The effect of long wave ultraviolet radiation (UV-A) on the photosynthetic activity of natural population of freshwater phytoplankton

The effect of ultraviolet radiation on diel changes and depth profiles of phytoplankton photosynthesis was studied in four temperate freshwater lakes. Photosynthetic oxygen production was determined by incubating lake water in light and dark bottles under various weather conditions. Half the light bottles were wrapped with sheets of vinyl chloride film to exclude light with wavelengths shorter than 400 nm. The inhibition of photosynthesis due to UV-A (320–400 nm) was observed during most of the daytime and was very strong around noon on both sunny and cloudy days. On sunny days, when the surface waters of the highly eutrophic Lake Suwa and Senzoku Pond were dominated by denseMicrocystis populations, cumulative daily production at the surface, estimated from the incubation of bottles from which UV-A was excluded by the vinyl film, were about double the rates obtained from glass bottles in which UV-A was present. The UV-A inhibition was detected from the surface toca 20 cm depth in hypereutrophic lakes and at depths greater than 50 cm in mesotrophic lakes. Analysis of the photosynthesis-irradiance (P-I) relationship obtained in the present study shows β, a parameter that describes photo-inhibition, is higher in the presence of UV-A than in its absence. This indicates that UV-A is the major cause of photo-inhibition of phytoplankton photosynthesis.     

Effect of different light intensities on growth,reproduction, amino acid synthesis,fat and sugar contents in ulva fasciata delile

Various physiological processes of Ulva fasciata were investigated in the laboratory under light intensities of 1500, 2500 and 3500 lux respectively.It was shown that there is a strong correlation between light intensity and growth rate, which increased with the increase in light intensity up till 2500 lux. Light intensities above 3000 lux resulted in bleaching of the algal thalli.In no instance there was any discharge of swarmers in total darkness nor at very reduced light intensities of about 100 lux.Zoospores were always negatively phototactic, while gametes were positively phototactic, appearing always on the well-illuminated sides of the culture bottles.The maximum yield of total nitrogen, dry weight, and amino acid content coincides with the optimum light intensity. Under such conditions leucine, valine, -alanine and glutamic acids are found in abundance, while phenyl alanine, -aminobutyric and glycine are moderately represented.The amount of total fat content increases with the increse in light intensity up till 3500 lux. This might refer to a strong correlation between the rate of photosynthesis and the fat synthesis.It was found that fructose and raffinose were present in negligible amounts under reduced light intensities (1500 lux), while sucrose was found in rather higher quantities. The quantity of glucose is higher than that of fructose and raffinose but much less than that of sucrose under the same light intensity.Alexandria UniversityKuwait UniversityKuwait University     

Influence of carbon supply on the circadian rhythmicity of photosynthesis and its stimulation by blue light in Ectocarpus siliculosus: clues to the mechanism of inorganic carbon acquisition in lower brown algae

Stimulation or light-saturated rates of photosynthesis in Ectocarpus siliculosus (Dillwyn) Lyngb. by blue light was eliminated by increasing dissolved inorganic carbon (DIC) or by lowering pH in natural seawater. The amplitude of the circadian rhythm of photosynthesis was also diminished under these conditions, and the pH compensation points in a closed system were higher in the presence of blue light and during the circadian day. These observations suggest that blue light and the circadian clock regulate the activity of a carbon acquisition system in these plants. The inhibitor of external carbonic anhydrase, acetazolamide, reduced overall rates of photosynthesis by only about 30%, but ethoxyzolamide suppressed the circadian rhythm of photosynthesis almost completely and markedly reduced the duration of responses to blue light pulses. Similar patterns were obtained when photosynthesis was measured in strongly limiting DIC concentrations (0–0.5 mol m^?3). Since blue light stimulated photosynthesis under these conditions of strong carbon limitation, we suggest that blue light activates the release of CO₂ from an internal CO₂ store. We propose a metabolic pathway with similarities to that of CAM plants. Non-photosynthetic fixation leads to the accumulation of a storage metabolite. The circadian clock and blue light control the mobilization of CO₂ at the site of decarboxylation of this metabolite. In the presence of continuous blue light the pathway is proposed to cycle and act as a pump for CO₂ into the chloroplasts. This hypothesis helps to explain a number of previously reported peculiarities of brown algal photosynthesis.     

