Structural aspects of akinete germination in the cyanobacterium Anabaena variabilis

Electronmicroscopical investigations of light activated akinetes in different phases before outgrowth of the germinating cell showed two alterations in the akinete envelope, obviously in connection with the germination process. After induction of germination the akinetes show formation of an expanding more or less electron dense layer between the outer cell wall layer (outer membrane, L_IV) and the condensed part of the akinete coat (the transformed sheath of the vegetative cell). Between this new formed layer and the mentioned part of the akinete coat thick laminar layers are deposited which contain alternately electron dense and electron transparent strata. The expanding layer is assumed to be a mucous layer which acts as swelling body causing, after bursting of the layered shell, the expulsion of the germinating cell in the manner characteristic for Anabaena variabilis.     

Effects of Ekalux Ec-25 on akinete germination and sporulation ofPithophora kewensis

The effect of the organophosphorus insecticide Ekalux Ec-25 (quinalphos) was studied on akinete germination, and sporulation of the green algaPithophora kewensis (Cladophorales). Initiation of akinete germination was delayed by 1–3 d with 0.025–0.1% concentrations of the pesticide used. The percentage of akinete germination was found to be markedly affected by the treatment with insecticide, except with the concentration of 0.25% Ekalux Ec-25, where percent germination was observed to be higher than in the control. Generally, the time taken for the initiation of akinete formation increased and percent sporulation decreased with the increase of concentrations from 0.025 to 0.1% of Ekalux Ec-25.     

Environmental influences on akinete germination of Anabaena circinalis and implications for management of cyanobacterial blooms

Certain cyanobacteria, including the noxious bloom-forming species Anabaena circinalis Rabenhorst, produce thick-walled reproductive structures (akinetes) which may serve as a resting stage and ensure survival during adverse growth conditions. The effect of certain environmental variables (temperature, salinity and desiccation) on akinete germination of A. circinalis was investigated under laboratory conditions, to determine the conditions under which germination was inhibited. The overall aims were to provide a broader understanding of the life history and ecology of this species and to assess suppression of akinete germination as a potential management strategy for control of cyanobacterial blooms in the lower Murray River, Australia. The results indicated a marked threshold of temperature and salinity tolerance for germination of A. circinalis, but the latter was not within a range that could be successfully manipulated in a natural ecosystem. However, it was found that desiccation of akinetes for moderately short periods can significantly impair their capacity to germinate. It is, therefore, speculated that allowing periodic drying of shallow wetlands adjacent to the Murray River and in other areas may reduce the size of the inoculum for population growth by reducing viability of akinetes in surface sediments.     

The relative importance of photoperiod and temperature as cues for seasonal acclimation of thermoregulation in pouched mice (Saccostomus campestris: Cricetidae) from southern Africa

Summary The effect of short photoperiod and cold on metabolism and thermoregulation was investigated in pouched mice (Saccostomus campestris: Cricetidae) from three localities in southern Africa which experience contrasting climatic conditions. Mice were initially acclimated to long photoperiod (14L: 10D) at 25°C, followed first by a decline in photoperiod (to 10L: 14D) and then by a fall in temperature (to 10°C). Minimum observed metabolic rate (basal metabolic rate) was unaffected by the decline in photoperiod but increased significantly following cold acclimation. Because minimal thermal conductance remained constant throughout the study the increase in minimum observed metabolic rate caused a decline in lower critical temperature to around 26°C. In contrast to minimum observed metabolic rate, regulatory non-shivering thermogenesis improved significantly following the decline in both photoperiod and temperature. However, pouched mice from the warmest locality were significantly less responsive to photoperiod than those from the other two localities whose survival might depend upon their ability to accurately predict seasonal changes in temperature. Neither photoperiod nor temperature had any effect on body mass, yet pouched mice from the most arid locality, where food supply might be unpredictable, were significantly smaller and had lower total energy requirements than those from areas experiencing higher annual rainfall. These results indicate that S. campestris displays considerable geographical variation in energy requirements together with differences in the use of photoperiod as an anticipatory cue for predicting the onset of winter. These differences appear to be related to the availability of energy and the relative severity of climatic conditions in each locality.Abbreviations ANOVA analysis of variance - BMR basal metabolic rate - C _m minimal thermal conductance - M _b body mass - MOMR minimum observed metabolic rate - MWU Mann-Whitney U-test - NA noradrenaline - NST non-shivering thermogenesis - RMR resting metabolic rate - RQ respiratory quotient - T _a ambient temperature - T _b body temperature - T _1c lower critical temperature - oxygen consumption - maximum - following NA injection     

