The long-term organ culture of tissues from adult Necturus maculosus,the mud puppy

Fragments of Necturus maculosus liver, spleen and kidney were cultured at 25°C in 50% Minimal Essential Medium (MEM) or 50% Leibovitz L-15 Medium (L-15) for up to 49 days. The integrity of tissue structure was evaluated, hepatocyte cell and nuclear volumes were measured, the respiration rates of freshly-isolated and cultured liver fragments were determined, and the mitotic incidences in cultured liver, spleen and kidney were estimated. The addition of adrenalin caused a reduction in the glycogen content of liver cultures, and the subsequent addition of insulin resulted in a net increase in glycogen synthesis. Glycogen levels fell in fragments cultured in L-15, but rose in cultures in MEM. Arginase and ornithine transcarbamylase levels fell gradually throughout a 49-day culture period in L-15. Evidence presented supports the position that the survival of tissues in vitro is related to cell size and respiration rate. These experiments show that N. maculosus is a suitable donor of tissues for long-term organ culture studies on the maintenance and control of tissue-type specific structure and function.     

Cell proliferation in Xenopus tissues: A comparison of mitotic incidence in vivo and in organ culture

Using the Colcemid technique, the mitotic incidence (MI) was measured in the epidermis, lung, spleen, liver, kidney and ovarian follicular cells of metamorphosed, immature Xenopus laevis laevis. The MI was higher at 25°C than at 20°C, and there was a significant ranking correlation between organs in respect of the MI in different animals. With the exception of the liver and kidney, organ cultures showed good preservation for up to six days in vitro using a medium supplemented with 10% fetal calf serum, and values for MI comparable with or even higher than in vivo were obtained.     

Thermal effects on the biological parameters of the predatory mirid Pilophorus gallicus (Hemiptera: Miridae)

Pilophorus gallicus Remane (Hemiptera: Miridae) is a predatory mirid reported in deciduous trees in the western Mediterranean area. This work aimed to determine the biological and demographic parameters for this species at different temperatures (15, 20, 25 and 30°C). Egg hatching times shortened from 57.8 days at 15°C to 9.2 days at 30°C, and nymphal development times declined from 62.8 days at 15°C to 11.1 days at 30°C. The hatching and nymphal survival rates were low at 15 and 30°C. The lower thermal thresholds for the egg and nymphal development were 12.4 and 12.0°C, respectively. These high thermal thresholds could be a safety mechanism to avoid the emergence of nymphs in the unfavorable winter period. Female weight increased between 15 and 25°C and decreased at 30°C. The fecundity increased from 70.2 eggs per female at 15°C to 212.4 eggs per female at 25°C, and decreased to 88.5 eggs per female at 30°C. Fertility ranged from 9.4% at 15°C to 40.9% at 25°C, being 24.9% at 30°C. The intrinsic rate of natural increase (r_m) rose from 0.001 to 0.081 between 15 and 25°C and decreased to 0.05 at 30°C. In summary, this species performs poorly at low temperatures and has a relative tolerance of high temperatures (30°C); its performance was best at 25°C. Knowledge of the variation in the biological parameters with temperature may be very useful for the understanding of its ecology and population dynamics.     

The development of Puccinia hordei on barley cv. Zephyr

Germination of uredospores of Puccinia hordei was similar on cover-slips and on the first leaves of barley seedlings (cv. Zephyr) at 100 % r.h. over the range 5–25 °C, being greatest at 20 °C. At 15, 20 and 25 °C maximum germination was attained in 6 h. No uredospores germinated on coverslips in humidities below saturation. The numbers of pustules which subsequently developed on plants incubated at 5, 10, 15 or 18 °C and 100 % r.h. for varying periods up to 24 h, were directly related to rise in temperature and length of incubation. The time from inoculation to eruption of pustules (generation time) was 6 days at 25 °C, 8 days at 20 °C, 10 days at 15 °C, 15 days at 10 °C and 60 days at 5 °C. Pustule production on inoculated plants which had been kept at 5 °C was rapidly accelerated when they were transferred to 20 °C. Data obtained at constant temperatures were used to predict generation times of the fungus in the field. The productivity of pustules, determined as weight of uredospores, was examined at 10, 15 and 20 °C. Significantly more spores were produced at 15 than at 10 °C and most were produced at 20 °C. The results are discussed in relation to those obtained by other workers and to the development of brown rust in the field.     

