First estimates of productivity in Lessonia trabeculata and Lessonia nigrescens (Phaeophyceae, Laminariales) from the southeast Pacific

Lessonia is the main Laminariales found along the southeast Pacific coast. Lessonia nigrescens Bory de Saint‐Vincent in the intertidal and Lessonia trabeculata Villouta et Santelices in the subtidal, are the most important habitat constructors in rocky coastal communities in northern and central Chile. In both species, the seasonal production and erosion of distal tissue were estimated in biomass units using the Area of Constant Biomass Model that combined the individual blade elongation, obtained with the traditional hole‐punching method, with the blade length and biomass distribution along the blade. In austral late spring (December 96) and autumn (May 97), blade production and erosion were transformed to the level of population from standing stock measurements (number and biomass of blades and plants per substrate area), considering that previous blade weight analysis showed the highest and lowest values at these times, as well as the population parameter extremes that were expected to occur. Both species displayed a seasonal pattern, with a production increase in later winter and spring and decrease towards the end of summer that coincided with higher distal tissue erosion. At the level of individual blades, Lessonia trabeculata showed higher mean production (0.026 g dw d⁻¹) and erosion (0.01 g dw d⁻¹) than L. nigrescens (production 0.01 g dw d⁻¹ and loss 0.002 g dw d⁻¹). The standing stocks, with respect to density and biomass, were similar in spring and autumn for both populations. Nevertheless, the net productivity (production minus erosion) of the intertidal L. nigrescens showed greater values due to the greater density of blades (2112 ± 1360 (SE) blades m⁻²) compared with the subtidal L. trabeculata (527 ± 151 (SE) blades m⁻²). Spring net productivities of 42 g dw m⁻²d⁻¹ (254 g ww m⁻²d⁻¹; 11.46 gC m⁻²d⁻¹) for L. nigrescens and 11 g dw m⁻² d⁻¹ (64 g ww m⁻² d⁻¹; 2.46 gC m⁻²d⁻¹) for L. trabeculata were estimated. A preliminary model of production and biomass fate for Lessonia populations is proposed.     

A comparison between two populations of Lessonia trabeculata (Phaeophyta: Laminariales) microscopic stages

Lessonia trabeculata is the most important brown seaweed in northern Chilean subtidal environments. It usually grows in areas with high to moderately high water movement, and only occasionally in sheltered habitats. These different hydrodynamic environments produce morphologically and physiologically distinct plants. The purpose of this study was to determine probable differences in reproductive features between two L. trabeculata populations exposed to different water movement intensities. The zoospore production, settlement, germination, gametophyte fertility and survival were studied.Zoospore release and spore attachment capacity were higher from blades from El Francés Beach, the locality with higher water movement energy. Germination at 24 hours was higher in the germlings from El Francés Beach than those from La Herradura Bay. However, after 4 days of culture germination rates were similar. Average survival rates of gametophytes were similar for both populations. After 20 days culturing gametophytes from El Francés Beach fertilization was observed, while in samples from La Herradura Bay the sporophytes were seen at 27d.The results suggest a higher reproductive vitality in plants of Lessonia trabeculata living in an environment with an active water movement, than plants growing in a sheltered environment.     

Parthenogenesis in the brown alga Lessonia nigrescens (Laminariales,Phaeophyceae) from central Chile1*

Parthenogenesis, the development of female gametes without fertilization, is relatively common in brown algae, although limited quantitative information on the phenomenon is available. Its occurrence is reported for the first time in Lessonia nigrescens Bory, a member of the Laminariales and a key ecological component of the benthic algal communities along the Chilean coast. Isolated female gametophytes developed into parthenosporophytes throughout the year, with a maximum in spring to early summer. Isolated male gametophytes, on the other hand, never developed fronds. Parthenosporophytes obtained in the laboratory developed normally when cultivated under greenhouse conditions, and the resulting individuals were indistinguishable in size, shape, texture, and color from heterozygous sporophytes. Quantification of DNA of various tissues demonstrated that early during their development, parthenosporophytes duplicated their DNA content, displaying levels similar to heterozygous sporophytes and almost twice the level found in gametophytes. One out of 45 individuals from a field population yielded only female gametophytes, strongly suggesting that parthenogenesis does occur in wild stands of L. nigrescens.     

