The effects of different types of management,functions, and characteristics of stands in Polish forests on the amount of coarse woody debris

In the modern forestry paradigm, many factors influence the amount of coarse woody debris (CWD). The present paper analyzes the effects of both local (national) programs (special functions of forests) and European programs (Natura 2000 sites), as well as the individual characteristics of forest stands. The study was conducted on 2,752 sampling plots distributed over an area of about 17,500 ha and located in lowland stands having a species composition typical of large areas in central Europe. Natura 2000 areas contained significantly more CWD (8.4 m³/ha) than areas not covered by the program (4.8 m³/ha). However, this is due to the fact that Natura 2000 sites involve well-preserved forest areas, such as nature reserves (26.6 m³/ha). In the managed forests that have been covered by the Natura 2000 program over the past several years, the volume of CWD has not increased. Forests with ecological and social functions differed slightly in the amount of CWD. More CWD occurred in protected animal areas (8.7 m³/ha) than in stands damaged by industry (3.9 m³/ha). Intermediate CWD levels were found in water-protection forests and in forests located around cities and military facilities. In managed forests, the lowest CWD volume was observed in middle-aged stands. The species composition of the stand had little effect on the volume of CWD. Only stands with a predominance of ash and alder had higher CWD levels (13.5 m³/ha). More CWD was found in stands whose species composition did not represent the potential site quality (6.4 m³/ha) than in habitats with the optimum species composition (3.8 m³/ha). CWD volume should be systematically increased taking into consideration local natural conditions. Such efforts should be focused on particularly valuable regions, and especially on Natura 2000 sites, where the threshold values reported from other European forests should be reached. Leaving some trees to die naturally and retaining reasonable amounts of such trees ought to be incorporated into CWD management practice in Poland.     

Response of a Juvenile Cyprinid,Lake Minnow <Emphasis Type="Italic">Eupallasella perenurus</Emphasis> (Pallas), to Different Diets

Three commercial starters (Carp Starter, Uni Starter and Perla Plus) and one non-commercial, with frozen Chironomidae larvae as a reference diet, were evaluated for the intensive rearing of juvenile lake minnow Eupallasella perenurus, a cyprinid fish that is critically endangered in Poland. The growth, condition, survival, body deformities, and chemical body composition were studied. The 90-day laboratory experiment was performed at 22 °C with fish that were initially 24.6 mm (mean total lenth (TL)) and 0.11 g (mean body weight (BW)). Satisfactory fish growth was attained with all of the diets; however, the largest (p ≤ 0.05) final size (48.5 mm TL, 1.55 g BW) and the lowest condition coefficient (K = 1.34) were noted in fish fed the non-commercial starter. The final survival rates were very high (97.5–100%). Skeletal deformities (in 74 to 92% fish) were recorded exclusively in fish fed commercial starters. All commercial starters resulted in considerably higher lipid content and lower ash content than did the non-commercial starter and the reference diet. This suggests that both these factors might be responsible for body deformities. The present results proved that only the non-commercial starter is suitable for juvenile E. perenurus rearing under controlled condition, and that none of the commercial starters can be recommended.     

The involvement of the anterior cingulate cortex in remote contextual fear memory

Although the molecular, cellular, and systems mechanisms required for initial memory processing have been intensively investigated, those underlying permanent memory storage remain elusive. We present neuroanatomical, pharmacological, and genetic results demonstrating that the anterior cingulate cortex plays a critical role in remote memory for contextual fear conditioning. Imaging of activity-dependent genes shows that the anterior cingulate is activated by remote memory and that this activation is impaired by a null alpha-CaMKII mutation that blocks remote memory. Accordingly, reversible inactivation of this structure in normal mice disrupts remote memory without affecting recent memory.     

Diversity of ectomycorrhizal fungus assemblages on nursery grown European beech seedlings

? Context

An important feature of forest nursery management is the production of first-rate planting stock, which is closely connected with an adequate development of ectomycorrhizas on seedling roots.

? Aims

The aim of this study was to provide a detailed assessment of the ectomycorrhizal (ECM) fungal diversity of European beech grown in bare-root forest nursery conditions.

? Methods

Morphological and molecular analyzes were used to identify ECM fungi, quantify fungal richness, and record differences in the relative abundance of individual taxa.