Effects of low temperature on diapause termination and body colour change in adults of a stink bug, Plautia stali

Abstract. Diapause adults of Plautia stali Scott maintained at 20°C under short day conditions (LD 12:12 h) were exposed to four temperatures of 5–20°C to examine the effect on diapause development which was assessed in terms of oviposition. Diapause adults kept at 20°C under short day conditions changed their body colour gradually from brown to green and started egg laying after a prolonged preoviposition period. Those transferred to either 10 or 15°C also showed colour change but did not lay eggs. Bugs exposed to 5°C underwent neither body colour change nor oviposition and died more rapidly than those kept at higher temperatures. When 30-day-old diapause adults were chilled at 5, 10 or 15°C for 30 or 60 days and returned to 20°C and long day conditions (LD 16:8 h), the preoviposition period varied primarily depending on the chilling, but not on the temperature. On the other hand, when 60day-old diapause adults chilled for 30 days were observed at 20°C and long day conditions, their preoviposition period tended to be longer as the chilling temperature was lower In this case, a temperature of 10°C appeared to intensify diapause. Therefore, the effect of chilling on diapause development varied depending on the age at which insects were chilled. When chilled bugs were transferred to short day conditions at 20°C, most females failed to lay any eggs and some turned green, then after a while, some green bugs changed to brown again. These results indicate that bugs remained sensitive to short day conditions even after a 60-day chilling at 10 or 15°C.     

Effects of temperature on biology and life table parameters of the Asian citrus psyllid, Diaphorina citri Kuwayama (Homoptera: Psyllidae)

The development, survivorship, longevity, reproduction, and life table parameters of the Asian citrus psyllid, Diaphorina citri Kuwayama were evaluated at 10°C, 15°C, 20°C, 25°C, 28°C, 30°C and 33°C. The populations reared at 10°C and 33°C failed to develop. Between 15°C and 30°C, mean developmental period from egg to adult varied from 49.3 days at 15°C to 14.1 days at 28°C. The low‐temperature developmental thresholds for 1st through 5th instars were estimated at 11.7°C, 10.7°C, 10.1°C, 10.5°C and 10.9°C, respectively. A modified Logan model was used to describe the relationship between developmental rate and temperature. The survival of the 3rd through 5th nymphal instars at 15–28°C was essentially the same. The mean longevity of females increased with decreasing temperature within 15–30°C. The maximal longevity of individual females was recorded 117, 60, 56, 52 and 51 days at 15°C, 20°C, 25°C, 28°C and 30°C, respectively. The average number of eggs produced per female significantly increased with increasing temperature and reached a maximum of 748.3 eggs at 28°C (P<0.001). The population reared at 28°C had the highest intrinsic rate of increased (0.199) and net reproductive rate (292.2); and the shortest population doubling time (3.5 days) and mean generation time (28.6 days) compared with populations reared at 15–25°C. The optimum range of temperatures for D. citri population growth was 25–28°C.     

Effects of assay and acclimation temperatures on incorporation of amino acids into protein of isolated hepatocytes from the european eel, Anguilla anguilla L.

Hepatocytes of adult eels acclimated to 5° C, 10° C and 20° C, respectively were isolated by perfusion of the liver with collagenase. The liver-somatic index and the protein content of liver cells showed significantly higher values in fish kept at the lower temperatures. However, in the adenine nucleotide content and energy charge no significant differences were observed between the 5° C and the 20° C acclimation groups. The incorporation of radioactivity from a ¹⁴C-labelled amino acid mixture into perchloric acid precipitates was used as an estimate of over-all protein synthesis. When eel hepatocytes were incubated in Hanks' solution containing tracer amounts of amino acids, labelling of perchloric acid precipitates showed linear time courses over at least 60 min at 10° C and 20° C assay temperatures. The total cellular radioactivity, however, exhibited non-linear time courses. In the measurement range from 5° C to 25° C Arrhenius plots of protein labelling exhibited a discontinuity in both groups of fish. Hepatocytes from 10° C-acclimated eel showed almost twice the incorporation rates of amino acids as those from the 20° C-acclimated fish. It is concluded that high temperature dependencies in the low temperature range require an increase in the capacity of the apparatus for protein synthesis during cold acclimation.     

