Endogenous ascorbic acid modulates meristem reactivation in white spruce somatic embryos and affects thymidine and uridine metabolism

Previously, we demonstrated that, in some cell lines, exogenous applications of ascorbic acid (ASC) enhance the conversion frequency of white spruce (Picea glauca (Moench) Voss) somatic embryos, by stimulating mitotic activity in the apical meristems. To examine this event in more detail, we investigated the effects of ASC on de novo, salvage and degradation pathways of pyrimidine metabolism by following the metabolic fate of (14)C-labeled orotic acid, thymidine, uridine and uracil in shoot and root poles of germinating embryos, after altering the cellular ASC content of the embryos. Alterations in endogenous ASC content did not affect the utilization of either orotic acid or uracil, but affected the metabolism of thymidine and uridine. Specifically, a lowering of endogenous ASC content by applications of lycorine (L), an inhibitor of the last enzyme of the ASC de novo biosynthetic pathway, resulted in a lower embryo conversion frequency, as well as a reduced percentage of thymidine and uridine incorporated into nucleotides and nucleic acids. The reduction in thymidine and uridine anabolism was mainly ascribed to the decreased activities of thymidine kinase (TRK) and uridine kinase (URK), the respective salvage enzymes of thymidine and uridine, measured in L-treated embryos. These effects were solely a result of a decrease in endogenous ASC content because applications of ascorbic acid plus lycorine (ASC + L) increased embryo conversion frequency, thymidine and uridine salvage activities, and TRK and URK activities to near control values. Inclusion of exogenous ASC in the germination medium did not affect the percentage of embryos able to convert to viable plantlets, although it increased thymidine and uridine utilization for nucleic acid synthesis in the shoot and root poles of the embryos. Taken together, these findings confirm that cellular ASC plays a key role in the reactivation of the apical meristems of germinating white spruce somatic embryos.     

In vivo growth and tolerance to lime-induced iron chlorosis of leaf-derived cvs. Tomuri and Hayward kiwifruit (Actinidia deliciosa) somaclones

Summary

The present paper reports on somaclonal variation of leaf-derived ‘Tomuri’ and ‘Hayward’ somaclones regenerated at variable pH, and the effect of the in vitro selection pressure on their tolerance to lime-induced iron chlorosis as adult plants in pots and in the field. In most plants grown in pots, the above ground component length (AGL) and the node number (NN), and also root weight of ‘Hayward’, tended to increase with the regeneration pH (RpH). The higher vigour of ‘Hayward’ somaclones regenerated at high pH seemed to be confirmed in the field. In contrast, the pH selection pressure during regeneration had no clear effect on plant tolerance to lime. For ‘Hayward’, the group of somaclones obtained at pH 5.7 showed at high lime levels higher average AGL and NN than those of plants regenerated at pH 7; while clone 7.5-1, the only surviving somaclone among those obtained at the highest pH, was apparently the most tolerant to lime. The percentage of ‘Tomuri’ somaclones obtained at pH 7.5 that seemed less sensitive to lime was slightly higher than those of somaclones regenerated at pH 5.7 and 7. ‘Tomuri’ and ‘Hayward’ controls were both very sensitive to lime. Some somaclones which had been able to grow at high pH in vitro were also fairly tolerant to lime when cultured in pots and/or in the field, especially ‘Tomuri’ 5.7-5, -6; 7-6, -8, -14 and 7.5-6, and ‘Hayward’ 5.7-5; 7-2, -3, -5 and 7.5-1. The in vitro regeneration and selection methods here described therefore seem useful for creating and selecting new variability in kiwifruit and promising for obtaining plants tolerant to high pH and lime levels in the soil.     

Somatic embryo,adventitious root and shoot regeneration in in vitro grown quince leaves as influenced by treatments of different length with growth regulators

The purpose of this work was to acquire more information on the capacity of in vitro grown quince (Cydonia oblonga Mill.) leaves to simultaneously regenerate somatic embryos, adventitious roots and shoots, and to evaluate the variations induced on regeneration response by treatments of different length with growth regulators. After 2 days of liquid treatment with 2,4-dichlorophenoxyacetic acid, the leaves were cultured for 0, 3, 6, 9, 12, 15, 18 and 21 days on gelled growth medium containing the basal components of Murashige and Skoog and kinetin (Kin) 4.5 μM + naphthaleneacetic acid (NAA) 0.5 μM. At the end of each treatment period, the leaves were cultured on a transfer medium in the absence or in the presence of a growth regulator combination represented by N⁶-benzylaminopurine (BA) 2.66 μM + gibberellic acid 0.58 μM + indole-3-butyric acid 0.3 μM. The culture period for all the treatments was fixed to 52 days.     

Effect of interstock (M.9 and M.27) on vegetative growth and yield of apple trees (cv “Annurca”)

With the purpose of obtaining apple plants with reduced vigour but deep and expanded root apparatus, more adapted for establishment of apple orchards in dry, windy areas and higher altitudes, different interstem/rootstock combinations were compared, using seedling as rootstock and, alternatively, two weak interstocks (M.9 and M.27). For all treatments the plants were grafted at two height from soil (10 and 20 cm), interposing interstock 10 or 20 cm long. Plants with interstock showed lower vegetative growth in comparison with control plants with a reduction of 80% (M.27) and of 50% (M.9) of canopy growth. Moreover, use of interstock determined, as compared with the control plants, an increase of fruit production and average fruit weight. It was noted that increasing the interstock length, caused reduction of plants vigour and fruit production. The results showed that, the combination “Annurca Rossa del Sud”/M.9 (10 cm long)/seedling rootstock (20 cm high) was the most efficient for productivity, early maturing and plant vigour control.     

