The circadian rhythm of a behavioral photoresponse in the dinoflagellate Gyrodinium dorsum

Summary The circadian rhythm of the photoresponse to blue light in the dinoflagellate Gyrodinium dorsum Kofoid was investigated by the use of a closed circuit television system. The initial cessation of movement upon stimulation (stop-response) was used as the index of light reception. Under constant dark conditions cells grown on a 12L:12D regime show an endogenous circadian rhythm in their stop-response with maximum responsiveness occurring approximately one hour before the beginning of the expected light phase. This rhythmic response was only observed if the cells were irradiated with red light (620 nm) prior to stimulation with blue light. After preirradiation both far-red reversibility and the shift in the stop-response action spectrum from 470 nm to 490 nm could also be demonstrated. These findings may be related to the diurnal migration of marine dinoflagellates.This study was supported by National Science Foundation grant GB 5137.     

Photoresponses of Chaoborus larvae

The diel vertical migration of Chaoborus larvae is a well known phenomenon. In order to quantify the ability of larvae to utilize underwater light cues in their migration, we measured photoresponses of fourth-instar Chaoborus punctipennis larvae in the laboratory. The action spectrum for these larvae was characterized by a maximum in sensitivity at 400 nm, a plateau at a lower sensitivity from 480 to 560 nm, and a region of much lower sensitivity at wavelengths longer than 620 nm. Dark-adapted larvae exhibited a positive phototaxis at low light intensity which shifted to a negative phototaxix as light intensity increased. At 540 nm the threshold intensity was 1.5 × 10^?9 W/m² for positive phototaxis and about 10^?6 W/m² for negative phototaxis. Light adaptation decreased sensitivity and altered the phototactic pattern. Larvae have a clear circadian rhythm in negative phototaxis, in which greatest responsiveness occurs early in the day. We suggest that the rhythm in photoresponsiveness primarily controls the timing of the downward migration at dawn.     

铜绿丽金龟对不同光谱的行为反应

【目的】旨在获得铜绿丽金龟 Anomala corpulenta Motschulsky敏感的波谱范围及性别差异。【方法】本研究利用室内行为学的方法,测试了在波长为340~610 nm的14个单色光刺激下铜绿丽金龟的趋、避光行为反应,并计算趋、避光反应率曲线。【结果】观测显示,各单色光均能引起铜绿丽金龟产生一定的趋、避光反应,其雌雄虫的趋光敏感光区位于紫外光（405 nm）、蓝光（460 nm）和绿光区（505和570 nm）,性别对趋光行为有一定的影响,但是仅存在于趋光反应率曲线波峰的大小,而对波峰的位置没有影响。铜绿丽金龟的避光行为反应,在雌雄之间无明显的性别分化,其避光敏感光谱为紫外光（380 nm）、蓝光（440 nm）和绿光区（492和505 nm）。【结论】铜绿丽金龟成虫对不同波长光的趋性存在差异,性别对其光谱行为反应有一定的影响。该研究结果为铜绿丽金龟光视觉的深入研究奠定必要的理论基础,也为利用其趋光性对该金龟甲进行综合治理提供科学依据。     

Phototactic behaviour of Pachyneuron aphidis (Hymenoptera: Pteromalidae) – hyperparasitoid of Myzus persicae (Hemiptera: Aphidiae)

We investigated the spectral sensitivity and response to light intensity of Pachyneuron aphidis (Hymenoptera: Pteromalidae), which is a common hyperparasitoid of Aphidius gifuensis (Hymenoptera: Braconidae), a key natural enemy of the green peach aphid, Myzus persicae (Hemiptera: Aphididae). To do so, we used 15 monochromatic lights (emitting various specific wavelengths from 340 to 649 nm) and white light. P. aphidis adults are diurnal insects that show a positive phototaxis to a broad spectrum of light. Significant differences were found between sexes in the phototactic responses of P. aphidis to different monochromatic lights. Female P. aphidis showed four peaks of sensitivity at 380, 450, 504 and 589 nm. Male P. aphidis show two peaks of sensitivity, one at 450 nm and second at 628 nm. P. aphidis adults showed an increased phototactic response at low intensities and a decreased phototactic response at high intensities for both UV light and blue light. This experiment will help provide a scientific basis for the development of colour traps for insect pest management.     