Culture studies on early development of Lessonia trabeculata (Phaeophyceae,Laminariales): Seasonality and acclimation to light and temperature

Lessonia trabeculata is an important economic and ecological algal resource of Chile. Due to intense use in abalone and alginate industries, severe problems of over‐harvesting are emerging. We compared sporophyte‐initiation and ‐growth in two populations from northern and southern Chile (Bahía Inglesa and Maicolpué) under laboratory conditions. Irradiance and temperature were the most important factors affecting gametophyte development. Meiospores harvested in spring exhibited maximum reproductive and growth potential, while spores released during autumn were moribund, and died within a few days. In both study sites, we found evidence for acclimation: Meiospores collected in summer required higher levels of irradiance and temperature for maximum development than winter spores. Juvenile sporophytes from both localities responded similarly to temperature and/or irradiance. The best conditions for recruitment of sporophytes from both localities were 15°C and white fluorescent light of 40–70 μmol m^?2 s^?1. Although due to its low growth potential L. trabeculata is not a good candidate for mariculture, our results provide the necessary knowledge for laboratory‐based seedling production, which is needed for restoration and repopulation projects in damaged areas.     

Culture and growth of Lessonia trabeculata (Phaeophyta,Laminariales) juvenile sporophytes in La Herradura de Guayacan Bay,northern Chile

Lessonia trabeculata is one of the major kelps found along the northern coast of Chile. In addition to its ecological and economic importance, L. trabeculata may be severely affected by environmental disturbances such as El Níño, which during 1982–1983 cleared wide areas along the coast of Peru and Chile. The main goal of this work was to mass culture L. trabeculata and to observe the growth of sporophytes obtained in the laboratory and cultured in the sea. Juvenile sporophytes obtained in the laboratory were attached between 1 and 6 m in depth. The linear growth rate, as blade elongation, was recorded weekly for seven months. No significant differences (p < 0.05) were found in sporophyte blade linear growth at different depths. The best elongation growth rate was 7.5 ± 1.6 mm d^–1 at 3 m during March. This preliminary work suggests that L. trabeculata follows an annual growth cycle similar to that of other Laminariales with a high rate of blade elongation during the summer and decreasing towards autumn. This species can be considered a potential candidate for aquaculture to increase the availability of raw material and aid in repopulation of overexploited areas.     

Variation in Patterns of Metal Accumulation in Thallus Parts of Lessonia trabeculata (Laminariales; Phaeophyceae): Implications for Biomonitoring

Seaweeds are well known to concentrate metals from seawater and have been employed as monitors of metal pollution in coastal waters and estuaries. However, research showing that various intrinsic and extrinsic factors can influence metal accumulation, raises doubts about the basis for using seaweeds in biomonitoring programmes. The thallus of brown seaweeds of the order Laminariales (kelps) is morphologically complex but there is limited information about the variation in metal accumulation between the different parts, which might result in erroneous conclusions being drawn if not accounted for in the biomonitoring protocol. To assess patterns of individual metals in the differentiated parts of the thallus (blade, stipe, holdfast), concentrations of a wide range of essential and non-essential metals (Fe, Cr, Cu, Zn, Mn, Pb, Cd, Ni and Al) were measured in the kelp Lessonia trabeculata. Seaweeds were collected from three sampling stations located at 5, 30 and 60 m from an illegal sewage outfall close to Ventanas, Chile and from a pristine location at Faro Curaumilla. For the majority of metals the highest concentrations in bottom sediment and seaweed samples were found at the site closest to the outfall, with concentrations decreasing with distance from the outfall and at control stations; the exception was Cd, concentrations of which were higher at control stations. The patterns of metal concentrations in different thallus parts were metal specific and independent of sampling station. These results and the available literature suggest that biomonitoring of metals using seaweeds must take account of differences in the accumulation of metals in thallus parts of complex seaweeds.     

