Responses of Acilius sulcatus (Coleoptera: Dytiscidae) to chemical cues from perch (Perca fluviatilis)

In this study we tested the hypothesis that the presence of chemical stimuli from a hungry predator would initiate anti-predator responses, while stimuli from a satiated predator would not. We used chemical stimuli released from starved perch (Perca fluviatilis) and from satiated perch (predator). As prey we used adult Acilius sulcatus (Coleoptera: Dytiscidae). The reaction of the beetles to different predator conditions was tested during daytime. We also tested the reaction to starved perch during the night. A. sulcatus activity decreased when it was exposed to stimuli released from starved perch during daytime when visibility was poor, due to the presence of artificial vegetation. There was, however, no reaction to satiated perch under the same experimental conditions. These results indicate that A. sulcatus can discriminate between chemical cues from hungry and satiated fish predators. When visibility was good and the concentration of chemical cues was constant, the beetles did not react to starved perch in the daytime, but their activity decreased at night in response to stimuli released from starved perch. Visual as well as chemical cues seem to be important for detecting a potential predator. When visibility is good, beetles seem to rely on visual stimuli, while in darkness they seem to use chemical stimuli to detect the presence of predators. Received: 4 October 1996 / Accepted: 7 February 1997     

Role of visual and olfactory cues from agricultural hedgerows in the orientation behavior of multicolored Asian lady beetle (Coleoptera: Coccinellidae)

Harmonia axyridis Pallas is an introduced lady beetle common in eastern North American agroecosystems. Two-choice behavioral bioassays were performed to determine whether visual and olfactory stimuli from prey and host habitats could elicit taxis in wild-collected H. axyridis adults and whether beetles exhibit a preference among stimuli. Soybean aphid (Aphis glycines Matsumura) spends much of the year in agricultural hedgerows residing on buckthorn (Rhamnus cathartica L), and H. axyridis is frequently observed feeding on aphids in this habitat. Olfactory bioassays were performed in a Y-tube olfactometer and tested the response of beetles to the odor of buckthorn leaves, apple leaves (Malus domestica Borkh.), and buckthorn leaves both naturally and artificially infested with A. glycines. No differences were observed between the numbers of beetles moving toward the odor of buckthorn artificially infested with A. glycines and uninfested buckthorn, but more beetles preferred naturally infested buckthorn over uninfested buckthorn. Visual bioassays were performed in an acrylic tube arena,and tested beetle response to silhouettes and to apple and buckhorn leaves. Beetles were significantly more likely to choose silhouettes over blank space in visual trials. Significantly more beetles moved toward buckthorn leaves than blank space, but beetles did not discern between apple and buckthorn until olfactory cues were also included. This study lays the foundation for future work examining the response of H. axyridis to visual and olfactory cues in Ontario agroecosystems, which could help enhance effectiveness of H. axyridis as a biological control and mitigate its impacts as a pest species.     

Olfactory <Emphasis Type="Italic">versus</Emphasis> Contact Cues in Host Plant Recognition of a Monophagous Chrysomelid Beetle

The importance of olfactory versus contact cues for host plant recognition was investigated in the tortoise beetle Cassida canaliculata Laich. (Coleoptera: Chrysomelidae), which is strictly monophagous on meadow sage. The reaction of adult beetles to olfactory and contact host cues was tested using three bioassays (locomotion compensator, six-chamber-olfactometer, ‘stem arena’) to account for different behavioral contexts. Bioassay-guided fractionation of plant extracts was elaborated to characterize the nature of contact stimuli. The beetles were only slightly attracted to odors from small amounts of leaf material. However, when contact cues were provided additionally, the beetles showed strong preferences for samples of their host plant over controls. Bioassay-guided fractionation led to isolation of at least two non-polar contact stimuli acting in concert that are sufficient for host plant identification in C. canaliculata.     

