Jessica Beltrán

Spore release in Acrochaetium sp. (Rhodophyta) is bacterially controlled

The facultative red algal epiphyte Acrochaetium sp. liberated spores preferentially and recruited more successfully in laboratory cultures when its host Gracilaria chilensis C. J. Bird, McLachlan et E. C. Oliveira was present. The same effect was also induced by cell‐free medium from G. chilensis, suggesting it contained a molecular signal. Antibiotics prevented spore release in Acrochaetium sp., even when G. chilensis was present, suggesting a prokaryotic origin of the signal. Simultaneous application of N‐butyl‐homoserine‐lactone (BHL) restored the spore‐release capacity, which demonstrated that spore release was not directly inhibited by the antibiotics and indicated that bacterially generated N‐acyl‐homoserine‐lactones (AHLs) regulate spore release. An involvement of AHL was further indicated by the fact that two different halofuranone inhibitors of AHL receptors also inhibited spore release when they were applied at relatively low concentrations. Of seven different AHLs tested, only BHL induced the effect. However, BHL was only active at relatively high concentrations (100 μM), and it was not detected in spore‐release‐inducing medium of G. chilensis. Another water‐soluble AHL or an AHL structure analog is therefore probably the active compound in G. chilensis cultures. The data presented demonstrate that life cycle completion in Acrochaetium sp. strongly depends on bacteria, which are not always present in sufficient numbers on the alga itself. Exogenous bacteria that are associated with G. chilensis or with other potential substrates may therefore trigger timely spore liberation in Acrochaetium sp., provided that the necessary concentration of AHL is reached. This first finding of AHL perception in a red alga confirms that AHL signalling is more widespread among eukaryotes than was thought until recently. However, spore release of a second red alga, Sahlingia subintegra (Rosenv.) Kornmann, was unaffected by AHL, and the reaction observed is therefore not universal.

Phospholipases and Galactolipases Trigger Oxylipin-Mediated Wound-Activated Defence in the Red Alga Gracilaria chilensis against Epiphytes

We investigated the wound response of the commercially important red alga, Gracilaria chilensis, in order to obtain insight into its interaction with epiphytic pests. After wounding, the host releases free fatty acids as well as the hydroxylated eicosanoids, 8R‐hydroxy eicosatetraenoic acid (8‐HETE) and 7S,8R‐dihydroxy eicosatetraenoic acid (7,8‐di‐HETE). While the release of free arachidonic acid and subsequent formation of 8‐HETE is controlled by phospholipase A, 7,8‐di‐HETE production is independent of this lipase. This dihydroxylated fatty acid might be directly released from galactolipids. Physiologically relevant concentrations of oxylipins reduced spore settlement of Acrochaetium sp. (Rhodophyta, Acrochaetiaceae) and suppressed the development of hapteria in Ceramium rubrum (Rhodophyta, Ceramiaceae) when these model epiphytes were exposed to artificial surfaces that contained 8‐HETE or 7,8‐di‐HETE. Thus, the immediate release of oxylipins can be seen as G. chilensis defence against epiphytes.

Diversity, phenomenology and epidemiology of epiphytism in farmed Gracilaria chilensis (Rhodophyta) in northern Chile

This study identified the most common epiphytes infecting the algal host Gracilaria chilensis on a farm in northern Chile. Simultaneously, the types of host–epiphyte interfaces were characterized and their relative abundance and temporal variability were monitored. Five types of anatomical relationships were detected. Infection type I included the epiphytes weakly attached to the surface of the host and not associated with damage of host tissues (i.e. Hincksia mitchelliae, H. granulosa and Ectocarpus acutus). Infection type II included those epiphytes strongly attached to the surface of the host but not associated with any host tissue damage (i.e. Acrochaetium sp., Antithamnionella sp. and Colpomenia sinuosa). Infection type III included all the epiphytes that penetrated the outer layer of the host wall without damaging its cortical cells (i.e. Xenococcus sp. and Sahlingia subintegra). Infection type IV included epiphytes penetrating deep into the host cell wall, disorganizing the cortical tissue (i.e. Ulva lactuca and Acrosorium corallinarum). Infection type V included epiphytes that penetrated deeply into the cortex, reached the medullary tissue and caused destruction of the hosts cells in the area around the infection (i.e. Ceramium rubrum and Polysiphonia harveyi). Prevalence varied with time and with infection type, with types II and III reaching up to 80% and 90% of the thalli respectively. Severity of epiphyte infection was similar to the distribution of infection prevalence, with crustose epiphytes colonizing up to 80% of the host surface.

