Botanical pesticides and their human health safety on the example of Citrus sinensis essential oil and Oulema melanopus under laboratory conditions

Biopesticides derived from plants have recently received increasing interest as an alternative to conventional chemicals suitable for integrated pest management and organic farming. In this study, Citrus sinensis (Osbeck) essential oil was tested for its potential for wheat protection against Oulema melanopus (L.). The chemical composition of the essential oil was analyzed by gas chromatography-mass spectrometry (GC-MS). Twenty-four compounds were identified in the oil among which d-limonene was a predominant component, followed by myrcene, α-pinene, linalool, sabinene, decanal, and other minor components. Direct contact toxicity assay showed the effectiveness of the essential oil against O. melanopus larvae causing the mortality of 85% during 48 hours. The persistence study revealed high dissipation rate of the oil from treated plants, whereas concentrations lower than 0.01 g/kg were detected by GC-MS 5 min after the application of the oil, suggesting its environmental and food safety. These results, together with previous literature reports reviewed in this study, indicate the great potential of the C. sinensis essential oil for future use in crop protection against insect pests.     

Invasive Chinese pond mussel Sinanodonta woodiana threatens native mussel reproduction by inducing cross‐resistance of host fish

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Development of alder carr after the abandonment of wet grasslands during the last 70 years

? The secondary succession of wet grasslands to communities of alder carr dominated by Alnus glutinosa was recorded in different parts of Europe during the 20th century. However, knowledge of such development of alder carr remains insufficient.

? The development of alder carr was reconstructed at five sites in the Czech Republic, using historical aerial photographs and methods of dendrochronology. The aims were to investigate the succession from wet grasslands to alder carr at sites previously intensively managed for agriculture and to find out the role of fluctuations in the groundwater table, caused by artificial drainage channels, in the observed stand dynamics and tree growth.

? The spread of forest (i.e., an increase in forest cover) predominated until the 1970s at all sites. This trend was disrupted by a large-scale dieback of forest stands in four of the five sites after the 1970s, followed by an increase in patch heterogeneity, as indicated by landscape metrics. The radial growth increment in Alnus glutinosa has been affected predominately by local environmental factors, probably including the changing degree of waterlogging. Forest dieback was presumably connected with a lesser extent of drainage channels.

? Our results indicate that observed successional pathways at sites of alder carr were probably caused by local changes in the groundwater table.

     

Distribution,hybridisation and morphological variation in Alnus rohlenae (Betulaceae) an endemic species of the Balkan Peninsula

The diploid A. glutinosa (L.) Gaertn. is widespread throughout the European continent, except in the Iberian and Balkan Peninsulas where tetraploid populations have been discovered. We focused on the tetraploid species described as A. rohlenae Vít, Douda and Mandák that occupies the western part of the Balkan Peninsula, where it has likely completely replaced the diploid species. While the distribution range of the diploid A. glutinosa s. str. is well known, the exact distribution range of the tetraploid A. rohlenae is unknown. Here, we report the first exact distribution of the tetraploid A. rohlenae and the anticipated hybrid zones in which it is in contact with diploid populations using flow cytometry and morphometrics. Tetraploids are located primarily in the mountainous parts of the study area and towards the lowlands are gradually being replaced by diploids, forming a contact zone. We compare the main morphological characteristics of both species. Due to the geographical proximity of the study species, the morphological differences between them are clear outside the contact zones. However, within the contact zones, we recorded hybridisations that obscure the morphological differences between species, probably due to the presence of triploid hybrids.