Germination behaviour of seeds of Withania somnifera (L.) Dunal: a high value medicinal plant

In order to evolve a quick method for smooth and optimum germination for Withania somnifera- a medicinally efficacious multipurpose plant, present investigation was carried to study the effect of physico-chemical treatments, storage, temperature, photoperiod and growth regulators (GA₃, IAA, IBA, 2–4 D and BA) on germinability. The most effective treatment is GA₃ at 150 μg/ml concentration at 25 °C. The optimal temperature for germination is 25 °C and continuous light favored germination showing that photoperiod has a significant role. The seedlings derived from seeds performed well when grown in a glasshouse. The data have implications for conservation and cultivation of the species studied.     

AKINETE GERMINATION IN ANABAENA CIRCINALIS (CYANOPHTA)

Akinetes of a clonal culture of Anabaena circinalis Rabenhorst from Mt. Bold reservoir (eutrophic), South Australia, were isolated and the effects of light, phosphorus, and nitrogen availability on their germination were investigated. Light was required but there was no significant difference in percentage of germination after 72 h if akinetes were incubated in ASM-1 medium at irradiances of 15, 30, or 50 μmol.m^-2.s^-1. Maximum akinete germination occurred by 48 h. Nitrogen was not required, as 88% of akinetes germinated in the flasks without combined nitrogen added to the medium and without N₂ in the air. NH₄⁺-N at 28 mg N.L^-1 completely suppressed germination, whereas 28 mg NO₃ N.L^-1 had no effect relative to the controls without nitrogen. Phosphorus was required, and at 48 h percentage of germination in the flasks with 0.6 mg P.L^-1 added (78%) was significantly greater than in the flasks with 0.06 P.L^-1 (58%) and 0 mgP.L^-1 (24%) added. Germlings in the 0 mg P.L^-1 flasks were only 2–4 cells long and stunted in appearance, whereas germlings at all other P concentrations were 8–16 cells long. It is likely that the isolation process exposed some akinetes to intracellular phosphorus released from lysing vegetative cells, but this was insufficient to allow normal development in the 0 mg P.L^-1 flasks. The plot of percentage of germination vs. initial phosphorus concentration, in the medium showed a relationship analogous to Michaelis-Menten nutrient uptake kinetics, suggesting that a specific membrane-bound enzyme system(s) is involved, with phosphorus as the substrate. The half saturation value (K_S) for germination was 50 μg P.L^-1.     

Factors regulating recruitment from the sediment to the water column in the bloom-forming cyanobacterium Gloeotrichia echinulata

1. The influence of light, temperature, sediment mixing and sediment origin (water depth) on the recruitment of the cyanobacterium Gloeotrichia echinulata was examined in the laboratory. 2. Light and temperature were the most important factors initiating germination in G. echinulata. 3. The extent of germination (recruited biovolume) was mainly regulated by temperature and sediment mixing. Furthermore, sediment mixing significantly enhanced the frequency of observed heterocysts and colonies. 4. Despite the fact that the deep and shallow sediments contained a similar number of akinete colonies, the highest recruitment occurred from shallow sediments, indicating that akinetes from shallow sediments have a higher viability than those from deeper parts of the lake. 5. Our results support the hypothesis that shallow sediments are more important than profundal sediments for the recruitment of G. echinulata to the pelagic zone.     

Akinetes of the cyanobacteriumNostoc PCC 7524: morphological changes during synchronous germination

Following dilution into fresh medium in the light, akinetes ofNostoc PCC 7524 germinated synchronously. Synchrony was maintained at a high level during the first 24 h, at which time the young filaments were composed either of three cells (with N₂ as nitrogen source) or four cells (with NO ₃ ^- or NH ₄ ⁺ ), and at a slightly lower level during the next 24 h of growth. The pattern of cell division was similar in media containing the different nitrogen sources although the timing of the major events varied. In the presence of N₂ or NO ₃ ^- , heterocysts differentiated synchronously; the first developed invariably from a terminal cell of the young filament at approximately 19 h, the second from the other terminal cell after further vegetative cell division. Heterocyst differentiation did not occur in the presence of NH ₄ ⁺ . In the absence of nitrogen (gas phase argon: CO₂) akinete germination initially followed the same pattern as that observed in N₂, this early stage probably occurring at the expense of intracellular reserve materials.During germination, a new laminated layer, similar in structure and position to that found in the heterocyst envelope, appeared in the akinete envelope. This layer was not present in the germinating akinetes of a mutant which was incapable of forming heterocysts.     