Ecological studies of Chloroflexis,a gliding photosynthetic bacterium

Summary Chloroflexis, a gliding, filamentous, photosynthetic bacterium, is present in the stratified algal-bacterial mats which occur in the 50°–70°C temperature range of alkaline hot spring effluents. The organism is in association with the alga in the upper, algal layer, and also forms thick, orange mats beneath the algal layer. Natural populations of Chloroflexis from these mats demonstrated light-stimulated uptake of some ¹⁴C-labelled organic compounds. Photosynthetic ¹⁴CO₂ fixation by natural samples of Chloroflexis was investigated with respect to temperature, light intensity and mat depth. Bacterial photosynthesis was determined in samples in which algae were present by use of the inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Bacterial photosynthesis was maximal at depths down to about 3 mm and then decreased rapidly to very low levels at greater depths. The greatest amounts of bacteriochlorophyll pigments were also concentrated in the top 3–4 mm of the mat. The optimum light intensity for bacterial photosynthesis (about 400 ft-c) was considerably lower than the normal summer light intensity at the surface of the mat (5000-8000 ft-c).The temperature optima for photosynthesis by the bacterial component of natural mat samples from several sites of different temperatures in a hot spring thermal gradient were determined. Temperature optima approximated the environmental temperatures, indicative of the occurrence of strains of Chloroflexis adapted to different temperatures. Although bacterial standing crop was greatest in the temperature range 50°–55°C, maximum photosynthetic efficiency was observed at about 45°C. Sulfide was stimulatory to photosynthetic ¹⁴CO₂ fixation by naturally occurring populations of Chloroflexis under field conditions. These data are consistent with the hypothesis that Chloroflexis may utilize sulfide as an electron donor for photosynthetic CO₂ reduction. However, it is also likely that Chloroflexis grows photoheterotrophically in these mats, obtaining organic compounds from algal excretory products.     

Modification of the interaction between<Emphasis Type="Italic">Escherichia coli</Emphasis> and bacteriophage in saline sediment

The effects of various combinations of light intensity, oxygen concentration, and CO₂ concentration on photosynthesis and growth in several algal types were studied. The results suggest the following. (1) Different algae show different responses to high oxygen concentrations and high light intensities. (2) Inhibition of photosynthesis (CO₂ fixation and growth), if seen, increases with increasing oxygen concentration and with increasing light intensity (at light intensities greater than saturation). (3) The inhibition of net photosynthesis observed cannot be attributed to high light intensity alone. (4) The inhibition cannot be attributed to increased rates of excretion of organic materials under conditions of high oxygen concentration and high light intensity. (5) Increased concentrations of CO₂ can decrease the effect of high oxygen and light in some algae. (6) The decrease in net photosynthesis observed is probably the result of photorespiration. (7) The effect of light intensity, oxygen concentration, or CO₂ concentration on algal photosynthesis should not be studied without considering the effect of the other factors. Some implications of these results, as related to primary productivity measurements, are also discussed.     

BENTHIC MICROALGAL COLONIZATION IN STREAMS OF DIFFERING RIPARIAN COVER AND LIGHT AVAILABILITY1

Benthic microalgal colonization, light climate, and algal photosynthesis–irradiance relationships were analyzed in two vegetated creeks in southeast Australia, differing in their degree of riparian cover and hence light availability. Diatoms were the main component of the benthic microalgal communities in both creeks. The light received was three times higher in the partly shaded than in the completely shaded creek throughout the 45‐day study period. The algal community in the most shaded stream (Dandenong Creek [DG]) was almost permanently under subsaturating light, with a range of 0.06–0.25 light h a day above saturation irradiance (I_k), whereas in the partly shaded stream (Deep Creek [DC]) a higher period of saturating light, 1.2 light h a day above I_k, was seen. The initial slopes (α) of photosynthesis versus irradiance relationships were similar in the two creeks during the initial stages of colonization, indicating that early algal colonizers could behave as generalists with respect to light. In later stages of colonization, however, α and I_k diverged in the two streams. In DG the α values remained high, up to the end of the colonization, whereas in DC α substantially decreased. In DG there was a higher chl concentration per surface area, and algae had a higher content of chl per cell than in the partly shaded creek. In DC the algal community became adapted to increasing light availability during the course of the colonization, but changes in the photosynthesis–irradiance parameters were accompanied by a shift in the composition of the diatom community. We concluded that the algal community is highly influenced by adaptation of the algal species to governing light conditions throughout colonization.     

