of bisphenol A, has been reported to be a potential TDC. In this study, zebrafish (F0) were exposed to environmentally relevant concentrations (1, 10, and 100 μg/L) of BPS from 2 h post-fertilization to 120 days post-fertilization and then paired to spawn. Plasma levels of thyroxine (T4) were significantly decreased in F0 females while 3,5,3′-triiodothyronine (T3) plasma levels were significantly increased in F0 females and males; moreover, TH content in eggs (F1) spawned by exposed F0 generation exhibited similar changes as the F0 females, with significant decreases in T4 and increases in T3, demonstrating BPS-induced maternal transfer of thyroid endocrine disruption. Further, excessive levels of maternal T3 in the offspring resulted in delayed embryonic development and hatching, swim bladder inflation defect, reduction in motility, developmental neurotoxicity, and lateral stripe hypopigmentation in non-exposed F1 embryos and larvae. These results highlight the adverse effects on the early development of offspring induced by transgenerational thyroid endocrine disruption, which have been ignored by previous studies. Therefore, these results can further improve our understanding of the ecological risks of TDCs.

Genetics and Mechanisms of Permethrin Resistance in the Santa Luiza Strain of <I>Boophilus microplus</I> (Acari: Ixodidae)

The Santa Luiza strain of the southern cattle tick, Boophilus microplus (Canestrini) (Acari: Ixodidae), is resistant to both permethrin and amitraz. A study was conducted at the USDA Cattle Fever Tick Research Laboratory in Texas to investigate the genetic basis of permethrin resistance with cross-mating experiments, and to determine the mechanisms of permethrin resistance through synergist bioassays and biochemical analysis of esterase profiles. The Muñoz strain, an acaricide-susceptible reference strain, was used as the susceptible parent and the Santa Luiza strain, originating in Brazil, was used as the resistant parent. The Food and Agriculture Organization larval packet test was used to measure the levels of susceptibility of larvae of the parental strains, F1, backcross, F2, and F3 generations to permethrin. Results of reciprocal crossing experiments suggested that permethrin resistance was inherited as an incomplete recessive trait. There was no significant maternal effect on larval progeny's susceptibility to permethrin in the F1 and subsequent generations. The values of the degree of dominance were estimated at -0.700 and -0.522 for the F1 larvae with resistant and susceptible female parents, respectively. Results of bioassays on larval progeny of the F1 backcrossed with the resistant parent strain and of the F2 generations suggested that one major gene was responsible for permethrin resistance in the Santa Luiza strain. Selection of F3 larvae with either permethrin or amitraz led to significantly increased resistance to both permethrin and amitraz, indicating a close linkage between genes responsible for permethrin and amitraz resistance. The possible involvement of metabolic enzymes in permethrin resistance in the Santa Luiza strain of B. microplus was dismissed by the lack of enhanced synergism by TPP or PBO, as observed in synergist bioassays, as well as by the lack of enhanced esterase activity in the Santa Luiza strain relative to the susceptible strain. The results of this study suggest that other mechanisms, including a possible new sodium channel mutation that is different from the one currently known, may be responsible for permethrin resistance in the Santa Luiza strain of B. microplus.

Segregation of European Corn Borer,Ostrinia nubilalis, Aminopeptidase 1, Cadherin, and Bre5-Like Alleles, from a Colony Resistant toBacillus thuringiensisCry1Ab Toxins, are not Associated with F2Larval Weights when Fed a Diet Containing Cry1Ab

Protein receptors may be required for activated Bacillus thuringiensis Cry toxins (Cry1Ab) to bind midgut epithelium prior to pore formation. Single nucleotide polymorphism markers from two Ostrinia nubilalis Hübner (Lepidoptera: Crambidae) midgut peptide receptors, cadherin (OnCad), aminopeptidase N 1 (OnAPN1), and OnBre5 (Onb3GalT5; a β-1,3-galactosyltransferase family 5 member) were used to examine segregation in F2 families derived from paired matings of Cry1Ab-resistant females and Cry1Ab-susceptible males. Genotypic frequencies for these markers did not deviate from Mendelian expectations. Analysis of F2 larvae indicate the segregation of single nucleotide pores in OnAPN1, OnBre5 (Onb3GalT5), and OnCad marker loci were independent of the segregation of logio weights of larvae feeding on Cry1Ab diet.

Acetylcholinesterase in the sea urchin Lytechinus variegatus: characterization and developmental expression in larvae.