AKINETE DIFFERENTIATION IN ANABAENA CIRCINALIS (CYANOPHYTA)1

Three lines of evidence established conclusively that phosphorus limitation triggered akinetes to differentiate in Anabaena circinalis Rabenhorst. First, akinetes differentiated when phosphorus was limited, but not when nitrogen, inorganic carbon, iron, trace elements, or light were limited, or when dissolved oxygen concentration was increased. In the phosphorus limitation experiment, akinetes appeared first in the 0 mg P-L^?1 cultures, and the higher the initial concentration of phosphorus was, the longer it took for akinetes to differentiate. Second, akinete differentiation commenced when Q_p fell to the same critical concentration in all cultures. The critical Q_p for akinete differentiation in A. circinalis was 0.3-0.45 pg P·cell^?1, and there was no significant difference between cultures grown with 0.6, 0.2, 0.06, or 0 mg P · L^?1 (F= 5.48, of = 3, P > 0.05). Similarly, there were no significant differences between P cultures in internal cellular soluble reactive phosphorus (SRP) concentration (F= 0.63, df = 3, P > 0.05) or external SRP per cell in the medium (F= 5.16, df= 3, P > 0.05) when akinete differentiation commenced. Both were between 0.01 and 0.07 pg SRP-cell^?1. A thorough literature search indicates that this information has not been reported previously. The third line of evidence came from electron micrographs, which illustrated that polyphosphate was present in trichomes prior to akinete differentiation but was absent in trichomes with akinetes indicating that phosphorus reserves were depleted when akinetes differentiated. Lipid globules (carbon reserve) and cyanophycin granules (nitrogen reserve) increased in number in trichomes with akinetes, compared to trichomes without akinetes. Thus, the ratio of internal P:C:N was different in trichomes with akinetes compared to trichomes without akinetes and may be important in activating akinete-differentiating genes.     

Interactive effects of temperature and photoperiod on the daily activity and energy metabolism of pouched mice (Saccostomus campestris: Cricetidae) from southern Africa

The daily activity and energy metabolism of pouched mice (Saccostomus campestris) from two localities in southern Africa was examined following warm (25 °C) and cold (10 °C) acclimation under long (LD 14:10) and short (LD 10:14) photoperiol. There was no differential effect of photoperiod on the daily activity or metabolism of pouched mice from the two localities examined, which suggests that reported differences in photoresponsivity between these two populations were not the result of differences in daily organisation. Neverthe-less, there was a significant increase in metabolism at 10 °C, irrespective of photoperiod, even though seven cold-acclimated animals displayed bouts of spontaneous torpor and saved 16.4–36.2% of their daily energy expenditure. All but one of these bouts occurred under short photoperiod, which suggests that short photoperiod facilitated the expression of torpor and influenced the daily energy metabolism of these individuals. As expected for a noctureal species, the amount of time spent active increased following acclimation to short photoperiod at 25 °C. However, there was a reduction in mean activity levels under short photoperiod at 10 °C, possibly because the stimulation of activity by short photoperiod was masked by a reduction in activity during bouts of spontaneous torpor. Cold temperature clearly had an overriding effect on the daily activity and metabolism of this species by necessitating an increase in metabolic heat production and eliciting spontaneous torpor which overrode the effect of short photoperiod on activity at an ambient temperature of 10 °C.Abbreviations 3-ANOVA three-way analysis of variance - %ACT percentage of time spent active - ADMR average daily metabolic rate - M _b body mass - MR metabolic rate - MR_dark metabolic rate recorded during the dark phase - MR_light metabolic rate recorded during the light phase - NST non-shivering thermogenesis - RQ respiratory quotient - STPD standard temperature and pressure, dry - T _a ambient temperature - T _b body temperature - VO₂ oxygen consumption     

Thermal niche for seed germination of Xyris species from Brazilian montane vegetation: Implications for climate change

