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.     

MORPHO‐FUNCTIONAL PATTERNS OF PHOTOSYNTHESIS IN THE SOUTH PACIFIC KELP LESSONIA NIGRESCENS: EFFECTS OF UV RADIATION ON 14C FIXATION AND PRIMARY PHOTOCHEMICAL REACTIONS1

The morpho‐functional patterns of photosynthesis, measured as ¹⁴C‐fixation and chl fluorescence of PSII, also as affected by different doses of UV radiation in the laboratory were examined in the South Pacific kelp Lessonia nigrescens Bory of the coast of Valdivia, Chile (40°S). The results indicated the existence of longitudinal thallus profiles in physiological performance. In general, blades exhibited higher rates of carbon fixation and pigmentation as compared with stipes and holdfasts. Light‐independent ¹⁴C fixation (LICF) was high in meristematic zones of the blades (3.5 μmol ¹⁴C·g^?1 fresh weight [FW]·h^?1), representing 2%–16% (percentage ratio) of the photosynthetic ¹⁴C fixation (20 μmol ¹⁴C·g^?1 FW·h^?1). Exposures to UV radiation indicated that biologically effective UV‐B doses (BED_{photoinhibition300}) of 200–400 kJ·m^?2 (corresponding to current daily doses measured in Valdivia on cloudless summer days) inhibit photosynthetic ¹⁴C fixation of blades by 90%, while LICF was reduced by 70%. The percentage ratio of LICF to photosynthetic ¹⁴C fixation increased under UV exposure to 45%. Primary light reactions measured as maximum quantum yield (F_v/F_m) and electron transport rate (ETR) indicated a higher UV susceptibility of blades as compared with stipes and holdfasts: after a 48 h exposure to UV‐B, the decrease in the blades was close to 30%, while in the stipes and holdfasts it was <20%. The existence of translocation of labeled carbon along the blades suggests that growth at the meristem may be powered by nonphotosynthetic processes. A possible functional role of LIFC, such as during reduction of photosynthetic carbon fixation due to enhanced UV radiation, is discussed. These results in general support the idea that the UV‐related responses in Lessonia are integrated in the suite of morpho‐functional adaptations of the alga.     

Experimental transplants of the large kelp Lessonia nigrescens (Phaeophyceae) in high-energy wave exposed rocky intertidal habitats of northern Chile: Experimental, restoration and management applications

Potential for addressing ecological and physiological issues becomes severely limited when the organisms required to experimentally test specific hypotheses are absent from the study areas. This report describes a simple and inexpensive device for re-planting kelps into the lower intertidal zone of wave-swept rocky habitats, using Lessonia nigrescens Bory as a model organism. The device allows a wide range of plant sizes to be anchored by the holdfast. Transplanted kelps regenerated, re-attached to the substratum and overgrew the transplanting device. These results confirm the feasibility of using this technique to tackle a variety of highly relevant questions involving ecological, physiological, conservation, restoration and management issues.     

FACTORS ASSOCIATED WITH HOST SPECIFICITY IN SPOROCLADOPSIS NOVAE-ZELANDIAE (CHLOROPHYTA)1

This study tests the hypotheses that substrate stability and rugose microtopography are the main factors that determine the tissue-specific epiphytism in the chlorophyte Sporocladopsis novae-zelandiae Chapman. Both in vitro and field experiments showed that the epiphyte did not develop on stable algal surfaces nor on artificial (smooth or rugose) substrata. In field experiments, however, other macroalgae settled more abundantly on rugose substrata. Similarly, our field experiments did not support the hypothesis that the physical environment provided by the spatial location of the host in the intertidal or subtidal was a relevant factor associated with specificity. In contrast, our laboratory experiments suggested that some component of the host cell wall might trigger the formation of penetrating rhizoids. These did not develop in isolated cultures of the epiphyte but were detected when the epiphyte contacted its natural host. Also, preliminary results showed that rhizoid-like structures developed, although at very low frequency, only in plants grown in culture medium containing a crude extract of sorus cortical tissue.     