IDENTIFICATION OF CRYPTIC SPECIES IN THE LESSONIA NIGRESCENS COMPLEX (PHAEOPHYCEAE, LAMINARIALES)1

The kelp Lessonia nigrescens Bory is the most ecologically and economically important seaweed in rocky intertidal and shallow subtidal habitats along the temperate Pacific South American coasts. Recent molecular studies suggest the existence of two lineages, one (northern lineage) from 17° S to 30° S and a second (central lineage) from 29° S to 41° S. To identify and name these lineages we performed morphological, nomenclatural and field studies. Four external and three internal anatomical traits permitted a morphological separation of the two lineages. The internal structure of both lineages was different from the isolectotype of Lessonia nigrescens. It is therefore concluded that the name Lessonia nigrescens should not be used for the Chilean material. Chordaria spicata Suhr appears as the oldest available name for the central lineage, while Lessonia berteroana Montagne is the oldest name for the northern lineage. In both cases, the type material consisted of small‐sized, apical branches of larger plants. The new combination Lessonia spicata (Suhr) Santelices is proposed for the central lineage and we reinstate Lessonia berteroana for the northern lineage. Laminaria scissa Suhr is reduced to synonym of L. spicata. Representative specimens of Lessonia nigrescens were not found during new visits to its type locality in Cape Horn and along Chile. Future studies should verify the status of this species.

Up-Regulation of Lipoxygenase, Phospholipase, and Oxylipin-Production in the Induced Chemical Defense of the Red Alga Gracilaria chilensis against Epiphytes

The red alga Gracilaria chilensis is commercially farmed for the production of agar hydrocolloids, but some susceptible algae in farms suffer from intense epiphyte growth. We investigated the induced chemical defense response of G. chilensis against epiphytes and demonstrated that an extract of an epiphyte-challenged alga can trigger a defense response. The hormonally active metabolites were purified by RP-HPLC. Treatment with the extract or the purified fraction changed the chemical profile of the alga and increased resistance against epiphyte spores. Semi-quantitative RT-PCR and enzyme assays demonstrated that this metabolic response occurs after an increase in lipoxygenase and phospholipase A2 activity. Although this suggests the involvement of regulatory oxylipins, neither jasmonic acid nor the algal metabolite prostaglandin E2 triggers comparable defense responses.

SEX RATIO VARIATION IN THE LESSONIA NIGRESCENS COMPLEX (LAMINARIALES, PHAEOPHYCEAE): EFFECT OF LATITUDE, TEMPERATURE, AND MARGINALITY1

Little is known about variation of sex ratio, the proportion of males to females, in natural populations of seaweed, though it is a major determinant of the mating system. The observation of sexual chromosomes in kelps suggested that sex is partly genetically determined. However, it is probably not purely genetic since the sex ratio can be modified by environmental factors such as salinity or temperature. In this paper, sex ratio variation was studied in the kelp Lessonia nigrescens Bory complex, recently identified as two cryptic species occurring along the Chilean coast: one located north and the other south of the biogeographic boundary at latitude 29°–30° S. The life cycle of L. nigrescens is characterized by an alternation of microscopic haploid gametophytic individuals and large macroscopic fronds of diploid sporophytes. The sex ratio was recorded in progenies from 241 sporophytic individuals collected from 13 populations distributed along the Chilean coast in order (i) to examine the effect of an environmental gradient coupled with latitude, and (ii) to compare marginal populations to central populations of the two species. In addition, we tested the hypothesis that the sex ratios of the two cryptic species would be affected differently by temperature. First, our results demonstrate that sex ratio seems to be mainly genetically determined and temperature can significantly modify it. Populations of the northern species showed a lower frequency of males at 14°C than at 10°C, whereas populations of the southern species showed the opposite pattern. Second, both species displayed an increased variation in sex ratio at the range limits. This greater variation at the margins could be due either to differential mortality between sexes or to geographic parthenogenesis (asexual reproduction).

