Lack of glyphosate resistance gene transfer from Roundup Ready soybean to Bradyrhizobium japonicum under field and laboratory conditions

A field study was conducted at the Russell E. Larson Agricultural Research Center to determine the effect of transgenic glyphosate-resistant soybean in combination with herbicide (Roundup) application on its endosymbiont Bradyrhizobium japonicum. DNA of bacteroids from isolated nodules was analysed for the presence of the transgenic 5-enolpyruvylshikimate-3-phosphate synthase (CP4-EPSPS) DNA sequence using polymerase chain reaction (PCR). To further assess the likelihood that the EPSPS gene may be transferred from the Roundup Ready (RR) soybean to B. japonicum, we have examined the natural transformation efficiency of B. japonicum strain 110spc4. Analyses of nodules showed the presence of the transgenic EPSPS DNA sequence. In bacteroids that were isolated from nodules of transgenic soybean plants and then cultivated in the presence of glyphosate this sequence could not be detected. This indicates that no stable horizontal gene transfer (HGT) of the EPSPS gene had occurred under field conditions. Under laboratory conditions, no natural transformation was detected in B. japonicum strain 110spc4 in the presence of various amounts of recombinant plasmid DNA. Our results indicate that no natural competence state exists in B. japonicum 110spc4. Results from field and laboratory studies indicate the lack of functional transfer of the CP4-EPSPS gene from glyphosate-tolerant soybean treated with glyphosate to root-associated B. japonicum.     

Physiological responses of Corythucha ciliata adults to high temperatures under laboratory and field conditions

Under high temperature conditions, insects can tolerate to survive through various physiological mechanisms, which have been well documented in laboratory studies. However, it is still unclear as to whether these laboratory data can scale up to those in the field. Here we studied dynamics of heat-induced metabolites in Corythucha ciliata adults under both laboratory and field conditions to examine their significance in thermal tolerance of the species. We compared the effects of controlled thermal treatments (2 h at 33–43 °C at 2 °C intervals in the laboratory) and naturally increasing thermal conditions (10:00–14:00 at 2-h intervals (33.5–37.2 °C) on a hot summer day in a field in Shanghai, China) on water content and levels of water-soluble protein, triglycerides, mannitol, and sorbitol in the adult bodies. The results showed that water content significantly decreased and all other metabolic parameters significantly increased in response to temperature stresses with similar patterns in both the laboratory and field, although the respective threshold temperatures were different under the two conditions. The close linkage observed in the two conditions suggests that a short period of heat stress induces water loss and accumulation of thermal metabolites in C. ciliata adults. This heat-resistance provides a defense mechanism counteracting thermal damage in C. ciliata.     

Some metabolical aspects of physical exercise under laboratory and field conditions

Parameters of exercise physiology were studied in nearly 300 subjects to resolve whether these indices were equally suitable under laboratory and field conditions to assess the level of physical fitness and optimum work load. Respiratory gas exchange, heart rate and exercise acidosis parameters were studied. The inference drawn on the basis of the obtained data has been that both the mode and the intensity of the imposed exercise exert significant influence on the variation of physiological parameters. During running either on the treadmill or in the field test, blood lactate levels were comparable, but performances related to these concentrations were not the same. When different modes of exercise were employed, also lactate levels differed between the laboratory and field studies. The performance of patients under or after exercise rehabilitation following acute myocardial infarction by using instrumental monitoring in the laboratory was found to excel that attained during rehabilitation exercise training. Any change in the level of physical fitness can only be reliably followed when physiological parameters are obtained with the same mode of exercise and intensity under the same environmental conditions. Modern training planning of sports and exercise should take into account the data derived from both the laboratory and the field studies concerning cardiorespiratory system and metabolism.     