? Results

Twenty-seven different mycorrhizal fungal taxa were found in four tested nurseries. Of these 27, only 6 were present in more than one nursery. Taxa richness in individual nurseries ranged from 6 to 13, indicating that ECM colonization potential is nursery-dependent. The most frequent taxa were: Tuber sp. 1, Cadophora sp., Cenococcum geophilum agg., Hebeloma sacchariolens, and Laccaria tortilis. According to the analysis of similarity, ECM fungal communities were significantly different between nurseries (R?=?0.7741; p?<?0.0001).

? Conclusion

Beech seedlings grown in bare-root nurseries are colonized by a highly diverse suite of ECM fungi; thus, artificial inoculation of seedlings is generally not necessary in nursery practice.     

A simple and efficient CRISPR/Cas9 platform for induction of single and multiple,heritable mutations in barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.)

Background

Genome editing of monocot plants can be accomplished by using the components of the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/CRISPR associated Cas9) technology specifically optimized for these types of plants. Here, we present the development of RNA-guided Cas9 system for simplex and multiplex genome editing in barley.

Results

We developed a set of customizable RNA-guided Cas9 binary vectors and sgRNA modules for simplex and multiplex editing in barley. To facilitate the design of RNA-guided Cas9 constructs, the pBract derived binary vectors were adapted to Gateway cloning and only one restriction enzyme was required for construction of the sgRNA. We designed a synthetic, codon optimized Cas9 gene containing the N terminal SV40 nuclear localization signal and the UBQ10 Arabidopsis 1st intron. Two different sgRNAs were constructed for simplex editing and one polycistronic tRNA-gRNA construct (PTG) for multiplex editing using an endogenous tRNA processing system. The RNA-guided Cas9 constructs were validated in transgenic barley plants produced by Agrobacterium-mediated transformation. The highest mutation rate was observed in simplex editing of the cytokinin oxidase/dehydrogenase HvCKX1 gene, where mutations at the hvckx1 locus were detected in 88% of the screened T₀ plants. We also proved the efficacy of the PTG construct in the multiplex editing of two CKX genes by obtaining 9 plants (21% of all edited plants) with mutations induced in both HvCKX1 and HvCKX3. Analysis of the T₁ lines revealed that mutations in the HvCKX1 gene were transmitted to the next generation of plants. Among 220 screened T₁ plants we identified 85 heterozygous and 28 homozygous mutants, most of them bearing frameshift mutations in the HvCKX1 gene. We also observed independent segregation of mutations and the Cas9-sgRNA T-DNA insert in several T₁ plants. Moreover, the knockout mutations of the Nud gene generated phenotype mutants with naked grains, and the phenotypic changes were identifiable in T₀ plants.

Conclusions

We demonstrated the effectiveness of an optimized RNA-guided Cas9 system that can be used for generating homozygous knockout mutants in the progeny of transgenic barely plants. This is also the first report of successful multiplex editing in barley using a tRNA processing system.

     

Deadwood volume in strictly protected,natural, and primeval forests in Poland

Standing and downed deadwood at different stages of decay provides a crucial habitat for a wide range of organisms. It is particularly abundant in unmanaged forests, such as strictly protected areas of national parks and nature reserves. The present work used the available data for such sites in Poland, analyzing a total of 113 studies concerning 79 sites to determine the causes contributing to variation in deadwood volume based on the duration of conservation, changes in deadwood volume over time (for those sites which were examined multiple times), elevation above sea level, forest type, stage of forest development, input of dead trees from the years preceding deadwood measurements, live tree volume, and the proportion of downed to standing deadwood). Depending on species composition and site altitude, most tree stands fell into one of four categories: subalpine spruce forests, montane beech-fir forests, low altitude beech-fir forests, or oak-hornbeam and riparian forests. The mean deadwood volume for all forest types amounted to 172.0 m³/ha. The mean volume of deadwood in montane beech-fir forests (223.9 m³/ha) was statistically significantly greater than in the other three forest types, for which it ranged from 103.5 to 142.5 m³/ha. A direct effect of the duration of conservation on deadwood volume was not identified. Nevertheless, analysis of repeated measurements on the same sample plots at 10-year intervals showed a consistent rise in mean deadwood volume. A linear regression model for all the analyzed factors reported from montane beech-fir forests and subalpine spruce forests showed that in addition to site altitude, another statistically significant variable was the input of dead trees (R²?=?63.54%).     