Artificial induction of sporulation in Lessonia (Phaeophyta, Laminariales)

The effects of three physical variables on sporulation, germination and gametophyte survival were evaluated on two species of Laminariales at Coquimbo, Chile (30° S). Parameters were evaluated using reproductive fronds of Lessonia nigrescens Bory and Lessonia trabeculata Villouta and Santelices with treatment variables including type of pre-rinse, dehydration, and temperature. Although the response was slightly different in the two species, it was found that pre-rinsing the blades with tap water produced marked sporulation without affecting germination and survival. Minor effects were observed with other variables, including positive effects of lower temperature and negative effects of longer dehydration periods. The high commercial value of these two species has promoted much interest in their artificial propagation in Chile. This revised version was published online in June 2006 with corrections to the Cover Date.     

Molecular phylogeny and timing of radiation in Lessonia (Phaeophyceae,Laminariales)

The genus Lessonia Bory de Saint‐Vincent is distributed solely in the Southern Hemisphere, with four species found in South America and five in Australasia. Our goal was to determine the evolutionary relationships between the Lessonia species of the two disjunct areas and hypothesize dispersal patterns and timing of radiation. We combined mitochondria, plastid and nuclear markers in a comprehensive dataset from multiple individuals per known species. Furthermore, for some species we added samples from multiple populations to take account of their widespread distribution over known bioregions. In all analyses the Australasian Lessonia formed a monophyletic group and in most analyses the South American species form a paraphyletic grade. Delimitations of the accepted species are highly supported except for L. variegata J. Agardh and L. nigrescens Bory de Saint‐Vincent. We showed for the first time four lineages for the New Zealand endemic Lessonia variegata with an unexpected high level of genetic differentiation. Our analysis cannot resolve whether the dispersal of the common ancestor of this genus was from the Americas to Australasia or vice versa. Molecular clock analysis suggested that a sudden radiation took place in Australasia c. 3.5 Mya when almost all Australasian species diverged within a time frame of only 35 000 years.     

Ecological effects of harvesting Lessonia (Laminariales,Phaeophyta) in central Chile

Lessonia nigrescens and L. trabeculata are economically important canopy-forming kelps in Chile. Experimental harvesting of stipes above the first dichotomy reduces stipe movement and inter-stipe friction, allowing the development of a heavy epiphytic load and increased grazing. Complete stipe removal leads to holdfast death as neither species is able to simultaneously regenerate all stipes. The invertebrate fauna inside the holdfast does not respond to upper canopy changes, but mortality does occur in partial or complete plant removals. Kelp removal also affects inter-plant distances, results in increased access of grazers to the outside and inside of kelp holdfasts, reduces recruitment of other algal species, and modifies the morphology of L. trabeculata such that the plants become more susceptible to removal by water movement.     

The genus Lessonia Bory (Phaeophyta,Laminariales) in Southern Chile and Argentina

Study of Lessonia in southern Chile and Argentina indicates that it is represented there by three species: L. nigrescens Bory, L. flavicans Bory and L. vadosa Searles sp. nov. Plants of these three species have been confused with each other in the literature, but appear clearly separable. Lessonia nigrescens is distinguished from the others by its massive, solid holdfast and numerous stipes. In Lessonia flavicans and L. vadosa a single stipe arises from a branched holdfast. These two species are distinguishable from each other in details of blade morphology, anatomy, and ecology. The taxonomic status of plants in South America outside of the study area remains unresolved. Populations in the Falkland Islands and in northern Chile in particular are in need of further study.     

