Horizontal or vertical photobioreactors? How to improve microalgae photosynthetic efficiency

The productivity of a vertical outdoor photobioreactor was quantitatively assessed and compared to a horizontal reactor. Daily light cycles in southern Spain were simulated and applied to grow the microalgae Chlorella sorokiniana in a flat panel photobioreactor.The maximal irradiance around noon differs from 400 μmol photons m⁻² s⁻¹ in the vertical position to 1800 μmol photons m⁻² s⁻¹ in the horizontal position. The highest volumetric productivity was achieved in the simulated horizontal position, 4 g kg culture⁻¹ d⁻¹. The highest photosynthetic efficiency was found for the vertical simulation, 1.3 g of biomass produced per mol of PAR photons supplied, which compares favorably to the horizontal position (0.85 g mol⁻¹) and to the theoretical maximal yield (1.8 g mol⁻¹). These results prove that productivity per unit of ground area could be greatly enhanced by placing the photobioreactors vertically.     

Production of rhizoids by Caulerpa prolifera in culture

Studies on Caulerpa prolifera rhizoids were carried out to determine the possibility of mass culture, because the rhizoids produce a bio-adhesive. Rhizoids can be induced by cutting the base of a blade and floating it in a media or planting it in sand. Measurement of rhizoid production included determination of number, length, and the weight of attached sand grains. The growth experiments were for 1–2 weeks and fronds growth was compared to rhizoid production. Optimal conditions for rhizoid growth included low levels of nitrogen and phosphate (less than 5 and 2 μM, respectively), low irradiance (30 μmol photon m⁻² s⁻¹), moderate temperature (22–28°), continuous shaking, addition of microelements and auxin (1 ppm) and initially detached fronds followed by attachment. Under these optimal conditions maximal weekly growth reached 70–170 rhizoids per blade, 7–11 mm length and maximal attachment to sand grains. Blade growth of C. prolifera responded similarly to rhizoid production and reached a weekly growth rate of 30–130%.     

Diurnal movements of chloroplasts in Halophila stipulacea and their effect on PAM fluorometric measurements of photosynthetic rates

Since diurnal chloroplast movements in Halophila stipulacea were described by Drew in 1979, this phenomenon has not been studied further for seagrasses. In addition to an apparent photoprotective role, such movements may affect the measurements of photosynthetic rates based on pulse amplitude modulated (PAM) fluorometry. This is because calculations of electron transport rates (ETR) are directly affected by the light absorption of the leaves (or the so-called absorption factor, AF), the latter of which changes with the movements of the chloroplasts. In this work, we therefore determined chloroplast clumping and dispersal, and measured AFs, chlorophyll contents and PAM fluorescence diurnally for H. stipulacea grown under two irradiance regimes. Diurnal chloroplast clumping occurred in high-light grown (HL) plants (∼450 μmol photons m⁻² s⁻¹ during midday), which was accompanied by a decrease in AF values (from 0.56 in the early morning to 0.34 at midday) but not in the chlorophyll content. Also, non-photochemical quenching (measured as NPQ) increased during the day in these plants. No such chloroplast movements and, thus, no diurnal changes in AF values (0.60 ± 0.04 throughout the day), and no changes in NPQ, were found in low-light grown (LL) plants (∼150 μmol photons m⁻² s⁻¹ during midday). As a consequence of the chloroplast clumping in HL plants, and its effect on AF values, maximal ETRs did not differ significantly between HL and LL plants. This finding thus shows the importance of taking into account changing AF values along the day when calculating ETRs of H. stipulacea, and other seagrasses potentially featuring diurnally changing AFs, under high-irradiance conditions.     