PHOTOPHYSIOLOGY OF A TURF ALGAL COMMUNITY: INTEGRATED RESPONSES TO AMBIENT LIGHT AND STANDING BIOMASS1

This study investigated the variation in the relationship between photosynthesis and ambient light (P‐E curves) for turf algal communities on a temperate reef off the coast of South Australia, analyzing the integrated effects of ambient light and standing biomass. The photophysiology of turfs was studied in situ on a seasonal basis, examining algal communities growing on artificial substrate (plates) at depths of 4 m and 10 m. P‐E curves and estimates for the photokinetic parameters (P_m, R_d, α, E_k, and E_c) were obtained through oxygen evolution methods, using an automated underwater respirometer. Photoacclimation responses to changes in ambient light were strongly affected by the biomass of the community. P_m showed an inverse relationship to standing biomass, irrespective of depth and season, which was considered to be a response to self‐shading and boundary layer effects. Biomass effects imposed a high variance on estimates for all photosynthetic parameters, overshadowing differences observed for season and depth. Biomass also affected photoinhibition on turf communities, where significant afternoon depression of photosynthesis was observed in sparse turf patches when compared to denser patches. High areal productivity rates were maintained across all seasons with a significant decrease only being observed during winter.     

A theoretical consideration on oxygen production rate in microalgal cultures

Because algal cells are so efficient at absorbing incoming light energy, providing more light energy to photobioreactors would simply decrease energy conversion efficiency. Furthermore, the algal biomass productivity in photobioreactor is always proportional to the total photosynthetic rate. In order to optimize the productivity of algal photobioreactors (PBRs), the oxygen production rate should be estimated. Based on a simple model of light penetration depth and algal photosynthesis, the oxygen production rate in high-density microalgal cultures could be calculated. The estimated values and profiles of oxygen production rate by this model were found to be in accordance with the experimental data. Optimal parameters for PBR operations were also calculated using the model.     

Influence of Algal Photosynthesis on Biofilm Bacterial Production and Associated Glucosidase and Xylosidase Activities

Natural photosynthetic biofilms were incubated under light (100 mmol m-2 s-1) and dark conditions to elucidate the impact of photosynthesis on bacterial production, abundance, biovolume, biomass, and enzyme activities over 24 h. Use of organic carbon-free media limited carbon sources to algal photosynthesis and possibly the polysaccharides of the biofilm matrix. Bacterial production of biofilm communities was significantly higher in light incubations (p <0.001). The greatest differences in production rates between light and dark incubations occurred between 8 and 24 h. Biomass-specific a- and b-glucosidase and b-xylosidase activities were stimulated by photosynthesis, with significantly greater activities occurring at hours 16 and 24 in the light treatment (p <0.01). The results indicate that algal photosynthesis can have a significant impact on bacterial productivity, biomass, biovolume, and enzyme production over longer time periods at low photon flux densities (?100 mmol m-2 s-1).     

Calculation of light penetration depth in photobioreactors

Light penetration depth in high-densityChlorella cultures can be successfully estimated by Beer-Lambert's law. The efficiency of light energy absorption by algal cultures was so high that algal cells near the illuminating surface shade the cells deep in the culture. To exploit the potential of high-density algal cultures, this mutual shading should be eliminated or minimized. However, providing more light energy will not ease the situation and it will simply drop the overall light utilization efficiency.     

Responses of Two Coastal Algae (Skeletonema costatum and Chlorella vulgaris) to Changes in Light and Iron Levels1

Iron (Fe) is essential for phytoplankton growth and photosynthesis, and is proposed to be an important factor regulating algal blooms under replete major nutrients in coastal environments. Here, Skeletonema costatum, a typical red-tide diatom species, and Chlorella vulgaris, a widely distributed Chlorella, were chosen to examine carbon fixation and Fe uptake by coastal algae under dark and light conditions with different Fe levels. The cellular carbon fixation and intracellular Fe uptake were measured via ¹⁴C and ⁵⁵Fe tracer assay, respectively. Cell growth, cell size, and chlorophyll-α concentration were measured to investigate the algal physiological variation in different treatments. Our results showed that cellular Fe uptake proceeds under dark and the uptake rates were comparable to or even higher than those in the light for both algal species. Fe requirements per unit carbon fixation were also higher in the dark resulting in higher Fe: C ratios. During the experimental period, high Fe addition significantly enhanced cellular carbon fixation and Fe uptake. Compared to C. vulgaris, S. costatum was the common dominant bloom species because of its lower Fe demand but higher Fe uptake rate. This study provides some of the first measurements of Fe quotas in coastal phytoplankton cells, and implies that light and Fe concentrations may influence the phytoplankton community succession when blooms occur in coastal ecosystems.     