Understanding how climate change will affect regeneration from seeds is important for developing conservation strategies. We evaluated seed germination requirements for sympatric species of Xyris from montane rupestrian grasslands (campo rupestre) in Brazil to determine their thermal niche and thermal requirements for seed germination. We also assessed whether projected temperature increases would affect seed germination of the species. Seed germination was evaluated at a wide range of constant temperatures (10–40°C) under light (12-hr photoperiod) and dark conditions. Base temperatures (T_b) and thermal times for 50% germination (θ₅₀) were calculated for three species. The effects of projected mean temperature increase on seed germination percentage and timing were evaluated. All species revealed an absolute light requirement for germination. Thermal germination niche breadth was greatest for X. asperula (15 to 35°C) and narrowest for X. seubertii (20 and 25°C). Base temperatures for X. asperula, X. pilosa and X. trachyphylla were 9.0, 12.8 and 11.1°C, respectively. In the scenario with the highest temperature increase (A2), the greatest reductions in seed germination are observed for X. pilosa and X. seubertii. The lowest projected temperature increase (2°C) was sufficient to decrease by 1 day the germination time of X. asperula and X. pilosa. Species of Xyris do not present a pattern for thermal germination niche and thermal requirements values, indicating that the effects of climate warming on the regeneration of these seeds will probably vary among species.     

Germination responses to GA3 and short‐time chilling of three endemic species: Tripleurospermum pichleri,Cirsium leucopsis and Senecio olympicus (Asteraceae)

In the present study, the germination characteristics of three endemic species from Turkey, Tripleurospermum pichleri (Boiss.) Bornm., Cirsium leucopsis D.C and Senecio olympicus Boiss. (Asteraceae), were investigated. Germination was studied for fresh seeds, for seeds subjected to short‐time chilling (15 days, moist +4°C), to GA₃ (100, 150 and 250 ppm) and a combination of chilling and GA₃; in all cases seeds were incubated either at 20/10°C day/night with light daytime or at 20°C in darkness with daily short‐time dim light (DSDL). In C. leucopsis seeds, all GA₃ treatments enhanced the final germination percentages. The mean germination time (MGT) of C. leucopsis was lower under DSDL than with photoperiod. The chilling treatment with GA₃ in DSDL significantly increased germination in S. olympicus seeds (from 45 to 87%). Germination increased to 55% in T. pichleri by chilling under photoperiod compared with 32% by chilling followed by DSDL. In conclusion, these three co‐existing endemic Asteraceae species have different germination behaviours; something that should be taken into account for ex situ propagation. However, an efficient way to germinate all species is to use 250 ppm GA₃ and 20/10°C with photoperiod.     

Photoperiod buffers responses to salt and temperature during germination of two coastal salt marsh colonizers Juncus acutus and Juncus maritimus

Juncus acutus and J. maritimus are two colonizers of coastal marsh rangeland with worldwide distribution. We tested the effect of salt, temperature, and photoperiod on the germination capacity of seeds of the two Juncus from the Rhône delta (south of France). We measured the first day of germination, the mean time, speed, and rate of germination on seeds subjected to five salinity levels, three Dark–Light temperatures and two D-L photoperiods (12–12 and 10–14). The 10–14 D-L photoperiod, although it corresponds to one of the two main germination periods of J. acutus and J. maritimus in the northern Mediterranean, has never been previously used to study their germination capacity. Analyses showed significant effects of salinity and temperature on the germination parameters tested, and for these two factors we found results comparable to those of previous studies. Salinity slowed down and reduced the germination process while spring temperatures had a positive effect. A surprising result was obtained by changing the D-L photoperiod from 12–12 to 10–14 which then largely buffered the negative effect of salt and high temperatures on germination. This capacity of J. acutus and J. maritimus to withstand salty conditions during the germination phase due to the spring photoperiod could be decisive in the ability of both species to colonize saline environments.     