Comparison of sexual compatibility between laboratory and wild Mexican fruit flies under laboratory and field conditions

The sexual compatibility between laboratory (LF) and wild (WF) strains of the Mexican fruit fly, Anastrepha ludens (Loew), was analyzed using analogous methodologies and experimental arenas under both laboratory and field conditions. Sexual compatibility was quantified with the following indices: the isolation index (ISI), male relative performance index (MRP), female relative performance index (FRPI), and the relative sterility index (RSI). ISI detected a certain level of incompatibility between strains under both laboratory and field conditions, because LF females tended to mate with LF males. LF mating performance was higher under laboratory than under field conditions. The relative performance indices for LF and the relative sterility index were higher in the laboratory than in the field. Differences between LF and WF in the times that males started calling and mating were observed in both environments. Importantly, WF males reduced their sexual activity under laboratory environments, whereas LF maintained similar activity levels in both conditions. The possible applications of the above-mentioned methods, not only to assess fly quality but also to determine the suitability of conditions in mass-rearing facilities, are discussed. Correlating laboratory quality to sexual behavior may contribute in the development of environmental parameters for mass-rearing facilities.     

Clutch size decisions of a gregarious parasitoid under laboratory and field conditions

Under field conditions, insect parasitoids probably experience lower rates of host encounter and life expectancy than under optimal conditions in laboratory studies. We examined the clutch size response of Mastrus ridibundus, a gregarious idiobiont parasitoid of codling moth, Cydia pomonella, cocoons, to variation in both host encounter rate and life expectancy as possible explanatory variables in a comparison of brood size in the field and laboratory. Under laboratory conditions, mean clutch size (number of eggs laid) declined from 5.8 to 3.4 as host encounter rate increased from one to eight cocoons per day. In contrast, when we reduced life expectancy by withholding honey as a food source, females did not adjust clutch size. Mean brood size (number of progeny surviving to pupation) of females foraging in walnut orchards (3.9) was significantly greater than that under laboratory conditions with excess hosts (3.1). Brood size also increased with host size in the field, but not under laboratory conditions. Brood size fitness curves were derived using both host-finding ability in the field and lifetime fecundity under laboratory conditions as indices of female fitness. Host-finding ability increased exponentially with body size, generating an estimated Lack brood size of 4.3, but lifetime fecundity increased linearly with body size, giving a Lack brood size estimate of 5.5. Under field conditions, female M. ridibundus produced brood sizes that closely approximated the Lack brood size estimated from host-finding ability, but that were significantly smaller than that estimated from lifetime fecundity. These observations suggest that, in contrast to lifetime fecundity measures from the laboratory, host-finding ability in the field provides a more accurate estimate of lifetime reproductive success for parasitoids with a low expectation of future reproduction. Copyright 2003 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.      

Equivalent inbreeding depression under laboratory and field conditions in a tree-hole-breeding mosquito

Understanding the consequences of inbreeding has important implications for a wide variety of topics in population biology. However, most studies quantifying the effects of inbreeding are performed under artificial farm, greenhouse, laboratory or zoo conditions. Although several authors have argued that the deleterious effects of inbreeding (inbreeding depression) are likely to be more severe under natural field conditions than in artificial experimental environments, these arguments are usually speculative or based on indirect comparisons. We quantified the effects of inbreeding on fitness traits in a tree-hole-breeding mosquito Aedes geniculatus) under near-optimal laboratory conditions and in three natural tree holes. Our index of fitness (Ro) was lower in the field than in the laboratory and declined due to inbreeding in both However, environments, we found no significant interactions between inbreeding depression and environmental conditions. In both the field and laboratory a 10% increase in the inbreeding coefflicient (F) led to a 12-15) decline in fitness (Ro) These results suggest that inbreeding depression will not necessarily be more extreme under natural field conditions than in the laboratory.     

Susceptibility of Cydia pomonella to Bacillus thuringiensis under laboratory and field conditions

The delta-endotoxin of 12 strains in 10 subspecies of Bacillus thuringiensis Berliner was highly active against Cydia pomonella (L.) when assayed under laboratory conditions on artificial diet. These results could not be confirmed in the field.The disappointing results obtained under field conditions are due to the behaviour of the target insect. C. pomonella larvae do not ingest food during penetration of the fruit. The larva bites pieces of the epidermis and deposits them without ingestion on top of the entry hole.