Cultivation of Gigartina skottsbergii (Gigartinales, Rhodophyta): Recent advances and challenges for the future

This study integrates landings statistics and biological studies of the red algaGigartina skottsbergii Setchell & Gardner. The analysis of the landings and carrageenan production in Chile suggeststhat this resource will suffer a strong harvesting pressure during the nextyears. Biological results on sporulation, germination, sporeling growth and survivorship in laboratory,indoor tanks and field conditions, indicated that cultivation of this species istechnically feasible, as spores can be seeded on ropes and other substrata. Vegetative propagation of this species through tissue fragmentationis also possible. Vegetative fragments of this carrageenophyte have 20 to30% higher growth rates than whole fronds in suspended culture systems. Protoplast production can be also explored for bypassing restrictions inspore availability. Major advantages that encourage the cultivation of G. skottsbergii include its gel quantity and quality, its pathogen-freecondition, a high reproduction potential and its regeneration capacity. Onthe other hand, the major constraints are related to its relatively slowgrowth as compared to other carrageenophytes, limited availability ofspores and high mortality during juvenile stages.

Healing and regeneration responses in Gigartina skottsbergii (Rhodophyta, Gigartinales): optimization of vegetative propagation for cultivation

The red alga Gigartina skottsbergii is becoming increasingly valuable as a resource to providing the raw material for the carrageenan industry established in Chile and elsewhere. As a result, wild stocks of the species are subject to intense harvesting by local fishermen. With the current levels of harvesting, it seems likely that natural stands of G. skottsbergii will soon collapse. Although cultivation seems an alternative, knowledge regarding the biology of the species is exceedingly limited. This study reports the first attempt to determine the optimal conditions for vegetative propagation of this species in the laboratory. For this purpose, the processes of wound healing and regeneration of frond fragments and haptera were studied under controlled conditions of temperature, light, and media strength. Our results demonstrated that excised tissues of G. skottsbergii were able to seal the exposed areas in approximately 20 days, by a wound healing process characterized by a re-differentiation of medullary cells into a normal cortex. Our data also demonstrated that frond fragments are better than haptera for propagation purposes. The development of new cortical tissue at the cut surface is followed, within 60 days, by localized blade-like outgrowths along the repaired area. Furthermore, the healing and regenerative responses in both frond fragments and haptera differed in efficiency according to the various combinations of factors, with optimum of 10–15 °C, 5 ?mol m−2 s−1 and plain seawater or standard SFC medium for the fronds. The two types of responses were negatively affected by seawater enriched with a double concentration of nutrients.

INFECTIOUS DISEASES OF MAZZAELLA LAMINARIOIDES (RHODOPHYTA): CHANGES IN INFECTION PREVALENCE AND DISEASE EXPRESSION ASSOCIATED WITH SEASON, LOCALITY, AND WITHIN‐SITE LOCATION1

This study addresses the issues of infection prevalence and disease expression in two wild populations of the red algal host Mazzaella laminarioides and their variability associated with locality, season, and spatial location of the host in the intertidal zone. Our results demonstrated that Endophyton ramosum is the most frequent infective pathogen affecting M. laminarioides in Matanzas and Pucatrihue. This situation prevailed through the year and across the high‐to‐low intertidal gradient. Although there was a general trend for lower levels of infection in late winter and early spring, only in a few, cases was well‐defined seasonality detected. Furthermore, clear seasonal patterns, as displayed by deformative disease in the high intertidal zone of Pucatrihue, were attenuated in the middle and lower intertidal zones. Differences in levels of infection in M. laminarioides between the high intertidal zones of Matanzas and Pucatrihue diminished toward the low intertidal zone. Thus, effects of seasonality and locality on infection prevalence may be influenced, at least in part, by the position of the hosts an the intertidal zone.

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.