Summer acclimatization in the short-tailed field vole,Microtus agrestis

We investigated the changes that occurred in basal and noradrenaline-induced metabolic rate, body temperature and body mass in short-tailed field voles,Microtus agrestis, during exposure to naturally increasing photoperiod and ambient temperature. These parameters were first measured in winter-acclimatized voles (n=8) and then in the same voles which had been allowed to seasonally acclimatize to photoperiod and ambient temperature (6 months later). Noradrenaline induced metabolic rate, basal metabolic rate and nonshivering thermogenesis were significantly higher in winter-acclimatized compared to summer-acclimatized voles. There was a significant positive relationship between basal metabolic rate and noradrenaline-induced metabolic rate. Body mass was significantly higher in summer-acclimatized compared to winter-acclimatized voles. There was a significant positive relationship between body mass and noradrenaline-induced metabolic rate in both winter-acclimalized and summer-acclimatized voles; however, there was no relationship between basal metabolic rate and body mass in either seasonal group of voles. Body temperature after measurements of basal metabolic rate was not significantly different in the seasonal cohorts of voles. However, body temperature was significantly higher in winter-acclimatized compared to summer-acclimatized voles after injection of noradrenaline. Previously we have found that a long photoperiod was not a sufficient stimulus to reduce thermogenic capacity in winter-acclimatized voles during cold exposure, since basal metabolic rate increased to compensate for a reduction in regulatory nonshivering thermogenesis. Here we found that a combination of increased ambient temperature and photoperiod did significantly reduce thermogenic capacity in winter-acclimatized voles. This provided evidence that the two aspects of non-shivering thermogenesis, obligatory and regulatory, are stimulated by different exogenous cues. Summer acclimatization in the shorttailed field vole is manifest as a significant decrease in both basal and noradrenaline-induced metabolic rate, combined with a significant increase in body mass.Abbreviations ANCOV A analysis of covariance - BAT brown adipose tissue - BM body mass - BMR basal metabolic rate - NST non-shivering thermogenesis - NA noradrenaline - V the maximum V recorded following mass specific injection of noradrenaline - V the maximum V recorded following mass specific injection of saline - T _a ambient temperature - T _b rectal body temperature - T _1c lower critical temperature - UCP uncoupling protein - V oxygen consumption     

Shortening of the photoperiod affects sleep distribution, EEG and cortical temperature in the Djungarian hamster

To asses the influence of photoperiod on sleep regulation EEG, EMG, and cortical temperature were continuously recorded for two baseline days and after 4 h sleep deprivation in Djungarian hamsters (Phodopus sungorus) adapted to a short photoperiod (light dark 816). Comparison to previous data collected in a long photoperiod (lightdark 168) showed several major effects of photoperiod: 1. A prominent change in the 24-h distribution, duration and number of vigilance state episodes, whereas the total amount of sleep and waking was unchanged; 2. Cortical temperature was 0.7°C lower in the short photoperiod; 3. There was a significant negative correlation between cortical temperature and the frequency of REM sleep episodes; and 4. Absolute EEG power density showed a marked reduction in the short photoperiod. After sleep deprivation EEG slow-wave activity (mean power density 0.75–4.0 Hz) in NREM sleep showed a remarkably similar increase in both photoperiods demonstrating the robustness of the homeostatic regulation of sleep. Cortical temperature remained above baseline values after sleep deprivation in the short photoperiod whereas a negative rebound was present in the long photoperiod. Our results support the hypothesis that cortical temperature has a strong influence on REM sleep propensity and indicate the possibility of an optimum cortical temperature for recovery sleep after sleep deprivation. The lower EEG power density in the short photoperiod may contribute to energy conservation.Abbreviations LP long photoperiod - NREM non-rapid-eye-movement - REM rapid-eye-movement - SCN suprachiasmatic nucleus - SD sleep deprivation - SP short photoperiod - SWA slow-wave activity - T _CRT cortical temperature     

<Emphasis Type="Italic">In vitro</Emphasis> germination and axillary shoot propagation of <Emphasis Type="Italic">Centaurea zeybekii</Emphasis>

In vitro culture is an important aid for ex situ conservation of rare, endemic or threatened plants. In this work, we establish an efficient method for the seed germination, seedling development, and axillary shoot propagation of Centaurea zeybekii Wagenitz. The seeds, collected from a wild population, were surface sterilised and cultured on various in vitro germination media. The effects of photoperiod and temperature on seed germination were also investigated. Germinations were obtained after 6 weeks in culture and the radicle emergence was evaluated as a main indicator. A high frequency of germination was obtained on distilled water supplemented with vitamines and 1 mg/L GA₃. Although the seed germination frequencies were not affected by photoperiod, the highest germination frequency was obtained at 24 ± 2°C. A high frequency of axillary shoot proliferation was produced on MS medium supplemented with 1 mg/L BA. Then, the axillary shoots were separated and transferred to MS medium with or without plant growth regulators for rooting. Rhizogenezis was promoted after 6 weeks only in MS and 1/2 MS media containing 0.5 mg/L IBA. The rooting process was very slow and the percentage of shoot rooting was also very low (15%). The present study not only enables reinforcement of wild plant populations using ex situ growth of individuals, but it also helps to large number of aseptic seedling to use it in clonaly micropropagation studies.     

