Effects of photoacclimation on the light niche of corals: a process-based approach

The ecology of photosynthetic organisms is influenced by the need to adjust the photosynthetic apparatus to variable light environments (photoacclimation). In this study, we quantified different components of the photoacclimation process for a reef-building coral (Turbinaria mesenterina, Lamarck, 1816): including, variation in absorption cross-section, size of photosynthetic units, turnover time, chlorophyll content, and colony respiration. We used these calibrations to characterize this species’ light niche, and to determine the sensitivity of the niche boundaries to different processes of photoacclimation. Results showed that the breadth of the light niche was most sensitive to the size of the photosynthetic unit, absorption cross-section, and rates of respiration. Habitats with the highest light availability did not lead to maximal energy acquisition. This was because, although corals acclimated to high light have high rates of photosynthesis per unit chlorophyll, their chlorophyll content was strongly reduced. This suggests that potential energetic benefits that could be achieved through increased light harvesting (i.e., increased chlorophyll content) in high-light habitats are outweighed by costs associated with photoprotection. Such costs appear to place an upper bound on the habitat distributions of coral species. Our approach reveals how the photophysiological processes involved in photoacclimation interact to determine the light niche.     

High-temperature tolerance of photosynthesis in the red alga Chondrus crispus

Chondrus crispus (Stackhouse) is a perennial red seaweed, common in intertidal and shallow sublittoral communities throughout the North Atlantic Ocean. In the intertidal zone, C. crispus may experience rapid temperature changes of 10 to 20C° during a single immerison-emerision cycle, and may be exposed to temperatures that exceed the thermal limits for long-term survival. C. crispus collected year-round at Long Cove Point, Chamberlain, Maine, USA, during 1989 and 1990, underwent phenotypic acclimation to growth temperature in the laboratory. This phenotypic acclimation enhanced its ability to withstand brief exposure to extreme temperature. Plants grown at summer seawater temperature (20°C) were able to maintain constant rates of lightsaturated photosynthesis at 30°C for 9 h. In contrast, light-saturated photosynthetic rates of plants grown at winter seawater temperature (5°C) declined rapidly following exposure to 30°C, reached 20 to 25% of initial values within 10 min, and then remained constant at this level for 9 h. The degree of inhibition of photosynthesis at 30°C was also dependent upon light intensity. Inhibition was greatest in plants exposed to 30°C in darkness or high light (600 mol photons m^-2s^-1) than in plants maintained under moderate light levels (70 to 100 mol photons m^-2s^-1). Photosynthesis of 20°C-acclimated plants was inhibited by exposure to 30°C in darkness or high light, but the degree of inhibition was less than that exhibited by 5°C-grown plants. Not only was light-saturated photosynthesis of 20°C plants less severely inhibited by exposure to 30°C than that of 5°C plants, but the former also recovered faster when they were returned to growth conditions. The mechanistic basis of this acclimation to growth temperature is not clear. Our results indicate that there were no differences between 5 and 20°C-grown plants in the thermal stability of respiration, electron transport associated with Photosystems I or II, Rubisco or energy transfer between the phycobilisomes and Photosystem II. Overall, our results suggest that phenotypic acclimation to seawater temperature allows plants to tolerate higher temperatures, and may play an important role in the success of C. crispus in the intertidal environment.     

Rapid growth of clones of the red alga Chondrus crispus: applications in assays of toxic substances and in physiological studies

Growth rates of excised apical segments from three Chondrus crispus Stackhouse clones were rapid, reproducible and easily measured using simple equipment. Clonal segments exhibited a high degree of phenotypic stability, with coefficients of variation in growth rates being typically less than 5%. Sensitivity of the assay was demonstrated using 24-h pulses of the toxicants Cu²⁺ (10 to 150 ppb) and the molluscicide Bayluscide (100 to 500 ppb). As a further demonstration of applicability of this assay system, growth rates were measured over two consecutive 24-h photoperiods using a normally pigmented red gametophytic clone and a green colored mutant derived from it. Growth rates were highest in the first hours of the light phase for both clones, with growth of the green mutant being consistently inferior to that of the normally pigmented one. Significant changes were measured with intervals as short as 4 h using only five apical segments for each treatment. The C. crispus assay described is sensitive, relatively rapid, and statistically robust when applied to toxicant testing in seawater and to physiological studies.     

