The use of RAPD technique for the identification and classification of Pisum sativum L. genotypes

Summary The genomic DNAs of 42 Pisum sativum genotypes, representing four wild and cultivated subspecies were used as templates in RAPD reactions. Amplification with eight decamer primers generated 149 polymorphic products. Genetic similarities of RAPD profiles were estimated via a coefficient of Jaccard and then the data were processed by cluster analysis (UPGMA). Each genotype was clearly identified and separated from the others. Our results show that RAPD technology is a rapid, precise and sensitive technique for identification of pea genotypes. However, the phylogenetic relationships within the Pisum sativum, which we tested by bootstrap analysis (Wagner parsimony), must be interpreted with caution.     

A practical and reliable procedure for in vitro induction of tetraploid tomato

A relatively rapid, practical and reliable procedure for in vitro production of tetraploid tomato using colchicine is described. Using this procedure 11.11% generation of tetraploids was obtained after exposure of seedling shoot meristem explants with 8 mM colchicine for 96 h. Confirmation of tetraploid production (2n = 48), was determined by flow cytometry and verified by cytogenetic analysis of root tip preparations. The results indicate that the procedure is adequate for induction and screening of tetraploid tomato plantlets.     

Response of gladiolus (Gladiolus spp) plants after exposure corms to chitosan and hot water treatments

The state of Morelos, Mexico has gradually become an important producer of gladiolus. Some preconditioning treatments of corms are empirically done causing uneven emergence and low quality of flowers. In this investigation, before planting, gladiolus corms var. ‘Blanca Borrego’ were dipped in chitosan (chitosan reagent and commercial chitosan Biorend^®), in hot water at various temperatures and in treatments combined with Biorend^® at 1.5% and hot water. Results indicated that the most influenced variables were corm germination, number of flowers per spike, number of cormlets and vase life. Overall, the commercial product Biorend^® at 1.5% accelerated corm emergence in approximately 4 days, the number of flowers increased by 2–7 and the vase life extended for 3 days. The number of cormlets was also duplicated. Corms dipped in the commercial chitosan Biorend^® at 1.5% at different intervals of time were not greatly affected except for the emergence and number of cormlets. However, for this experiment there were significant effects on the number of leaves and flowers because of the interactions between chitosan and the immersion time. The temperature of 55 °C affected plant development because emergence was delayed by 6 days; and there were less number of leaves, flowers and cormlets. On the other hand, the incidence of Fusarium oxysporum in naturally infected corms was 0% at temperatures of 55 °C and 50 °C. Immersion times (0, 10, 15 and 20 min) in hot water at 50 °C did not show significant effects on plant development and vase life. Corms dipped in Biorend^® at 1.5% and hot water at 50 °C accelerated their emergence for about 1–7 days, the number of flowers increased by two, extended the storage life for 1–3 days and increased the number of cormlets. The integration of these two treatments -Biorend^® and hot water- might be a good option for increasing the gladiolus plant quality and vase life.     

Estimation of actual evapotranspiration for a drip-irrigated Merlot vineyard using a three-source model

A study was performed in order to evaluate the three-source model (Clumped model) for direct estimation of actual evapotranspiration (ET_a) and latent heat flux (LE) over a drip-irrigated Merlot vineyard trained on a vertical shoot positioned system (VSP) under semi-arid conditions. The vineyard, with an average fractional cover of 30%, is located in the Talca Valley, Region del Maule, Chile. The performance of the Clumped model was evaluated using an eddy covariance system during the 2006/2007 and 2007/2008 growing seasons. Results indicate that the Clumped model was able to predict ET_a with a root mean square error (RMSE), mean bias error (MBE), and model efficiency (EF) of 0.33, −0.15 mm day⁻¹ and 74%, respectively. Also, the Clumped model simulated the daytime variation of LE with a RMSE of 36 W m⁻², MBE of −8 W m⁻², and EF of 83%. Major disagreement (underestimated values) between observed and estimated values of ET_a was found for clear days after rainfall or foggy days, but underestimated values were less than 10% of the data analysis. The results obtained in this study indicate that the Clumped model could be used to directly estimate vine water requirements for a drip-irrigated vineyard trained on a VSP. However, application of the Clumped model requires a good characterization of the drip-irrigated vineyard architecture.     