Multiple light inputs control phototaxis in Synechocystis sp. strain PCC6803

The phototactic behavior of individual cells of the cyanobacterium Synechocystis sp. strain PCC6803 was studied with a glass slide-based phototaxis assay. Data from fluence rate-response curves and action spectra suggested that there were at least two light input pathways regulating phototaxis. We observed that positive phototaxis in wild-type cells was a low fluence response, with peak spectral sensitivity at 645 and 704 nm. This red-light-induced phototaxis was inhibited or photoreversible by infrared light (760 nm). Previous work demonstrated that a taxD1 mutant (Cyanobase accession no. sll0041; also called pisJ1) lacked positive but maintained negative phototaxis. Therefore, the TaxD1 protein, which has domains that are similar to sequences found in both bacteriophytochrome and the methyl-accepting chemoreceptor protein, is likely to be the photoreceptor that mediates positive phototaxis. Wild-type cells exhibited negative phototaxis under high-intensity broad-spectrum light. This phenomenon is predominantly blue light responsive, with a maximum sensitivity at approximately 470 nm. A weakly negative phototactic response was also observed in the spectral region between 600 and 700 nm. A deltataxD1 mutant, which exhibits negative phototaxis even under low-fluence light, has a similar action maximum in the blue region of the spectrum, with minor peaks from green to infrared (500 to 740 nm). These results suggest that while positive phototaxis is controlled by the red light photoreceptor TaxD1, negative phototaxis in Synechocystis sp. strain PCC6803 is mediated by one or more (as yet) unidentified blue light photoreceptors.     

Proximate control of diel vertical migration in Phyllosoma larvae of the Caribbean spiny lobster Panulirus argus

Phyllosoma larvae of the spiny lobster Panulirus argus undergo diel vertical migration (DVM), in which they are at depth during the day and nearer the surface at night. This study determined the visual spectral sensitivity of Stage I larvae and investigated whether light plays a proximate role in DVM as an exogenous cue and as an entrainment cue for an endogenous rhythm in vertical migration. Under constant conditions, larvae have a circadian rhythm (24.5-h period) in vertical swimming that resulted in a twilight DVM pattern. The behavioral response spectrum and electroretinogram recording indicated two photoreceptor spectral classes with maxima at 360 and 486 nm. When stimulated in an apparatus that simulated the underwater angular light distribution, dark-adapted larvae showed only positive phototaxis, with a threshold intensity of 1.8 × 10(13) photons m(-2) s(-1) (3.0 × 10(-5) μmoles photons m(-2) s(-1)). They have an avoidance response to predator shadows in which they descend upon sudden decreases in light intensity of more than 69%. When stimulated with relative rates of decrease in light intensity as occur at sunset they ascended, whereas they descended upon relative rates of light intensity increase as occur at sunrise. Thus, the DVM pattern is controlled by both an endogenous circadian rhythm in swimming and behavioral responses to light at sunrise and sunset.     

光谱和光强度对西花蓟马雌虫趋光行为的影响

利用行为学方法研究了光谱、光强对西花蓟马Frankliniella occidentalis (Pergande)雌成虫的趋、避光行为的影响。结果显示：（1）在340~605nm波谱内14个波长其光谱趋光行为反应为多峰型,峰间主次较明显。趋光行为反应中,蓝绿区498~524nm有一较宽峰,趋光率20.31%;其它各峰依大小次序分别位于紫光380nm、蓝光440nm;（２）避光行为反应中,蓝光440nm处略高,避光率17.19%;紫外340nm处亦有一峰,避光率15.63%;（３）随光强增强其趋光反应率增大,白光、380nm和524nm刺激时其光强趋光行为反应呈一倒“L”型式样,498nm为峰型,440nm时为一较缓的平直线型;光强最弱时仍均有一定趋光率 ,最强时均未出现高端平台;（４）随光强增强其避光反应率增大,440nm为较平缓直线;340nm刺激时为较缓波动线。结果表明：光谱对其趋光行为有很大影响,光强度的影响较大且影响大小与波长因素有关。     

Action spectra of phototactic responses of the flea beetle, Phyllotreta striolata

Abstract.  Adult flea beetles, Phyllotreta striolata , show a strong positive phototactic response. The action spectrum of phototaxis of dark-adapted beetles was measured with minimal required light intensity between the wavelength range of 300 nm and 600 nm. Male and female flea beetles showed identical phototacitc behaviours. The beetles were most sensitive to light with peak wavelengths between 350 nm and 430 nm in the morning. In the afternoon and evening, the sensitivity to wavelengths shorter than 430 nm decreased, but they remained most sensitive to 430 nm. These results suggest that changes in sensitivity of the photoreceptors or nervous integration influence the phototactic responses, and that the blue wavelengths are more attractive than others.     