Effects of experimental thinning on the growth and maturation of the brown alga Undaria pinnatifida (Laminariales; Phaeophyta) cultivated in Matsushima Bay,northern Japan

In northern Japan, massive production of high-quality specimens of the kelp Undaria pinnatifida prior to the traditional harvest season is strongly needed to meet commercial demand. To address this need, we tested the effects of controlled thinning by cutting small plants on the growth and maturation of sporophytes cultivated in Matsushima Bay, northern Japan. In early December 2009, the stocking densities of cultivated kelps were thinned to 10 and 5 plants per 4 cm thread section in two experimental groups. In contrast, no thinning was conducted during cultivation until March 2010 in the control group (about 16–20 plants per 4 cm thread section). Morphological features, photosynthesis, and carbon and nitrogen contents were compared among the three groups. Compared to kelps of control group, total length, stipe width, the length and width of sporophyll, and dry weights of blade and sporophyll showed significantly higher values in kelps of the two experimental groups. However, stipe length of kelps of control group was significantly greater than those of the two experimental groups. The photosynthetic rates and nitrogen contents of kelps of the two experimental groups were significantly greater than those of the control from January to March. No significant differences occurred in all these parameters between kelps of the two experimental groups. Based on these results, the growth and maturation of U. pinnatifida sporophytes can be promoted greatly by experimental thinning, and this will improve production of high-quality specimens of kelps and increase economic returns before the traditional harvest season.     

Growth and reproductive patterns of <Emphasis Type="Italic">Undaria pinnatifida</Emphasis> sporophytes in a cultivation farm in Busan,Korea

Monthly growth and reproduction of Undaria pinnatifida sporophytes were examined over a period of 5 months in a cultivation farm in Korea. A total of 11 characters of Undaria were measured to determine a reliable morphological character representing its growth and reproduction. Plant weight of Undaria sporophytes increased steadily over the experimental period, but it increased in four different ways. Undaria pinnatifida increased body weight by growth in length and width (October–early December), and by growth in width with the thickening of blade and stipe when sporophytes began to be fertile (December–January). In the middle of January, growth in length and width had almost stopped with the maturation of Undaria sporophytes. Finally, the weight of Undaria increased again by growth in width at the end of February. Present results indicate that Undaria sporophytes increase body weight by growth in length and width at different times, and the relationship between reproduction and vegetative growth is exclusive. Plant weight was positively correlated and fitted well with stipe width and blade width. The blade of Undaria was very thin (ca. 254 μm) and breakable by wave action, but its stipe was strong and relatively thick (ca. 8.7 mm). Furthermore, the fertility of U. pinnatifida was fitted better with stipe width than blade width. Thus, we suggest that the stipe width is the most feasible character with which to estimate the growth and reproduction of U. pinnatifida sporophytes in the cultivation farm.     

Primary production,respiration and calcification of the temperate free-living coralline alga Lithothamnion corallioides

Calcification and primary production responses to irradiance in the temperate coralline alga Lithothamnion corallioides were measured in summer 2004 and winter 2005 in the Bay of Brest. Coralline algae were incubated in dark and clear bottles exposed to different irradiances. Net primary production reached 1.5 μmol C g⁻¹ dry wt h⁻¹ in August and was twice as high as in January–February. Dark respiration showed significant seasonal variations, being three-fold higher in summer. Maximum calcification varied from 0.6 μmol g⁻¹ dry wt h⁻¹ in summer 2004 to 0.4 μmol g⁻¹ dry wt h⁻¹ in winter 2005. According to P–E curves and the daily course of irradiance, estimated daily net production and calcification reached 131 μg C g⁻¹ dry wt and 970 μg CaCO₃ g⁻¹ dry wt in summer 2004, and 36 μg C g⁻¹ dry wt and 336 μg CaCO₃ g⁻¹ dry wt in winter 2005. The net primary production of natural L. corallioides populations in shallow waters was estimated at 10–600 g C m⁻² y⁻¹, depending on depth and algal biomass. The mean annual calcification of L. corallioides populations varied from 300 to 3000 g CaCO₃ m⁻². These results are similar to those reported for tropical coralline algae in terms of carbon and carbonate productivity. Therefore, L. corallioides can be considered as a key element of carbon and carbonate cycles in the shallow coastal waters where they live.     