EAG and behavioural responses of the larger grain borer, Prostephanus truncatus, and its predator, Teretriosoma nigrescens, to the borer‐produced aggregation pheromone

Abstract.Electroantennogram (EAG) and behavioural studies were conducted with Prostephanus truncatus (Horn) (Col.: Bostrichidae) and the predatory beetle, Teretriosoma nigrescens Lewis (Col: Histeridae) in regard to their responses to the components of the prey‐produced aggregation pheromone. There were hardly any differences between species or sexes regarding perception thresholds. In field and olfactometer experiments, female P. truncatus were more responsive to the pheromone than males, and both sexes reacted more strongly to the minor pheromone component, T2, than to T1. Sex ratios among trap catches of T. nigrescens were slightly male‐biased. The predator did not differentiate behaviourally between the pheromone components.     

Pi30 DNA probe may be useful for the identification of Prevotella intermedia at the species or strain level

Recently, we introduced a new method for the rapid screening of bacterial species-or subspecies-specific DNA probes, named the "inverted dot blot hybridization screening method." This method has subsequently been then applied to develop species-or strain-specific DNA probes for Prevotella intermedia and Prevotella nigrescens. In a previous study, the inverted dot blot hybridization data showed that a probe, Pi30, was specific for P. intermedia. In this study, the DNA probe Pi30 was evaluated by Southern blot analysis to determine if it could distinguish P. intermedia from P. nigrescens. The data showed that the probe Pi30 reacted with the genomic DNAs from the reference strains and clinical isolates of both P. intermedia and P. nigrescens, but the size of the signal bands was different. In addition, the probe Pi30 reacted with a 1.4 kbp fragment from the genomic DNAs digested with Pst I of the P. intermedia strains but not with any fragments of P. nigrescens strains. The result indicates that the probe Pi30 could be useful for the identification of P. intermedia by restriction fragment length polymorphism (RFLP) at the species or strain level.     

Incidence of Prevotella intermedia and Prevotella nigrescens in Periodontal Health and Disease

The incidence of black-pigmented rods (BPRs), especially Prevotella intermedia and Prevotella nigrescens, in periodontal health and disease were examined. Furthermore, the degradative enzyme activities of P. intermedia were compared among the strains from periodontal health and disease. Microbiological specimens were collected from subgingival crevice or periodontal pocket by paper point. The BPRs were found in 71.1% of periodontally healthy subjects (n = 45), and in 47.1% of healthy sites (n = 34) and 87.8% of active sites (n = 41) among periodontally diseased patients. Porphyromonas gingivalis was detected only in active sites of periodontally diseased patients (17.8% of 180 strains). P. intermedia was the predominant BPR in both healthy and active sites (37.3 and 41.7%, respectively) of the patients. However, P. nigrescens was the predominant BPR (70.5% of 173 strains) in periodontally healthy subjects. The enzyme activities of esterase, esterase-lipase, acid-phosphatase and α-fucosidase of P. intermedia strains isolated from active sites in patients were significantly higher (P < 0.05) than those of healthy subjects. The results suggest that P. intermedia might increase the activity of degradative enzymes under a certain condition and support the progression of periodontitis.     

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.     

Electrophoretic analysis of the prey spectrum of Teretrius nigrescens (Lewis) (Col., Histeridae), a predator of Prostephanus truncatus (Horn) (Col., Bostrichidae), in Mexico, Honduras, and Benin

Abstract: Extensive collections of Teretrius nigrescens in Mexico, Honduras, and Benin, were electrophoretically analysed to elucidate the prey spectrum of the predator. Both polyacrylamide gel electrophoresis and cellulose acetate electrophoresis were used. Beetles were sampled with pheromone traps, using the synthetic aggregation pheromone of Prostephanus truncatus , and directly from farmers' maize stores. The proportion of electrophoretically detected prey protein from adult T. nigrescens in pheromone traps was low: of the 1108 specimens analysed, only in 34 cases, prey protein could be clearly identified. More frequently, prey protein was detected in adult T. nigrescens sampled in maize stores in Mexico, Honduras, and Benin, with 87 samples showing distinct prey bands of the total 1214 predators analysed. Of the 241 T. nigrescens larvae sampled in maize stores in Benin, 136 showed distinct bands of prey protein. In all samples, P. truncatus was the most frequently detected prey species. The second most often identified prey species was Sitophilus zeamais . The results are discussed with regard to various methods for prey spectrum analysis and specifically the biology of T. nigrescens .