A highly accurate, single PCR reaction for parentage assignment in Senegal sole based on eight informative microsatellite loci

From a battery of microsatellite markers (100 loci), recently identified by our group, we have selected eight for parentage assignment in Senegal sole ( Solea senegalensis ). This tool is based on microsatellite loci obtained from four genomic DNA libraries and one cDNA library. Within the eight loci (six from anonymous genomic DNA sequences and two located in expressed sequence tags of known genes), we have found, in an analysis of a reproductive broodstock, between nine and 16 alleles. The expected heterozygosity was between 0.616 and 0.860. In addition, we have optimized the polymerase chain reaction (PCR) conditions to amplify all loci simultaneously in a single multiplex PCR reaction, and we have tested three lots of male and female (five to six individuals) and three offspring (50–60 larvae each). The use of the eight microsatellite loci, the possibility of amplifying them in a single PCR reaction and the high value of the exclusion probability (0.9992) make this multiplex PCR method a unique tool for parentage assignment.
Finally, analysing one meiotic gynogenetic progeny, we have determined the relative distance of six of these loci to the centromere, and we have also found that all of them are unlinked. All these characteristics confer this tool with a high accuracy for parentage studies and genetic population analyses of Senegal sole.     

Effects of two diets on lipid composition and reproductive performance of brill (Scophthalmus rhombus) eggs

The effects of two diets with different lipid composition (squid Loligo gahi and one commercial pellet for breeders) on reproductive performance and egg lipid composition of brill (Scophthalmus rhombus) were compared. A total of 36 adult brill weighing 1482 ± 432 g were randomly distributed in the experimental tanks. A flow‐through system composed of four tanks with 4.2 m² of bottom area (5.0 m³ × 1.2 m water depth) was used. The experiment lasted for approximately 5 months (168 days). From the four mature females fed squid, three ovulated. Five females fed pellets matured and ovulated. Total ovulations were of 10 and 44, by the three females fed squid and the five ovulating female fed pellets respectively. Egg viability was higher, although not significantly, for females fed pellets. This indicates that pellets could be a better food for breeding females. In general, the egg total lipid content and the lipid classes composition did not seem to be affected by diet. In contrast, results indicate that not only diets but also individual females, regardless of the diet consumed, are determinant for egg fatty acid composition, clearly modulating their composition.     

Water-use efficiency in cork oak (Quercus suber) is modified by the interaction of water and light availabilities

We studied the interaction of light and water on water-use efficiency in cork oak (Quercus suber L.) seedlings. One-year-old cork oak seedlings were grown in pots in a factorial experiment with four light treatments (68, 50, 15 and 5% of full sunlight) and two irrigation regimes: well watered (WW) and moderate drought stress (WS). Leaf predawn water potential, which was measured at the end of each of two cycles, did not differ among the light treatments. Water-use efficiency, assessed by carbon isotope composition (delta(13)C), tended to increase with increasing irradiance. The trend was similar in the WW and WS treatments, though with lower delta(13)C in all light treatments in the WW irrigation regime. Specific leaf area increased with decreasing irradiance, and was inversely correlated with delta(13)C. Thus, changes in delta(13)C could be explained in part by light-induced modifications in leaf morphology. The relationship between stomatal conductance to water vapor and net photosynthesis on a leaf area basis confirmed that seedlings in higher irradiances maintained a higher rate of carbon uptake at a particular stomatal conductance, implying that shaded seedlings have a lower water-use efficiency that is unrelated to water availability.     

Reproductive performance and seasonal plasma sex steroid and metabolite levels in a captive wild broodstock of brill Scophthalmus rhombus L.

This study reports egg production by captive wild brill Scophthalmus rhombus, a potential new flatfish species for Southern Europe‐Mediterranean mariculture, as well as seasonal plasma levels of 17β–estradiol, testosterone, 11–ketotestosterone, proteins, triglycerides, glucose and lactate. A mean egg production of 102 800 eggs kg body weight⁻¹ was achieved during the 2005 spawning period (January–March), although a continuous egg supply could only be obtained from some females, which had a higher relative fecundity (261 019±10 393 eggs kg⁻¹) with 12–17 eggs batches released at a mean interval of 3.4 days. Most eggs were obtained with water temperatures ranging from 12 to 14°C, and under increasing temperatures (up to 2.9°C). Potential egg viability (70.1±2.9%), fertilization (72.2±3.4%) and hatching rates (31.9±3.9%) showed high variability, with potential viability tending to decrease as the water temperature increased (mainly between 16 and 17°C) and 0% hatching above 16.6°C. The endocrine changes that brill underwent during late gametogenesis, spawning and postspawning periods were similar to those reported in other Pleuronectiformes. This study establishes an important basis for further research on the biology and physiology of brill reproduction, directed towards the optimization of the breeding techniques used currently.     