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Zusammenfassung Das delta-Endotoxin von B.t. war in Laborversuch auf Kunstmedium gegenüber den Larven des Apfelwicklers, C. pomonella, sehr aktiv. Die hohe Aktivität konnte aber unter Feldbedingungen nicht mehr bestätigt werden.Es wurde nachgewiesen, dass die unbefriedigenden Resultate von B. thuringiensis unter Feldbedingungen auf das Verhalten der Junglarven zurückzuführen sind: Die Larven nehmen während dem Eindringen in den Apfel keine Nahrung auf, sondern deponieren die herausgebissenen Epidermistücke über der Einbohrstelle.

     

Effect of pyramiding Bt and CpTI genes on resistance of cotton to Helicoverpa armigera (Lepidoptera: Noctuidae) under laboratory and field conditions

Transgenic cotton (Cossypium hirsutum L.) varieties, adapted to China, have been bred that express two genes for resistance to insects, the CrylAc gene from Bacillus thuringiensis (Berliner) (Bt), and a trypsin inhibitor gene from cowpea (CpTI). Effectiveness of the double gene modification in conferring resistance to cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), was studied in laboratory and field experiments. In each experiment, performance of Bt+CpTI cotton was compared with Bt cotton and to a conventional nontransgenic variety. Larval survival was lower on both types of transgenic variety, compared with the conventional cotton. Survival of first-, second-, and third-stage larvae was lower on Bt+CpTI cotton than on Bt cotton. Plant structures differed in level of resistance, and these differences were similar on Bt and Bt + CpTI cotton. Likewise, seasonal trends in level of resistance in different plant structures were similar in Bt and Bt+CpTI cotton. Both types of transgenic cotton interfered with development of sixth-stage larvae to adults, and no offspring was produced by H. armigera that fed on Bt or Bt+CpTI cotton from the sixth stage onward. First-, second-, and third-stage larvae spent significantly less time feeding on transgenic cotton than on conventional cotton, and the reduction in feeding time was significantly greater on Bt+CpTI cotton than on Bt cotton. Food conversion efficiency was lower on transgenic varieties than on conventional cotton, but there was no significant difference between Bt and Bt+CpTI cotton. In 3-yr field experimentation, bollworm densities were greatly suppressed on transgenic as compared with conventional cotton, but no significant differences between Bt and Bt+CpTI cotton were found. Overall, the results from laboratory work indicate that introduction of the CpTI gene in Bt cotton raises some components of resistance in cotton against H. armigera, but enhanced control of H. armigera under field conditions, due to expression of the CpTI gene, was not demonstrated.     

Effectiveness of Metarhizium anisopliae formulations against dengue vectors under laboratory and field conditions

The oviposition behaviour of Aedes aegypti and the effectiveness of Metarhizium anisopliae conidia formulated in water or oil-in-water against A. aegypti adults and eggs were tested in multi-choice and no-choice tests in oviposition devices under laboratory conditions. Both females and males rested in the devices, regardless of the formulation, and were not repelled by the presence of conidia (up to 10⁶?conidia/cm²) without oil or formulated with oil on treated filter paper arranged in the device. However, at higher oil concentrations (≥0.1?μl/cm²), regardless of the presence of conidia, the number of eggs laid by gravid females on the filter paper dropped. The susceptibility of adults, especially of males, to fungal infection increased up to a 15-day incubation. An elevated number of larvae (≥41%) eclosed from eggs laid on the moistened filter paper in the device even without submersion of eggs in water, and these larvae subsequently died. In the laboratory, 1?μl/cm² oil combined with 10⁶?conidia/cm² clearly reduced eclosion to 1.8% after submersion of eggs in water compared to ≥13% eclosion in the control. In field tests in Goiânia, Brazil, eclosion of aedine larvae from eggs laid on filter paper previously treated with oil-in-water formulated conidia dropped to between 0% and 36% compared to 22–50% in the control. Promising results of laboratory and field tests with M. anisopliae formulated in water or oil-in-water and tested in a device emphasised the effectiveness of a fungus-based formulation for aedine mosquitoes in peridomestic areas.     