Effects of shading on calcareous benthic periphyton in a short-hydroperiod oligotrophic wetland (Everglades, FL, USA)

The effects of shade on benthic calcareous periphyton were tested in a short-hydroperiod oligotrophic subtropical wetland (freshwater Everglades). The experiment was a split-plot design set in three sites with similar environmental characteristics. At each site, eight randomly selected 1-m² areas were isolated individually in a shade house, which did not spectrally change the incident irradiance but reduced it quantitatively by 0, 30, 50, 60, 70, 80, 90 and 98%. Periphyton mat was sampled monthly under each shade house for a 5 month period while the wetland was flooded. Periphyton was analyzed for thickness, DW, AFDW, chlorophyll a (chl a) and incubated in light and dark BOD bottles at five different irradiances to assess its photosynthesis–irradiance (PI) curve and respiration. The PI curves parameters P _max, I _k and eventually the photoinhibition slope (β) were determined following non-linear regression analyses. Taxonomic composition and total algal biovolume were determined at the end of the experiment. The periphyton composition did not change with shade but the PI curves were significantly affected by it. I _k increased linearly with increasing percent irradiance transmittance (%IT = 1−%shade). P _max could be fitted with a PI curve equation as it increased with %IT and leveled off after 10%IT. For each shade level, the PI curve was used to integrate daily photosynthesis for a day of average irradiance. The daily photosynthesis followed a PI curve equation with the same characteristics as P _max vs. %IT. Thus, periphyton exhibited a high irradiance plasticity under 0–80% shade but could not keep up the same photosynthetic level at higher shade, causing a decrease in daily GPP at 98% shade levels. The plasticity was linked to an increase in the chl a content per cell in the 60–80% shade, while this increase was not observed at lower shade likely because it was too demanding energetically. Thus, chl a is not a good metric for periphyton biomass assessment across variously shaded habitats. It is also hypothesized that irradiance plasticity is linked to photosynthetic coupling between differently comprised algal layers arranged vertically within periphyton mats that have different PI curves.     

SOME ASPECTS OF PHOTOSYNTHESIS IN CLADOPHORA GLOMERATA1

Photosynthetic rates in field-collected samples of Cladophora glomerata were measured using the oxygen electrode technique. Rates observed were three- to fourfold higher than literature values obtained with the light and dark bottle method. The discrepancy appears to be due to light-limiting conditions prevailing in the bottles. It was also found that photosynthesis in Cladophora does not saturate at low light intensities as reported by other workers.     

Thirty liter tower-type pilot plant for the mass cultivation of light- and motion-sensitive planktonic algae

A simple, low-priced 30 liter tower-type algal pilot plant for the cultivation of light- and motion-sensitive species is described. Two hundred g wet weight of Microcystis aeruginosa were obtained per harvest. Since the self-shading of denser cultures could be compensated for only to a limited extent by increasing the light intensity without damaging the cells, the efficiency of various culture-vessel widths was determined for the growth of Microcystis : the best results were obtained with a width of 3.5 cm. Light requirements of Microcystis were studied in shadowless suspensions. The compensation point of photosynthesis varied between 200 and 300 lx, depending on the preillumination, whereas the light saturation point was found to be near 4000 Ix. The light optimum for photosynthesis was not identical with that for good growth.     