Acetylcholinesterase (AChE) in the echinoid Lytechinus variegatus has been characterized. Kinetic parameters V(max), K(m), K(ss), and b were 2594+/-1048 nmol ATCh hydrolyzed/min/mg tissue wet weight, 185+/-11 microM, 308+/-100 mM, and 0.2, respectively for the substrate ATCh and 17.8+/-6.87 nmol BTCh hydrolyzed/min/mg tissue wet weight, 654+/-424 microM, 36+/-31 mM, and 0.6, respectively for BTCh. Pharmacologic analyses were performed with four inhibitors of cholinesterases, physostigmine, BW284c51, ethopropazine, and iso-OMPA, and yielded IC(50) values of 106+/-4 nM, 718+/-118 nM, 2.57+/-0.6 mM, and >0.0300 M, respectively. Both kinetic and pharmacologic results confirmed the existence of AChE in larval L. variegatus. Dimeric and tetrameric globular forms (determined by velocity sedimentation on sucrose gradients) were present in L. variegatus larvae. Activity of AChE increased significantly as larvae progressed in development from embryos to eight-arm larvae. Acetylcholinesterase activity of F1 larvae derived from sea urchins collected from wild populations and of F1 larvae derived from sea urchins cultured in the laboratory and fed two different diets suggest that the nutritional and/or environmental history of the adult sea urchin affect the developmental progression of AChE activity in the F1 offspring.

Local adaptation and maternal effects in two species of marine gastropod (genus Crepidula) that differ in dispersal potential

Adaptation to local conditions should be less likely in species with wide dispersal since periodic immigration from populations not under selection would counteract the short-term results of local selection. To test this hypothesis, we used 2 species of sympatric marine gastropods of the genus Crepidula that have vastly different early life dispersal stages. Offspring of C. fornicata emerge as long-lived free-swimming larvae, whereas offspring of C. convexa emerge as benthic juveniles. We investigated the local adaptation of each species at 2 locations by assessing their tolerance to the toxic metal copper in the laboratory. We conducted 96 h acute copper toxicity tests with juveniles of C. con- vexa and larvae of C. fornicata released in the laboratory by adults collected from a reference site (Lynn Harbor, Nahant, Massachusetts, USA) or a polluted site (New Bedford Harbor, Fairhaven, Massachusetts). To test for maternal effects, we also raised C. fornicata released in the laboratory (F1 generation) to sexual maturity and tested for differences in copper tolerance of their offspring (F2). As expected, juveniles of C. convexa from the polluted site were significantly more copper tolerant than juveniles from the reference site (p ≤ 0.05), suggesting local adaptation at the polluted site. Con- trary to expectations, F1 larvae of C. fornicata from the polluted site were not equally copper tolerant, but less copper tolerant than F1 larvae from the reference site. However, copper tolerance did not differ for F2 larvae of C. fornicata from the 2 sites. Our results suggest that populations of C. convexa can adapt to locally high metal concentrations, while populations of C. fornicata cannot, probably because of the periodic input of C. fornicata larvae from distant populations that do not experience high metal stress. Also, long-term exposure of C. fornicata adults to pollutants apparently decreased the copper tolerance of their offspring (F1 generation), an effect not seen in the laboratory-propa- gated F2 generation.

Ionizing Radiation for Quarantine Control of <I>Opogona sacchari</I> (Lepidoptera: Tineidae)

A discriminating irradiation dose of 150 gray (Gy) was used to determine the most tolerant immature stages of Opogona sacchari (Bojer) (Lepidoptera: Tineidae). Based on adult emergence, early and late pupae were determined to be the most tolerant stages, and they were significantly more tolerant than eggs, neonate larvae, and larvae that were 1, 2, or 3 wk old. Irradiation treatment of eggs, neonates, 1-wk-old larvae, 2-wk-old larvae, 3-wk-old larvae, early pupae, and late pupae at 150 Gy resulted in a 96, 96, 95, 73, 61, 8, and 9% reduction in adult emergence, respectively. Pupae were treated with irradiation doses between 60 and 400 Gy. Emergence to the adult stage was significantly reduced by irradiation, averaging 90% in experimental controls and 29% in the 400-Gy treatment. Egg production was also reduced by irradiation, although the average age of pupae at the time of irradiation had a larger effect on fecundity. In total, 2,527 pupae treated with 120 Gy eclosed and produced 47,221 eggs and three F1 larvae. In the 150-Gy treatment, 2,927 adults in total emerged from the 4,626 insects treated as pupae. These adults laid 62,878 eggs, none of which hatched. We conclude that a minimum dose of 150 Gy should be sufficient for sterilization of immature O. sacchari infesting export commodities.