Study of embryo rescue in floribunda rose

In the past few decades, breeders have faced a lot of problems in rose improvement due to low sexual reproduction and poor germination because of embryo abortion. Immature embryos may be recovered in vitro. An efficient protocol for embryo rescue in two floribunda roses (Arunima and Shocking Blue) was developed. The germination of immature embryos was achieved by manipulating the growth media, growth hormones and culture conditions. The embryos (rescued) germinated and grew considerably on Murashige and Skoog basal medium supplemented with 2.5 mg l^–l BA (6-benzylaminopurine), 0.5 mg l^–l GA₃ (gibberellic acid) and 3 (w/v) sucrose under 16-h photoperiod. A higher rate of germination was observed in cultures incubated 2 weeks in dark and subsequently transferred to 2 weeks in light at 16-h photoperiod. The embryo derived plantlets were successfully transferred to greenhouse and produced flowers. Embryo rescue technique in floribunda roses has great potential in floriculture industry.     

Factors affecting akinete differentiation in Cylindrospermopsis raciborskii (Nostocales, Cyanobacteria)

1. Cylindrospermopsis raciborskii is a potentially toxic freshwater cyanobacterium which can produce akinetes (reproductive spores) that on germinating can contribute to future populations. To further understand factors controlling the formation of these specialised cells, the effects of diurnal temperature fluctuations (magnitude and frequency), in combination with different light intensities and phosphorus concentrations were investigated under laboratory conditions. 2. Akinete differentiation was affected by the frequency of temperature fluctuations. Maximum akinete concentrations were observed in cultures that experienced multiple diurnal temperature fluctuations. 3. Akinete concentrations increased with increasing magnitude of temperature fluctuation. A maximum akinete concentration was achieved under multiple diurnal temperature fluctuations with a magnitude of 10 °C (25 °C to 15 °C). 4. A fourfold increase in light intensity (25–100 μmol m^?2 s^?1) resulted in an approximate 14‐fold increase in akinete concentration. 5. High filterable reactive phosphorus (FRP) concentrations (>70 μg L^?1) in the medium, combined with a multiple diurnal temperature fluctuation of 10 °C, supported the development of the highest akinete concentration.     

AKINETE FORMATION IN PLANKTONIC ANABAENA SPP. (CYANOBACTEFUA) BY TREATMENT WITH LOW TEMPERATURE1

The effects of temperature, light intensity and nutrient depletion on akinete formation in seven strains of planktonic Anabaena spp.: A. mucosa TAC426; A. crassa TAC436; A. spiroides TAC443 and TAC444; A. flosaquae TAC446; and A. ucrainica TAC448 and TAC449 were examined. A Marked Pfft of temperature on akinete formation was observed at 40 μmol photons·m^?2·sec^?1 and nutrient-sufficient conditions. At 20° C, akinetes did not develop in A. mucosa TAC426, A. crassa TAC436, A. spiroides TAC443, A. flos-aquae TAC446, or A. ucrainica TAC449 but were formed at frequencies of a little over 11% (ratio of filaments with akinetes to total filaments) in A. spiroides TAC444 and A. ucrainica TAC448. None of the strains fmd akinetes or heterocysts at 30° C and 35° C. At lower temperature (10° C and 15° C), akinetes developed in all the strains at maximum frequencies of 13.4–77.4% during the late exponential phase or late exponential to stationary phases of growth. With only one exception, low light or nutrient deletion did not lead to the induction of akinete diferentiation at 20° C. Only akinete formation in A. flosaquae TAC446 was induced by nitrogen deletion with a frequency of 12.1%, similar to that induced by low temperature, but the initiation of akinete formation in the strain was delayed compared to treatment with low temperature. These results show that temperature was the most important environmental factor triggering akinete formation in these species. In A. crassa TAC436 and A. spiroides TAC443 and TAC444, akinetes developed during the late exponential growth phase even though heterocysts were formed at a 100% frequency (ratio of filaments with heterocysts to total filaments) throughout the entire growth phase. In A. mucosa TAC426, A. flos-aquae TAC446, and A. ucrainica TAC448 and TAC449, there was a positive correlation between heterocyst and akinete formation, suggesting that the presence of a heterocyst may play a role in akinete formation.