富营养化水体中菹草光合荧光特性研究

用NH4Cl、NaNO3、KH2PO4配置不同氮磷浓度的试验水,将菹草(Potamogeton cripus)分别移栽于上述水体中,观测菹草在各水体中的生长发育状况,并用水下饱和脉冲叶绿素荧光仪(Diving-PAM)测定菹草光合作用PSII有效荧光产量(Y)、光化学荧光淬灭系数(qP)、非光化学荧光淬灭系数(qN)等指标。结果表明:在TN<2mg·L-1,TP<0.4mg·L-1,Chla<118mg·m-3试验水体中,菹草长势良好,株高增加了51.2%、46.6%,叶片数增大了57.7%、58.1%;在营养盐质量浓度很高(TN8mg·L-1,TP0.8mg·L-1)、藻类生物量极高(Chla421mg·m-3)的水体中,菹草的生长受到抑制,并出现死亡。Y、qP、qN的值说明了在TN<2mg·L-1,TP<0.4mg·L-1,Chla<118mg·m-3水体中,菹草叶片吸收的光能更多的参与光化学反应,光合作用增大。高营养盐质量浓度(TN8mg·L-1,TP0.8mg·L-1,Chla421mg·m-3)水体中的菹草叶片将大部分光能用于热耗散,光合作用降低。快速光响应曲线同时也说明各水体中,菹草的最小饱和光强(Ek)都为176μmol·m-2s-1,但高营养盐质量浓度(TN8mg·L-1,TP0.8mg·L-1,Chla421mg·m-3)水体中的菹草叶片的最大光电子传递速率(rETR-max)明显低于其他水体,随着水体中营养盐浓度的升高,菹草耐光抑制能力显著降低。     

Inorganic carbon uptake by photosynthetically active protoplasts of the red macroalga Chondrus crispus

Photosynthetically active protoplasts were isolated from Chondrus crispus Stackh. by treating thalli with -carrageenase produced from batch culture of Pseudomonas carrageenovora. Using the silicone oil centrifugation technique, it was found that the protoplasts: (1) did not generally accumulate inorganic carbon (C_i) above the concentration in their incubation medium; (2) were saturated at C_i concentrations of 3 to 4 mM; (3) had an intracellular pH of 7.50 when incubated at pH 7.5; and (4) their initial carbon fixation rate was reduced by carbonic anhydrase inhibitors. Although the carbon fixation rate of the protoplasts was about 30% that of thallus fragments, presumably due to the relatively harsh protoplast isolation treatment, the behavior of the protoplasts was similar to that of fragments. This similarity indicates that the protoplasts are photosynthetically active and behave as thallus fragments. Further, the data are consistent with the hypothesis that C. crispus acquires C_i for photosynthesis by the diffusion of CO₂ across the plasma membrane.     

冬季菹草对悬浮泥沙的影响

利用冬季沉水植物菹草构建由浅水到深水的串联系统,含沙水依次流经浅水区和深水区,测定各级菹草对水体悬浮泥沙含量和沉降量的影响.结果显示,菹草系统可以降低水体中悬浮泥沙含量,并随水力停留时间延长而降低程度加大.各运行条件下,菹草系统悬浮泥沙平均沉降量65.7 g·m-2,比对照组高39.4%.菹草表面泥沙的吸附是一个"吸附-沉降-再吸附" 的动态过程,吸附量为3.26～5.69 g·kg-1.菹草对冬季水生态系统的自净功能起着十分重要的作用.     

泥沙淤积对菹草萌发与初期生长的影响

泥沙淤积对菹草萌发与初期生长影响的试验研究结果表明,泥沙淤积会延迟菹草石芽的萌发,但泥沙淤积不会明显影响菹草石芽最终的萌发率;淤积厚度对菹草萌发苗的叶片数影响不明显,叶片长度、宽度和叶片面积随泥沙淤积厚度增加而减小,叶片的长/宽比值则随泥沙淤积厚度的增加而增加;菹草单株平均生物量随淤积厚度的增加而减小.     