Proteomic analysis of the eyespot of Chlamydomonas reinhardtii provides novel insights into its components and tactic movements

Flagellate green algae have developed a visual system, the eyespot apparatus, which allows the cell to phototax. To further understand the molecular organization of the eyespot apparatus and the phototactic movement that is controlled by light and the circadian clock, a detailed understanding of all components of the eyespot apparatus is needed. We developed a procedure to purify the eyespot apparatus from the green model alga Chlamydomonas reinhardtii. Its proteomic analysis resulted in the identification of 202 different proteins with at least two different peptides (984 in total). These data provide new insights into structural components of the eyespot apparatus, photoreceptors, retina(l)-related proteins, members of putative signaling pathways for phototaxis and chemotaxis, and metabolic pathways within an algal visual system. In addition, we have performed a functional analysis of one of the identified putative components of the phototactic signaling pathway, casein kinase 1 (CK1). CK1 is also present in the flagella and thus is a promising candidate for controlling behavioral responses to light. We demonstrate that silencing CK1 by RNA interference reduces its level in both flagella and eyespot. In addition, we show that silencing of CK1 results in severe disturbances in hatching, flagellum formation, and circadian control of phototaxis.     

光谱对东亚小花蝽趋光行为的影响

利用行为学方法研究了340—605nm波谱内14个单色光对东亚小花蝽Orius sauteri(Poppius)成虫的趋、避光行为的影响。结果显示:(1)各单色光均能引起东亚小花蝽成虫趋光行为反应,雌、雄虫的趋光行为反应曲线均为多峰型,峰间主次明显。趋光行为反应率较高的区域有3个,分别是紫外区(340、360 nm和380 nm)、绿光区(524 nm)和橙红光区(582 nm雄虫此处不高、605 nm);(2)各单色光都能引起东亚小花蝽成虫避光行为反应,雌虫避光行为反应曲线为多峰型,避光行为反应率最高的区域为蓝光近紫外区(400 nm),雄虫避光行为曲线为波浪形,避光行为反应率最高的区域为蓝光区(483 nm)。(3)性别对趋光行为反应趋势影响不大,但对趋、避光反应率有较大影响。雌虫趋光反应率最高为47.50%(380 nm),雄虫最高为35.00%(360 nm);雌虫避反应率最高为19.17%(400 nm),雄虫避光反应率最高为11.25%(483 nm)。结果表明:光谱对其趋光行为有一定影响,不同波长之间有较大差异;性别对其光谱行为反应有较大影响。     