Ionizing Radiation for Quarantine Control of Opogona sacchari (Lepidoptera: Tineidae)

A discriminating irradiation dose of 150 gray (Gy) was used to determine the most tolerant immature stages of Opogona sacchari (Bojer) (Lepidoptera: Tineidae). Based on adult emergence, early and late pupae were determined to be the most tolerant stages, and they were significantly more tolerant than eggs, neonate larvae, and larvae that were 1, 2, or 3 wk old. Irradiation treatment of eggs, neonates, 1-wk-old larvae, 2-wk-old larvae, 3-wk-old larvae, early pupae, and late pupae at 150 Gy resulted in a 96, 96, 95, 73, 61, 8, and 9% reduction in adult emergence, respectively. Pupae were treated with irradiation doses between 60 and 400 Gy. Emergence to the adult stage was significantly reduced by irradiation, averaging 90% in experimental controls and 29% in the 400-Gy treatment. Egg production was also reduced by irradiation, although the average age of pupae at the time of irradiation had a larger effect on fecundity. In total, 2,527 pupae treated with 120 Gy eclosed and produced 47,221 eggs and three F1 larvae. In the 150-Gy treatment, 2,927 adults in total emerged from the 4,626 insects treated as pupae. These adults laid 62,878 eggs, none of which hatched. We conclude that a minimum dose of 150 Gy should be sufficient for sterilization of immature O. sacchari infesting export commodities.

Development of Culex quinquefasciatus resistance to Bacillus sphaericus strain IAB59 needs long term selection pressure

Abstract Culex quinquefasciatus may exhibit resistance to the biolarvicide Bacillus sphaericus (Bsp), when subjected to high selection pressure. The major insecticidal factor present in most Bsp products is the binary (Bin) toxin. The IAB59 strain, in particular, has attracted attention, since it shows another toxic factor, besides the Bin toxin, and this feature is likely to decrease the risk for selecting resistance. The aim of this study was to investigate whether it was possible to select a Cx. quinquefasciatus colony (CqRL2/IAB59) for high level resistance towards Bsp strain IAB59, under laboratory conditions. Selection performed by treating groups of about 15,000 third and fourth instar larvae from each generation with high concentrations of Bsp IAB59 yielded a 70% average pre-imaginal mortality. The CqRL2/IAB59 colony reached a high level of resistance (≈40,000-fold), after 72 generations. This colony also displayed a high level of cross-resistance to strain 2362, close to 70,000-fold, while larvae remained susceptible to Bti. Cross-resistance to strain 2362 was observed before the colony had attained a high level of resistance to strain IAB59. Data from the reciprocal crosses, between individuals from the susceptible and the resistant colonies, showed that resistance is inherited as a recessive trait, since the LC50 of Bsp IAB59 towards F1 larvae were similar to that of the susceptible parental colony. Data from this study showed that continuous selection pressure with Bsp IAB59 can select Culex resistant populations, although the evolution of resistance is much slower, than formerly observed for the commercial 2362 strain.

Frequency of Resistance to <I>Bacillus thuringiensis</I> Toxin Cry1Ab in Greek and Spanish Population of <I>Sesamia nonagrioides</I> (Lepidoptera: Noctuidae)

The high-dose/refuge strategy is considered as the main strategy for delaying resistance in target pests to genetically modified crops that produce insecticidal proteins derived from Bacillus thuringiensis Berliner. This strategy is based on a key assumption that resistance alleles are initially rare (<10(-3)). To test this assumption, we used an F2 screen on natural populations of Sesamia nonagrioides Lefebvre (Lepidoptera: Noctuidae) from Greece and Spain. In total, 75 lines from Greece and 85 lines from Spain were screened for survival of F2 larvae on Cry1Ab corn, Zea mays L., leaves. No major resistance alleles were found. The frequency of resistance alleles in the Greek population was <9.7 x 10(-3) with 95% probability, which was very similar to that of the Spanish population (<8.6 x 10(-3) with 95% probability), and the expected frequencies were 3.2 x 10(-3) (0-0.0097) and 2.9 x 10(-3) (0-0.0086) in Greece and Spain (pooled 1.5 x 10(-3)). The experiment-wise detection probability of resistance was 94.0 and 97.5% for the Greek and the Spanish population, respectively. Evidence of alleles conferring partial resistance to Cry1Ab was found only for the Greek population. The frequency of alleles for partial resistance was estimated as 6.5 x 10(-3) with a 95% credibility interval between 8 x 10(-4) and 17.8 x 10(-3) and a detection probability of 94%. Our results suggest that the frequency of alleles conferring resistance to CrylAb, regarding the population of S. nonagrioides, may be rare enough so that the high-dose/refuge strategy could be applied with success for resistance management.