Freezing tolerance in the intertidal red algaeChondrus crispus andMastocarpus stellatus: Relative importance of acclimation and adaptation

The effect of repeated daily freezing on photosynthesis, growth and phenotypic acclimation to freezing was studied in the red algaeChondrus crispus Stackhouse andMastocarpus stellatus (Stackhouse in With.) Guiry. Algae used for experiments were collected from Chamberlain, Maine, between March and August 1987, and field observations and experiments were carried out at Chamberlain and Kresge Point, Maine between March 1987 and March 1989. After ca 30 d of daily freezing for 3 h at –5°C photosynthesis ofC. crispus was reduced to 55% of control values. Growth rates ofC. crispus were also reduced in fronds frozen daily compared to unfrozen controls, and eventually fronds became bleached and fragmented resulting in biomass losses. Fronds ofC. crispus, frozen daily, had higher photosynthetic rates following freezing events than unfrozen controls indicating that this species can acclimate to freezing conditions. Acclimation to freezing involves the light-harvesting reactions of photosynthesis. In contrast, photosynthesis and growth inM. stellatus were unaffected by repeated daily freezing for 3 h at –5°C for 36 d. No differences in photosynthesis following freezing were observed between frozen and control fronds suggesting thatM. stellatus does not phenotypically acclimate to freezing. The greater freezing tolerance ofM. stellatus relative toC. crispus results, in part, from genetic adaptations associated with plasma membranes and the light-harvesting reactions of photosynthesis.     

Effects of dissolved ammonium addition and host feeding with Artemia salina on photoacclimation of the hermatypic coral Stylophora pistillata

 Effects of nutrient treatments on photoacclimation of the hermatypic coral Stylophora pistillata (Esper) were studied. Studies on photoacclimation of colonies from different light regimes in the field were evaluated and used to design laboratory experiments. Coral colonies were collected in the Gulf of Eilat (Israel) from January to March 1993. Exterior branches of colonies from different depths (1 to 40 m) displayed different trends in production characteristics at reduced and very low levels of illumination. From 24 ± 3% to 12 ± 2% of incident surface photosynthetic active radiation (PAR_o), zooxanthella population density and chlorophyll a+c per 10⁶ zooxanthellae increased, a trend seen in the range of light levels optimal for coral growth (90 to 30% PAR_o). The P _max of CO₂ per 10⁶ zooxanthellae decreased, while P _max of CO₂ per 10³ polyps increased, indicating an increase in zooxanthella population density at low light levels. Proliferous zooxanthella frequency (PZF, a measure of zooxanthella division) declined significantly at light levels <18 ± 3% PAR_o. At the lowest levels of illumination (<5% PAR_o), zooxanthella population density decreased, as did the PZF; chl a+c per 10⁶ zooxanthellae was unchanged. In 28-d experiments, exterior coral branches from the upper surfaces of colonies from 3 m depth (65 ± 4% PAR_o) were incubated in aquaria under bright (80 to 90% PAR_o), reduced (20 to 30% PAR_o), and extremely low (2 to 4% PAR_o) light intensities. At each light intensity, the corals were maintained in three feeding treatments: sea water (SW); ammonium enriched SW (SW + N); SW with Artemia salina nauplii (SW + A). An increase in P _max of CO₂ per 10³ polyps was found in corals acclimated to reduced light (20 to 30% PAR_o) in nutrient-enriched SW, while in SW, where the increase in zooxanthella population density was smaller, it did not occur. Nutrient enrichments (SW + N at 2 to 4% PAR_o and SW + A at 20 to 30% PAR_o) increased zooxanthella population density, but had no effect on chl a+c per 10⁶ zooxanthellae. Acclimation for 14 d to reduced (10 to 20% PAR_o) and extremely low (1 to 3% PAR_o) light intensities shifted ¹⁴C photoassimilation into glycerol and other compounds (probably glycerides), rather than sugars. Both ammonium addition and feeding with Artemia salina nauplii resulted in an increase in photosynthetic assimilation of ¹⁴C into amino acids. We conclude that acclimation to reduced light consists of two processes: an increase in photosynthetic pigments and in zooxanthella population density. Both processes require nitrogen, the increase in zooxanthella population density needing more; this adaptation is therefore limited in nitrogen-poor sea water. Received: 19 June 1998 / Accepted: 13 June 2000     

Reproduction,growth and photosynthesis of gametophytes of Laminaria saccharina grown in blue and red light