Consequences of sequential attack for resistance to herbivores when plants have specific induced responses

Plants in nature are attacked sequentially by herbivores, and theory predicts that herbivore-specific responses allow plants to tailor their defenses. We present a novel field test of this hypothesis, and find that specific responses of Solanum dulcamara lead to season-long consequences for two naturally colonizing herbivores, irrespective of the second herbivore to attack plants. This result indicates that responses induced by the initial herbivore made plants less responsive to subsequent attack. We show that initial herbivory by flea beetles and tortoise beetles induce distinct plant chemical responses. Initial herbivory by flea beetles lowered the occurrence of conspecifics and tortoise beetles relative to controls. Conversely, initial herbivory by tortoise beetles did not influence future herbivory. Remarkably, the experimentally imposed second herbivore to feed on plants did not modify consequences (induced resistance or lack thereof) of the first attacker. Induction of plant chemical responses was consistent with these ecological effects; i.e. the second herbivore did not modify the plant's initial induced response. Thus, canalization of the plant resistance phenotype may constrain defensive responses in a rapidly changing environment.     

Suppressive effect of exposure to Chenopodium album var. centrorubrum on the feeding response of the tortoise beetle, Cassida nebulosa, to spinach

Depending upon preceding dietary exposure, the tortoise beetle, Cassida nebulosa L. (Coleoptera: Chrysomelidae), exhibits various feeding responses to spinach, Spinacia oleracea L. (Chenopodiaceae). Adult tortoise beetles reared as adults on the main host Chenopodium album var. centrorubrum Makino (CAC) (Chenopodiaceae) eat little spinach, whereas those reared on spinach accept this plant as food. In this study, we investigated the suppressive effect of exposure to CAC on the feeding response of tortoise beetles to spinach. To investigate the effect of chemical factors in CAC, we provided beetles with Gomphrena globosa L. (Amaranthaceae) as the base food and exposed them to a CAC extract, because bioassays have indicated that exposure to G. globosa did not affect the feeding response of beetles to spinach. Adults reared on G. globosa treated with CAC extract consumed significantly less than adults reared on untreated G. globosa . Thus, we determined that exposure to CAC components is responsible for the suppressive effect on the feeding response to spinach observed in the beetles. In a choice feeding test with membrane filter discs, we found that adults reared on G. globosa treated with CAC extract exhibited a greater feeding deterrent response to the spinach deterrent than adults reared on untreated G. globosa leaves. Therefore, the suppression of spinach consumption due to exposure to CAC is probably caused by the development of a response to the feeding deterrents present in spinach.     

Chemical stimuli from corn plants affect host selection and oviposition behavior of Sesamia nonagrioides (Lepidoptera: Noctuidae)

The role of chemical stimuli in host selection and oviposition of Sesamia nonagrioides (Lef) was studied in the laboratory using eight corn hybrids. Female oviposition preference was evaluated on plants of the eight hybrids, their extracts in pentane and methanol, and volatiles collected by steam distillation in ether. Results from two-choice bioassays indicate various degrees of female preference to the hybrids. Four hybrids received more eggs than control plants, whereas the remainder received fewer eggs. Relatively similar results were obtained when filter paper sticks baited with pentane and steam distillation extracts were evaluated. Methanol extracts from all eight hybrids received fewer eggs than the methanol controls. Results showed that pentane and steam distillation extracts contain chemicals that elicit female oviposition response, while methanol extracts may contain chemicals that deter or repel females to oviposition.     

Genetic activity profiles and pattern recognition in test battery selection

Computer-generated genetic activity profiles and pairwise matching procedures may aid in the selection of the most appropriate short-term bioassays to be used in test batteries for the evaluation of the genotoxicity of a given chemical or group of chemicals. Selection of test batteries would be based on a quantitative comparative assessment of the past performance of similar tests applied to other chemicals of the same structural group. The information potentially available for test-battery selection through the use of this pattern-recognition technique is considerably greater than the qualitative results obtained from individual short-term tests. Application of the method should further our understanding of the relationships between chemical properties and genotoxic responses obtained in short-term bioassays and also may contribute to our knowledge of the mechanisms of complex processes such as carcinogenesis. This approach to battery selection should be augmented by careful consideration of established principles of genetic toxicity testing; that is, a chemical should be evaluated in a battery of tests representing the full range of relevant genetic endpoints.     

Responses to olfactory and visual cues by over-wintered and summer generations of the pollen beetle, Meligethes aeneus

Abstract.  Behavioural responses to odours of oilseed rape in bud and flower stage and to green and yellow colours are studied for the two main phenological stages of pollen beetles, Meligethes aeneus , a major pest of oilseed rape, Brassica napus . Over-wintered individuals oviposit in buds of oilseed rape. Adults of the new generation (i.e. the summer generation) feed on flowers of different plant species before over wintering. In olfactometer experiments, the over-wintered beetles display a higher preference for odour of oilseed rape at the bud stage compared with the summer generation, both with and without colour stimuli. Flower odours are preferred in combination with yellow colour. Colour stimuli presented alone do not affect the behaviour. The summer generation beetles respond to both bud and flower odour. Adding colour stimuli changes the summer generations preference towards yellow and flower odour.     