Ammonium uptake by field-grown Eriophorum vaginatum roots under laboratory and simulated field conditions

Nitrogen (N) deficiencies in tundra ecosystems could be caused, in part, by the kinetics of root N uptake. The objectives of this study were to quantify NH₄ uptake by field-grown excised roots of Eriophorum vaginatum I. under controlled NH₄ concentrations (0-250 μmol I^-1) and temperatures (5-20°C) and to evaluate this laboratory derived model as a means of estimating field NH₄ uptake. There was no consistent temperature effect on root NH₄ uptake which suggests a relative in-sensitivity of E. vaginatum roots to short-term temperature fluctuations. The Michaelis-Menten equation parameters for NH₄ uptake were V_max= 22.1 μmol h^-1 g^-1 and K_m= 191 μmol I^-1. Using field NH₄ concentrations, field E. vaginatum root biomass data, and the Michaelis-Menten equation, an estimate was made of NH₄ uptake over a 42 day period; this estimate of NH₄ uptake accounted for 28% of the net incorporation of N into leaves and roots which is a reasonable estimate for E. vaginatum which relies primarily on N retranslocation for supplying new leaves and roots. Major uncertainties in field N uptake rates, model parameterization, and site characterization preclude an accurate model validation and indicate research areas most in need of future study.     

Calling behaviour of the potato aphid Macrosiphum euphorbiae oviparae under laboratory and field conditions

Abstract.  1. The calling behaviour of virgin oviparae of the potato aphid Macrosiphum euphorbiae (Thomas) (Homoptera: Aphididae) was studied at three different constant temperatures under laboratory conditions. The mean age of calling for the first time decreased with a decrease in temperature from 2.9 days at 20 °C to 2.1 days at 10 °C. At all temperature regimes, the mean onset time of calling advanced from about 5 to 3 h after the onset of the photophase, and the mean time spent calling increased by > 4 h over the 8 days.
2. Cohorts of oviparae were also observed at two different periods in late summer–early autumn in the field, to examine the effects of fluctuating abiotic conditions (temperatures, wind velocity, rain) and age on calling behaviour. As under constant laboratory conditions, the mean age of calling for the first time declined with declining temperature, from 3.7 days in early September to 1.6 days at the end of September. Age-related changes in the mean onset time of calling and the mean time spent calling were much less evident under field conditions, due to the inhibitory effects of low temperatures, high winds, and rain on female calling activity.
3. The results are discussed within the context of reproductive success and address a previously proposed hypothesis suggesting that species-specific calling windows may serve as a reproductive isolating mechanism for sympatric aphid species.     

Gas exchange responses of Chesapeake Bay tidal marsh species under field and laboratory conditions

Summary Laboratory and field gas exchange measurements were made on C₃ (Scirpus olneyi Gray) and C₄ (Spartina patens (Ait.) Mahl., Distichlis spicata (L.) Green) species from an irregularly flooded tidal marsh on the Chesapeake Bay. Laboratory measurements were made on plants grown from root stocks that were transplanted to a greenhouse and grown under high light and high nutrient conditions. The two C₄ species were similar in their laboratory gas exchange characteristics: both had higher net carbon exchange rates, higher mesophyll conductances, higher photosynthetic temperature optima and lower leaf conductances than the C₃ species. The laboratory photosynthetic water use efficiency of the C₄ species was approximately three times that of the C₃ species.Field gas exchange responses of the above species were measured in situ a Chesapeake Bay tidal marsh. Despite differences in biological potential measured in the laboratory, all three species had similar in situ carbon exchange rates on a leaf area basis. On a dry weight basis, leaves of the two C₄ species had about 1.4 times higher light saturated CO₂ assimilation rates than the C₃ species. Light saturation of CO₂ exchange occurred at photosynthetic photon flux densities of 80 n Einstein cm^-2s^-1, compared with 160 n Einstein cm ^-2s^-1 in the laboratory grown plants. Spartina patens and Scirpus olneyi had similar daily CO₂ assimilation rates, but the daily transpiration rate of the C₃ species was almost twice that of the C₄ species. Spartina patens showed greater seasonal decrease in photosynthesis than Distichlis spicata and Scirpus olneyi. The two C₄ grass species maintained higher mesophyll conductances and photosynthetic water use efficiencies than the C₄ sedge.     