Responses to ocean acidification and diurnal temperature variation in a commercially farmed seaweed,Pyropia haitanensis (Rhodophyta)

To investigate carbon and nitrogen metabolism in Pyropia haitanensis in response to the combined conditions of ocean acidification and diurnal temperature variation, maricultured thalli were tested in acidified culture under different temperature treatments. The results showed a combined effect of ocean acidification and diurnal temperature difference on the C and N metabolism and growth of P. haitanensis. In acidifed culture, algal growth, maximum photosynthetic rate, nitrate reductase (NR) activity, amino acid (AA) content and AA score (AAS) were more significantly enhanced in seaweed under diurnal temperature variation than in seaweed at constant temperature. In acidified seawater, soluble carbohydrates in P. haitanensis increased due to greater dissolved inorganic carbon (DIC), whereas soluble proteins decreased. Under the diurnal temperature treatment, higher temperature during the light period enhanced accumulation of algal photosynthates, whereas lower temperature in the dark period reduced energy consumption, resulting in enhanced algal growth, AA content and AAS. We concluded that suitable diurnal temperature difference would be conducive to C fixation and N assimilation under ocean acidification. However, excessively high temperatures would depress algal photosynthesis and increase energy consumption, thereby exerting a negative effect on algal growth.     

Polymixis and algal production in a tropical lake: latitudinal effects on the seasonality of photosynthesis

SUMMARY. 1. Phytoplankton production was measured during 1981–82 in Bahia de Puno. a large polymictic system that is part of LakeTiticaca in the tropical Andes (Peru-Bolivia), Photosynthesis followed aweak seasonal pattern through each 12-month cycle but the largestvariations occurrud between years. 2. The large temporal variations in productivity appeared to becontrolled by biological events rather than by the direct effeets ofseasonal or aseasonal physical processes. Major shifts in production overthe 2-year period were attributable to the proportional abundance andactivity of N₂-fixing blue-green algae. Production was uncorrelated withthe seasonal radiation cycle. 3. In vivo fluorescence and photosynthetic assays revealed largechanges in phytoplankton community structure over time and depth.The shifts in vertical structure were associated with changes in transparency and water column stabilily.Physiological as well as floristiceffects dictated the variations in fluorescence per unit chlorophyll a. 4. An analysis of twenty-seven primary productivity data sets fromdifferent latitudes shtiwed that Bahia dc Puno resembled other tropicallakes in its relatively weak coupling between algal photosynthesis andseasonal variations in light. Productivity in the temperate zone correlated with the annual cycle of incoming radiation while the tropicalproduction data did not. For many of the tropical lakes, but fewtemperate lakes, the between-month variance in primary production wassignificantly higher than the between-month variance for incomingradiation. There was a significant positive relationship between maximum production variance explained hy light (maximum r² for each dataset) and latitude. 5. As in many tropical lakes. Bahia de Puno experiences moderatelylarge variations in algal photosynthesis.The muted seasonal cycle of light explains only a small portion of the total variation In productivity, and other factors such as weuther-related events are disproportionately important.     

PHOTOINHIBITION AND PHOTOACCLIMATION OF TURF ALGAL COMMUNITIES ON A TEMPERATE REEF,AFTER IN SITU TRANSPLANTATION EXPERIMENTS1

The photophysiology of turf algal communities was studied in situ on a temperate reef off the coast of South Australia. Algal communities were grown on artificial substrate at depths of 2, 4, and 10 m. To investigate the response of the algal communities to changing light environments in both the short and long term, reciprocal transplantation experiments were conducted among these depths on a seasonal basis. The extent of photoinhibition was assessed every 3 h for the first 2 days following transplantation and then on a daily basis for 16 days after transplantation. Photosynthetic acclimation was assessed using photosynthesis–light curves obtained from transplanted and non‐transplanted turfs after the acclimation period. Transplanted turfs responded very quickly to the light shift. Algae acclimated to low light (10 m depth) were highly susceptible to photoinhibition and photodamage, having greater decreases in maximum and effective quantum yields than turfs from shallower depths. Yield recovery and acclimation usually occurred very rapidly in algae from all depths (3–5 days), but were faster in spring and summer compared with winter. Changes in photosynthetic capacity (across seasons, depths, and after transplantation to a different depth) were accompanied by changes in respiration, so that the ratio of net to gross photosynthetic capacity (P_mnet : P_mgross) remained high and constant over the whole range of light levels. We discuss the possible acclimation strategies of turfs, taking into account the balance between photoacclimation, production, and growth strategy.     