The induction of reproductive activity by blue light in female gametophytes of the brown alga Lamnaria saccharina has been investigated and related to other effects of red and blue light on these plants. Although germination of zoo-spores is slightly delayed in red light, female gametophytes grow at similar rates in blue and red light of the same quantum irradiance up to an age of 8 to 10 days, when plants in blue light begin to form eggs. The percentage of plants forming eggs is proportional to the total quanta of blue wavelengths received up to a saturating value of about 400 μE·cm^-2; a 50% response is induced by 200 μE·cm^-2. The action spectrum for the induction of fertility has a main peak at 430 to 450 nm, with two subsidiary peaks in the blue and one in the near-ultraviolet light, but the action spectrum for photosynthesis is quite different, with peaks in the red, green and blue regions. These results indicate that egg formation in female gametophytes occurs as a specific morphogenetic response to blue light, which is independent of photosynthesis and growth, and trial experiments with male gametophytes of L. saccharina and gametophytes of other species of Laminariales indicate that these plants react similarly.     

Effects of UV-A LED light irradiation on growth of cultured RAW 264.7 cells

The aim of this study was to examine the effects of ultraviolet A (UV-A) irradiation-induced damage on cultured macrophage RAW 264.7 cells and determine which components produced these manifestations. RAW 264.7 cells were irradiated with 365 nm UV-A using a light-emitting diode (LED). Cell viability and damage were determined using a calcein-AM and propidium iodide dual-staining assay and lactate dehydrogenase leakage, respectively. Intracellular reactive oxygen species (ROS) were measured by H₂DCF-DA. The components of ROS in each medium were measured using an electron paramagnetic resonance (EPR) spectrometer in the presence of 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) and 2,2,5,5-tetramethyl-3-pyrroline-3-carboxamide (TPC). While UV-A irradiation for 2 min significantly suppressed cell growth, LDH leakage did not occur. Addition of N-acetyl cysteine restored inhibition of cell proliferation, and reduced intracellular ROS levels. The EPR signal in the presence of TPC increased with time but was decreased by sodium azide. In addition, a typical EPR spectrum was obtained in the presence of DMPO, indicating the presence of a hydroxyradical. The spectrum was diminished by L-histidine. Data suggest that ROS generated in cells or culture medium by UV-A irradiation is predominantly singlet oxygen, and this singlet oxygen suppressed cell proliferation.     

Effects of non-circadian light/dark cycles on the growth and moulting of Palaemon elegans reared in the laboratory

The growth and moulting of Palaemon elegans Rathke has been compared under a circadian (12 h light: 12 h dark) and two non-circadian (8 h light:8 h dark and random light:dark) light/dark cycles. In prawns reared individually from hatching to the late juvenile phase, growth, measured as increase in total length, was significantly retarded in the non-circadian regimes, during zoeal, post-larval and early juvenile development. This effect was primarily the result of reduced increments at moulting in the non-circadian regimes. Growth of prawns reared from hatching to the post-larval phase, measured as wet and dry weights, was similarly reduced in a non-circadian regime. These effects support the hypothesis that the integrity of an animal's physiology is partially dependent on maintaining, through the action of daily environmental cycles, correct timing relationships between its oscillating sub-systems.     

有机氮比例和光强对赤潮藻球形棕囊藻生长和光合作用的影响

采用实验室一次性培养的方法,研究了有机氮比率和光照强度相互作用对赤潮藻球形棕囊藻（Phaeocystis globosa）生长和光合作用的影响。结果表明,在高光条件下（120μmol.m-2.s-1）,有机氮含量为20%时,其细胞密度、比生长率均显著高于对照组和其他处理组,球形棕囊藻生长最好;在低光条件下（10μmol.m-2.s-1）,有机氮含量大于50%时,其细胞密度要高于硝态氮和有机氮为20%的实验组。无论是高光还是低光条件下,有机氮含量变化对球形棕囊藻Fv/Fm影响不大。但在高光条件下,不同氮源对其Fv/Fm有显著的影响,硝氮组显著高于尿素组而在低光照条件下,以尿素为唯一氮源的对照组达到最大光能转化效率0.752,但与硝氮组差别不显著。对于既能利用无机氮又能利用有机氮的海洋微藻来说,有机氮的存在可能显著促进其增殖,从而改变浮游植物群落结构,并可能在适宜的环境条件下形成赤潮。     