Elk Kelp, Pelagophycus porra, distribution limited due to susceptibility of microscopic stages to high light

Elk Kelp, Pelagophycus porra (Leman) Setchell, is commonly observed in deep (20-50 m) water along the outer edge of Giant Kelp, Macrocystis pyrifera (C. Agardh), forests in southern California, USA and northern Baja California, Mexico, but rarely occurs in shallower water or within the adjacent M. pyrifera beds. To investigate the factors that limit P. porra from establishing populations within these shallower habitats, juvenile P. porra sporophytes were transplanted from a deep (20 m) water P. porra zone to shallower (15 and 8 m) sites within and inside an adjacent M. pyrifera beds. Transplanted P. porra exhibited no differences in survival or growth among depths, although reproductive maturity was observed only at the two shallower sites. When P. porra propagules were experimentally introduced to areas under dense M. pyrifera canopies and areas cleared of the M. pyrifera canopies within the shallower M. pyrifera bed, P. porra still failed to recruit even though this procedure resulted in a four-fold increase in recruitment in the natural P. porra zone. Laboratory-based culturing of P. porra microscopic stages revealed that they grew and survived better under low light conditions characteristic of the natural P. porra zone (2-4 μmol photons m^− 2 s^− 1) than under higher light conditions characteristic of the M. pyrifera beds(18-20 μmol photons m^− 2 s^− 1). Pulse-amplitude modulated (PAM) fluorometry indicated that while naturally-occurring P. porra's adult sporophytes were able to photoacclimate to increasing irradiance as they grew from the benthos towards the surface, P. porra's microscopic stages were unable to photoacclimate to increased irradiances, and subsequently exhibited 100% mortality under these higher light conditions. Altogether, our study suggests that vulnerability of P. porra microscopic stages to higher irradiances appears to be the primary factor inhibiting P. porra from establishing populations in shallower water and stresses the importance of a multiple life-history approach when investigating species distributions.     

Influence of ammonium sulphate feeding time on fed-batch Arthrospira (Spirulina) platensis cultivation and biomass composition with and without pH control

Previous work demonstrated that a mixture of NH₄Cl and KNO₃ as nitrogen source was beneficial to fed-batch Arthrospira (Spirulina) platensis cultivation, in terms of either lower costs or higher cell concentration. On the basis of those results, this study focused on the use of a cheaper nitrogen source mixture, namely (NH₄)₂SO₄ plus NaNO₃, varying the ammonium feeding time (T = 7-15 days), either controlling the pH by CO₂ addition or not. A. platensis was cultivated in mini-tanks at 30 °C, 156 μmol photons m⁻² s⁻¹, and starting cell concentration of 400 mg L⁻¹, on a modified Schlösser medium. T = 13 days under pH control were selected as optimum conditions, ensuring the best results in terms of biomass production (maximum cell concentration of 2911 mg L⁻¹, cell productivity of 179 mg L⁻¹ d⁻¹ and specific growth rate of 0.77 d⁻¹) and satisfactory protein and lipid contents (around 30% each).     

Ovalbumin labeling with p-hydroxymercurybenzoate: The effect of different denaturing agents and the kinetics of reaction

The aim of our study was to investigate how denaturing agents commonly used in protein analysis influence the labeling between a reactive molecule and proteins. For this reason, we investigated the labeling of ovalbumin (OVA) as a globular model protein with p-hydroxymercurybenzoate (pHMB) in its native state (phosphate buffer solution) and in different denaturing conditions (8 mol L⁻¹ urea, 3 mol L⁻¹ guanidinium thiocyanate, 6 mol L⁻¹ guanidinium chloride, 0.2% sodium dodecyl sulfate, and 20% methanol). In addition to chemical denaturation, thermal denaturation was also tested. The protein was pre-column simultaneously denatured and derivatized, and the pHMB-labeled denatured OVA complexes were analyzed by size exclusion chromatography (SEC) coupled online with chemical vapor generation–atomic fluorescence spectrometry (CVG–AFS). The number of –SH groups titrated greatly depends on the protein structure in solution. Indeed, we found that, depending on the adopted denaturing conditions, OVA gave different aggregate species that influence the complexation process. The results were compared with those obtained by a common alternative procedure for the titration of –SH groups that employs monobromobimane (mBBr) as tagging molecule and molecular fluorescence spectroscopy as detection technique.     