Common Leafspot of Alfalfa: Ascospore Germination and Disease Development in Relation to Moisture and Temperature

In a moist chamber Pseudopeziza medicaginis ascospores infected alfalfa (Medi sativa L.) moderately to abundantly within 6–10 h at 10–20 °C and within a longer time-span outside this temperature range. Approximate limits of the range were 2.5 and 28 °C; no infection took place at 30 °C. At 14°C ascospores infected alfalfa abundantly at 98 %relative humidity (RH) and above, moderately at 97%, sparsely at 95 and 96%, but not at 94% and below. Ascospores were hydrophilic, germinating best at or near 100%, RH but did not germinate at or below 93 % RH. After infection was established, tiny leafspots became visible within 6–7 days at constant temperatures of 15–25°, 10 days of 10°C, 13 days of 5 °C, and 25 days of 2.5 °C. They failed to develop into normal size spots within 4 weeks at constant temperatures near 30 °C, or near 10 °C and lower. Temporary exposure of incipiently diseased plants 1–6 days to 30–38 °C adversely affected subsequent leafspot development at 20–24°C. Inhibition depended on temperature and on the extent of post-infection disease development.     

Cross-mating experiments with geographically different populations of <Emphasis Type="Italic">Amblyomma cajennense</Emphasis> (Acari: Ixodidae)

The effect of temperature on the development and fecundity of Sancassania polyphyllae fed on tissues of Polyphylla fullo larvae was studied at 15, 20, 25, 30, and 35 ± 1°C and 65 ± 10% RH in a dark incubator. Mean developmental period of immature stages decreased significantly with increasing temperatures from 15 to 30°C. Developmental periods at 30–35°C were not significantly different. The estimated lower developmental thresholds of the various immature stages ranged between 10.1 and 11.5°C. The thermal constant for the egg-to-female adult was 93.5 degree-days. The pre-oviposition, oviposition, and post-oviposition periods and female longevity were significantly longer at 15°C than at higher temperatures. Mean total and daily fecundity were the highest at 25°C, which were significantly different from those obtained at 15, 20 and 30°C. The net reproductive rate (R ₀) was the highest at 25°C (588.3 ♀/♀). The longest mean generation time (T ₀) occurred at 15°C (36 days) and the shortest occurred at 30°C (9.2 days). The highest intrinsic rate of increase (r _m) for S. polyphyllae was observed at 25 (0.61 ♀/♀/day) and 30°C (0.62 ♀/♀/day).     

Life table and heat tolerance of Acyrthosiphon pisum (Hemiptera: Aphididae) in subtropical Taiwan

The effect of temperature on the life table of Acyrthosiphon pisum reared on Pisum sativum was evaluated under laboratory conditions using temperatures of 10, 15, 20, 25, 30, and 35°C. The development time of juvenile A. pisum decreased with increasing temperature (from 21.3 days at 10°C to 4.7 days at 35°C). Adult longevity also decreased with increasing temperature (from 53.2 days at 10°C to 2.3 days at 35°C). Interestingly, 70% and 25% of A. pisum nymphs reared at 30°C and 35°C, respectively, successfully developed into adults. These temperatures have previously been considered unsuitable for A. pisum development. However, adult aphids reared at 30°C and 35°C failed to reproduce. Linear regression analysis revealed that the lower development threshold of A. pisum was 153.1 degree‐days above 1.9°C. Maximal average reproductive capability was observed at 10°C for A. pisum adults, with each adult producing more than 120 nymphs. The intrinsic rate of increase (r_m) of A. pisum increased from 0.124/day at 10°C to 0.337/day at 25°C, whereas opposite trends were observed for the net reproductive rate (R₀) and the mean generation time (GT). At 20°C and 25°C, the intrinsic rate of increase of A. pisum was significantly higher than at 10°C and 15°C (P < 0.0001), indicating that 20°C and 25°C are within the optimal range for the growth of A. pisum, and that 30°C is beyond the upper threshold limit for reproduction, which involves a temperature range that is narrower than that of the survival range (upper limit is unknown, but above 35°C).     