Capturing Community Context of Human Response to Forest Disturbance by Insects: A Multi-Method Assessment

The socioeconomic and environmental features of local places (community context) influence the relationship between humans and their physical environment. In times of environmental disturbance, this community context is expected to influence human perceptual and behavioral responses. Residents from nine Colorado communities experiencing a large outbreak of mountain pine beetles (Dendroctonus ponderosae) were surveyed in 2007. Multiple analytic methods including ordinary least squares regression and multilevel modeling techniques were used to evaluate a community-context conceptual model of factors influencing individual actions in response to forest disturbance by beetles. Results indicated that community biophysical and socioeconomic characteristics had important impacts on participation in beetle-related actions and influenced the relationships of individual-level variables in the conceptual model with beetle-related activities. Our findings have implications for natural resource management and policy related to forest disturbances, and for developing a methodology appropriate to measure the general community context of human-environment interactions.     

Leaf volatiles as attractants for neonate Helicoverpa armigera Hbn. (Lep., Noctuidae) larvae

The 1st instar Helicoverpa armigera larvae were bioassayed in the laboratory to study their orientational responses towards leaf volatiles of four leguminous crops: chickpea, Cicer arietinum L.; pigeonpea, Cajanus cajan Millsp.; blackgram, Vigna mungo L.; and cowpea, Vigna unguiculata L. (Walp.). The gram podborer larvae showed positive orientational responses towards leaves of all four test plants. Whole leaves of chickpea, pigeonpea and blackgram were more attractive for gram podborer larvae than cowpea whole leaves. Larval attraction for crushed (damaged) leaves of chickpea, blackgram and cowpea was significantly higher than the attraction for pigeonpea crushed leaves. The orientational responses of gram podborer larvae for crushed leaves of cowpea were significantly higher compared to whole leaves. However, the whole pigeonpea leaves elicited higher orientational responses than the crushed leaves. Maceration was not observed to affect the attractancy of chickpea and blackgram leaves. Further, the leaves were extracted in n-hexane and methanol. It was observed that the crude extracts of all the test leaves elicited positive orientational responses of larvae. In no-choice tests, the orientational preference of the larvae for the hexane extracts of all the test leaves was statistically equal. Similarly, the methanol extracts of leaves of all the test plants also attracted a greater percentage of larvae in no-choice tests. However, under two-choice bioassays, hexane foliage extract of all the test plants elicited higher orientational responses of larvae compared to the methanolic extracts of same leaves. The results of these bioassays clearly indicate that all the test leaves emit kairomones for gram podborer larvae. Moreover, kairomonal components of these leaves are, at least in part, extractable in hexane and methanol, which are higher in hexane than methanol.     

The effect of human activities and their associated noise on ungulate behavior

Background

The effect of anthropogenic noise on terrestrial wildlife is a relatively new area of study with broad ranging management implications. Noise has been identified as a disturbance that has the potential to induce behavioral responses in animals similar to those associated with predation risk. This study investigated potential impacts of a variety of human activities and their associated noise on the behavior of elk (Cervus elaphus) and pronghorn (Antilocapra americana) along a transportation corridor in Grand Teton National Park.

Methodology/Principal Findings

We conducted roadside scan surveys and focal observations of ungulate behavior while concurrently recording human activity and anthropogenic noise. Although we expected ungulates to be more responsive with greater human activity and noise, as predicted by the risk disturbance hypothesis, they were actually less responsive (less likely to perform vigilant, flight, traveling and defensive behaviors) with increasing levels of vehicle traffic, the human activity most closely associated with noise. Noise levels themselves had relatively little effect on ungulate behavior, although there was a weak negative relationship between noise and responsiveness in our scan samples. In contrast, ungulates did increase their responsiveness with other forms of anthropogenic disturbance; they reacted to the presence of pedestrians (in our scan samples) and to passing motorcycles (in our focal observations).

