Influence of spermine and nitrogen deficiency on growth and secondary metabolites accumulation in Castilleja tenuiflora Benth. cultured in a RITA® temporary immersion system

The effect of exogenous spermine (SPM) on Castilleja tenuiflora shoots developing under nitrogen deficiency (ND) stress was evaluated. Shoots cultivated in a temporary immersion system were subjected to four experimental treatments: (1) control; (2) exogenous SPM; (3) ND; and (4) ND+SPM. Shoots were longer in the ND+SPM treatment (6.3 ± 0.5 cm) than in the ND treatment (4.2 ± 0.5 cm). The total chlorophyll content was similar in the control and SPM treatments (0.41 µg mg^?1 FM) and the highest values of total phenolic content were detected at 21 days in the ND+SPM treatment (84.1 ± 0.05 GAE g^?1 DM). In the ND+SPM treatment, the phenylalanine ammonia lyase activity increased earlier than in ND treatment, and reached its maximum at day 21 (3.9 ± 0.2 µmol E‐CIN h^?1 mg^?1 protein). Compared with the control, the ND and ND+SPM treatments resulted in increased secondary metabolites contents in both root and aerial parts. The strongest effect was in the roots, where the SPM and ND+SPM treatments both resulted in increased quercetin content (4.3‐fold that in the control). Our results showed that SPM partially counteract the damage caused by ND and results in increased contents of valuable bioactive compounds.     

Airborne ascospores in Mérida (SW Spain) and the effect of rain and other meteorological parameters on their concentration

Airborne ascospores have been reported to be allergenic or plant pathogens, and their presence has traditionally been associated with rainfall events. The aim of the present study was to analyze the presence of airborne ascospores in relation to weather parameters in a town in SW Spain. A seven-day recording spore trap (Burkard) was used to sample the air over 2 years at 15 m above ground level on the terrace roof of the hospital in Mérida (SW Spain). Fungal spores were identified and counted by means of two longitudinal scans over the slides under ×1000 microscopy. A correlation analysis was made of the daily meteorological data and the airborne ascospore concentrations, and t-tests were used to compare data between the 2 years. Nineteen ascospore types were defined, including one-cell ascospores (Chaetomium, Diatrype, Helvella, Xylaria), tow-cell ascospores (Diaporthe, Mycosphaerella, Nectria, Venturia), transversally septate ascospores (Melanomma, Leptosphaeria, Paraphaeosphaeria, Sporormiella, Massaria), transversally and longitudinally septate ascospores (Pleospora), and ascospores within asci (Sordaria). Leptosphaeria consisted of a group of four types described according to the number of cells, hyaline grade, wall thickness, and ornamentation, and other ascospores comprised one last additional type. The average airborne ascospore concentration was 153 ascospores/m3. One-third each of this total were from the Leptosphaeria group, with an average 54 ascospores/m3, and the two-cell ascospores or Venturia-like group (Diaporthe, Mycosphaerella, Nectria, Venturia) with 51 ascospores/m3 on average. In third position was Pleospora with 27 ascospores/m3 on average. The month with highest concentration was September, with 238 ascospores/m3, and the lowest March, with 56 ascospores/m3. By seasons, autumn had the highest concentrations, followed by winter, spring, and summer. The maximum daily concentration reached was 3,371 ascospores/m3. Daily rainfall was significantly correlated with the ascospore types Diatrype, Mycosphaerella, Nectria, two subtypes of Leptosphaeria, and Pleospora. Relative humidity was positively correlated with those ascospore types and also with Diaporthe and Paraphaeosphaeria, and negatively with Chaetomium and Melanomma. The concentration was higher on rainy days than on days without rain for Pleospora, Leptosphaeria (3 subtypes), Diatrype, Diaporthe, Nectria, Mycosphaerella, and Paraphaeosphaeria. The daily temperatures were in general correlated with the same types as the relative humidity, but with the opposite sign. For the monthly data, there were no statistically significant differences between the 2 years studied.     

Oxidative stress induces alkaloid production in Uncaria tomentosa root and cell cultures in bioreactors