Life history parameters for Nesidiocoris tenuis (Reuter) (Het., Miridae) under different temperature regimes

The development time for eggs and nymphs and female fertility were determined for Nesidiocoris tenuis Reuter (Het., Miridae: Dicyphini) at 15, 20, 25, 30, 35 and 40 ± 1°C, using tomato, Solanum esculentum (Miller), as substrate and eggs of Ephestia kuehniella Zeller as substitute prey. At 40°C, N. tenuis was unable to develop and barely reproduced. Egg development ranged from 30.8 days at 15°C to 6.3 days at 35°C. The cumulative thermal requirements for the eggs were 148.6 degree days (°d) and the lower thermal threshold, 10.3°C. The duration of the nymphal instar decreased from 55.9 days at 15°C to 8.6 days at 35°C. The thermal constant for the nymphs was 182.3 °d and the lower thermal threshold 11.7°C. No nymphs survived at 40°C, and the highest mortalities were at extreme temperatures (15 and 35°C). Female and male weights were influenced significantly by temperature. The fertility of N. tenuis females was reduced greatly at 15 and 40°C. The highest fertility during an observation period of 18 days following female emergence (79.5–60.0 nymphs per female) was within the temperature range of 20 to 35°C. Fertility was related directly to female weight and temperature (r² = 0.932). Based on development, reproduction data and thermal requirements, the optimum temperature range for N. tenuis was established as being between 20 and 30°C. Overall, N. tenuis is the most thermophilous of all dicyphines from vegetable crops in the Mediterranean area studied so far.     

Development times and fecundity of three important arthropod pests of strawberry in the United Kingdom

The development rates and fecundity of three important pests of strawberry in the UK were determined over a range of temperatures. Development time of the strawberry tarsonemid mite, Phytonemus pallidus, from egg lay to adult, ranged from a mean of 28.4 days at 12.5°C to 8.8 days at 25°C. No nymphs developed to adult at 10°C. Females lived for up to 45 days and laid a mean of 24.3 and 28.5 eggs at 20°C and 25°C respectively. Total development time from egg lay to adult for the strawberry blossom weevil, Anthonomus rubi, ranged from a mean of 95.7 days at 10°C to 18.2 days at 25°C. Mean fecundity at 20°C was 157.6 eggs, and the oviposition period averaged 71.6 days. When nymphs were reared on strawberry, development of the European tarnished plant bug, Lygus rugulipennis, from egg lay to adult, ranged from 83.8 days at 15°C to 28.8 days at 25°C. Development times on groundsel were shorter and ranged from 65.6 to 22.2 days at 15°C and 25°C. Only two nymphs developed to adults at 10°C; no eggs hatched at that temperature. Mean fecundity at 20°C was 75.4 eggs, but ranged from 23 to 179. Under a fluctuating temperature regime of 10°C for 12 h:20°C for 12 h, nymphs of L. rugulipennis took 40.3 days to become adult on strawberry, and 33.4 days on groundsel. Simple linear models fitted the developmental rate ‐ constant temperature relationship well for all species, accounting for 95–98% of the total variation in observed developmental rates. Development under fluctuating temperatures illustrated the potential problem of extrapolating linear models beyond the conditions of the experiment.     

Effects of photoperiod and temperature on diapause induction and termination in the swallowtail,Sericinus montelus

Abstract Sericinus montelus overwinters as diapausing pupae. In the present study, the effects of photoperiod and temperature on diapause induction and termination of diapause are investigated. The results obtained demonstrate that high temperature can reverse the effect of short day‐lengths on diapause induction. Under an LD 12 : 12 h photoperiod, all pupae enter diapause at 15, 20 and 25 °C, whereas all pupae develop without diapause at 35 °C. No pupae enter diapause under an LD 14 : 10 h photoperiod when the temperature is above 20 °C. Photoperiodic response curves obtained at 25 and 30 °C indicate that S. montelus is a long‐day species and the critical day‐length is approximately 13 h at 25 °C. At 25 °C, the duration of diapause is shortest when the diapausing pupae are maintained under an LD 16 : 8 h photoperiod and increases under LD 14 : 10 h and LD 12 : 12 h photoperiods. Under an LD 16 : 8 h photoperiod, the duration of diapause is shortest when the diapausing pupae are maintained at 25 °C, followed by 20 and 30 °C, and then at 15 °C. These results suggest that a moderate temperature favours diapause development under a diapause‐averting photoperiod in this species. The duration of diapause induced by an LD 12 : 12 h photoperiod is significantly longer at 25 °C than those at 15, 20 and 30 °C, and is shortest at 15 °C. At 25 °C, the duration of diapause induced by LD 6 : 18, LD 12 : 12 and LD 13 : 11 h photoperiods is similar and longer than 90 days. Thus, the diapause‐inducing conditions may affect diapause intensity and a photoperiod close to the critical day‐length has significant influence on diapause intensity in S. montelus.     