Repopulation of intertidal areas with Lessonia nigrescens in northern Chile

Intertidal rocky areas in northern Chile were repopulated experimentally with the brown alga Lessonia nigrescens using spore seeding and placement of reproductive fronds. The results were successful, and it is suggested that methods developed in the field can be done by people without special training.     

Comparative analysis of peroxiredoxin activation in the brown macroalgae Scytosiphon gracilis and Lessonia nigrescens (Phaeophyceae) under copper stress

Among thiol‐dependent peroxidases (TDPs) peroxiredoxins (PRXs) standout, since they are enzymes capable of reducing hydrogen peroxide, alkylhydroperoxides and peroxynitrite, and have been detected in a proteomic study of the copper‐tolerant species Scytosiphon gracilis. In order to determine the importance of these enzymes in copper‐stress tolerance, TDP activity and type II peroxiredoxin (II PRX) protein expression were compared between the opportunistic S. gracilis and the brown kelp Lessonia nigrescens, a species absent from copper‐impacted sites due to insufficient copper‐tolerance mechanisms. Individuals of both species were cultured with increasing copper concentrations (0–300 µg l^?1 Cu) for 96 h and TDP activity and lipoperoxides (LPXs) were determined together with II PRX expression by immunofluorescence and Western blot analysis. The results showed that TDP activity was higher in S. gracilis than L. nigrescens in all copper concentrations, independent of the reducing agent used (dithiothreitol, thioredoxin or glutaredoxin). This activity was copper inhibited in L. nigrescens at lower copper concentrations (20 µg l^?1 Cu) compared to S. gracilis (100 µg l^?1 Cu). The loss of activity coincided in both species with an increase in LPX, which suggests that TDP may control LPX production. Moreover, II PRX protein levels increased under copper stress only in S. gracilis. These results suggest that in S. gracilis TDP, particularly type II peroxiredoxin (II PRX), acts as an active antioxidant barrier attenuating the LPX levels generated by copper, which is not the case in L. nigrescens. Thus, from an ecological point of view these results help explaining the inability of L. nigrescens to flourish in copper‐enriched environments.     

Micropopulation differentiation in phenol content and susceptibility to herbivory in the Chilean kelp Lessonia nigrescenss (Phaeophyta,Laminariales)

Micropopulation differences in phenol content between intertidal and subtidal individuals of the kelp Lessonia nigrescens were found. Subtidal plants showed: (1) significantly higher phenol content than intertidal individuals, in vegetative and reproductive tissues, (2) intra-plant differences, with higher content in apical frond tissues, (3) higher resistance to consumption by herbivorous fishes. The microscopic progeny of subtidal plants showed the same trend as adult plants: (1) haploid spores from subtidal plants had higher phenol content than spores from intertidal individuals, and (2) the microscopic sporophytes derived from subtidal spores and gametophytes were less consumed by herbivorous snails (Tegula tridentata) than those derived from intertidal plant propagules. No increase in phenol content was detected after mechanical injury to experimental fronds, or after transplantation to the subtidal environment.In addition to the absence of inducible responses, the different phenol content between intertidal and subtidal individuals, in adult diploid plants and also in the haploid progeny, suggests that both environments differ someway enough to fix the mentioned features on the plants of Lessonia nigrescens. It is likely that the differences in herbivory between the two distributional extremes contributed to the observed pattern.     

Development and characterization of nine polymorphic microsatellite markers in the Chilean kelp Lessonia nigrescens

A total of nine microsatellite loci were isolated and characterized in the Chilean kelp Lessonia nigrescens Bory. Using two different enriched libraries, we observed 1-14 alleles per locus in two samples of 21 kelp individuals each. The observed heterozygosities ranged from 0.05 to 0.80 and all loci are in Hardy-Weinberg equilibrium for one or both samples. Seventeen samples collected from different sites showed high allele diversity along the species distribution. The variation detected at these markers is currently being used for the study of populations of Lessonia nigrescens at different geographical scales.     