Comparative scanning electron microscope analysis of the enamel of permanent human,bovine and porcine teeth

BackgroundBovine and porcine teeth are often used in in vitro experiments as substitutes of human teeth.ObjectivesThe aim of the present study was to perform a comparative analysis of enamel morphology of permanent human, bovine and porcine teeth under the scanning electron microscope.MethodsAs many as 10 human, 10 bovine, and 10 porcine teeth were studied. All the teeth were sectioned and the halves were randomly divided into 2 groups according to the examined tissue (vestibular enamel at the mid-height of the dental crown and in the cervical area). Human and bovine enamel was etched for 15 sec and porcine enamel for 30 sec. The scanning electron microscope analysis was performed. The length and width of enamel prisms were determined with the “Met-Ilo” 1.1 computer program.ResultsAll enamel samples revealed the same etching pattern—Silverstone''s type 2. Bovine enamel showed a similar porosity and the amount of interprismatic enamel compared to human enamel while the amount and width of interprismatic enamel bands in porcine enamel were evidently greater. The shape of the porcine prisms was visually similar to human prisms, although dimensions were significantly different. However, bovine prisms differed in form and appeared to be distinctly elongated.ConclusionsReported findings indicate that the results of experimental studies carried out on bovine and porcine enamel should not be compared with the results obtained on human enamel.     

GLP1 and GIP are involved in the action of synbiotics in broiler chickens

Background: In order to discover new strategies to replace antibiotics in the post-antibiotic era in meat-type chicken production, two new synbiotics were tested:(Lactobacillus salivarius IBB3154 plus galactooligosaccharide(Syn1) and Lactobacillus plantarum IBB3036 plus raffinose family oligosaccharides(Syn2).Methods: The synbiotics were administered via syringe, using a special automatic system, into the egg air chamber of Cobb 500 broiler chicks on the 12 thday of egg incubation(2 mg of prebiotics + 105 cfu bacteria per egg).Hatched roosters(total 2,400) were reared on an experimental farm, kept in pens(75 animals per pen), with free access to feed and water. After 42 d animals were slaughtered. Blood serum, pancreas, duodenum and duodenum content were collected.Results: Syn2 increased trypsin activity by 2.5-fold in the pancreas and 1.5-fold in the duodenal content. In the duodenum content, Syn2 resulted in ca 30% elevation in lipase activity and 70% reduction in amylase activity.Syn1 and Syn2 strongly decreased expression of m RNA for GLP-1 and GIP in the duodenum and for GLP-1 receptors in the pancreas. Simultaneously, concentrations of the incretins significantly diminished in the blood serum(P 0.05). The decreased expression of incretins coincides with changed activity of digestive enzymes in the pancreas and in the duodenal content. The results indicate that incretins are involved in the action of Syn1 and Syn2 or that they may even be their target. No changes were observed in key hormones regulating metabolism(insulin, glucagon, corticosterone, thyroid hormones, and leptin) or in metabolic indices(glucose,NEFA, triglycerides, cholesterol). Additionally, synbiotics did not cause significant changes in the activities of alanine and aspartate aminotransferases in broiler chickens. Simultaneously, the activity of alkaline phosphatase and gamma glutamyl transferase diminished after Syn2 and Syn1, respectively.Conclusion: The selected synbiotics may be used as in ovo additives for broiler chickens, and Syn2 seems to improve their potential digestive proteolytic and lipolytic ability. Our results suggest that synbiotics can be directly or indirectly involved in incretin secretion and reception.     

2-Trichloromethylbenzimidazole,a new selective chromogenic reagent for detection of some heteroaromatic pesticides on thin-layer chromatographic plates

A highly selective method for the detection of several pesticides containing a heteroaromatic ring in the molecule (e.g. nicotine, pyrifenox or prochloraz) on thin-layer chromatographic (TLC) plates is described. The necessary detection condition is the presence in the ring of at least one tertiary nitrogen atom without substitutents on the two adjacent carbon atoms and, in the case of azoles, the protection of a secondary NH group by an alkyl or acyl group. The detection is realized by spraying the developed chromatogram with an acetone solution of 2-trichloromethylbenzimidazole and subsequently heating at 100-150°C in a drier. The detection limit is 20 ng to 10 μg depending on the detected compound. Some pesticide formulations may be analyzed using this method.