A high-throughput method for isolation of salicylic acid metabolic mutants

Background  

Salicylic acid (SA) is a key defense signal molecule against biotrophic pathogens in plants. Quantification of SA levels in plants is critical for dissecting the SA-mediated immune response. Although HPLC and GC/MS are routinely used to determine SA concentrations, they are expensive and time-consuming. We recently described a rapid method for a bacterial biosensor Acinetobacter sp. ADPWH_lux-based SA quantification, which enables high-throughput analysis. In this study we describe an improved method for fast sample preparation, and present a high-throughput strategy for isolation of SA metabolic mutants.     

Novel biological properties of Oenothera paradoxa defatted seed extracts: effects on metallopeptidase activity

In this study, for the first time, we used the in vitro metallopeptidase model for the identification of a potential novel activity of defatted evening primrose seed extracts. Prepared extracts of different polarity (aqueous, 60% ethanolic, isopropanolic, and 30% isopropanolic) at concentrations of 1.5-100 microg/mL exhibited a significant and dose dependent inhibition of three tested enzymes. The 50% inhibition of enzymes activity showed that aminopeptidase N (APN) was the enzyme affected to the greatest extent with IC50 at the level of 2.8 microg/mL and 2.9 microg/mL for aqueous and 30% isopropanolic extracts, respectively. The activity of neutral endopeptidase (NEP) was quite strongly inhibited by the extracts as well. The HPLC-DAD analysis and bioguided fractionation led to the identification of four active compounds: (-)-epicatechin gallate, proanthocyanidin B3, oenothein B, and penta-O-galloyl-beta-D-glucose (PGG). Oenothein B has been shown previously to inhibit metallopeptidases. The three other compounds are known to inhibit angiotensin-converting enzyme (ACE), but they have not been previously reported to inhibit the NEP and APN activity. PGG and procyanidins with different degrees of polymerization, as the dominating compounds in O. paradoxa seeds, seemed to play a role in the crude extract activity.     

Tree,stand and site characteristics affecting the occurrence of lammas growth and multiple tops in field-grown Norway spruce

New Forests - In the Nordic-Baltic region, there has been a growing concern about an increasing occurrence of multiple tops in young stands of Norway spruce. There is however a lack of...     

The effect of biostimulators on the content of crude oil and total protein in winter oilseed rape (Brassica napus L.) seeds

The objective of the study was to determine the effect of the experimental factors on the seed quality of three winter oilseed rape cultivars. The following factors were studied: I-three winter oilseed rape morphotypes: population, hybrid restored characterised by a semi-dwarf growth pattern, hybrid restored characterised by a conventional growth pattern; II-two sowing methods: a spacing of 22.5?cm, a spacing of 45.0?cm; III-four types of biostimulants: control-no biostimulants, the biostimulant Tytanit^®, Asahi^®SL, Silvit^®. Seed samples were subjected to chemical analysis to determine: crude oil, total protein and crude fibre. Natural stimulants of plant growth and development contributed to an increase in crude oil ranging from 7.4 to 9.5?g?kg^–1 d.m., on average, compared with control. The biostimulants applied did not affect total protein content which was, however, affected by meteorological conditions in individual study years as well as the genetic factor. Biostimulants increased crude fibre content compared with control.     

The effects of fertilization on the health status,nutrition and growth of Norway spruce forests with yellowing symptoms