Susceptibility of the cactus weevil Metamasius spinolae to Beauveria bassiana and Metarhizium anisopliae under laboratory and field conditions

Aims: To evaluate different entomopathogenic fungal isolates against the cactus weevil Metamasius spinolae under laboratory and field conditions, and select an isolate to be used as a tool in the management of this insect pest. Methods and Results: Four experiments were carried out. The effect of temperature on the in vitro growth of eight isolates of entomopathogenic fungi (two Metarhizium anisopliae and six Beauveria bassiana) was assessed. The susceptibility of adult M. spinolae to the same isolates was evaluated. Using three selected isolates, the interaction between susceptibility and sex of the insect was studied. Finally, a field experiment was carried out to evaluate infection of adult M. spinolae by the same three isolates under natural abiotic conditions. Overall, growth rate was greatest at 25°C for all the isolates. In vitro growth of M. anisopliae was greater than B. bassiana. Mortality of adult M. spinolae was greater when inoculated with B. bassiana compared with isolates of M. anisopliae. Susceptibility had no interaction with the sex of the insect. The proportion of insects succumbing to infection was smaller when incubated under field conditions than when incubated under laboratory conditions. Conclusions: The experiments described here showed a complex interaction between entomopathogenic fungi and M. spinolae, and these data allows us to select isolate Bb107 as a first step towards its use in the management of this pest insect. Significance and Impact of the Study: Our results showed that entomopathogenic fungi can be used for the control of M. spinolae, which may help reduce the use of chemical insecticides and, therefore, the exposure of Opuntia ficus‐indica producers to pesticides.     

Stability of spinosad resistance in Frankliniella occidentalis (Pergande) under laboratory conditions

The stability of spinosad resistance in western flower thrips (WFT), Frankliniella occidentalis (Pergande), populations with differing initial frequencies of resistance was studied in laboratory conditions. The stability of resistance was assessed in bimonthly residual bioassays in five populations with initial frequencies of 100, 75, 50, 25 and 0% of resistant individuals. There were no consistent changes in susceptibility of the susceptible strain after eight months without insecticide pressure. In the resistant strain, very highly resistant to spinosad (RF50>23,000-fold), resistance was maintained up to eight months without further exposure to spinosad. In the absence of any immigration of susceptible genes into the population, resistance was stable. In the case of the population with different initial frequency of resistant thrips, spinosad resistance declined significantly two months later in the absence of selection pressure. With successive generations, these strains did not change significantly in sensitivity. Spinosad resistance in F. occidentalis declined significantly in the absence of selection pressure and the presence of susceptible WFT. These results suggest that spinosad resistance probably is unstable under field conditions, primarily due to the immigration of susceptible WFT. Factors influencing stability or reversion of spinosad resistance are discussed.     

