EFFECT OF ROCKY INTERTIDAL AMPHIPODS ON ALGAL RECRUITMENT: A FIELD STUDY1

Evidence about the potential of mobile marine invertebrates to act as algal spore dispersal agents is presently circumstantial. Using a field correlational and experimental protocol, our study tested the hypothesis that amphipods can increase the spore recruitment of the red alga Iridaea laminarioides Bory. Iridaea laminarioides spore recruitment onto glass slides was measured at a site with high amphipod abundance and a site with low density of amphipods. To evaluate the effect of an Ulva canopy on recruitment, replicated glass slides with and without a surrounding Ulva canopy were installed at both sites. The number of I. laminarioides spores recruited on the glass slides was four to eight times higher at the high amphipod abundance site than at the low density site. However, the presence of an Ulva canopy covering the glass slides did not significantly increase the recruitment of I. laminarioides. Because the abundance of I. laminarioides, the proportion of cystocarpic plants, and the percentage of open cystocarps only differed slightly between the low and high abundance amphipod sites, we suggest that the variation in recruitment between the sites is due to the differences in amphipod abundance (and their movements) and not to differences in spore production. Moreover, the presence of I. laminarioides cystocarps showing amphipod grazing scars was significantly higher at the high amphipod density site than at the low density site.     

A new subfamily of sucrose transporters, SUT4, with low affinity/high capacity localized in enucleate sieve elements of plants

A new subfamily of sucrose transporters from Arabidopsis (AtSUT4), tomato (LeSUT4), and potato (StSUT4) was isolated, demonstrating only 47% similarity to the previously characterized SUT1. SUT4 from two plant species conferred sucrose uptake activity when expressed in yeast. The K(m) for sucrose uptake by AtSUT4 of 11.6 +/- 0.6 mM was approximately 10-fold greater than for all other plant sucrose transporters characterized to date. An ortholog from potato had similar kinetic properties. Thus, SUT4 corresponds to the low-affinity/high-capacity saturable component of sucrose uptake found in leaves. In contrast to SUT1, SUT4 is expressed predominantly in minor veins in source leaves, where high-capacity sucrose transport is needed for phloem loading. In potato and tomato, SUT4 was immunolocalized specifically to enucleate sieve elements, indicating that like SUT1, macromolecular trafficking is required to transport the mRNA or the protein from companion cells through plasmodesmata into the sieve elements.     

How to correctly determine the different chlorophyll fluorescence parameters and the chlorophyll fluorescence decrease ratio R<Subscript>Fd</Subscript> of leaves with the PAM fluorometer

This contribution is a practical guide to the measurement of the different chlorophyll (Chl) fluorescence parameters and gives examples of their development under high-irradiance stress. From the Chl fluorescence induction kinetics upon irradiation of dark-adapted leaves, measured with the PAM fluorometer, various Chl fluorescence parameters, ratios, and quenching coefficients can be determined, which provide information on the functionality of the photosystem 2 (PS2) and the photosynthetic apparatus. These are the parameters F_v, F_m, F₀, F_m′, F_v′, NF, and ΔF, the Chl fluorescence ratios F_v/F_m, F_v/F₀, ΔF/F_m′, as well as the photochemical (q_P) and non-photochemical quenching coefficients (q_N, q_CN, and NPQ). q_N consists of three components (q_N = q_E + q_T + q_I), the contribution of which can be determined via Chl fluorescence relaxation kinetics measured in the dark period after the induction kinetics. The above Chl fluorescence parameters and ratios, many of which are measured in the dark-adapted state of leaves, primarily provide information on the functionality of PS2. In fully developed green and dark-green leaves these Chl fluorescence parameters, measured at the upper adaxial leaf side, only reflect the Chl fluorescence of a small portion of the leaf chloroplasts of the green palisade parenchyma cells at the upper outer leaf half. Thus, PAM fluorometer measurements have to be performed at both leaf sides to obtain information on all chloroplasts of the whole leaf. Combined high irradiance (HI) and heat stress, applied at the upper leaf side, strongly reduced the quantum yield of the photochemical energy conversion at the upper leaf half to nearly zero, whereas the Chl fluorescence signals measured at the lower leaf side were not or only little affected. During this HL-stress treatment, q_N, q_CN, and NPQ increased in both leaf sides, but to a much higher extent at the lower compared to the upper leaf side. q_N was the best indicator for non-photochemical quenching even during a stronger HL-stress, whereas q_CN and NPQ decreased with progressive stress even though non-photochemical quenching still continued. It is strongly recommended to determine, in addition to the classical fluorescence parameters, via the PAM fluorometer also the Chl fluorescence decrease ratio R_Fd (F_d/F_s), which, when measured at saturation irradiance is directly correlated to the net CO₂ assimilation rate (_{P N}) of leaves. This R_Fd-ratio can be determined from the Chl fluorescence induction kinetics measured with the PAM fluorometer using continuous saturating light (cSL) during 4–5 min. As the R_Fd-values are fast measurable indicators correlating with the photosynthetic activity of whole leaves, they should always be determined via the PAM fluorometer parallel to the other Chl fluorescence coefficients and ratios.     

