Mapping of QTLs in tomato line FLA456 associated with resistance to a virus causing tomato yellow leaf curl disease

Tomato (yellow) leaf curl disease (TYLCD) is a serious threat to tomato production in the tropics and subtropics. The genetics of resistance to Tomato yellow leaf curl Thailand virus Taiwan strain (TYLCTHV-[TW]) in a highly resistant tomato line FLA456 was studied through quantitative trait loci (QTL) analysis. Four QTLs named qTy4.1, qTy6.1, qTy10.1 and qTy11.1 were detected on chromosomes 4, 6, 10, and 11, respectively, through evaluation of an F₆ recombinant inbred line (RIL) population derived from a cross between FLA456 (resistant) and CLN1621L (susceptible). Gene action of all QTLs was recessive based on disease reaction of the F₁. The markers SINAC1 and SLM4-34 flanked qTy4.1 on chromosome 4, and SLM11-12 and SLM11-17 defined qTy11.1, which co-located with the previously identified Ty-5 and Ty-2 loci, respectively. qTy6.1 was flanked by the markers SLM6-55 and TES-0014, and qTy10.1 by the markers SLM10-80-SLM10-46 on chromosomes 6 and 10. The LOD values of the putative QTLs ranged from 2.79 to 13.76. The phenotypic variance explained by each QTL ranged from 7.1 to 31.9 %. The four QTLs collectively contributed about 60.5 % of the phenotypic variation in resistance against TYLCTHV-[TW]. Group mean severity scores of those RILs possessing three or four qTy were generally lower than RIL groups with only one or no qTy. Given the diversity of begomoviruses that cause TYLCD across the regions, the new QTLs from FLA456 would be valuable in tomato breeding for developing varieties with durable resistance. Two QTL intervals (qTy4.1 and qTy10.1) contained virus resistance candidate genes such as CTV.22 and eukaryotic translation initiation factor 4E.     

Molecular changes during intra and inter recurrent selection of two populations of maize: one adapted and one non adapted to the selection environment

In several reciprocal recurrent selection (RRS) programs heterosis and favorable characteristics are achieved by means of one adapted and one non adapted population. We evaluated with molecular markers, an intrapopulation selection followed by RRS of one Spanish population adapted to Mediterranean Spain and one US Corn Belt population non adapted to Spanish conditions. Results from other authors suggest that during recurrent selection, non adapted populations have higher loss of variability, genetic differentiation and lower effective population size than expected according to the number of families selected each generation. This could be due to natural selection which is not under the breeder’s control and is expected to mainly act on non adapted populations. The number of markers with convergent allelic change was similar to the number of markers with divergent allelic change which explains the lack of genetic differentiation and the failure to increase heterosis during RRS because the effects of both types of changes compensate. It seems that the predominant mode of gene action depends on the particular germplasm involved in the RRS. By evaluating the allelic changes during selection, we identified four regions (2.04, 4.06, 6.03, 9.02) that significantly changed during selection in our selection experiment and that have been associated to selection in other selection experiments and to multiple traits in QTL experiments.     

Pseudorabies virus infection in mink: a host-specific pathogenesis

Pseudorabies virus (PRV) is an alphaherpesvirus that causes a neurological disease in many wild and domestic animals. The neuropathology elicited by PRV is quite consistent regardless of the host with the only exception of mink, in which it is characterized by a vasculopathy rather than by an encephalitis. In this study, we aimed to investigate the underlying pathogenic mechanism(s) of PRV infection in mink by using immunohistochemistry and laser capture microdissection (LCM) on material from naturally and experimentally infected animals. The inflammatory reaction induced by PRV was minimal or absent not only in the nervous system, where we identified a low number of macrophages and a few T lymphocytes, but also in the primary replication site, the oropharyngeal mucosa; however, the number of PRV-infected cells detected by immunohistochemistry was extremely high both in the peripheral mucosa and in the nervous tissue. On the other hand, the vascular pathology included parenchymal hemorrhages of various degrees and, in specific cortical areas of the brain, fibrinoid degeneration of the capillary walls. Detection of viral antigens by immunohistochemistry revealed infection of endothelial cells of capillaries situated both in the oropharyngeal mucosa and in the brain stem; the presence of PRV DNA in vessels was further demonstrated by PCR performed on LCM samples of brain capillaries. These results can be interpreted as supporting the idea that the different pathology of the disease in mink may be the consequence of an increased endotheliotropism of PRV in this species. Infection of the vessel wall may then lead to vascular pathology and impairment in endothelial cell function, resulting in a weak immune response to infection.     