Recruitment of the intertidal kelp Lessonia nigrescens Bory in northern Chile: successional constraints and opportunities

The strong 1982/83 El Niño event caused local extinction in populations of the low intertidal kelp Lessonia nigrescens Bory on northern Chilean coasts. The kelp has partially recolonized, but its recovery has apparently been low. This study documents the effect of biological factors that potentially decrease the velocity of kelp recolonization in northern Chile. A removal experiment showed that encrusting coralline algae, which dominate the lower intertidal, significantly reduce the recruitment of L. nigrescens in the presence of herbivores. Epithallium shedding by encrusting corallines is the most probable cause for this inhibition process. On the other hand, grazing on encrusting corallines by the chiton Enoplochiton niger (Barnes), a large-sized herbivore (10–20 cm length), could also affect kelp recruitment. The intestinal content of E. niger, where encrusting corallines are the main item (84.2%), revealed the presence of L. nigrescens. Among the natural substrata on which kelp recruits, measured at five different localities, the coarsely-branched alga Corallina officinalis (Decaisne) Kützing reached the highest frequency, despite its extremely low cover (< 1%) in the field. This suggests that settlement on turfs of C. officinalis allows the kelp to escape from herbivory, thus facilitating its recruitment. The roles of timing of kelp recruitment and seasonal grazing are also discussed.     

Antioxidant enzymes gene expression and antihypertensive effects of seaweeds Ulva linza and Lessonia trabeculata in rats fed a high-fat and high-sucrose diet

The objective of this work was to evaluate the antihypertensive and antioxidant effect of seaweeds (Ulva linza and Lessonia trabeculata) in rats which were fed a hypercaloric diet. Seaweed at 400 mg kg^?1 of body weight was administered for 8 weeks to Wistar rats that were fed with a standard diet or a hypercaloric diet. Intra-abdominal fat, insulin resistance, and lipid profile of the rats were determined. Liver was isolated to determine antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx)] activity and gene expression. The administration of seaweed to the rats reduced the levels of intra-abdominal fat, arterial blood pressure, insulin resistance, and cholesterol and triglyceride serum levels. U. linza reduced the GPx activity in control animals but increased it in animals with MS, which could be reduced by using L. trabeculata. Both seaweeds diminished the SOD and GPx expression and increased CAT in control group. Both seaweeds reduced the CAT expression in animals with metabolic syndrome. Combined effects of the different compounds found in the seaweeds explain the regulating effect over different antioxidant enzymes and metabolic pathways that protect the animals fed a hypercaloric diet against the development of hypertension, hyperglycemia, and obesity.     

Morphological,ultrastructural, and genetic characterization of coalescence in the intertidal and shallow subtidal kelps Lessonia spicata and L. berteroana (Laminariales,Heterokontophyta)

Coalescing macroalgae may fuse with conspecifics, forming genetically heterogeneous entities known as chimera. This process has been shown in taxa from roughly half the red algal orders and in the Codium species, a green alga. Field observations indicate that common and dominant kelps along central Chile exhibit a fused holdfast. We evaluated whether such fusions are true coalescence processes in Lessonia spicata and Lessonia berteroana. To this end, we characterized the ultrastructural event involved in holdfast fusion in the laboratory. Additionally, coalescence in natural populations was quantified by measuring the frequency of individuals with genetically heterogenic stipes within the same holdfast. Results indicate that coalescence appears as a frequent process in laboratory, mostly restricted to intraspecific fusions. During fusion, the meristodermatic cells located in the contact area modify their morphology and reduce the number of plastids, mitochondria, and cell inclusions. The cell wall becomes much thinner and develops plasmodesmata, enhancing communication with equivalent cells of the other coalescencing individual. Stipe genotyping indicates that there is a widespread occurrence of chimerism in both species and genetic heterogeneity is increasing directly with the increasing number of stipes. The combination of results suggests that kelp frequently coalesce in the field, and the histological response observed approaches that of red algae. Since kelps are part of the dominant vegetation in low intertidal and shallow subtidal beds, the adaptive values of coalescence in these species should be evaluated. It is concluded that coalescence and chimerism are evolutionary convergent processes, occurring in all three major groups of seaweeds.     