光质对烤烟生长发育、主要经济性状和品质特征的影响

以烤烟（Nicotiana tabacum）云烟87为材料,在烤烟团棵期和打顶期用透光率相近的有色薄膜分别对其进行遮光处理直到采收结束,研究光质对烤烟生长发育、主要经济性状和品质特征的影响。结果表明：蓝光有促进烟株生殖生长和延缓烟叶成熟的作用,红光、黄光和白光对烟株的生育进程无明显影响,打顶期遮光有延长烟株大田生育期的趋势。光质对叶片生长发育影响最大,红光和蓝光的影响效应主要表现在烟叶生育前期,黄光和白光的影响效应主要体现在烟叶生育后期。自然光和红膜下烟株的农艺性状综合表现最好,红光有促进烟茎生长和叶片发育的作用,蓝光和黄光则抑制茎干生长和降低叶面积。烟株生长前期增加红光比例有助于初烤烟叶主要经济性状的改善,后期蓝光的作用逐渐凸显。烟株不同生长时期红光和黄光的适当组合在改善烟叶品质中起着重要作用,烟株生长前期增加红光比例有利于降低初烤烟叶烟碱和蛋白质含量,提高糖类物质含量和评吸质量,但对氧化钾含量作负效应贡献;烟株生长后期提高黄光比例可使初烤烟叶的化学成分更为协调,对吸食品质有较好的提升作用。     

外源脱落酸(ABA)增强菹草抗镉(Cd2+)胁迫能力

以高等沉水植物菹草(Potamogen crispus)为试验材料,观察4 mg·L-1 Cd2 胁迫条件下外施脱落酸(abscisic acid,ABA)对菹草某些生理生化指标的影响.结果表明,外施ABA可以明显减轻Cd2 毒害作用,使Cd2 胁迫下菹草叶片中叶绿素、可溶性蛋白、脯氨酸含量增加,超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)活性提高,过氧化物酶(POD)活性和O(-)/(·)2产生速率降低,且以施用2.5～5 μmol·L-1 ABA效果最好.     

铝对土壤微生物区系组成的影响

通过大豆盆栽试验,研究了金属铝对土壤微生物区系组成的影响。结果表明,当铝质量分数<1.1 g/kg时,土壤细菌、放线菌的数量随着铝质量分数的增加而增加;当铝质量分数>1.1 g/kg时,土壤细菌、放线菌的数量随着铝质量分数的增加而减少。铝对丝状真菌的生长具有明显的促进作用。在低质量分数下,铝对土壤氨化细菌、硝化细菌、反硝化细菌具有一定的刺激作用;当质量分数>1.1 g/kg时,则表现为抑制作用,最大抑制率分别达48%、96%和95%;土壤硝化细菌、反硝化细菌可以作为铝污染土壤生态环境的指示菌。自生固氮菌的数量随铝质量分数的升高而逐渐下降。铝对土壤微生物区系产生抑制作用的临界质量分数在1.3 g/kg左右。     

菹草生长对沉积物间隙水中各形态磷浓度的影响

通过人工构建湖泊生态系统,用湖泊沉积物的上层泥和下层泥分别代表清淤前后的2种基质,研究菹草(Potamogeton crispus)生长对沉积物不同层次间隙水中各形态磷浓度的影响.垂直方向上,将2种基质中的间隙水分为表层、中层和底层3个层次.结果表明,菹草生长能够不同程度地降低各层间隙水中磷浓度,上层泥和下层泥2种基质均表现为表层间隙水溶解性总磷(DTP)和溶解性活性磷(SRP)含量降低幅度最大,DTP含量分别降低68.97％和83.09％,SRP含量分别降低54.57％和96.02％.菹草的吸收作用使得各层间隙水中磷浓度均低于无沉水植物的对照组.试验结束时,2种沉积物中TP含量降低幅度差异不显著,说明疏浚前后种植菹草均可有效控制湖泊内源磷污染.     