Effects of maternal diet quality on offspring performance in the rove beetle Tachyporus hypnorum

Abstract1. The reproductive output of an individual is known to be influenced by diet quality, but the quality of the parent's diet can also influence the performance of the offspring. Dietary maternal effects may interact with the effects of the offspring's diet to produce a variety of response patterns.2. Maternal effects were investigated in a polyphagous predator, the rove beetleTachyporus hypnorum, using three single‐species diets: two low‐quality diets consisting of the aphidsSitobion avenaeandRhopalosiphum padi, and the high‐quality controlDrosophila melanogasterdiet. Offspring of females fed these diets were raised on the same monotypic diets and allowed to reproduce. Several fitness parameters were measured to indicate possible maternal effects in both F1 and F2 generations.3. Maternal diet effects in F1 were found in egg size, hatching success, time to hatching, larval development time, larval survival, and sex ratio. Both aphid diets resulted in smaller eggs. A diet ofR. padiresulted in reduced hatching success, longer time spent in the egg stage, and a female‐biased sex ratio. A maternal diet ofR. padialso prolonged larval development onS. avenaediet, while a maternal diet ofS. avenaedecreased survival on theR. padidiet. These effects were independent of egg size.4. A maternal diet ofR. padienhanced the survival of F1 larvae raised on the same diet. Developmental selection operating through a high egg mortality may be the explanation for this seemingly positive effect.5. Sitobion avenaealone caused a significant reduction in the hatching success of F2 eggs, thus revealing grandmaternal effects.6. The prediction that polyphagous predators are less likely to evolve adaptive maternal effects is supported by the fact that none of the documented maternal effects could be interpreted as adaptive.

Mode of Inheritance of Amitraz Resistance in a Brazilian Strain of the Southern Cattle Tick, Boophilus microplus (Acari: Ixodidae)*

The southern cattle tick, Boophilus microplus (Canestrini), has developed resistance to amitraz in several countries in recent years. A study was conducted at the USDA Cattle Fever Tick Research Laboratory in Texas to investigate the mode of inheritance of amitraz resistance with cross-mating experiments. The Muñoz strain, a laboratory reared acaricide-susceptible reference strain, was used as the susceptible parent and the Santa Luiza strain, originating in Brazil, was used as the resistant parent. A modified Food and Agriculture Organization Larval Packet Test was used to measure the levels of susceptibility of larvae of the parental strains, F1, backcross, F2, and F3 generations. Results of reciprocal crossing experiments suggested that amitraz resistance was inherited as an incomplete recessive trait. There was a strong maternal effect on larval progeny's susceptibility to amitraz in both the F1 and the subsequent generations. The values of the degree of dominance were estimated at -0.156 and -0.500 for the F1 larvae with resistant and susceptible female parents, respectively. Results of bioassays on larval progeny of the F1 backcrossed with the resistant parent strain and that of the F2 generations suggested that more than one gene was responsible for amitraz resistance in the Santa Luiza strain. Comparisons of biological parameters (engorged female weight, egg mass weight, and female-to-egg weight conversion efficiency index) indicated significant differences between different genotypes. The differences appeared to be heritable, but not related to amitraz resistance. Results from this study may have significant implications for the management of amitraz resistance.

Fitness of Cry1A-Resistant and -Susceptible &lt;I&gt;Helicoverpa armigera&lt;/I&gt; (Lepidoptera: Noctuidae) on Transgenic Cotton with Reduced Levels of Cry1Ac

The performance of Helicoverpa armigera (Hübner) on 15-wk-old cotton plants was compared for a susceptible strain, a near-isogenic laboratory-selected strain, and F1 progeny of the two strains. Glasshouse experiments were conducted to test the three insect types on conventional plants and transgenic plants that produced the Bacillus thuringiensis (Bt) toxin Cry1Ac. At the time of testing (15 wk), the Cry1Ac concentration in cotton leaves was 75% lower than at 4 wk. On these plants, < 10% of susceptible larvae reached the fifth instar, and none survived to pupation. In contrast, survival to adulthood on Cry1Ac cotton was 62% for resistant larvae and 39% for F1 larvae. These results show that inheritance of resistance to 15-wk-old Cry1Ac cotton is partially dominant, in contrast to results previously obtained on 4-wk-old Cry1Ac cotton. Growth and survival of resistant insects were similar on Cry1Ac cotton and on non-Bt cotton, but F1 insects developed more slowly on Cry1Ac cotton than on non-Bt cotton. Survival was lower and development was slower for resistant larvae than for susceptible and F1 larvae on non-Bt cotton. These results show recessive fitness costs are associated with resistance to Cry1Ac.