Genetic variability for responses to short‐ and long‐term salt stress in vegetative sunflower plants

Sunflower (Helianthus annuus L.) has been rated as moderately salt‐resistant, and variability for salt resistance has been detected within this crop. However, variability in salt‐resistance mechanisms has not been assessed. Osmotic tolerance, the relation of salt resistance with whole‐plant Na⁺ and K⁺ distribution and tissue Na⁺ tolerance were investigated in several sunflower inbred lines. Plants were grown under controlled conditions, in pots with sand and perlite irrigated with salinized (NaCl, –0.65 MPa) nutrient solution. Osmotic tolerance was assessed from the initial effects of the salt treatment on plant elongation in eleven sunflower lines. Long‐term salinity responses were evaluated in four of those lines, by assessing whole‐plant growth. A principal components analysis (PCA) was run on relative‐to‐control growth data, and this information was used to establish a relative resistance ranking, which indicated lines HAR2 > HAR1 > HA64 > HAR5. Osmotic tolerance was observed in HA64 and HAR2. The lines showed different degrees of Na⁺ accumulation, it was very low in some of them, but relative salt resistance was not associated to this trait. Tissue Na⁺ tolerance was deduced by comparing the percentage of dead leaves as a function of leaf blade Na⁺ accumulation, and it was higher in HAR1 than in the rest. These results indicate that variability for salt‐resistance mechanisms exists in sunflower. Osmotic tolerance and tissue Na⁺ tolerance were detected in different lines, highlighting that such variability may be exploited for increasing salt resistance in this crop.     

Effect of dietary protein level and source on digestive proteolytic enzyme activity in juvenile Senegalese sole, Solea senegalensis Kaup 1850

The effect of dietary protein level and protein source on growth and proteolytic activity of juvenile Solea senegalensis was studied. In Experiment 1, fish were fed on four experimental diets containing increased protein levels (36, 46, 56 and 67%). In Experiment 2, Senegalese soles were fed on five diets with partial substitution of fish meal by soybean meal, soybean protein concentrate, soybean protein isolate, wheat gluten meal or pea protein concentrate. Results prove that growth and proteolytic activity in the distal intestine of fish were affected by the quantitative increase in dietary protein. The origin of protein source used in the elaboration of experimental diets affected both the amount and composition of the alkaline proteases secreted into the intestinal lumen; however, it did not decrease animal growth. Juvenile Senegalese sole showed capability to modulate digestive protease secretion when the concentration and/or source of dietary protein were modified. Quantity and quality of dietary protein affected protein hydrolysis in Senegalese sole intestine. This study establishes that 30% fish meal protein can be replaced by soybean derivatives without affecting intestinal proteases. Replacement with wheat gluten meal or pea protein concentrate should be taken cautiously, but further research is needed to establish whether growth performance and digestive enzyme physiology of Senegalese sole are affected by plant protein-supplemented diets in a long-term trial.     

Effect of diet on the lipid composition of the commercial clam Donax trunculus (Mollusca: bivalvia): sex‐related differences

This paper describes the lipid composition of the commercial bivalve Donax trunculus and the differences originated when the animals were fed with two phytoplankton species (Tetraselmis suecica and Chaetoceros sp.) in a hatchery. We also analysed sex‐related differences in lipid classes and fatty acid profile. Total lipids were higher in females than in males. Triglycerides and phospholipids were the major lipid components, and the former dominated in females and the latter in males. The main fatty acids in both sexes were 16:0, 20:5n‐3 (eicosapentaenoic acid) and 22:6n‐3 (docosahexaenoic acid). Females showed higher percentages of saturated and monounsaturated fatty acids and lower levels of polyunsaturated fatty acids than males. Significant differences in total lipid, lipid classes and fatty acid profiles were also found due to diet. Docosahexaenoic acid decreased and total lipids, free fatty acids, arachidonic acid and EPA increased in both sexes. Despite these differences, the condition of the species was maintained and the broodstock even maturated.     

Kinetics of Lead Bioaccumulation from a Hydroponic Medium by Aquatic Macrophytes Pistia stratiotes

The goal of this work was to study quantitatively lead bioaccumulation from a lead-doped nutrient medium by using a living aquatic macrophytes Pistia stratiotes. Several sets of aquatic plants with approximately 30 g weight were grown in greenhouse conditions and in hydroponic solutions supplied with a non-toxic Pb²⁺ concentration. The synchrotron radiation total X-ray fluorescence spectrometry was used to determine the metal concentrations in dry plants and hydroponic media as a function of time. Four different non-structural bioaccumulation models were applied to describe the process dynamics and to estimate the accumulated lead maximum capacity and rate constants. According to the experimental data, both biosorption and bioaccumulation mechanisms can be considered. Due to the low desorption rate constant, the experimental data were well described by the irreversible kinetic model. The results concerning modeling of living macrophytes’ metal bioaccumulation kinetics can be used to predict the heavy metal removal dynamics from wastewaters in artificial wetlands.     

Amplified Fragment Length Polymorphism Diversity in Cephalosporium maydis from Egypt

ABSTRACT Cephalosporium maydis, the causal agent of late wilt of maize, was first described in Egypt in the 1960s, where it can cause yield losses of up to 40% in susceptible plantings. We characterized 866 isolates of C. maydis collected from 14 governates in Egypt, 7 in the Nile River Delta and 7 in southern (Middle and Upper) Egypt, with amplified fragment length polymorphism (AFLP) markers. The four AFLP primer-pair combinations generated 68 bands, 25 of which were polymorphic, resulting in 52 clonal haplotypes that clustered the 866 isolates into four phylogenetic lineages. Three lineages were found in both the Nile River Delta and southern Egypt. Lineage IV, the most diverse group (20 haplotypes), was recovered only from governates in the Nile River Delta. In some locations, one lineage dominated (up to 98% of the isolates recovered) and, from some fields, only a single haplotype was recovered. Under field conditions in Egypt, there is no evidence that C. maydis reproduces sexually. The nonuniform geographic distribution of the pathogen lineages within the country could be due to differences in climate or in the farming system, because host material differs in susceptibility and C. maydis lineages differ in pathogenicity.