The roles ofDrosophila ocelli and compound eyes in phototaxis

Summary Drosophila have three types of photoreceptors in their compound eyes: R1–6, R7, and R8. In addition they have simple eyes, ocelli, with another type of photoreceptor. The role of each type of receptor and the possible interaction of their inputs were examined in an innate visual preference task, fast walking phototaxis. Flies were found to be attracted to light, i.e., positively phototactic. We compared the strength of the photopositive response and the spectral preference of normal fly strains and mutant fly strains lacking functional ocelli, R1–6, or R7, singly or in combination. Electroretinographic measures were used to confirm the specificity of deficits in visual mutant strains and the normal functioning of intact receptors.The strength of the photopositive response was strong, as indicated by the high correlation between increases in the intensity of the variable stimulus and increasing numbers of flies attracted toward it. Nearly all strains with or without intact receptor types showed high correlations whether the constant intensity stimulus offered as the alternative choice was bright 467 nm light (Figs. 1 and 2) or dim 572 nm light (Figs. 3 and 4). These constant stimuli were selected so that data in relevant intensity ranges of receptor function would be obtained. An important exception to the high correlations in the intensityresponse functions occurred with flies lacking function in all receptor types except R8; their positive phototaxis was extremely weak in dim light (Fig. 3).Analyses of the phototactic spectral sensitivities (Figs. 5 and 6), as well as comparisons with known electrophysiological spectral sensitivities, were used to determine the inputs from compound eye receptors and to demonstrate central interaction of these inputs with ocellar input. Several experiments with converging evidence suggest that R7 (when present) and R8 dominate fast phototaxis in the conditions of our experiment. R1–6 is the predominant compound eye receptor type in ERG measures; however, its behavioral input is clearly demonstrated only as enhancing R8 dominance of phototaxis in experiments using a dim constant stimulus and as enhancing R7 dominance of phototaxis in experiments using a bright constant stimulus. Similarly, the presence of ocellar receptors also facilitates R8 input in dim light and R7 input in bright light. The data substantiating these respective conclusions are: (1) a lack of dim light phototaxis in a mutant strain with only R8 functional (Fig. 3); and (2) a lack of an ultraviolet (UV) maximum from R7 in bright light phototaxis in a mutant strain with only R7 and R8 functional (Fig. 5c).Generally, absence of the ocelli and R1–6 had remarkably little effect on fast phototactic behavior except for the interaction with R7 and R8 inputs. This interaction is consistent with a theory that ocelli serve to modulate compound eye sensitivity.Abbreviations ERG electroretinogram - PDA prolonged depolarizing afterpotential - R (1–6, 7, 8) retinular cell(s) - UV ultraviolet We thank K. Frayer, F. Garfinkel, K. Hansen, M. Johnson, R. Srygley, and G. Sullivan for technical assistance; K. Hansen was instrumental in running the experiments at extremely dim conditions. Supported by grants NSF-BNS-76-11921 and NIH-1-RO1-EY-02487-01A1 (to W.S.S.). Experiments reported in this paper were included in a dissertation (Karin G. Hu) submitted in partial fulfillment of the requirements of the Ph.D. degree to the Department of Psychology, The Johns Hopkins University, Baltimore, Maryland 21218. We thank members of the Graduate Board Dissertation Examining Committee for their comments: Drs. E. Blass, R. DeVoe, K. Muller and W. Sofer.     

Circadian rhythm of gravitaxis in Euglena gracilis.

Euglena gracilis, a unicellular, photosynthetic flagellate is a model system for environmentally controlled behavioral reactions. One pronounced reaction is the orientation with respect to gravity. In synchronized cultures with no cell growth a distinct circadian rhythm of negative gravitactic orientation could be observed. The main maximum of sensitivity was detected 5 h after the beginning of the subjective day, the main minimum 5 h before the beginning of the subjective day. Transferring synchronized cultures to continuous light resulted in an almost instantaneous loss of rhythmicity. In contrast, after transfer to permanent darkness cells exhibited a circadian rhythm with a progressive shortening of the period for more than 5 days. These findings are in contrast to the circadian rhythm of phototaxis in Euglena, where a free-running period of 24 h was observed. Parallel measurements of negative gravitactic orientation, velocity, cell shape as well as cAMP concentration in synchronized cultures revealed a circadian rhythm of all reactions. The results are discussed with regard to the possible role of cell shape and cAMP in gravitactic orientation.     

Gametic behavior in a marine green alga, Monostroma angicava: an effect of phototaxis on mating efficiency

The role of phototactic behavior of gametes was tested experimentally in the slightly anisogamous marine green alga Monostroma angicava Kjellman, and the effect of phototaxis on mating efficiency was discovered. Both male and female gametes showed positive phototaxis in response to a white light source. In contrast, they did not respond to a red light source. Their swimming velocity did not differ between these two illuminating light sources. It was, therefore, suggested that the search ability of the gamete itself might not vary between phototactic and non-phototactic conditions. The number of zygotes formed during the mating process may be expressed as the product of the number of encounters between male and female gametes and the fraction of encounters that result in sexual fusion. In this study, with high densities of male and female gametes mixed in test tubes, almost all minor (fewer in number) gametes fused sexually within 10 min. After dilution of the gamete suspensions by half, mating efficiency in test tubes illuminated by white light from above was higher than that in dark controls. This suggests that male and female gametes gathered at the water surface through their positive phototaxis, thus increasing the rate of encounters. Mating efficiency also decreased if the test tubes were illuminated from above by white light and also shaken. Since negative phototaxis is clearly shown in planozygotes, we suggest that positive phototaxis of male and female gametes in M. angicava is an adaptive trait for increasing the rate of gametic encounters rather than for the dispersal of zygotes as previously reported for zoospores of some marine algae. Received: 12 February 1999 / Revision accepted: 24 May 1999     