Exposure times in rapid light curves affect photosynthetic parameters in algae

Short-and long-duration light curves were applied to four macroalgae (Ulva sp., Codium fragile, Ecklonia radiata and Lessonia variegata), and two microalgal species (Chlorella emersonii and Chaetoceros muellerii). With increasing light curve duration, the maximal relative electron transport rate increased by a factor of three in E. radiata, and by factors of 1.25 and 1.23 in C. emersonii and L. variegata, respectively, but did not change in C. fragile and Ch. muellerii. The light saturation point E_k increased by 26 μmol photons m⁻² s⁻¹ in C. emersonii and 20 μmol photons m⁻² s⁻¹ in Ch. muellerii and E. radiata with elevated light curve exposure times. Oscillatory patterns of the continuous fluorescence readings reflect accumulation of Q_A. Continuous fluorescence values increased, or decreased, by approximately 10% within light curve increments. However, oscillations of 25% were not uncommon, which shows that cells are changing their photo-physiological response state during steady light conditions. Increasing dark acclimation times prior to light curve application lowered maximal relative electron transport rates in the C. emersonii (from 28 ± 1.7 to 25 ± 1.2 for 15 and 95 min dark acclimation in short-duration light curves respectively). This effect was counterbalanced by longer light curve application. It can therefore be concluded that manipulation of light exposure and dark incubation prior to the experiment affects the photosynthetic response, presumably due to different activation states of photosynthetic and photoprotective mechanisms. The highly species-specific photo-response patterns imply that a common rapid light curve protocol will generate artefacts in some species.     

Cryopreservation of Greenshell™ mussel (Perna canaliculus) trochophore larvae

The Greenshell™ mussel (Perna canaliculus) is the main shellfish species farmed in New Zealand. The aim of this study was to evaluate the effects of cryoprotectant concentration, loading and unloading strategy as well as freezing and thawing method in order to develop a protocol for cryopreservation of trochophore larvae (16–20 h old). Toxicity tests showed that levels of 10–15% ethylene glycol (EG) were not toxic to larvae and could be loaded and unloaded in a single step. Through cryopreservation experiments, we designed a cryopreservation protocol that enabled 40–60% of trochophores to develop to D-larvae when normalized to controls. The protocol involved: holding at 0 °C for 5 min, then cooling at 1 °C min⁻¹ to −10 °C, holding for a further 5 min, then cooling at 0.5 °C min⁻¹ to −35 °C followed by a 5 min hold and then plunging into liquid nitrogen. A final larval rearing experiment of 18 days was conducted to assess the ability of these frozen larvae to develop further. Results showed that only 2.8% of the frozen trochophores were able to develop to competent pediveligers.     

Protective effects of propolis on cryopreservation of common carp (Cyprinus carpio) sperm

Cryopreservation of sperm is common procedures in aquaculture, particularly used for routine in artificial insemination. However, these application cause damages and adversely affected sperm motility, viability and consequently lower hatching rates. The objective of this study is to determine whether propolis has an effect on cryopreservation and fertilization ability and to investigate the potential protective effect of propolis on spermatozoa of Cyprinus carpio. Many studies have been done in cryopreservation offish spermatozoa, but none of them contain propolis in extender composition. The extenders were prepared by using modified Kurokura Solution to which 10% Me₂SO added with different levels of propolis (0.2, 0.4, 0.6, 0.8 and 1 mg ml⁻¹) and 10% egg yolk (as a control without propolis). The pooled semen samples diluted at the ratio of 1:9 by the extenders were subjected to cryopreservation. The percentage and duration of motility and fertilization tests of cryopreserved sperm samples have been done immediately after thawing and compared with control and fresh semen. The extenders containing propolis exhibited higher percentage motility and motility duration than control group (P < 0.05). Especially the group IV (0.8 mg ml⁻¹ propolis) and the group V (1 mg ml⁻¹ propolis) showed significant positive effects on both post thaw motility and hatching ability. The propolis maintained the integrity of the spermatozoa during the cryopreservation process. Evaluating with its contents, it has been shown that propolis is an appropriate cryoprotective agent in fish semen.     

