Chlorophyll catabolism pathway in fruits of Capsicum annuum (L.): stay-green versus red fruits

The aim of the present study is to investigate the chlorophyll catabolism pathway of wild-type red and stay-green mutants of Capiscum annuum (L.) fruits. In the wild-type red lines chlorophyll catabolism is concomitant with the start of carotenogenesis, whereas in the stay-green mutant lines the chlorophylls coexist with that process, even in over-ripe fruit. During the first stages of ripening, the chlorophyll a/chlorophyll b ratio is similar for both genotypes, but as ripening proceeds, the ratio in the stay-green lines becomes very high as a result of a blocked degradation of chlorophyll a while chlorophyll b is degraded at a normal rate. The absence of dephytylated chlorophylls in the wild-type lines distinguishes these from the mutant lines, in which there is a sequential accumulation of chlorophyllide a and pheophorbide a. Allomerized chlorophylls (13(2)-OH-chlorophyll a and b) have also been identified in the catabolic process of the mutant lines, but are absent from the wild type. Consequently, an alteration in pheophorbide a oxygenase (PaO) activity seems to be responsible for the stay-green genotype in the lines of pepper analyzed in this study.     

Oxytocin in pig seminal plasma is positively related with in vivo fertility of inseminated sowsOA

Background: Identification of relevant in vivo biomarkers for fertility remains a challenge for the livestock industry.Concentrations of the small peptide hormone oxytocin(OXT), involved in male reproductive function and present in the seminal plasma(SP) of several species could be a robust one. This study characterized concentrations of SPOXT in ejaculates from boars used in artificial insemination(AI) programs aiming to evaluate its relationship with sperm quality variables and in vivo fertili...     

Pigment metabolism of 'Sikitita' olive ( Olea europaea L.): a new cultivar obtained by cross-breeding

The new olive cultivar 'Sikitita' was obtained from a cross between the 'Picual' and 'Arbequina' varieties. 'Sikitita' was selected for its features, making it particularly suited to high-density olive hedgerow orchards. From the standpoint of chloroplast pigment metabolism, the fruits of the 'Picual' and 'Arbequina' varieties have significant differences. It is therefore extremely interesting to analyze the descendants of both cultivars. With regard to chlorophyll catabolism, 'Sikitita' has proven to be a cultivar with low pigmentation and low levels of chlorophyllase activity. This is contrary to the findings obtained to date, where varieties with low pigmentation are a consequence of high chlorophyllase activity ('Arbequina') and highly pigmented fruits are due to low chlorophyllase activity ('Picual'). 'Arbequina' was, until recently, the only cultivar described that had developed a carotenogenic process, despite its anthocyanic ripening. However, from its father ('Arbequina'), the 'Sikitita' cultivar has inherited the pool of enzymes necessary to esterify xanthophylls at the chromoplast level. This makes 'Sikitita' a very interesting cultivar, with potential chemotaxonomic differences (such as esterified xanthophylls in the olive oils), and demonstrates the interest in genetic improvement programs for olive cultivars with different organoleptic characteristics.     

Characterization of Vibrio tapetis strains isolated from diseased cultured Wedge sole (Dicologoglossa cuneata Moreau)

The first isolation of Vibrio tapetis from Wedge sole (Dicologoglossa cuneata) is reported. The bacterium was recovered from ulcers of ailing cultured fish, from two different outbreaks occurred in spring 2005. The four isolates found (a200, a201, a204 and a255) were biochemically, genetically and serologically characterized and diagnosis was confirmed by PCR V. tapetis specific primers and multilocus sequencing analysis (MLSA). The isolates constituted a homogeneous phenotypic and genotypic group, being distinct to the already serological and genetic groups defined within the species. A virulence evaluation of the isolate a255 was also carried out; however this strain was unable to induce disease in fry and juvenile Wedge sole.     

Acid PAGE procedure for detection of phaseolins in field beans (Phaseolus vulgaris L.)

Summary A procedure based on acid polyacrylamide gel electrophoresis (acid PAGE) has been described. Resolution of phaseolin bands appears to be relatively superior to that obtained with basic PAGE system. The procedure discriminates S, T, C, M and B types of phaseolins; S, T and C have been reported previously while M and B types were identified in the present study.     

Motility Characteristics and Fertilizing Capacity of Boar Spermatozoa Stained with Hoechst 33342

Flow cytometry sorting of spermatozoa using fluorescence dye Hoechst 33342 is the only effective sex selection methodology validated in numerous laboratories. This study was carried out to determine the effect of Hoechst 33342 on the motility and fertility of stained boar spermatozoa. Experiment 1 evaluated motility parameters (percentage of motile spermatozoa, velocity, angularity and oscillation) of boar spermatozoa stained with Hoechst 33342 by a computer‐aided sperm analysis (CASA) instrument. Spermatozoa (30 million/ml) were divided into five treatment groups and stained during 1 h at 35°C with 9, 18, 27, 60 and 90 μM of H33342. There were no differences in sperm motility patterns nor percentages of motile spermatozoa incubated in the presence of 9, 18 or 27 μM. Percentage of motile spermatozoa and motility parameters decreased significantly (p < 0.05) at 60 μM of Hoechst 33342. Spermatozoa were immotile at concentration of 90 μM. In experiment 2, pregnancy rates, farrowing rates and litter size from sows (n = 275) artificially inseminated (AI) with either Hoechst 33342 stained (27 μM) or unstained (control) spermatozoa were determined. Sows inseminated with stained spermatozoa had no significant lower pregnancy rate (88.33%) as compared with controls (90.32%). Staining neither affected farrowing rates (85.0 vs 87.7%) nor total number of piglets born (10.56 ± 0.32 vs 10.47 ± 0.24, stained and controls, respectively). No phenotypical abnormalities were registered among the newborn piglets. The data suggest that incubating spermatozoa with Hoechst 33342 at levels required for X‐ and Y‐bearing chromosome sperm sorting, does not impair sperm viability or their fertility after AI.     