In vitro and in vivo effect of flutamide on steroid hormone secretion in canine and human inflammatory breast cancer cell lines

The aim was to study the effects of flutamide on cell proliferation, in vivo tumour growth and steroid production in canine and human IBC cell lines. IPC‐366 and SUM149 cell cultures were exposed to flutamide concentrations for 72 hours. Additionally, IPC‐366 and SUM149 xenotransplanted mice were treated subcutaneously with flutamide 3 times a week for 2 weeks. Steroid hormones determination in culture media, serum and tumour homogenates (pregnenolone, progesterone, androstenedione, testosterone, dihydrotestosterone, 17β‐oestradiol and oestrone sulphate) were assayed by EIA. in vitro cell proliferation percentages showed a decrease in all flutamide dosages in IPC‐366 and SUM149. in vivo flutamide reduced tumour size by 55% to 65%, and metastasis rates decreased. In treated groups, androgen levels in culture media, serum and tumour homogenates were increased as oestrogen levels decreased. These results suggest that flutamide treatment inhibits cell proliferation and promotes tumour reduction by increasing androgen levels and also support future therapy approaches.     

Modelling the Phenological Relationships of Questing Immature Ixodes Ricinus (Ixodidae) Using Temperature and NDVI Data

All active stages of the tick Ixodes ricinus were collected monthly at two sites in northern Spain between the years 2000 and 2007. We used percentile accumulation of the active stage in the environment to evaluate simple and coherent correlations between accumulation of the active stages of larvae and nymphs and medium‐resolution MODIS satellite‐derived information on the climate, including monthly and accumulated temperature and the Normalized Difference Vegetation Index (NDVI). This framework is not intended to predict the actual abundance of ticks in the field as a measure of the hazard to humans, but to provide a basic structure for addressing the phenology of the tick in its geographic range. We demonstrated that the accumulation of larval ticks in the active stage is a sigmoid function of the accumulated temperature from the beginning of the calendar year. We also demonstrated that the accumulated temperature necessary to recruit nymphs from the questing larval stage is a function of the changes in accumulated larvae and nymphs and the accumulated temperature and NDVI recorded by the Aqua sensor. The low p‐values obtained in the regressions confirmed that such recruitment can be calculated using time intervals to estimate, for example, the beginning of the questing period or the time of the year when a population peak can be expected. The comparison among predicted and actual accumulated temperatures between larvae and nymph recruitment had an averaged error of ±20 days in one complete year. The use of accumulated temperature and NDVI proposed in this study opens up the re‐evaluation of reports on the phenology of the tick in Europe. This framework is intended to evaluate the same correlations along the tick's range and predict its phenological patterns in areas of pathogen transmission risk for humans.     

Evaluation of a fluorescence-polarization assay for the diagnosis of bovine brucellosis in México.

A homogeneous fluorescence-polarization assay (FPA) was used for the serological diagnosis of bovine brucellosis in México. The assay uses O-polysaccharide prepared from Brucella abortus lipoplysaccharide (20-30 kDa) conjugated with fluorescein isothiocyanate as a tracer. To measure the fluorescence polarization, a FPM-1 fluorescence-polarization analyzer was used with the procedure described by Nielsen et al. (1996b). A cut-off value of 90 millipolarization (mP) units was used for testing 560 bovine sera from different areas of México. (305 positive sera and 255 negative sera according to the complement fixation test; CFT.) Some were tested with the Rose Bengal plate (RB) test (n = 490) and some with the rivanol-agglutination (RIV) test (n = 190). Sensitivities were 98.3%, 99.3% and 99.0%, and specificities were 68.8%, 55.4% and 96.9%, respectively, for RB, RIV and FPA. The FPA gave a kappa coefficient of agreement with respect to CFT of 0.96, while RB and RIV (relative to the CFT) gave coefficients of 0.70 and 0.61, respectively. Finally, ROC analysis suggested a cut-off value which agreed with the one recommended in the test procedure. We concluded that FPA is a suitable test to be used instead of the CFT in Mexican conditions.     