Demography and feeding behavior of the kelp crab Taliepus marginatus in subtidal habitats dominated by the kelps Macrocystis pyrifera or Lessonia trabeculata

We studied the population and feeding ecology of the kelp crab Taliepus marginatus in subtidal kelp forests dominated by either of two morphologically different kelp species (Macrocystis pyrifera or Lessonia trabeculata) in northern Chile. The sizes and abundances of T. marginatus differed between the two kelp habitats. Kelp crabs were more abundant in the M. pyrifera forest than in the L. trabeculata forest. Size‐frequency distributions showed that juvenile and immature crabs were more common in the M. pyrifera forest than in the L. trabeculata forest, where reproductive adults predominated. The smaller crabs in the M. pyrifera habitat also consumed a higher proportion of kelp tissues than the larger crabs in the L. trabeculata habitat, which had a higher proportion of animal food in their diet. In both kelp forests, individuals of T. marginatus showed a similar pattern of nocturnal feeding over a 24‐h period, consuming more food at night than during the day. The more complex and dense forests of M. pyrifera appear to present better nursery habitats for juvenile kelp crabs than the more open and less dense forests dominated by L. trabeculata. These results suggest that the role of the two kelp habitats for T. marginatus varies during the life cycle of the kelp crabs, with M. pyrifera tending to have nursery function and L. trabeculata being more suitable as a reproductive habitat.     

Growth and reproductive responses of Laminaria longicruris (Laminariales,Phaeophyta) to nutrient enrichment

A series of comparative culture experiments were conducted in order to determine responses of Laminaria longicruris male and female gametophytes and juvenile sporophytes to several temperatures (5, 10, 15, 20 °C), light levels (10, 35, 75 µmol m^–2 s^–1) and media nitrogen concentrations (0, 20, 100 µM ammonium-nitrogen). Responses were measured as numbers of male and female gametophytes producing gametangia and number of sporophytes produced following fertilization. Both male and female gametogenesis was reduced at 5 and 20 °C versus 10 and 15 °C. At 20 °C gametogenesis inhibition was greater with higher levels of ammonium-nitrogen concentration (100 µM). Sporophyte production was more sensitive to light, temperature and nitrogen concentration than gametogenesis. Production of sporophytes was inhibited completely at 20 °C. At lower temperatures, increasingly higher nutrient concentrations produced greater inhibition of production of sporophytes.     

Inshore spawning grounds of the squid Doryteuthis gahi suggest the consistent use of defoliated kelp Lessonia trabeculata in Central Chilean waters

Coastal spawning grounds of the squid Doryteuthis gahi were identified for the first time at three sites in Central Chile during 2014 and 2015. A total of 15 egg masses were collected from between 10 and 15?m depth and brought to the laboratory for evaluating capsular, embryonic and when possible, paralarval characters. Egg capsules from all spawning areas were similar in size (～25?mm in length) and number of embryos per capsule (～15). Egg-laying patterns, in addition to field observations, suggest that relatively small D. gahi may be using shallow waters in semi-protected environments to reproduce and spawn. Additionally, the differential use of healthy versus defoliated kelp Lessonia trabeculata suggests that egg-laying females selectively choose substrata that limit mechanical damage to the fragile egg capsules and that also allow for adequate water flow, which could be reduced in a mass of kelp fronds. These findings highlight the opportunistic behaviour of D. gahi which use overgrazed L. trabeculata, a condition that is widespread in areas with high herbivory pressure.