Changes in spleen histology in response to antigenic stimulation in the snapping turtle,Chelydra serpentine

Proliferative and migratory changes of lymphoid cells in the spleen were observed in turtles stimulated with KLH and maintained at 30°C. Small foci of pyroninophilic lymphoblasts were first seen in the white pulp at about day 5. Progressive enlargement of these centers continued and peaked by days 8-12. By days 15-20 the white pulp returned to a normal (unimmunized) state, while the number of pyroninophilic cells, primarily plasma cells, increased markedly in the red pulp. At days 22-25, the number of plasma cells returned to normal levels and the spleen appeared normal for the remainder of the 60 day observation period. These events suggest that at 30°C,-the turtle is capable of a strong and prompt proliferative response in the white pulp sheaths, followed by migration and differentiation of lymphoblasts into plasma cells, n the red pulp., Observations of pyroninophilic cells in sinuses, venules and veins of the spleen and a concomitant depletion of cells in red pulp, further suggest a migration from the spleen to other sites. Following a second antigenic challenge, at day 60, no significant histological changes were observed at 30°C. Nor were any changes observed following primary or secondary antigenic challenge, in animals maintained at 10°C. These findings are discussed with regard to immunological memory and low temperature immunosuppression in ectothermic vertebrates.     

Thermal responses in the citrus fruit fly,Dacus tsuneonis: evidence for a pupal diapause

Thermal responses controlling pupariation and adult eclosion in a citrus fruit fly,Dacus tsuneonis (Miyake), were studied to understand the winter biology of this species. When mature larvae were exposed to various temperature conditions, the highest percentage of pupariation was obtained at 15 °C, although the variance at this temperature was greater than at 20 °C or 25 °C. Pupariation occurred most rapidly at 20 °C and an alternating temperature with a mean of 15 °C. At constant 15 °C, pupae failed to emerge as adults. Pupae were characterized by a reduced respiration rate, which is typical of a diapausing pupa. When insects were stored at different temperatures for 45 days after pupariation, and then transferred to 25 °C, adult eclosion occurred earlier when the initial temperature was 10 °C than when it was 5 °C or 15 °C. Adult eclosion occurred most synchronously and pupal mortality was lowest when insects were stored at 15 °C for 90 days before incubation at 25 °C. These results strongly suggest thatD. tsuneonis enters a pupal diapause.     

The effect of temperature on the population growth potential of Culex annulirostris Skuse (Diptera: Culicidae)

The effect of temperature on the population growth potential of Culex annulirostris was determined by studying larval growth rate and survival at seven temperatures between 10 and 40°C, and adult survival and fecundity at 20, 25 and 30°C. All larvae died at Wand 40°C; survival was greatest at 25°C. The period for complete juvenile development ranged from 8.57 days (35°C) to 37 days (15°C). Development from egg to adult required 196 day-degrees above 9.7°C with incubation temperatures between 15 and 30°C. Population growth potential was positive at 20, 25 and 30°C, greatest at 25°C, but negative at 15°C. The minimum temperature for population growth was estimated as 17.5°C.     

Presence of larval and adult diapauses in a subtropical scarab beetle: graded thermal response for synchronized sexual maturation and reproduction

Abstract The white grub Dasylepida ishigakiensis has a 2‐year life cycle and spends approximately 9 months as a nonfeeding larva, pupa and adult on a subtropical island. Evidence is presented indicating that this beetle has two diapauses that appear to synchronize this long life cycle with the seasons. Larvae exposed to 20, 22.5, 25 and 27.5 °C late in the third (last) stadium pupate rapidly except for some individuals kept at the highest temperature. The latter pupate upon transfer to 22.5 °C, indicating that larval diapause is maintained at high temperature but terminates upon transfer to a lower temperature. Pupal development is directly temperature‐dependent in the range 20–30 °C. Adults develop reproductive organs (i.e. the ovary in females and the seminal vesicles and accessory glands in males) rapidly at 15 and 20 °C, whereas those kept at 25 °C take a long time to do so. Ovarian development is completely suppressed at 30 °C but initiated upon transfer to 20 °C. In the laboratory, males with well‐developed reproductive organs mate even with sexually immature females , whereas females with undeveloped ovaries show no sexual behaviour. Although the two diapauses of this species are thermally regulated (i.e. a characteristic commonly expressed by insects in summer diapause), adults of this beetle emerge from pupae late in the autumn and remain in the soil for 2 months. Adult diapause effectively serves to synchronize the time of sexual maturation with the coldest month of the year.     