     

Using plant essences as alternative mean for northern root-knot nematode (<Emphasis Type="Italic">Meloidogyne hapla</Emphasis>) management

The northern root-knot nematode (Meloidogyne hapla) is a prominent root vegetable pest in the Czech Republic. Effects of several plant essences for M. hapla management on carrot (Daucus carota) were tested. The tested compounds included commercially available plant essences from Ocimum basilicum (sweet basil), Mentha arvensis (field mint), Tagetes erecta (Mexican marigold), and commercial product of neem seed (Azadirachta indica). Two further essences from Eugenia caryophyllata (clove) and Origanum majorana (marjoram) were obtained by distillation. Tests were conducted on container-grown carrots maintained under field conditions. The delivery method involved absorbing the tested essences in perlite particles placed in a substrate. The results showed a negative correlation between gall numbers of the carrots and treatment concentrations, especially for clove and majoram. Similar results were achieved when considering M. hapla egg numbers. Furthermore, the same treatments showed significant differences (P ≥ 0.05) in fresh and dry root weights. Based on these results the management of M. hapla using plant essences seems to be possible, however, further research is desirable to determine the appropriate dosage needed for efficient and most appropriate delivery method of these substances under field conditions.     

The relationship between water quality and indigenous and alien crayfish distribution in the Czech Republic: patterns and conservation implications

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Early responses of in vitro cultured fish‐growing glochidia: The effects of taurine,lipids and sera on Margaritifera margaritifera

In vitro culture has great potential for the propagation of freshwater mussels in both commercial and conservation aquaculture. The use of in vitro techniques precludes the need for host fish, thus decreasing costs and increasing efficiency. However, protocols are still lacking for many species that grow substantially during the parasitic stage. In this study, we tested the effects of taurine addition, serum type and source of lipids on the survival rate and increase in length of Margaritifera margaritifera larvae during the initial stage of culture (first 11 days of exposure to media). Our results show that taurine has no significant effect on the early survival rate of glochidia; however, the possible importance of this amino acid in subsequent stages is discussed. The use of an emulsified lipid mixture instead of traditional fish oil showed significantly higher rates of survival. Finally, the addition of serum showed variable effects, with both horse serum and newborn calf serum having higher survival in trials using mussel populations from different sources. These findings can contribute to the invention or improvement of in vitro protocols for species growing during infection and at the same time show the potential that early survival assessments could have for the development of in vitro methods in species with long parasitic stages.     

Comparative molecular analysis of Bursaphelenchus vallesianus,a wood‐inhabiting nematode isolated from declining pine trees in the Czech Republic

The Bursaphelenchus genus (Nematoda: Parasitaphelenchidae) comprises mostly wood‐inhabiting nematodes that feed on various tree‐colonizing fungi. One species of the genus, B. xylophilus, has been proven as an agent causing pine wilt disease (PWD). However, involvement of other Bursaphelenchus species in the PWD remains enigmatic. In the current paper, comparative molecular analysis is performed based on nuclear ribosomal DNA (rDNA) of B. vallesianus, a species that was recently isolated from pine trees (Pinus sylvestris) exhibiting wilting and declining symptoms in the Czech Republic. Sequencing of the nuclear‐encoded ITS1–5·8S–ITS2 rDNA region confirmed previous taxonomic conclusions based on morphology. Evolutionary reconstructions resulted in a phylogenetic tree, where the Czech isolate of B. vallesianus occupied a common clade together with other species belonging to the so‐called B. sexdentati group. Unexpectedly, comprehensive analysis of the sequence data revealed a genetic variation distinguishing the Czech isolate of B. vallesianus from all other species of the B. sexdentati group. This dissimilarity consists of the presence of a four nucleotide exchange found in the 5·8S rRNA‐coding gene. The newly identified genetic variation appears to affect the 5·8S rRNA folding, as deduced from secondary structure models. Additionally, it is shown that for the first time, to the authors’ knowledge, both bursaphelenchid internal transcribed spacers (ITS1 and ITS2) fold into the multibranched closed loops. While the ITS2 closed loop is formed with help of canonical 5·8S‐28S rRNA pairing, the ITS1 forms the thermodynamically stable closed loop with no support of flanking rRNA sequences. The current information on bursaphelenchid ITS rDNA sequence diversity and structure is further discussed.