Improved automated monitoring and new analysis algorithm for circadian phototaxis rhythms in Chlamydomonas

Automated monitoring of circadian rhythms is an efficient way of gaining insight into oscillation parameters like period and phase for the underlying pacemaker of the circadian clock. Measurement of the circadian rhythm of phototaxis (swimming towards light) exhibited by the green alga Chlamydomonas reinhardtii has been automated by directing a narrow and dim light beam through a culture at regular intervals and determining the decrease in light transmittance due to the accumulation of cells in the beam. In this study, the monitoring process was optimized by constructing a new computer-controlled measuring machine that limits the test beam to wavelengths reported to be specific for phototaxis and by choosing an algal strain, which does not need background illumination between test light cycles for proper expression of the rhythm. As a result, period and phase of the rhythm are now unaffected by the time a culture is placed into the machine. Analysis of the rhythm data was also optimized through a new algorithm, whose robustness was demonstrated using virtual rhythms with various noises. The algorithm differs in particular from other reported algorithms by maximizing the fit of the data to a sinusoidal curve that dampens exponentially. The algorithm was also used to confirm the reproducibility of rhythm monitoring by the machine. Machine and algorithm can now be used for a multitude of circadian clock studies that require unambiguous period and phase determinations such as light pulse experiments to identify the photoreceptor(s) that reset the circadian clock in C. reinhardtii.     

Phototaxis in water fleas (Daphnia magna) is differently influenced by visible and UV light

The effects of vertical illumination with monochromatic lights on phototaxis of Daphnia magna in a test chamber were determined at five levels of equal quantal flux density (between 188 and 6.42 · 10⁻⁵ nEinstein). Visible adaptation light (500 nm) and subsequent spectral test light had the same quantal flux density. The animals reacted to ultraviolet light (260–380 nm) with negative phototaxis, whereas visible light (420–600 nm) caused positive phototaxis. Action spectra were determined, based on the evaluation of different parameters of phototactic behavior. The maximum spectral sensitivity in the ultraviolet was found at 340 nm. The maximum spectral efficiency in the visible varied in dependence on light intensity. Ecological consequences of the results are discussed. Accepted: 3 August 1998     

Ciliary behavior of a negatively phototactic Chlamydomonas reinhardtii

With an instrument that can record the motion of both cilia of the unicellular alga Chlamydomonas reinhardtii for many hours, the behavioral differences of its two cilia have been studied to determine their specific role in phototaxis. The organism was held on a fixed micropipette with the plane of ciliary beating rotated into the imaging plane of a quadrant photodetector. The responses to square-wave light patterns of a wide range of temporal frequencies were used to characterize the responses of each cilium. Eighty-one cells were examined showing an unexpectedly diverse range of responses. Plausible common signals for the linear and nonlinear signals from the cell body are suggested. Three independent ciliary measures--the beat frequency, stroke velocity, and phasing of the two cilia--have been identified. The cell body communicates to the cilia the direction of phototaxis the cell desires to go, the absolute light intensity, and the appropriate graded transient response for tracking the light source. The complexity revealed by each measure of the ciliary response indicates many independent variables are involved in the net phototactic response. In spite of their morphological similarity, the two cilia of Chlamydomonas respond uniquely. Probably the signals from the cell body fan out to independent pathways in the cilia. Each cilium modifies the input in its own way. The change in the pattern of the effective and recovery strokes of each cilium associated with negative phototaxis has been demonstrated and its involvement in phototactic turning is described.     

Automated Recording of Vertical Negative Phototactic Behaviour in Daphnia magna Straus (Crustacea)

Diurnal vertical migration is a well-known phenomenon in the circadian activity rhythms of zooplankton. Our goal was to test whether negative phototaxis in Daphnia magna clone BEAK (provoked by artificially induced light stress, alternating light and dark phases in 2 h intervals), and its interference with the endogenous rhythm of diurnal vertical migration, can be automatically registered with a biomonitor. For the first time the vertical swimming behaviour of D. magna was recorded quantitatively based on non-optical data recording in a fully automated biotest system, the Multispecies Freshwater Biomonitor in a new experimental setup consisting of a column of three recording units (3-level chambers). Circadian vertical migration was clearly recorded in the 3-level chambers and the rhythm was more clear with 5 than with 1 organism per chamber. The organisms clearly responded to induced light stress with negative phototaxis, however best in larger chambers. The artificially induced rhythm was influenced by the endogenous rhythm. This approach may facilitate long-term observations of vertical swimming activity of zooplankton in the future.     