Bicarbonate use in three aquatic plants

Our study aimed to test the ability of aquatic plants to use bicarbonate when acclimated to three different bicarbonate concentrations. To this end, we performed experiments with the three species Ceratophyllum demersum, Egeria densa, Lagarosiphon major to determine photosynthetic rates under varying bicarbonate concentrations. We measured bicarbonate use efficiency, photosynthetic performance and respiration. For all species, our results revealed that photosynthetic rates were highest in replicates grown at low alkalinity. Thus, E. densa had approx. five times higher rates at low (264 ± 15 μmol O₂ g⁻¹ DW h⁻¹) than at high alkalinity (50 ± 27 μmol O₂ g⁻¹ DW h⁻¹), C. demersum had three times higher rates (336 ± 95 and 120 ± 31 μmol O₂ g⁻¹ DW h⁻¹), and L. major doubled its rates at low alkalinity (634 ± 114 and 322 ± 119 μmol O₂ g⁻¹ DW h⁻¹). Similar results were obtained for bicarbonate use efficiency by E. densa (136 ± 44 and 43 ± 10 μmol O₂ mequiv. L⁻¹ g⁻¹ DW h⁻¹) and L. major (244 ± 29 and 82 ± 24 μmol O₂ mequiv. L⁻¹ g⁻¹ DW h⁻¹). As to C. demersum, efficiency was high but unaffected by alkalinity, indicating high adaptation ability to varied alkalinities. A pH drift experiment supported these results. Overall, our results suggest that the three globally widespread worldwide species of our study adapt to low inorganic carbon availability by increasing their efficiency of bicarbonate use.     

Contribution of fern gametophytes to the growth of produced sporophytes on the basis of carbon gain

Sporophytes appeared on most gametophytes of Thelypteris palustris (Salisb.) Schott that reached a certain size, which is interpreted to be a critical size of gametophytes for the production of sporophytes. After sporophytes were produced, attached gametophytes ceased dry weight growth, but the gametophytes which did not produce sporophytes grew successively. It was hypothesized that matter produced by gametophytes was being supplied to young sporophytes. Photosynthesis and respiration of gametophytes with attached sporophytes were not significantly different from that of gametophytes without sporophytes. Photosynthetic activity of gametophytes dropped from 0.18 to 0.03 mol CO₂ g^–1 s^–1 during the growth period. The higher photosynthetic rates of gametophytes in the early growth stage were important for reaching the critical size for sporophyte production in a short time. Sporophytes in the one leaf stage averaged 0.14 mol CO₂ g^–1 s^–1 of photosynthetic activity. The results show that sporophytes that had expanded the first leaf grow by their own photosynthetic production. Gametophytes allocated the photosynthate for sporophytes and it was an important aid before the one-leaf stage. The supportive role of gametophytes ended at that stage.     

Light-dependent phosphate uptake of a submersed macrophyte Myriophyllum spicatum L.

The uptake kinetics of phosphate (Pi) by Myriophyllum spicatum was determined from adsorption and absorption under light and dark conditions. Pi uptake was light dependent and showed saturation following the Michaelis-Menten relation (in light: V = 16.91 × [Pi](1.335 + [Pi]), R² = 0.90, p < 0.001; in the dark: V = 5.13 × [Pi](0.351 + [Pi]), R² = 0.77, p < 0.001). Around 77% of the loss of Pi in the water column was absorbed into the tissue of M. spicatum, and only 23% was adsorbed on the surface of the plant shoots. Our study shows that M. spicatum shoots have a much higher affinity (in light: 3.9 μmol g⁻¹ dw h⁻¹ μM⁻¹; in the dark: 3.7 μmol g⁻¹ dw h⁻¹ μM⁻¹) and V_max (maximum uptake rate, shoot light) for Pi uptake than many other aquatic macrophytes (in light: 0.002-0.23 μmol g⁻¹ dw h⁻¹ μM⁻¹; in the dark: 0.002-0.19 μmol g⁻¹ dw h⁻¹ μM⁻¹), which may provide a competitive advantage over other macrophytes across a wide range of Pi concentrations.     