We studied the effects of magnesium (Mg) NPK fertilizer applied in 2000 to maturing Norway spruce (Picea abies /L./ Karst.) stands with yellowing symptoms and situated on poor acidified soils in the ?umava mountains (the Czech Republic). Long-term defoliation, nutrient content in the foliage and diameter increment were evaluated in three specific variants: yellowing trees, trees without yellowing symptoms and fertilized trees. The results indicate differences in soil characteristics, nutrient content and radial growth with response to climate. Fertilization increased Mg content and base saturation in the soil and decreased the amount of exchangeable aluminum and hydrolytic acidity. Differences were significant especially in magnesium, calcium and phosphorus contents in the foliage of yellowing trees. There was also a significant difference in manganese content; yellowing trees showed deficiency in comparison with both fertilized and unfertilized healthy trees. Two years after the initial fertilization (to present), defoliation and yellowing symptoms in the unfertilized yellowing trees were more pronounced compared to other variants. These symptoms were more influenced by fertilization than foliation. The duration of significant positive effects of fertilization on diameter increment lasted for a shorter time than the effect on foliage. Simultaneously, higher sensitivity to climatic factors was observed in the fertilized trees.     

Impact of water scarcity on spruce and beech forests

One of the greatest threats posed by ongoing climate change may be regarded the drought caused by changes in precipitation distribution. The aim of presented study was to characterize reactions to dry conditions and conditions without drought stress on gross primary production (GPP) and net ecosystem production (NEP) of spruce and beech forests, as these two species dominate within the European continent. Daily courses of GPP and NEP of these two species were evaluated in relation to an expected decrease in CO2 uptake during dry days. The occurrence of CO2 uptake hysteresis in daily production was also investigated. Our study was performed at Bily Kriz(spruce) and Stitna(beech) mountain forest sites during 2010–2012 period. We applied eddy covariance technique for the estimation of carbon fluxes, vapor pressure deficit and precipitation characteristics together with the SoilClim model for the determination of drought conditions, and the inverse of the Penman–Monteith equation to compute canopy conductance. Significant differences were found in response to reduced water supply for both species. Spruce reacts by closing its stomata before noon and maintaining a reduced photosynthetic activity for the rest of the day, while beech keeps its stomata open as long as possible and slightly reduces photosynthetic activity evenly throughout the entire day. In the spruce forest, we found substantial hysteresis in the light response curve of GPP. In the beech forest, the shape of this curve was different: evening values exceeded morning values.     

Evaluation of silicon as a nutritional supplement for greenhouse zinnia production

Greenhouse production of floricultural crops uses soilless substrates, which have limited amounts of plant-available silicon (Si) compared to mineral soils. The inclusion of Si supplements in greenhouse production has proven beneficial for several crops. The goal of this study was to evaluate the incorporation of Si supplements for improved flower quality of greenhouse-produced zinnia (Zinnia elegans). In the first study, quantitative horticultural traits and leaf Si, macronutrient and micronutrient concentrations were compared between Si-treated and control plants. The Si supplements evaluated were rice husk ash substrate incorporation, hydrous potassium silicate (KSiO₃) substrate incorporation or weekly KSiO₃ substrate drenches, and weekly sodium silicate (NaSiO₃) foliar sprays. Several horticultural traits differed in Si-treated plants. The flower and stem diameters were larger in plants supplemented with at least one rate of each Si supplement. Reduced plant heights and 3–5-day delays in anthesis occurred in plants supplemented with weekly KSiO₃ drenches. Leaf Si concentrations were 1.2–3.3-fold higher in Si-supplemented plants, while the macronutrients, N, S, Mg, and Ca, were lower and K was higher only in KSiO₃-supplemented plants. In a second study, the effects of weekly KSiO₃ drenches and weekly NaSiO₃ foliar sprays on the leaf resistance of zinnia plants were evaluated. Only the NaSiO₃ foliar sprays moderately increased leaf resistances. While not supporting an active role for Si in reducing transpiration, NaSiO₃ foliar sprays may form antitranspirant film that increases leaf resistance. Thus, the beneficial effects of Si supplementation on greenhouse-produced zinnia depend on the form and rate of Si used.     