Phototaxis in the magnetotactic bacterium Magnetospirillum magneticum strain AMB-1 is independent of magnetic fields

Magnetotactic bacteria (MTB) can rapidly relocate to optimal habitats by magneto-aerotaxis. Little is known about MTB phototaxis, a response that might also aid navigation. In this study, we analyzed the relationship between phototaxis and magnetotaxis in Magnetospirillum magneticum strain AMB-1. Magnotactic AMB-1 cells migrated toward light, and migration increased with higher light intensity. This response was independent of wavelength, as AMB-1 cells migrated equally toward light from 400 to 750 nm. When AMB-1 cells were exposed to zero magnetic fields or to 0.2 mT magnetic fields that were opposite or orthogonal to the light beam, cells still migrated toward the light, indicating that phototaxis was independent of magnetotaxis. The R _mag value and coercive force (H _c) of AMB-1 increased when the bacteria were illuminated for 20 h, consistent with an increase in magnetosome synthesis or in magnetosome-containing cells. These results demonstrated that the M. magneticum AMB-1 responded to light as well as other environmental factors. To our knowledge, this is the first report of phototactic behavior in the bacteria of Magnetospirillum.     

Phototactic behaviour of the parasitoid Encarsia formosa (Hymenoptera: Aphelinidae)

We investigated the spectral sensitivity and response to light intensity of Encarsia formosa (Hymenoptera: Aphelinidae), which is a key natural enemy of the greenhouse whitefly, Trialeurodes vaporariorum, and the tobacco whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae). To do so, we used 15 monochromatic lights (emitting various specific wavelengths from 340 to 649?nm) and white light. E. formosa adults, which are diurnal insects, showed a positive phototaxis to a broad spectrum of light, with peaks of sensitivity at 414, 340, 450, and 504?nm. These results show that this parasitoid is generally more sensitive to short wavelength lights than long wavelength lights across all spectral ranges tested. Furthermore, E. formosa adults showed an increased phototactic response at low intensities and a decreased response at high intensities, for both ultraviolet light and violet light. Thus, E. formosa showed both colour and intensity preferences. This experiment provides a scientific basis for the development of colour traps for insect pest management and improves understanding of the ecological significance of colour vision by E. formosa.     

Characterization of the Eyespot Regions of "Blind"Chlamydomonas Mutants after Restoration of Photophobic Responses

Chlamydomonas reinhardtii exhibits photophobic and positive and negative phototactic responses that can be defined for cell populations using computerized cell tracking and motion analysis. Mutants CC-2359 and FN68 are pigment deficient mutants that are blocked in carotenoid synthesis and lack these photo responses. In particular, neither mutant exhibits flash-induced photophobic responses to visible light stimuli to which wild-type gametic cells exhibit a strong response, with several behavioral stages. Upon addition of all-trans retinal to these mutants, the photophobic responses are restored with minor quantitative differences from wild-type populations. Using both light and electron microscopy, we have compared the ultrastructural characteristics of wild-type C. reinhardtii to those of both mutants. As previously described, wild-type cells contain an eyespot consisting of 2–4 layers of pigmented granules encased within thylakoid membranes, located between the distal extremities of the flagellar root. This structure is also visible as an orange-red spot in light microscopy. The photoreceptor is thought to be concentrated in the plasma membrane above the eyespot. The mutant, CC-2359, lacks this eyespot as seen by both light and electron microscopy, even when the photophobic response has been restored. FN68-like mutants studied earlier by Morel-Laurens and Feinlieb and others contain an eyespot which can be seen only by electron microscopy. In FN-68, the eyespot generally has the same dimensions as in wt cells, differing mainly in pigment granule appearance. Consistent with these findings, several laboratories have shown that the full range of phototactic responses can be reconstituted in FN68 and CC-2359, but that negative phototaxis requires a significantly stronger light stimulus in the latter strain. We confirm the suggestion that the eyespot is not necessary for the photophobic response, and is not critical for the appropriate assembly and function of the photophobic response receptor in the membrane. Furthermore, the locus of reconstitution of the functional receptor is not the eyespot. Because of the definitive demonstration of the absence of the eyespot in CC-2359, however, the eyespot may play a role in negative phototaxis.