Pacific sardine (Sardinops sagax, Jenyns 1842) landings prediction. A neural network ecosystemic approach

In this study the performances of computational neural networks (CNNs), multiple linear regressions (MLRs) and generalised additive models (GAMs) to predict Pacific sardine (Sardinops sagax) landings and to analyse their relationships with environmental factors in the north area off Chile were studied. For this purpose several local and global environmental variables and indexes (sea surface temperature, sea level and Ekman transport index in the Chilean coast and, sea surface temperature in the area Niño 3 + 4 and Niño 1 + 2, and the south oscillation index) were considered as inputs or independent variables. Additionally, several CNNs were calibrated and validated adding the anchovy (Engraulis ringens) landings in the same area as model inputs. The time lags of the variables considered were selected through analysis of the non-linear cross-correlation functions and an alternative form of sensitivity analysis based on the approach of the missing value problem. The analysis of error measures with validation data set showed that the best results were obtained when local and global variables were used separately and combined with anchovy landings. Globally, the best result was given by a CNN with 18 input variables (model CNN 6(II) which only considered global variables and anchovy landings) and 10 neurons in a hidden layer. For this configuration the explained variance was slightly higher to 86% which supposed a standard error of prediction of 7.66%. These results were significantly better than those obtained with MLRs and GAMs. The strong correlation between predicted and observed sardine landings suggests that CNNs captured the trend of the historical data. Also, the generalisation capacity together the sensitivity analysis allowed us to identify the variables with a high weight in the model and partially to interpret the statistical functional relationships between these environmental variables and sardine landings.     

Myelopathy in Holstein x Gir calves in Brazil

This case report contains clinical and pathologic features of a degenerative myelopathy in Holstein X Gir crossbred calves in Brazil. The bilateral and symmetrical spinal cord white matter lesions were interpreted as a primary axonopathy that may be of the dying-back type.     

Efficacy of dinotefuran, permethrin and pyriproxyfen combination spot-on against Aedes aegypti mosquitoes on dogs

A spot-on formulation combining permethrin, dinotefuran and pyriproxyfen (Vectra 3D? spot-on solution for dogs - one 10-25kg pipette contains 196mg dinotefuran, 1429mg permethrin and 17mg pyriproxyfen) was evaluated in adult Beagle dogs in a study designed to measure its efficacy to control Aedes aegypti (anti-feeding effect and mortality effect). The trial was performed according to Animal Welfare and Good Clinical Practice. Twelve dogs (five males and seven female, >3 years old, weighing 8.8-13.0kg) were randomly allocated to treatment groups on pre-treatment mosquito counts: six dogs served as untreated controls, and six dogs were treated with the test formulation. Treatment consisted of applying a combination formulation to deliver at least 46.6mgkg(-1) permethrin, 6.40mgkg(-1) dinotefuran and 0.57mgkg(-1) pyriproxyfen. The combination is designed to control fleas, ticks, sand flies and mosquitoes. Each dog was infested with approximately 100 adult unfed A. aegypti once before treatment (day 6) then at 1, 7, 14, 21 and 28 days post-treatment. Counts and engorgement determination of dead and live mosquitoes were performed after 1h exposure period. In the treated group (group A), the repellency effect of the product based on engorgement status (anti-feeding effect), was 91.5%, 94%, 94.7%, 94% and 87% at 1, 7, 14, 21 and 28 days post-treatment. Mortality effect or insecticidal efficacy calculated at the end of the 1-h exposure was almost identical when calculated 24h after the 1-h exposure and remained above 93% until the end of the in-life phase. No adverse events were observed following treatment, including observations conducted 2, 4 and 24h after the last dog was treated.     

Increased apoptosis of CD4 and CD8 T lymphocytes in the airways of horses with recurrent airway obstruction

Recurrent airway obstruction (RAO, also known as equine heaves) is an inflammatory condition similar to human asthma caused by exposure of susceptible horses to poorly ventilated stable environments. The disease is characterized by neutrophilic airway inflammation, mucus hypersecretion and reversible bronchoconstriction. This inflammatory process is mediated by several factors, including antibodies, cytokines, resident cells of the airway and inflammatory cellular components that arrive in the respiratory tract. An increasing body of evidence has lent support to the concept that a dysregulation of T cell apoptosis may play a central role in the development of airway inflammation and the associated asthma. Therefore, the aim of this study was to investigate early and late apoptosis of CD4 and CD8 T cell subpopulations obtained from the airways of acute RAO-positive animals after exposure to hay/straw. The percentages of CD4 and CD8 T cells and their associated frequencies of apoptosis were quantified using flow cytometry. Hay/straw exposure induced clinical airway obstruction, airway neutrophilia and increased airway mucus production in RAO-positive horses. In addition, allergen exposure increased the percentage of CD4 T cells in RAO-positive horses as well as the frequency of early and late apoptosis in both CD4 and CD8 lymphocyte subpopulations. These results suggest that the higher frequency of lymphocyte apoptosis may play a role in disease progression of horses afflicted with RAO and may partially explain the characteristic remission of this pathological condition once the allergen source is removed. However, further studies are needed to clarify the role of T cell apoptosis in RAO-affected horses.