Comparison of Vernalization Requirements and Frost Resistance of Winter Rape Lines Derived from Double Haploids

Vernalization requirements and cold resistance of 13 lines of winter rape derived from doubled haploids obtained by androgenesis were investigated. The degree of vernalization requirements was examined in two parallel experiments. In the first experiment seedlings aged 2 weeks were vernalized during 63 and 35 days under controlled conditions, at 5 °C. In the second experiment the natural conditions of vernalization were differentiated by sowing the plants at four different dates in spring: March 15th, April 15th, May 15th, and at the latest date excluding vernalization, June 5th. The lines examined revealed differentiation of vernalization requirements that were similar in both experiments; all the lines required a period of exposure to cold before flowering. Cold resistance of lines was estimated in two successive series at −15 °C and −17 °C. Before testing the plants were kept in natural field conditions from the beginning of September until the middle of November. Testing of cold resistance was preceded by hardening under controlled conditions. Considerable differences in cold resistance was found in the forms investigated.
The results obtained do not show any relation between frost resistance and the degree of vernalization requirements, as the line characterized by the strongest winter habit of growth showed low cold resistance, whereas the lines which were closer to spring growth habit showed the highest level of cold resistance; the lines with the least vernalization requirements revealed the highest level of cold resistance. This conclusion can be confirmed by the significance of the correlation coefficients between the various indices defining the frost resistance and the indices of vernalization requirements.     

Microstructure of the small intestine in broiler chickens fed a diet with probiotic or synbiotic supplementation

The aim of the study was to determine the effect of dietary supplementation of a probiotic and a synbiotic on the morphometric parameters of the small intestine of broiler chickens. The experiment was conducted on three hundred sixty, one‐day‐old female Ross 308 chicks, which were randomly selected from 20,000 birds and divided into three treatment groups (n = 120) with ten replicates per treatment. The control group (C) was fed a commercial diet, the probiotic group (PRO) was fed the same diet with an added 1% of the probiotic Lavipan® (Lactococcus lactis, Carnobacterium divergens, Lactobacillus casei, Lactobacillus plantarum and Saccharomyces cerevisiae), and the synbiotic group (SYN) was fed the commercial diet with an added synbiotic: 0.8% of the prebiotic RFO (extracted from lupin seeds) and 1% Lavipan®. According to the manufacturer's data, apart from the typical probiotic action,microorganisms contained in the preparation release anti‐bacterial substances (hydrogen peroxide and bacteriocins) and, therefore, are antagonistic towards pathogenic bacteria present in the gut of animals. Supplementation took place in the first seven days of rearing, and all birds had ad libitum access to water and feed during the 42 days of the experiment. On the last day, all birds were slaughtered and samples from three segments of the small intestine were taken. Villi area, height, width and crypt depth ratios were read using Multiscan software. Synbiotic supplementation increased the BWG of broilers from first to tenth day of rearing, compared to the control group. The PRO group had improved villi morphometric parameters of the duodenum. In the jejunum and ileum, both bioactive substances improved villus width and villus surface area. Crypts were deeper in the small intestine of birds supplemented with bioactive substances, which allows greater renewal of the villi. As expected, the intestinal morphometric parameters of broiler chickens benefited from bioactive substance supplementation.     

Quality of Wine Grape Cultivars Grown in Czech Republic

This research aims to make an analysis of significant parameters in wine grape from Czech Republic. It was evaluated pH, content of sugar, ascorbic acid, titratable acidity, total phenolic content and texture properties. For analyzes 5 samples of wine grape were used, harvested during the three periods. The results show that during the ripening of wine grapes there were changes of selected qualitative parameters, particularly of total polyphenols, sugar and some varieties of wine grape also ascorbic acid (significantly in the variety ’Grüner Veltliner’, ’Chardonnay’, ’Pinot Blanc’). Pronounced changes qualitative parameters were among wine grapes harvested the 1st and 2nd seasons. Changes in textural properties also occurred during the harvest and for most varieties between 2nd and 3rd period of harvest. The PCA grouped chemical parameters of wine grape in independent sets and can be used as assessment tool of wine cultivars.