Diurnal courses of net photosynthesis and photosystem II quantum efficiency of submerged Lagarosiphon major under natural light conditions

An optode device for net-photosynthesis measurements, based on oxygen-depending quenching of fluorescence from O₂-specific sensors, and PAM fluorometry have been used to study diurnal courses of net-photosynthesis and the F_v/F_m ratio of the submerged plant Lagarosiphon major. Plants were pre-cultivated and studied in large mesocosm flow-through outdoor tanks under 50% and 80% shade cloth, respectively. Growth under the different shade cloths resulted in similar light compensation points (∼20 μmol photons m⁻² s⁻¹), but strongly different light saturation levels, with about 150 μmol m⁻² s⁻¹ for plants grown under 80% shade cloth and about 350 μmol m⁻² s⁻¹ for plants grown under 50% shade cloth. Plants under both growth conditions showed a transient reduction of the maximum F_v/F_m value in the afternoon (down to 70% of the morning control values under 80% shade cloth and down to 85% under 50% shade cloth), which was not accompanied by a reduction of the net photosynthetic rate. This indicated that the fluorescence parameter F_v/F_m must not be a reliable indicator of the rate of photosynthesis under all conditions. The new photo-optical device became evidenced as a valuable tool not only for laboratory experiments, but also for field studies of gas exchange of submerged plants.     

Nitrogen nutrition of Canna indica: Effects of ammonium versus nitrate on growth, biomass allocation, photosynthesis, nitrate reductase activity and N uptake rates

The effects of inorganic nitrogen (N) source (NH₄⁺, NO₃⁻ or both) on growth, biomass allocation, photosynthesis, N uptake rate, nitrate reductase activity and mineral composition of Canna indica were studied in hydroponic culture. The relative growth rates (0.05-0.06 g g⁻¹ d⁻¹), biomass allocation and plant morphology of C. indica were indifferent to N nutrition. However, NH₄⁺ fed plants had higher concentrations of N in the tissues, lower concentrations of mineral cations and higher contents of chlorophylls in the leaves compared to NO₃⁻ fed plants suggesting a slight advantage of NH₄⁺ nutrition. The NO₃⁻ fed plants had lower light-saturated rates of photosynthesis (22.5 μmol m⁻² s⁻¹) than NH₄⁺ and NH₄⁺/NO₃⁻ fed plants (24.4-25.6 μmol m⁻² s⁻¹) when expressed per unit leaf area, but similar rates when expressed on a chlorophyll basis. Maximum uptake rates (V_max) of NO₃⁻ did not differ between treatments (24-35 μmol N g⁻¹ root DW h⁻¹), but V_max for NH₄⁺ was highest in NH₄⁺ fed plants (81 μmol N g⁻¹ root DW h⁻¹), intermediate in the NH₄NO₃ fed plants (52 μmol N g⁻¹ root DW h⁻¹), and lowest in the NO₃⁻ fed plants (28 μmol N g⁻¹ root DW h⁻¹). Nitrate reductase activity (NRA) was highest in leaves and was induced by NO₃⁻ in the culture solutions corresponding to the pattern seen in fast growing terrestrial species. Plants fed with only NO₃⁻ had high NRA (22 and 8 μmol NO₂⁻ g⁻¹ DW h⁻¹ in leaves and roots, respectively) whereas NRA in NH₄⁺ fed plants was close to zero. Plants supplied with both forms of N had intermediate NRA suggesting that C. indica takes up and assimilate NO₃⁻ in the presence of NH₄⁺. Our results show that C. indica is relatively indifferent to inorganic N source, which together with its high growth rate contributes to explain the occurrence of this species in flooded wetland soils as well as on terrestrial soils. Furthermore, it is concluded that C. indica is suitable for use in different types of constructed wetlands.     