Antioxidant activity and lipid peroxidation in Artemia nauplii enriched with DHA‐rich oil emulsion and the effect of adding an external antioxidant based on hydroxytyrosol

Artemia nauplii catabolize polyunsaturated fatty acids (PUFA); in particular, they retroconvert docosahexaenoic acid (DHA, 22:6n‐3), so enrichment is a continuous quest towards increasing PUFA through the use of PUFA‐rich enrichment products. However, optimal conditions during enrichment (aeration, illumination and temperatures around 28°C) tend to accelerate autoxidation of PUFA and the formation of potentially toxic oxidation products. Water‐soluble antioxidants like the polyphenolic compound hydroxytyrosol (3,4‐dihydroxyphenylethanol), a polar molecule found in the water fraction resulting after the milling process of olives, arise as promising compounds to prevent oxidation during Artemia enrichments. We investigated the antioxidant activity and lipid peroxidation in Artemia nauplii during enrichment and the effect of adding an external antioxidant based on hydroxytyrosol during the enrichment with a PUFA‐rich emulsion (M70). For this purpose, the activity of antioxidant enzymes (catalase, superoxide dismutase, glutathione‐S‐transferase, glutathione peroxidase), as well as lipid peroxidation, was determined in enriched and unenriched Artemia nauplii. To validate antioxidant activity and lipid peroxidation, in a first experiment, nauplii were enriched with microalgae (Tetraselmis suecica), yeast (Saccharomyces cerevisiae) and M70 emulsion. In a second experiment, enrichment with a commercial emulsion (DC Super Selco), M70, and a combination of M70 and hydroxytyrosol (Hytolive, HYT) added as an external antioxidant were performed. The combination of M70 with HYT produced the best results, in terms of activity of antioxidant enzymes. The analysis of the fatty acids from total lipids showed that the addition of hydroxytyrosol preserved the DHA percentage of enriched nauplii.     

Maguey (<Emphasis Type="Italic">Agave salmiana</Emphasis>) infructescence morphology and its relationship to yield components

Agaves are long-lived semelparous plants that produce a high number of seeds, in dehiscent capsules, on the apical section of a stalk, up to 5 m long, after 8–25 years. These and other characteristics such as yield and yield components are scarcely evaluated in the plants of the Agave genus. The objective of this study was to quantify the capsules and seeds yield of A. salmiana Otto ex Salm-Dyck plants simultaneously maturing and growing in the same region. Infructescences of three plants simultaneously growing at San Luis Potosí, Mexico, were harvested. On them we evaluated the number of umbels, capsules and seeds (normal and sterile) and their mass per plant. The study was developed on a completely randomized design with each infructescence as an experimental unit. Also, the type distribution of the number and mass of these yield components along the stalk was evaluated with the Chi square test for goodness of fit, the Shapiro–Wilks for normality tests, asymmetry and kurtosis. Data were analyzed with the ANOVA and multiple comparisons by the Tukey test (p?≤?0.05). The number of umbels per plant (17–25), capsule per umbel (2–179) and per plant (554–1990), normal seed per capsule (0–297), normal seeds per plant (30,610–186,209) and sterile seeds per plant (211,059–619,251) widely and significantly varied among infructescences. Biomass of capsules per plant umbel (3–795 g), biomass of sterile and normal seeds per capsule (0.071–1.449 and 0–3.320 g), per umbel (0.34–97.76 and 0.21–185.26 g) and per plant (182–1052 and 334–2069 g) also varied widely. Seed yield was statistically different between plants simultaneously growing and maturing at the same site.