Cellular prion protein acquires resistance to proteolytic degradation following copper ion binding

The conversion of cellular prion protein (PrP(C)) into its pathological isoform (PrP(Sc)) conveys an increase in hydrophobicity and induces a partial resistance to proteinase K (PK). Interestingly, co-incubation with high copper ion concentrations also modifies the solubility of PrP(c) and induces a partial PK resistance which was reminiscent of PrP(Sc). However, concerns were raised whether this effect was not due to a copper-induced inhibition of the PK itself. We have therefore analyzed the kinetics of the formation of PK-resistant PrP(C) and excluded possible interference effects by removing unbound copper ions prior to the addition of PK by methanol precipitation or immobilization of PrP(C) followed by washing steps. We found that preincubation of PrPc with copper ions at concentrations as low as 50 microM indeed rendered these proteins completely PK resistant, while control substrates were proteolyzed. No other divalent cations induced a similar effect. However, in addition to this specific stabilizing effect on PrP(C), higher copper ion concentrations in solution (>200 microM) directly blocked the enzymatic activity of PK, possibly by replacing the Ca2+ ions in the active center of the enzyme. Therefore, as a result of this inhibition the proteolytic degradation of PrP(C) as well as PrP(Sc) molecules was suppressed.     

REPRODUCTIVE, CULTURE AND REGENERATION STUDIES OF THE EDIBLE RED ALGA CALLOPHYLLIS IN CHILE

In Chile, the demand of edible seaweeds has increased during recent years and Callophyllis variegata is one of the most demanded species. This study summarizes information on phenology, aspects, in vitro culture and vegetative propagation methods for Callophyllis. Results indicate that spore production occurs mainly during winter, and recruitment of new plants appear in nature in the spring. Culture studies indicate that spores presented higher germination and growth rates at 8° C and 10 to 12 μmol m⁻² s⁻¹. Furthermore, these results indicate that this species presents a high potential for regeneration from its holdfast. The manipulation of temperature, light and culture medium enhances the regeneration process and growth of Callophyllis in the laboratory.     

Accumulation of Hydroxycoumarins During Post-harvest Deterioration of Tuberous Roots of Cassava (Manihot esculenta Crantz)

The use of the root crop Cassava (Manihot esculenta Crantz)is constrained by its rapid deterioration after harvest. Chemicaland spectroscopic examination revealed the accumulation of fourhydroxycoumarins (esculin, esculetin, scopolin and scopoletin),compounds derived from the phenylpropanoid pathway, during thetime course of post-harvest deterioration. Fluorescence-microscopyrevealed their localization in the apoplast of the parenchyma.Scopoletin and scopolin showed the most dramatic increases inconcentration, peaking by day 2 after harvesting. A smallersecondary peak of scopoletin tended to be more pronounced incultivars showing lower susceptibility to deterioration. Evidencefor the metabolism of scopoletin to an insoluble coloured productby means of a peroxidase is presented. This product may be thecause of the discolouration of the vascular tissue during storage.Copyright 2000 Annals of Botany Company Cassava, hydroxycoumarins, Manihot esculenta, peroxidases, post-harvest physiological deterioration, wound response