Relationships among soil characteristics,plant macronutrients,and cardenolide accumulation in natural populations of Digitalis obscura

In the present study, we have investigated relationships among several soil parameters (pH, organic matter, total carbonate, macronutrients, electrical conductivity, cation‐exchange capacity) and macronutrient and cardenolide contents in leaves of wild Digitalis obscura plants. Young and mature leaves and soil samples were collected in ten different areas, corresponding to three Mediterranean bioclimatic belts (thermo‐, meso‐, and supramediterranean belts). Soil and leaf macronutrient (N, P, K, Ca, and Mg) contents and leaf cardenolide contents were determined. Bioclimatic conditions influenced the development of D. obscura, biomass being lowest in plant populations of the supramediterranean belt, but they did not exert any relevant effect on the contents of macronutrients in soils and plants. Magnesium (total and EDTA‐extracted) was the only soil macronutrient significantly correlated with its content in the plant. Cardenolide contents were negatively correlated with the N, P, and K contents in young leaves, whereas such correlations were highly significant and positive for Mg.     

Phenotypic,molecular and pathogenic characterization of Phlyctema vagabunda,causal agent of olive leprosy

Olive leprosy, caused by the fungus Phlyctema vagabunda, is a classic fruit rot disease widespread in the Mediterranean basin. From 2009 to 2013, new disease symptoms consisting of small circular necrotic leaf lesions, coin branch canker and shoot dieback were observed in Spanish and Portuguese olive orchards showing intense defoliation. Phlyctema‐like anamorphs were consistently isolated from leaves and shoots with symptoms. Representative isolates from affected leaves, shoots and fruits were characterized based on morphology of colonies and conidia, optimum growth temperature and comparison of DNA sequence data from four regions: ITS, tub2, MIT and rpb2. In addition, pathogenicity tests were performed on apple and olive fruits, and on branches and leaves of olive trees. Maximum mycelial growth rate ranged between 0.54 and 0.73 mm per day. Conidia produced on inoculated apple fruits showed slight differences in morphology among the representative fungal isolates evaluated. Phylogenetic analysis clustered all of the Phlyctema‐like isolates in the same clade, identifying them as Phlyctema vagabunda. On fruits, influence of wounding, ripening and cultivar resistance was studied, with cv. Blanqueta being the most susceptible cultivar. On branches, a mycelial‐plug inoculation method reproduced olive leprosy symptoms and caused shoot dieback. On leaves, Koch's postulates were fulfilled and the pathogen caused characteristic necrotic spots and plant defoliation. This is the first time that the pathogenicity of P. vagabunda in olive leaves has been demonstrated.     

Assessment of heavy metal tolerance in two plant species growing in experimental disturbed polluted urban soil

Purpose

Urban soil, which is strongly influenced by anthropogenic activities, receives a major proportion of trace metal wastes. The aim of this work was to determine heavy metal concentration in (a) soil, to know the degree of the soil pollution; (b) roots and leaves of two plant species, Brassica juncea as an accumulator plant and Solanum lycopersicum as a crop plant; and (c) drainage water, to evaluate the heavy metal mobility.

Materials and methods

The study area is located in Sants, a neighborhood in Barcelona (Catalonia, Spain). Thirty kilograms of two representative soil depths (0–15 and 15–40 cm) was sampled and subsequently mixed. The two studied species were cultivated for 3 weeks in greenhouse conditions, and all pots were irrigated with water weekly to field capacity with a nutrient solution (pH = 6.5). If not otherwise stated, given results are means ± standard deviation of four replicated pots each with a composite sample of 12 individual plants per treatment. X-ray fluorescence (FRX) and diethylenetriaminepentaacetic acid soil extraction (DTPA) were used for total and available metal soil contents, respectively. Weekly cumulative drainage water of each pot was collected in polyethylene bottles and stored at 4 °C until analysis.

Results and discussion

The main pollutants are Cu, Pb, and Zn with topsoil total concentrations of approximately 1355, 2230, and 6239 mg kg^?1, respectively. The same soil elements for available fractions were slightly elevated (9.6, 5.8, and 6.7% of total concentration). The concentrations of Cu, Pb, and Zn in the plants’ leaves are greater in B. juncea than in S. lycopersicum. Furthermore, they are greater in the roots than in leaves. The Pb concentrations in a crop plant exceeded the 0.10 mg kg^?1 limit established for vegetables devoted for food in the European legislation. Unusually elevated concentrations of Pb (over 10 μg L^?1) were detected in the drainage water. These values exceeded the acceptable toxic concentrations in waters, according to the Spanish legislation.

Conclusions

The urban soil studied was highly contaminated by Cu, Pb, and Zn, and this pollution is more evident in the topsoil. A great part of these heavy metals was bioavailable for plants. Thus, the two plants (S. lycopersicum and B. juncea) had an ability to transport heavy metals from the roots to the shoots, especially for Zn. Great contents of heavy metals in the drainage water after the irrigation of plants were observed.