Conclusions

These findings suggest that ungulates did not consistently associate noise and human activity with an increase in predation risk or that they could not afford to maintain responsiveness to the most frequent human stimuli. Although reduced responsiveness to certain disturbances may allow for greater investment in fitness-enhancing activities, it may also decrease detections of predators and other environmental cues and increase conflict with humans.     

Olfactory host‐finding behaviour of Oulema melanopus (Coleoptera: Chrysomelidae) and its parasitoid,Tetrastichus julis (Hymenoptera: Eulophidae)

Behavioural responses to the host‐associated olfactory cues have not been completely understood for the cereal leaf beetle, Oulema melanopus, and its principal parasitoid, Tetrastichus julis. We, therefore, investigated the role of olfactory cues in the host‐finding behaviour of these species using olfactory bioassays. Behavioural responses of O. melanopus to odours emanating from intact host plants (wheat, oat, barley) vs. a clean‐air control were tested using multichoice and two‐choice bioassays. For T. julis, responses of naïve and experienced adult female wasps to odours associated with the faecal coat of O. melanopus larvae were measured under multichoice and two‐choice conditions. Our results indicate that olfactory cues are involved in the host‐finding behaviour of both O. melanopus and T. julis. Olfactory responses of O. melanopus were influenced by the sex of the beetle and the physiological stage of adults (reproductively active vs. in reproductive diapause). Females respond to olfactory cues in greater proportions than males, and reproductively active, overwintered adults show greater responsiveness than teneral adults in reproductive diapause. Behavioural responses to cues emanating from different crop species were different in multichoice bioassays but not in two‐choice bioassays. Further, we report for the first time that the olfactory cues associated with the faecal coat of O. melanopus evoke host‐finding behaviour of its parasitoid, T. julis. Naïve female wasps are more likely to use these cues to locate the potential host than experienced females. The results of this investigation provide insights into host finding by both the species and the nature of behavioural response brought about by olfactory stimuli, and the results can help to design strategies to improve parasitoid activity by enhancing the crop environment to generate cues for host finding and to manage O. melanopus populations.     

Effect of height and color on the efficiency of pole traps for Aethina tumida (Coleoptera: Nitidulidae)

Olfactory cues released by adult bees, brood, pollen, and honey from a honey bee, Apis mellifera L., colony are the primary stimuli that guide the beetle Aethina tumida Murray (Coleoptera: Nitidulidae) to host colonies. To investigate the response of adult A. tumida to visual stimuli, we tested the influence of color and height on trap efficiency. Two pole trap colors (black and white) were evaluated at three heights (46 cm, 1 m, and 3m) from October 2008 to December 2009. A. tumida were trapped in the greatest numbers between 17 April and 15 May 2009. The lowest numbers were captured during the winter and fall. The trapping results showed that both color and trap height significantly influenced capture. The average catch in the white traps (mean +/- SE, 2.47 +/- 0.30) was significantly higher than that of the black traps (1.53 +/- 0.29) probably because white is more reflective than black. Among the heights evaluated, there were more beetles caught when traps were positioned at 46 cm (the same height as the entrance of the hives) with 3.07 +/- 0.51 beetles compared with beetles captured at 1 m (1.88 +/- 0.30) or 3 m (1.06 +/- 0.18) high. Male and female beetles exhibited similar responses to trap color and height. The relationship between the numbers of beetles in colonies and capture rates in traps was very poor and did not provide a basis to evaluate trap efficiency. In addition, because capture rates seemed generally low in relationship to the number of beetles in the apiary, substantial improvements to the trap may be necessary.     

Background gamma activity in the rabbit electroencephalogram as an index of the level of sustained (tonic) attention in the active oddball paradigm in rabbits

Tonic brain activity substantially affects the character of subjects' responsiveness to sensory stimuli. The dynamics of background gamma-band activity in rabbit electroencephalogram was investigated in the active oddball paradigm modified for animal studies. It was shown that increase in the power and coherence of gamma activity reflects the target stimulus expectancy. Correct responses to stimuli occur at a particular level of background gamma activity, which is likely to correspond to the optimal level of sustained (tonic) attention. Decrease in the level of background gamma activity leads to omissions of responses to target stimuli, while it's excessive level results in erroneous responses to nontarget stimuli (false alarms). The observed dynamics of tonic gamma activity can be interpreted as the result of variations in the level of tonic cholinergic activation of the brain cortex.     