Weatherability of a steam-rolled oat groat chlorophacinone ground squirrel bait under field and laboratory conditions

Investigations of the weatherability of spot-baiting for ground squirrel control in northern CA field studies were conducted with a registered bait consisting of 0.01% chlorophacinone (an anticoagulant rodenticide) on steam-rolled oat groats. For reference purposes, a laboratory test was later conducted in an environmental chamber simulating some of the observed weather conditions. Three weathering plots were established in alfalfa for field tests. Each was baited with rodenticide fortified bait that was handled the same as for a simultaneous control project. Test areas were protected with wire mesh to prevent bait consumption by birds and mammals. Bait samples were collected daily over 7 days, then frozen, and shipped for analysis. Test No. 1 conducted under wet conditions showed a 71% loss of chlorophacinone after 1 week. Test No. 2 demonstrated a 57% loss of chlorophacinone under drier conditions. Test No. 3, a 24 h test under very wet conditions within the alfalfa field irrigated by overhead sprinklers, had a 92% loss of chlorophacinone. Laboratory studies using controlled environmental conditions: light (16 h light:8 h dark), with a mean relative humidity of 98%, and a range of temperature 11.1–27.8°C (52–82°F) showed ≈50% loss of the chlorophacinone. The magnitude was less (37% at 7 days) in the environmental chamber when corrected for water weight gain.     

Nitrogen storage and remobilization in ash (Fraxinus excelsior) under field and laboratory conditions

 Storage and remobilization of nitrogen (N) were studied in ash trees (Fraxinus excelsior) under both field and greenhouse conditions. Experiments in the greenhouse providing ¹⁵N labelled fertilizer to the trees showed that the major quantity of N remobilized during subsequent spring was from the roots, and only a small amount from the stem. This corresponded with a loss of soluble N (proteins and low-molecular-weight compounds) from both roots and stem. On the two field sites, which differed in water availability, there was a decrease in bark N content during leaf growth, but on the dry site net N export from the bark was sustained throughout the whole vegetation period. Remobilized N was derived from soluble proteins and low-molecular-weight compounds on the moist site, which was demonstrated by the seasonal dynamics of a 56 kDa polypeptide in bark and wood. On the dry site, lower contents of soluble proteins were associated with smaller amounts of N remobilized compared to the moist site. Uptake studies of ¹⁵N labelled fertilizer indicated a higher contribution of current uptake to leaf N increment during spring at the dry site compared to the moist site. Differential N availability during the season had a decisive effect on the nitrogen storage dynamics at the two sites. Thus the influence of current N supply on N remobilization and storage as found in the greenhouse-grown plants could be verified under field conditions. Received: 28 July 1995 / Accepted: 17 July 1996     

Development and efficacy of three poison baits against Blattella germanica under laboratory conditions

Bait formulations are widely used for controlling the German cockroach, but cockroaches increasingly come into aversion and resistance to bait formulations. To optimize baits and delay insecticides resistance to Blattella germanica, five insecticides with different mechanisms of action were selected to test the stomach toxicity of B. germanica, and twelve nutrient-rich foods were selected to test the palatability of B. germanica in the research. The results revealed that flufiprole and dinotefuran had a significant effect on the decrease of B. germanica population, whereas hydramethylnon, permethrin did not work well in 72 h stomach toxicity and acrinathrin had a short potency in pesticide formulation because of its semi-volatile physical properties. Furthermore, B. germanica prefers the poison bait to rat feed. This research represents the bait consisting of peanut powder and maltose (9:1) has the best palatability to B. germanica, and flufiprole and dinotefuran have a lot of potential in baits development and application.     

Variations of skull-body regressions of the lemming (Dicrostonyx groenlandicus) under laboratory and field conditions

Collared lemmings, Dicrostonyx groenlandicus , maintained in the laboratory under short-day photoperiods (2 h light: 22 h dark) grew faster than their cohorts kept under long-day photoperiods (22 h light: 2 h dark). Significant differences in cranial, visceral and body components were found in both sexes. Linear regressions of skull-body parameters were compared to those of wild lemmings trapped at Eskimo Point, Northwest Territories, Canada, during three consecutive summers from 1973 to 1975. The data support the conclusion that variation in body size and skull development associated with changes in population density are, in part, due to photoperiod influences. The importance of this facultative response is discussed in relation to current theories on thermal regulation and variations in subnivial breeding success observed in this species.