Effect of water flow velocity on growth and morphology of cultured <Emphasis Type="Italic">Undaria pinnatifida</Emphasis> sporophytes (Laminariales,Phaeophyceae) in Okirai Bay on the Sanriku coast,Northeast Japan

Undaria pinnatifida sporophytes, originating from the same strain, were cultured at the commercial cultivation site exposed to wave action and the uncultivated site protected from water action of Okirai Bay, Northeast Japan, from January to April 2007; simultaneously, water flow velocity, water temperature, salinity, NO₃ + NO₂, and chlorophyll a were monitored to investigate the effect of water environment on their growth and morphology. Water temperature and salinity fluctuated within the optimal range for their growth whereas water flow velocity at the cultivation site was greatly fast compared with that at the uncultivated site. Successive chlorophyll a increases synchronized with NO₃ + NO₂ decreases were observed only at the uncultivated site for over a month; indicating developments of phytoplankton blooms and their nutrient consumption under the low-flow condition. Meanwhile, blade growth rate of cultured sporophytes was higher at the cultivation site than at the uncultivated site. Their thallus size expressed by six morphological characters (blade length, stipe length, blade wet weight, stipe wet weight, blade width, and undivided blade width) at the cultivation site became large in comparison with that at the uncultivated site. Their three morphological correlations (correlations between blade length and thallus length; blade wet weight and thallus wet weight; and undivided blade width and blade width) differed between the sites. They produced a thick and flat blade at the cultivation site but formed a thin and wrinkled blade at the uncultivated site. These results show the significant impact of water flow velocity on their growth and morphology.     

Morphological and physiological adaptive traits of Mediterranean narrow endemic plants: The case of Centaurea gymnocarpa (Capraia Island,Italy)

A case study on Centaurea gymnocarpa Moris & De Not., a narrow endemic species, was carried out by analyzing its morphological, anatomical, and physiological traits in response to natural habitat stress factors under Mediterranean climate conditions. The results underline that the species is particularly adapted to the environment where it naturally grows. At the plant level, the above-ground/below-ground dry mass (1.73 ± 0.60) shows its investment predominately in the above-ground structure with a resulting total leaf area per plant of 1399 ± 94 cm². The senescent attached leaves at the base of the plant contribute to limit leaf transpiration by shading soil around the plant. Moreover, the dense C. gymnocarpa leaf pubescence, leaf rolling, the relatively high leaf mass area (LMA = 12.3 ± 1.3 mg cm⁻²) and leaf tissue density (LTD = 427 ± 44 mg cm⁻³) contribute to limit leaf transpiration, also postponing leaf death under dry conditions. At the physiological level, a relatively low respiration/photosynthesis ratio (R/P_N) in spring results from high R [2.26 ± 0.59 μmol (CO₂) m⁻² s⁻¹] and P_N [12.3 ± 1.5 μmol (CO₂) m⁻² s⁻¹]. The high photosynthetic nitrogen use efficiency [PNUE = 15.5 ± 0.4 μmol (CO₂) g⁻¹ (N) s⁻¹] shows the large amount of nitrogen (N) invested in the photosynthetic machinery of new leaves, associated to a high chlorophyll content (Chl = 35 ± 5 SPAD units). On the contrary, the highest R/P_N ratio (1.75 ± 0.19) in summer is due to a significant P_N decrease and increase of R in response to drought. The low PNUE [1.5 ± 0.2 μmol (CO₂) g⁻¹ (N) s⁻¹] in this season is indicative of a greater N investment in leaf cell walls which may contribute to limit transpiration. On the contrary, the low R/P_N ratio (0.05 ± 0.02) in winter is resulting from the limited enzyme activity of the respiratory apparatus [R = 0.23 ± 0.08 μmol (CO₂) m⁻² s⁻¹] while the low PNUE [3.5 ± 0.2 μmol (CO₂) g⁻¹ (N) s⁻¹] suggests that low temperatures additionally limit plant production. The experiment of the imposed water stress confirms that the C. gymnocarpa growth capability is in conformity with the severe conditions of its natural habitat, likewise as it may be the case with others narrow endemic species that have occupied niches with similar extreme conditions.