Optimization of pigment content and the limits of photoacclimation for Ulva rotundata (Chlorophyta)

Two vegetative clones (designated 11/85 and 7/86 in accordance with month/year of collection) of the chlorophyte macroalga Ulva rotundata were collected in the vicinity of Beaufort, North Carolina, USA. Each was grown in an outdoor continuous-flow system in summer (>-20°C) of 1986 and late winter (10° to 17°C) of 1987 in graded scalar quantum irradiances ranging from 9 to 100% of full sunlight, with and without NH ₄ ⁺ enrichment. The pigment content of plants from each irradiance was determined following 4 to 8 d sunny weather. Chlorophyll (chl) and carotenoid content were inverse curvilinear functions of irradiance. The chl a:b and carotenoid: chl ratios were positively related to irradiance. The close nonlinear relationship between chl (a+b) and the chl a:b ratio was independent of clone, temperature or NH ₄ ⁺ -enrichment. Chl (a+b) content was linearly correlated with light-regulated growth rate in the summer, but showed a marked hysteresis in the relationship in winter due to photoinhibition. The photon growth yield (PGY, i.e., the biomass yield per unit absorbed light) was maximal for plants grown at slightly subsaturating irradiances, and dropped off sharply at lower irradiances. At higher irradiances, PGY declined gradually in summer and markedly in winter. Light absorption exceeded growth needs at full sunlight, suggesting that U. rotundata was incapable of further reducing its pigment content when growth rate was light-saturated. This, along with the linear chlgrowth relationship, is consistent with photosynthetic feedback regulation of chl content. Regardless of the mechanism, chl regulation may operate within the constraints of a resource tradeoff between light harvesting and carboxylation capacities, such that pigmentation must be optimized rather than maximized.     

Effects of light intensity and nutrient availability on diel patterns of cell metabolism and growth in populations of Synechococcus spp.

Between July 21 and August 8, 1984, phytoplankton were collected from the surface (2 m) and/or chlorophyll maximum of a neritic front, warm-core eddy 84-E and Wilkinson's Basin in the Northwest Atlantic Ocean and incubated up to 38 h in 200-liter vats. Effects of light intensity and nutrient availability on diel patterns of cell metabolism were analyzed in a 0.6- to 1-m fraction, where Synechococcus spp. represented 80 to 100% of the total photoautotrophs. Populations held under in situ conditions exhibited daytime peaks in photosynthetic potential (P_max) that were an order of magnitude higher than nighttime P_max values. Daytime phasing of P_max peaks had no relationship to asynchronous fluctuations in cellular activities of ribulose 1,5 bisphosphate carboxylase (RUBPCase) or phosphoenol pyruvate carboxylase (PEPCase), or to variations in chlorophyll content. Daytime P_max peaks were about 12 h out of phase with nighttime maxima in the frequency of dividing cells (FDC). The phase relationship between P_max and FDC could be altered by manipulating environmental conditions. High light exposure of depp populations did not affect timing of the P_max peak, but its magnitude increased and coincided with increased RUBPCase activity and chlorophyll photobleaching. In the eddy population, a major shift in the timing of peak P_max was induced when increased light intensity was accompanied by nutrient enrichment. This change coincided with major increases in cellular chlorophyll and carboxylating enzyme activity. Lowering irradiance and/or increasing nutrient availability elicited different diel pattern in cellular metabolism in surface populations from the eddy and from Wilkinson's Basin that appeared linked to differences in the nutrient status of the cells. Rates of cell division estimated from the percentage of dividing cells in preserved samples were 0.83 divisions d^-1 in surface warm-core eddy populations, supporting the view that carbon and nitrogen turnover rates in oligotrophic waters can be sufficient to promote near optimal growth of Synechococcus spp.     

Photosynthetic and morphological photoacclimation of the seagrass Cymodocea nodosa to season, depth and leaf position

The photoacclimation capacity of the seagrass Cymodocea nodosa was evaluated considering temporal (i.e. seasonal) and spatial (i.e. depth and within-leaf position) factors of variation. Changes along the leaf were measured in a population growing along a depth gradient (from intertidal to subtidal) in Cadiz Bay (Southern Spain) from 2004 to 2005. Photoacclimation was evaluated by photosynthesis (P–E curves), pigment content and leaf morphology. Plants of Cymodocea nodosa showed large physiological and morphological plasticity (mean %CV = 35.8 ± 3.4) according to the three factors considered. Seasonal patterns appeared for photosynthesis, respiration, pigment content and morphology. Nevertheless, seasonal patterns were not consistent with depth or leaf portions. The resulting data set offered different information depending on the analysis conducted; when only one factor (season, depth or leaf portion) was considered, some tendencies observed in the 3-way full design were masked. Accordingly, considering spatio–temporal variability is crucial when describing photoacclimation and estimating productivity in seagrass meadows.