Disentangling effects of induced plant defenses and food quantity on herbivores by fitting nonlinear models

Abstract Plants can respond to herbivore damage through both broad-scale (systemic) and localized induced responses. While many studies have quantified the impact of systemic responses on herbivores, measuring the impact of localized changes is difficult because plant tissues that have suffered direct damage may represent both a lower quality and a lower quantity of food. This article uses nonlinear models to disentangle the confounding effects of prior herbivory on food quantity and quality. The first (null) model assumes that herbivore performance is determined only by the quantity of food available to an average herbivore. Modified models allow two distinct effects of damage-induced defenses: an increase in the amount of food each herbivore is required to consume in order to achieve maximum performance and a reduction in the maximum performance even when herbivores are fed ad lib. Maximum likelihood methods were used to fit the models to data from field experiments in which Colorado potato beetle (Leptinotarsa decemlineata) larvae were reared on three varieties of potatoes that had been damaged to varying degrees by adult beetles. Prior damage reduced the mean mass of beetles at pupation, and this effect was due to both a decrease in food quantity and induced changes in food quality. In contrast, beetle survival was affected in some cases by reduced food quantity but showed no responses that could be attributed to induced defenses. I discuss this result in the context of previous studies of induced (mostly systemic) responses in the potato-potato beetle system, and I suggest that detailed studies of particular chemical responses and the proposed method of combining bioassays with quantitative models should be used as complementary approaches in future studies of herbivore-induced defenses in plants.     

Thresholds for sweet, salt, and sour taste stimuli in cockatiels (Nymphicus hollandicus)

Little is known about avian taste perception and how taste affects food choice. We designed a study to determine the concentrations of aqueous solutions of common chemical taste stimuli that result in altered consumption patterns. Using two-choice taste-preference tests, we studied the taste thresholds of caged cockatiels (Nymphicus hollandicus) for aqueous solutions of potassium chloride, a phosphate buffer (to test pH), fructose, and glucose. First, the preferred and nonpreferred bottle locations were determined for each bird. Then, depending on the compound, the test solutions were placed in bottles in either the preferred or the nonpreferred locations and water was placed in the opposite bottle. Four parameters were measured at the end of 3-day test periods (total consumption, consumption from water side, consumption from test solution side, and proportion of nonpreferred side consumption to total consumption). Experiments were repeated with increasing concentrations of test flavors until intake variables were significantly affected (P < 0.05). Cockatiels distinguished (P < 0.05) between purified water and 0.16 mol L(-1) potassium chloride, 0.40 mol L(-1) fructose, or 0.16 mol L(-1) glucose. The test birds did not distinguish between water and 0.05 mol L(-1) mono- and dibasic sodium phosphate buffer solution at any tested pH within the range of 4.9-7.7. When these findings are compared to previous experiments with the same birds, it becomes clear that the gustatory reactions of cockatiels for two different stimuli (e.g., NaCl and KCl) from the same general taste category (salt, sweet, sour) can vary widely. This variation in the responses to related stimuli could be the result of a number of factors including anion effects (for salts and acids) as well as nongustatory physiological processes (e.g., as renal control of blood osmolarity). Zoo Biol 20:1-13, 2001. Copyright 2001 Wiley-Liss, Inc.     

Oviposition pheromone in the Simulium damnosum complex: biological activity of chemical fractions from gravid ovaries

Abstract. Communal oviposition in the Afrotropical blackfly species complex Simulium damnosum Theobald (Diptera: Simuliidae) is mediated by a pheromone emitted by freshly laid eggs. Previously, two compounds (designated peaks A and B) emanating from fresh eggs were shown to be associated with attractiveness to gravid blackflies in bioassay. The present study investigated the role of these compounds by testing the responses of wild-caught Simulium yahense in Ghana to fractionated hexane extracts of gravid ovaries prepared by gas chromatography (GC). Although the fractions were prepared from Sierra Leonean Simulium leonense , GC analysis of the emissions from fresh S.yahense eggs showed that the volatile blends of both species were similar. When tested in a two-choice bioassay, 66% of ovipositing blackflies chose the substrate baited with a mixture of the four fractions recombined. In a series of bioassays testing responses to the four individual fractions presented with a control in a multiple-choice arrangement, only fraction 3 (containing peaks A and B) attracted significantly more ovipositions than the other fractions and control. However, fraction 3 failed to elicit a significant response when presented as the sole attractant with a control in a two-choice bioassay. It was concluded that fraction 3 , though mainly responsible for mediating aggregated oviposition by S.yahense , was acting in tandem with additional cues, probably further chemicals, which remain to be isolated and characterized.     