The effect of oxidative stress on indole alkaloids accumulation by cell suspensions and root cultures of Uncaria tomentosa in bioreactors was investigated. Hydrogen peroxide (H₂O₂, 200 μM) added to U. tomentosa cell suspension cultures in shaken flasks induced the production of monoterpenoid oxindole alkaloids (MOA) up to 40.0 μg/L. In a stirred tank bioreactor, MOA were enhanced by exogenous H₂O₂ (200 μM) from no detection up to 59.3 μg/L. Root cultures grew linearly in shaken flasks with a μ=0.045 days^?1 and maximum biomass of 12.08±1.24 g DW/L (at day 30). Roots accumulated 3α‐dihydrocadambine (DHC) 2354.3±244.8 μg/g DW (at day 40) and MOA 348.2±32.1 μg/g DW (at day 18). Exogenous addition of H₂O₂ had a differential effect on DHC and MOA production in shaken flasks. At 200 μM H₂O₂, MOA were enhanced by 56% and DHC by 30%; while addition of 800 and 1000 μM H₂O₂, reduced by 30–40% DHC accumulation without change in MOA. Root cultures in the airlift reactor produced extracellular H₂O₂ with a characteristic biphasic profile after changing aeration. Maximum MOA was 9.06 mg/L at day 60 while at this time roots reached ca. 1 mg/L of DHC. Intracellular H₂O₂ in root cultures growing in the bioreactor was 0.87 μmol/g DW compared to 0.26 μmol/g DW of shaken flasks cultures. These results were in agreement with a higher activity of the antioxidant enzymes superoxide dismutase and peroxidase by 6‐ and 2‐times, respectively. U. tomentosa roots growing in the airlift bioreactor were exposed to an oxidative stress and their antioxidant system was active allowing them to produce oxindole alkaloids.     

Environment‐driven changes in diversity of riparian plant communities along a mountain river

The study of changes in species richness and composition along rivers has focused on large spatial scales. It has been ignored that in different sections of the river (high mountain area, middle zone, and mouth of the river) the specific environmental conditions can generate different longitudinal patterns of the species richness and composition. In this study, we determine whether species richness and composition of the riparian plant communities change along a mountain river and whether these changes are related to environmental variables. We expect an increase in species richness and turnover along the river, that the upstream communities would be a subset of the downstream communities, and that such would be related to edaphic and hydrologic conditions. To test this, we sampled three strata of the riparian vegetation (upper: individuals with <1 cm of ND, middle: individuals with >1 cm of ND, low: individuals with >1 m tall) in a set of 15 sites that we place along a mountain river. Additionally, we recorded topographic, hydrological, morphological, and soil variables. We performed correlation analyzes to determine whether changes in species richness and turnover were related to increased distance to the origin of the river. Also, we obtained the nestedness and evaluated the importance of environmental variables with GLM, LASSO regression, and CCA. With the increase in distance, the species richness decreases in the upper stratum, but not in the middle and the low stratum (although the highest values were observed near the origin of the river), the turnover increase in all strata and the upstream communities were not a subset of the downstream communities. The changes in species richness and composition were related to topographic (altitude), hydrological (flow), and edaphic (conductivity and pH) variables. Our results indicate that at small spatial scales the patterns of richness and composition differ from what has been found at larger spatial scales and that these patterns are associated with environmental changes in the strong altitude gradients of mountain rivers.     

Sequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from <Emphasis Type="Italic">Asclepias fruticosa</Emphasis> latex

Asclepain f is a papain-like protease previously isolated and characterized from latex of Asclepias fruticosa. This enzyme is a member of the C1 family of cysteine proteases that are synthesized as preproenzymes. The enzyme belongs to the alpha + beta class of proteins, with two disulfide bridges (Cys22-Cys63 and Cys56-Cys95) in the alpha domain, and another one (Cys150-Cys201) in the beta domain, as was determined by molecular modeling. A full-length 1,152 bp cDNA was cloned by RT-RACE-PCR from latex mRNA. The sequence was predicted as an open reading frame of 340 amino acid residues, of which 16 residues belong to the signal peptide, 113 to the propeptide and 211 to the mature enzyme. The full-length cDNA was ligated to pPICZα vector and expressed in Pichia pastoris. Recombinant asclepain f showed endopeptidase activity on pGlu-Phe-Leu-p-nitroanilide and was identified by PMF-MALDI-TOF MS. Asclepain f is the first peptidase cloned and expressed from mRNA isolated from plant latex, confirming the presence of the preprocysteine peptidase in the latex.     

Oxidative Stress Increased in Pregnant Women with Iodine Deficiency

Iodine is an essential element trace for the synthesis of maternal thyroid hormones needed to support normal fetal development; it also acts as an antioxidant directly or induce antioxidant enzymes indirectly. Iodine deficiency and oxidative stress are associated with pregnancy complications. This study aimed to assess the urinary iodine concentration and its relationship with the antioxidant and oxidative stress status during gestation. Pregnant women were consecutively recruited from an obstetric clinic during all gestation trimesters, and urinary iodine concentration, antioxidant, and oxidative stress were determined. Results showed that 70 % of pregnant women have optimal iodine levels (150–200 μg/L), while approximately 30 % showed mild iodine deficiency (50–99 μg/L). Oxidative stress was significantly higher, and the antioxidant status was also compromised as evidenced by decreased total antioxidant status and superoxide dismutase (SOD) activity in pregnant women with mild iodine deficiency than pregnant women with optimal iodine levels. Significant positive correlations were noted between optimal iodine levels and total antioxidant status. Oxidative stress was significantly correlated with mild iodine deficiency. However, no significant correlation was found between iodine levels and SOD and catalase activities. In conclusion, for the first time, these data suggest a correlation between iodine levels and the antioxidant status during pregnancy.