The pond's shape matters: differential growth,physiological condition and survival of epibenthic Farfantepenaeus aztecus postlarvae

As a function of the water quality provided by square, circular and oval experimental ponds, the growth, survival and oxygen requirements in epibenthic postlarvae of Farfantepenaeus aztecus were analysed in relation to their routine metabolism and apparent heat increment. Temperature, oxygen concentration, pH and salinity were measured daily in two experimental ponds of each shape. The postlarvae oxygen consumption during two 24‐h cycles, their growth, physiological condition and survival and the productivity in the ponds were estimated. Low values of pH, oxygen concentration and phytobenthos productivity, and reduced postlarvae relative growth and survival were observed in the square ponds. We suggest that the latter results from a deficient water circulation related to the effect of the pond's shape on dissolved oxygen levels and, consequently, on growth and survival. The postlarvae routine metabolism, including feeding, varied between 1.91 and 2.25 mg O₂ h^?1 g^?1 wet weight, whereas the minimum oxygen concentration needed in the ponds is approximately 4.25 mg O₂ L^?1. These conditions were achieved in the oval ponds concurrent with higher survival and growth values, in which individuals distributed randomly, for which we suggest that oval‐shaped ponds could be the most adequate for the culture of this and other penaeid species.     

Effects of formalin on total haemocytes count and histopathological changes in the shrimp Litopenaeus schmitti (Pérez‐Farfante & Kensley 1997)

Formalin is used in shrimp industry to eliminate ectoparasites and as a disinfectant. However, the effect of formalin on shrimp's defense mechanism is unknown. Several biomarkers were used to assess the immune response of juveniles of the shrimp Litopenaeus schmitti exposed to different concentrations of formalin (10, 25 and 50 mg L^?1) during 24 and 48 h. Formalin concentrations of 25 and 50 mg L^?1 produced a significant reduction in the total haemocyte count at both 24 and 48 h. Peroxidase and phenoloxidase activity did not show variations. The total protein concentration in haemolymph of shrimps was not significantly different between experimental groups. Histological analysis showed hyperplasia in the gills of animals exposed at 10 mg L^?1 for 24 h; however, the exposure at 25 and 50 mg L^?1 caused necrosis in gills, hepathopancreas and the antennal gland.     

Salmonid fish viruses and cell interactions at early steps of the infective cycle

A flow cytometric virus-binding assay that directly visualizes the binding and entry of infectious pancreatic necrosis virus (IPNV), infectious haematopoietic necrosis virus (IHNV) and virus haemorrhagic septicaemia virus (VHSV) to several cell lines was established. The highest efficiency of binding was shown by the BF-2 cell line and this was used to study, at the attachment level, the interactions of these cells with salmonid fish viruses in coinfections, and to further determine if the earliest stage of the viral growth cycle could explain the previously described loss of infectivity of IHNV when IPNV is present. Our results demonstrated that IPNV binds to around 88% of cells either in single or dual infections, whereas IHNV attachment always decreased in the presence of any of the other viruses. VHSV binding was not affected by IPNV, but coinfection with IHNV reduced the percentage of virus-binding cells, which suggests competition for viral receptors or co-receptors. Internalization of the adsorbed IHNV was not decreased by coinfection with IPNV, so the hypothetical competence could be restricted to the binding step. Treatment of the cells with antiviral agents, such as amantadine or chloroquine, did not affect the binding of IPNV and VHSV, but reduced IHNV binding by more than 30%. Tributylamine affected viral binding of the three viruses to different degrees and inhibited IPNV or IHNV entry in a large percentage of cells treated for 30 min. Tributylamine also inhibited IHNV cytopathic effects in a dose-dependent manner, decreasing the virus yield by 4 log of the 50% endpoint titre, at 10 mm concentration. IPNV was also inhibited, but at a lower level. The results of this study support the hypothesis that IHNV, in contrast to VHSV or IPNV, is less efficient at completing its growth cycle in cells with a simultaneous infection with IPNV. It can be affected at several stages of viral infection and is more sensitive to the action of antiviral compounds.     