How to determine the susceptibility of Meligethes aeneus to neonicotinoids?

Three different bioassays were used to determine the susceptibility to the neonicotinoid Biscaya^® of oilseed rape pollen beetles collected from fields in Bavaria. The one in which the test substance was applied to the inner wall of glass tubes is recommended for future studies on pollen beetles because it is not dependent on the availability of plant material and provides precise information on the amount of insecticide required per unit area.     

小兴安岭森林地表鞘翅目成虫对不同干扰梯度的响应

为探讨小兴安岭凉水自然保护区森林生态系统中地表鞘翅目成虫群落对不同人为干扰梯度的响应,于2015年7月、8月和10月分别对轻度干扰[原始阔叶红松林(KY)和谷地云冷杉林(YL)]、中度干扰[阔叶红松择伐林(ZF)和次生白桦林(BH)]及重度干扰[落叶松人工林(RL)和红松人工林(RHS)]的6个林型进行取样调查。结果表明:(1)整个采样周期共捕获地表鞘翅目成虫879只,隶属9科44物种;其中轻度干扰生境共捕获6科29种251只(KY捕获5科21种150只,YL捕获4科20种101只),中度干扰生境捕获6科27种276只(ZF捕获3科20种144只,BH捕获6科23种132只),重度干扰生境捕获6科29种352只(RL捕获4科22种232只,RHS捕获5科17种120只)。(2)7月和8月步甲科和葬甲科占据数量优势,10月步甲科和葬甲科成虫数量明显减少而隐翅虫科数量占优势;不同林型及不同干扰梯度地表鞘翅目成虫物种总数和总个体数于7、8、10月均表现为下降趋势,且群落多样性也呈不同程度下降。(3)林型和月份对地表鞘翅目成虫群落结构具有显著影响,干扰梯度对群落结构无显著影响;6个林型之间地表鞘翅目成虫个体数量具有显著差异,但在物种组成上无显著差异;不同干扰梯度间地表鞘翅目成虫个体数、物种数无显著差异,且随干扰梯度变化没有明显的梯度性规律;相似性系数和聚类分析表明,属于同一干扰梯度的两个林型没有表现出高度的相似性。本研究表明干扰梯度不是决定各林型间地表鞘翅目成虫群落多样性存在差异的主要原因,凉水森林生态系统地表鞘翅目成虫对不同干扰梯度的响应不符合中度干扰假说,林型和时间则是影响地表鞘翅目成虫群落组成的显著因素,本实验为地表生物多样性保护和森林生态系统管理提供数据支撑。     

Chemotactic behavior of mutants of the nematodeCaenorhabditis elegans that are defective in osmotic avoidance

Summary Wild-typeC. elegans and seven derivative strains previously selected for absence of the wild-type tendency to avoid highly concentrated solutions (Culotti and Russell, 1978) were tested for responsiveness in 11 assays of chemotaxis, including 6 attractant and 3 repellent stimuli. The two strains altered in the geneosm-1 (P808 and P816) did not respond in any test except, possibly, one or two weak responses. Strain P801 responded to one attractant and two repellents. Strain P802 made moderately strong responses to most stimuli and avoided CO₂ in phosphate buffer as strongly as the wild-type. Of particular interest, this strain avoided OH^– which is attractive to wild-type. Strain P821 avoided CO₂ in phosphate buffer weakly, if at all, but did respond to the attractants Na⁺ and Cl^–. Conversely, strains P813 and P811 made little if any response to any attractant but did respond to the two strong repellents. Taken together with other results, these findings suggest that the osmotic response has more gene requirements in common with both attractive and repellent chemical stimuli than with thermal or mechanical stimuli. In addition, they indicate that the known chemical stimuli and the osmotic stimulus are probably mediated by at least 9 different receptors.This research was supported by the National Science Foundation and the National Institutes of Health. I would also like to thank Ms. Georgann Hardin, Mr. Kirt Rusenko, and Ms. Deborah Higgins for their assistance in carrying out the experiments. Drs. J.G. Culotti and R.L. Russell generously provided the mutant strains they had isolated.