Conditional expression of a myocardium‐specific transgene in zebrafish transgenic lines

To develop the first heart-specific tetracycline (Tet)-On system in zebrafish, we constructed plasmids in which the cardiac myosin light chain 2 promoter of zebrafish was used to drive the reverse Tet-controlled transactivator (rtTA) and the green fluorescent protein (GFP) reporter gene was preceded by an rtTA-responsive element. In the zebrafish fibroblast cell-line, rtTA-M2, one of rtTA's derivatives, demonstrated the highest increase in luciferase activity upon doxycycline (Dox) induction. We then generated two germ lines of transgenic zebrafish: line T03 was derived from microinjection of a plasmid containing rtTA-M2 and a plasmid containing a responsive reporter gene, whereas line T21 was derived from microinjection of a single dual plasmid. Results showed that line T21 was superior to line T03 in terms of greater GFP intensity after induction and with of minimal leakiness before induction. The photographic images of induced GFP in the heart of F2 larvae showed that the fluorescent level of GFP was dose-responsive. The level of GFP expressed in the F3 3 days postfertilization larvae that were treated with Dox for 1 hr decreased gradually after the withdrawal of the inducer; and the fluorescent signal disappeared after 5 days. The GFP induction and reduction were also tightly controlled by Dox in the F3 adult fish from line T21. This Tet-On system developed in zebrafish shows much promise for the study of the gene function in a specific tissue at the later developmental stage.

Relative Fitness of Cry1A-Resistant and -Susceptible &lt;I&gt;Helicoverpa armigera&lt;/I&gt; (Lepidoptera: Noctuidae) on Conventional and Transgenic Cotton

Glasshouse and laboratory experiments were conducted to evaluate the relative fitness of Cry1A-susceptible and laboratory-selected resistant strains of Helicoverpa armigera (Hübner). Life history parameters of H. armigera larvae feeding on young cotton plants showed a significant developmental delay of up to 7 d for the resistant strain compared with the susceptible strain on non-Bacillus thuringiensis (Bt) cotton. This fitness cost was not evident on artificial diet. There was no developmental delay in the F1 hybrid progeny from the reciprocal backcross of the resistant and susceptible strains, indicating that the fitness cost is recessive. In two cohorts tested, survival to pupation of resistant larvae on Bt cotton expressing Cry1Ac was 54 and 51% lower than on non-Bt cotton, whereas all susceptible and F1 larvae tested on Cry1Ac cotton were killed. Mortality of susceptible larvae occurred in the first or second instar, whereas the F1 larvae were able to develop to later instars before dying, demonstrating that resistance is incompletely recessive. The intrinsic rate of increase was reduced by >50% in the resistant strain on Cry1Ac cotton compared with the susceptible strain on non-Bt cotton. There was a significant reduction in the survival of postdiapausal adults from the resistant strain and the F1 strains, indicating that there is a nonrecessive overwintering cost associated with Cry1A resistance in H. armigera.

Evidence for additive polygenic control of pupation height in Drosophila ananassae.

In order to study the mode of inheritance of pupation height in Drosophila ananassae, two mass culture stocks derived from ecogeographically different localities in India, were used to make a complete set of 16 crosses, which include parentals, F1, backcrosses, and F2. Pupation height defined as the distance a larva pupates over the surface of culture medium was scored in all 16 crosses. The two parental lines showed significant difference in pupation height. The F1 larvae in both reciprocal crosses had intermediate pupation height and there was no difference between two reciprocal crosses as well as between F1 and mid parent value. However, there was greater variance in the F2 generation. These findings provide evidence that the inheritance of pupation height fits a classical additive polygenic model and suggested that there is substantial amount of additive genetic variation in natural populations of D. ananassae. Furthermore, the analysis of reciprocal backcrosses shows significant maternal effect. Progeny with low pupating mothers showed lower pupation height than those with low pupating fathers and progeny with high pupating mothers had higher pupation height than those with high pupating fathers. Since the maternal effect was found only in backcrosses but not in the F1, it is suggested that this maternal effect on pupation height follows the pattern of inheritance of a transient maternal effect.