Why are higher plants green? Evolution of the higher plant photosynthetic pigment complement

The physiological reason that higher plants are green is unknown. Other photosynthetic organisms utilize pigments that strongly absorb green light; therefore, there must have been natural forces that ‘selected’ the photosynthetic pigments found in higher plants. Based on previously published data and our recent findings about green light and photosynthesis within leaves (Sun et al.), a specific functional role is described for the primary photosynthetic pigments of higher plants, that were derived from green algal progenitors. The particular absorptive characteristics of chlorophylls a and b appear to perform two contradictory, but necessary functions in higher plants. Firstly, chlorophylls a and b absorb light for maximum utilization under non‐saturating conditions, a function that is well understood. Secondly, they can act as protective pigments under over‐saturating light conditions, when absorbed light is dissipated as heat. Under such conditions, a significant portion of light can also be efficiently utilized, especially in the bottom portion of the leaf, that is mainly illuminated by green light and not down‐regulated. The second function may have been the selective force that gave rise to the extremely successful terrestrial plants, that evolved from green algae.     

Acclimation of brown algal photosynthesis to ultraviolet radiation in Arctic coastal waters (Spitsbergen, Norway)

In field studies conducted at the Kongsfjord (Spitsbergen) changes of the irradiance in the atmosphere and the sublittoral zone were monitored from the beginning of June until the end of August 1997, to register the minimum and maximum fluxes of ultraviolet and photosynthetically active radiation and to characterise the underwater light climate. Measurements of photosynthesis in three abundant brown algal species (Alaria esculenta, Laminaria saccharina, Saccorhiza dermatodea) were conducted to test whether their photosynthetic performance reflects changing light climate in accordance with depth. Plants sampled at various depths were exposed to controlled fluence rates of photosynthetically active radiation (400–700 nm), UV-A (320–400 nm) and UV-B (280–320 nm). Changes in photosynthetic performance during the treatments were monitored by measuring variable chlorophyll fluorescence of photosystem II. In each species, the degree of inhibition of photosynthesis was related to the original collection depth, i.e. shallow-water isolates were more resistant than plants from deeper waters. The results show that macroalgae acclimate effectively to increasing irradiance levels for both photosynthetically active and ultraviolet radiation. However, the kinetics of acclimation are different within the different species. It is shown that one important strategy to cope with higher irradiance levels in shallow waters is the capability for a faster recovery from high light stress compared to isolates from deeper waters. Received: 13 March 1998 / Accepted: 16 May 1998     

Photosynthetic performance of freshwater Rhodophyta in response to temperature, irradiance, pH and diurnal rhythm

Responses of net photosynthetic rates to temperature, irradiance, pH/inorganic carbon and diurnal rhythm were analyzed in 15 populations of eight freshwater red algal species in culture and natural conditions. Photosynthetic rates were determined by oxygen concentration using the light and dark bottles technique. Parameters derived from the photosynthesis–irradiance curves indicated adaptation to low irradiance for all freshwater red algae tested, confirming that they tend to occur under low light regimes. Some degree of photo‐inhibition (β= ‐0.33–0.01 mg O₂ g^?1 DW h^?1 (μmol photons m^?2 s^?1)^?1) was found for all species/populations analyzed, whereas light compensation points (Ic) were very low (≤ 2 μmol photons m‐ photons s^?1) for most algae tested. Saturation points were low for all algae tested (Ik = 6–54 μmol photons m^?2 s^?1; Is = 20–170 umol photons m^?2 s^?1). Rates of net photosynthesis and dark respiration responded to the variation in temperature. Optimum temperature values for net photosynthesis were variable among species and populations so that best performances were observed under distinct temperature conditions (10, 15, 20 or 25°C). Rates of dark respiration exhibited an increasing trend with temperature, with highest values under 20–25°C. Results from pH experiments showed best photosynthetic performances under pH 8.5 or 6.5 for all but one species, indicating higher affinity for inorganic carbon as bicarbonate or indistinct use of bicarbonate and free carbon dioxide. Diurnal changes in photosynthetic rates revealed a general pattern for all algae tested, which was characterized by two relatively clear peaks, with some variations around it: a first (higher) during the morning (07.00–11.00 hours.) and a second (lower) in the afternoon (14.00–18.00 hours). Comparative data between the ‘Chantransia’ stage and the respective gametophyte for one Batrachospermum population revealed higher values (ca 2‐times) in the latter, much lower than previously reported. The physiological role of the ‘Chantransia’ stage needs to be better analyzed.