Enriching rotifers with “premium” microalgae. Isochrysis aff. galbana clone T-ISO

The effect of semi-continuous culture on the nutritional value of microalgae was tested in the rotifer Brachionus plicatilis in short-term enrichment experiments. Isochrysis aff. galbana clone T-ISO was cultured semi-continuously with renewal rates from 10 to 50% of the volume of the culture per day and used to feed the rotifers. After 24 h, dramatic differences in dry weight and protein, lipid and carbohydrate contents were observed in the rotifers depending on the renewal rate applied to the microalgal culture. Rotifers fed T-ISO cultured with low renewal rates showed low dry weight and organic content, whereas rotifers fed microalgae from nutrient-sufficient, high renewal rate cultures showed higher dry weight and increases up to 60% in protein, 35% in lipid and 100% in carbohydrate contents. Feed conversion rate (FCR) and organic FCR decreased with increasing renewal rates, indicating a more efficient assimilation of the microalgal biomass obtained at high growth rates. The fatty acid profile of rotifers reflected that of T-ISO, with maximum content of polyunsaturated fatty acids (PUFAs), n-3 fatty acids and docosahexaenoic acid (DHA) being found in the rotifers fed microalgae from the renewal rate of 40%. Results demonstrate that the biochemical composition of B. plicatilis is strongly modified through the use of semi-continuous cultures of microalgae in short-term enrichment processes. This technique provides an excellent tool to improve the nutritional value of the live feed used in fish larvae cultures.     

Outdoor mass microalgae production in Bahia Kino, Sonora, NW Mexico

The diatom Chaetoceros muelleri was grown in outdoor mass cultures under the winter, spring and summer conditions of Bahia Kino, Sonora, NW Mexico. The solar irradiance in winter was close to 60% of that available in spring and summer, but the cell concentrations and the harvestable biomass were one order of magnitude higher in spring and summer than in winter. There was no difference between the biomass harvested after 2 and 3 days in winter and summer, whereas in spring it was higher after 3 days. The protein content was significantly lower in winter, and carbohydrates and lipids were higher in winter and spring. The number of cells and the amount of harvestable biomass of outdoor cultures of C. muelleri depend on the temperature prevailing in each season, which causes significant differences in growth rates and in biochemical composition.     

Characterization of the surface properties of cellulosic fibers in fibrous and ground forms by IGC and contact angle measurements

Surface properties of fibrous and ground cotton and linen were investigated by inverse gas chromatography (IGC) and the contact angle with different liquids was also measured on fabrics composed of both fibers. Results proved that dispersion component of surface tension (γ _s ^d ) determined by IGC depends not only on the surface energy, but also on several factors influencing the adsorbability of probe molecules on the cellulosic substrates. For cotton samples, the trapping of n-alkanes among waxy molecules in the outer layer of fibers can be presumed. This effect results in larger γ _s ^d for cotton fibers than for linen in spite of the higher wettability of the linen fabrics. Besides the surface energy and trapping effects, the grinding also influences the γ _s ^d values. Specific enthalpy of adsorption (ΔH _A ^ab ) of polar probes could be determined on all linen samples, but only on the ground cotton sample. Lewis acid-base character calculated for linen and ground cotton samples depends on the same effects as the γ _s ^d does. The similar ΔH _A ^ab values of chloroform (acidic) and THF (basic) measured on each of the samples support the conclusion that the surface character is amphoteric, which is also proved by the high ΔH _A ^ab values of the amphoteric ethyl acetate and acetone probes.