Virus Free Garlic (Allium Sativum L.) Plants Obtained by Thermotherapy and Meristem Tip Culture

Virus infection in garlic considerably reduces yield and quality in Argentina. The production of virus free “seed” was attempted by means of thermotherapy and meristem tip culture. A hot water treatment was employed to determine the lethal temperature/time combination for clonal type (c.t.) Blanco cloves. It was established that 50°C × 20 min, 50°C × 15 min and 55°C × 5 min were the limit thermal/time combinations which garlic could withstand. Those treatments were employed followed by meristem tip culture, however, none of the successfully developed plants after culture (only 13 %) were virus-free. Hot air treatments in a growth chamber at 36°C lasting for 30, 40 and 60 days, and at 25°–32° for 30 days in a greenhouse were tested on c.t. Blanco. Cloves kept at room temperature throughout the experiment were employed as controls. In the 25°–32°C treatment, 73% of meristems produced plants and, of these, 33 % were virus free. After 30 and 40 days at 36 °C, 62 % and 67 % of the meristems developed into plantlets, of which respectively 51 % and 50 % were virus-free. Very few meristems (10 %) developed into plants when cloves had been kept at 36°C for 60 days but the resulting plantlets were all virus free. Controls produced 78 % of plants, of which 14 % were virus free. Results of hot air treatments of 36 °C for 40 days performed on c.t. Colorado, Rosado, Paraguayo, Espaol and Hilario Ascasubi were similar to those obtained with c.t. Blanco. In Espaol and Hilario Ascasubi, no virus-free plants were detected among control specimens (no thermotherapy treatment). The only virus (from up to 3 that infected the plants) that persisted in some plants after themotherapy and meristem tip culture was garlic yellow streak.     

Life table parameters of an indigenous strain of Neoseiulus californicus McGregor (Acari: Phytoseiidae) when fed Tetranychus urticae Koch (Acari: Tetranychidae)

The life table of the indigenous Neoseiulus californicus was studied at different temperatures and 65 ± 5% relative humidity under conditions of 16 h light : 8 h dark (LD 16:8). The total developmental period from egg to adult varied from 3.0 to 14.0 days at 15 to 35°C. Survival to adulthood ranges from 86.21 to 93.94%, with the highest rate at 25°C. The lower threshold temperature from egg to adult stages of females and males was 10.84 and 10.72°C, respectively, and the thermal constant was 57.14 degree‐days (DD) for females and 56.18 DD for males. Total number of eggs laid by each female was the highest (70.38 eggs) at 25°C, whereas average daily fecundity was the highest (3.69 eggs/female/day) at 30°C. The net reproductive rate was the highest (48.49) at 25°C and lowest (26.18) at 30°C. Mean generation time decreased from 19.04 to 11.47 days with increasing temperature from 20 to 30°C. Both intrinsic rate of natural increase (0.284) and finite rate of increase (1.32) were maximum at 30°C. Adult longevity was the highest (42.75 days for females and 32.60 days for males) at 20°C and lowest (22.70 days for females and 15.30 days for males) at 30°C. Sex ratio was female biased and was the highest (78.08) at 25°C and lowest (70.24) at 30°C. Developmental data of five constant temperatures, temperature thresholds and thermal requirements may be used to predict the occurrence, number of generations and population dynamics of N. californicus as an important biocontrol agent of Tetranychus urticae.     

Studies on halimeda. V. effect of temperature on chloroplast migration in this Siphonous green alga&*

Abstract

The effect of temperatures between 15 and 30°C on the daily cycle of chloroplast migration in Halimeda distorta and H tuna was determined from changes in segment pigmentation recorded by time‐lapse videography throughout the experiments. An un‐named opuntioid species was also tested between 20 and 35°C. Both the daily pattern and the amplitude of change in surface pigmentation, which were sustained for at least 5 days at 25°C, were unchanged at the higher temperatures. At 20°C the amplitude was considerably reduced but cyclical changes in surface pigmentation continued to occur throughout the 3‐day low temperature period. However, at 15°C even greater reduction in amplitude was observed together with a reduced rate of paling at the onset of darkness and absence of pre‐dawn re‐greening. Furthermore, at 15°C all daily changes in surface pigmentation had ceased by the second day in H tuna and by the third day in H distorta. These effects of lower temperatures were reversed when the plants were returned to 25°C, although after the 15°C treatment of H tuna the amplitude of the change in surface pigmentation in two of the three replicate plants was small on the first night back at 25°C whilst the third plant lost pigmentation progressively and was completely white, and apparently dead, two days later.