Comparison of RAPD, STS, ISSR and AFLP molecular methods used for assessment of genetic diversity in hop (Humulus lupulus L.)

CoxI genomic and cDNA sequences from gymnosperms and angiosperms were used to study the effects of RNA editing on gene evolution and phylogenyre construction. In six gymnosperms harboring edited coxI gene the number of nucleotide substitutions at 1^st, 2^nd and 3^rd codon positions was similar. In contrast, in angiosperms, the number of nucleotide substitutions at 1^st and 2^nd codon positions was much lower than at the 3^rd. The coxI gene in long-lived gymnosperms evolved much faster than in annual angiosperms. This accelerated rate of nucleotide substitution in gymnosperms is due to accumulation of T-C substitutions at edited sites that can randomly appear at all three codon positions. Editing predominantly occurred at 1^st and 2^nd codon positions as a result of selection against non synonymous T-C substitutions and other types of mutations. The tree topologies for the investigated species based on genomic DNA data were in concordance with their taxonomic positions. The trees based on polymorphic edited sites agreed with trees derived from complete sequence information. This indicates that edited sites are reliable sources of phylogenetic information especially for species that contain many edited sites. However, the fast evolution rate of coxI gene in gymnosperms has caused the long branches in the phylogenetic trees. The inclusion of the species with a processed paralog i.e., edited form of the coxI gene, affected the topology of phylogenetic trees, especially when the taxon with a processed paralog was closely related to the other species analyzed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Production potential,biodiversity and soil properties of forest reclamations: Opportunities or risk of introduced coniferous tree species under climate change?

European Journal of Forest Research - In the time of ongoing climate change and the increasing area of post-mining landscape, the successful afforestation of reclamation sites by suitable adaptive...     

Molecular typing of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma mali’ and epidemic history tracing by a combined T-RFLP/VNTR analysis approach

Apple proliferation caused by ‘Candidatus Phytoplasma mali’ is a disease of apple trees gaining increasing importance in Europe. The present study describes a high-throughput method for simultaneous typing of ‘Ca. P. mali’ at two genetic loci. This novel approach combines terminal restriction fragment length polymorphism (T-RFLP) analysis of a putative rhodanese-like protein gene and the analysis of the variable number of tandem repeats (VNTR) of the ribosomal protein L22 gene. The typing approach was applied to analyse a collection of DNA isolates from 310 apple trees tested positive for ‘Ca. P. mali’. Samples were taken between 2002 and 2010 in South Tyrol (Northern Italy). In addition, 15 samples of Cacopsylla melanoneura and 19 of C. picta were typed. Seven combined genetic profiles were found in the samples of infected apple trees: AT-2/rpX-A (81.0%), AT-1/rpX-D (8.4%), AT-1/rpX-E (4.2%), AT-1/rpX-A (3.2%), AT-1/rpX-B (1.6%), AT-1/rpX-C (1.0%) and AP/rpX-A (0.3%), and one mixed infection AP + AT-1/rpX-A + rpX-D (0.3%). Subtype rpX-E was discovered for the first time. In C. melanoneura samples the most frequent subtype was AT-1/rpX-E, followed by AT-1/rpX-D and AT-1/rpX-C. All C. picta samples displayed subtype AT-2/rpX-A. Analysis of the temporal distribution of subtype frequencies in apple trees revealed that exclusively subtype AT-1 in combination with four rpX subtypes was present in South Tyrol in the period from 2002 to 2004. From 2006 onwards subtype AT-2/rpX-A became dominant with an average frequency of 90%. The data obtained suggest that there may be a co-adaptation of particular ‘Ca. P. mali’ subtypes with different insect vector species.     

Seasonal colonisation of apple trees by ‘Candidatus Phytoplasma mali’ revealed by a new quantitative TaqMan real-time PCR approach

Apple proliferation (AP), caused by ‘Candidatus Phytoplasma mali’, is an economically important disease affecting many apple-growing areas in Europe. A new TaqMan real-time PCR assay was established for absolute quantification of ‘Ca. P. mali’ by using a single-copy gene of the host plant as a reference, which is amplified with the pathogen DNA in a single-tube reaction. Normalised estimates of phytoplasma concentration are ultimately expressed as the number of phytoplasma cells per host plant cell. The assay was used to monitor the ‘Ca. P. mali’ titre over the course of two growing seasons in roots and branches of symptomatic and asymptomatic but AP-positive apple trees. All 252 root samples from symptomatic and asymptomatic trees tested positive, with an average number of 59.8 ± 5.68 (standard error) and 55.1 ± 9.83 ‘Ca. P. mali’ per host cell, respectively. From the 378 shoot samples analysed, 81% of the symptomatic and only 20% of the asymptomatic samples were AP-positive with an average number of 9.4 ± 1.04 and 0.7 ± 0.13 ‘Ca. P. mali’ per host cell, respectively. This strengthens evidence that not the pathogen occurrence alone but the presence of a certain quantity of ‘Ca. P. mali’ in the aerial tree sections is involved in symptom expression. In addition, pronounced seasonality of the phytoplasma concentration was found, not only in branches, but also for the first time in roots of symptomatic and asymptomatic apple trees. Highest phytoplasma levels in roots were detected from December to May.     

Epidemiological enquiries in two Q fever outbreaks in a community of Baden-Württemberg during 2008 and 2009

In 2008 and 2009, two consecutive outbreaks of Q fever in humans were recorded in the district of Freudenstadt, northern Black Forrest, Baden-Württemberg, Germany. In 2008, a total of 41 persons from a single local community fell ill and were found infected with Coxiella burnetii. Although comprehensive diagnostic and epidemiological outbreak investigations were conducted and control measures taken which included vaccination of ruminants at risk in three parts of the affected community, re-occurrence of the disease in 2009 with further 29 confirmed human Q fever cases could not be prevented. While the origin of infection of the first outbreak was probably a flock of 550 sheep moved in the surrounding of the affected villages, the source of infection for the consecutive outbreak in 2009 could not be identified. It seems possible that meadows contaminated with infectious placenta or birth fluids represented the sources of infection.     

Nitrate deficiency reduces cadmium and nickel accumulation in chamomile plants

The effect of nitrogen (nitrate) deficiency (-N) on the accumulation of cadmium (Cd) and nickel (Ni) in chamomile ( Matricaria chamomilla ) plants was studied. Elimination of N from the culture medium led to decreases in N-based compounds (free amino acids and soluble proteins) and increases in C-based compounds (reducing sugars, soluble phenols, coumarins, phenolic acids, and partially flavonoids and lignin), being considerably affected by the metal presence. Proline, a known stress-protective amino acid, decreased in all -N variants. The activity of phenylalanine ammonia-lyase was stimulated only in -N control plants, whereas the activities of polyphenol oxidase and guaiacol peroxidase were never reduced in -N variants in comparison with respective +N counterparts. Among detected phenolic acids, chlorogenic acid strongly accumulated in all N-deficient variants in the free fraction and caffeic acid in the cell wall-bound fraction. Mineral nutrients were rather affected by a given metal than by N deficiency. Shoot and total root Cd and Ni amounts decreased in -N variants. On the contrary, ammonium-fed plants exposed to N deficiency did not show similar changes in Cd and Ni contents. The present findings are discussed with respect to the role of phenols and mineral nutrition in metal uptake.     

Copper Uptake, Physiology and Cytogenetic Characteristics in Three Matricaria chamomilla Cultivars

Selected physiological parameters and copper uptake by three chamomile (Matricaria chamomilla) cultivars (two tetraploid????Lutea?? and ??Unknown?? and one diploid????Novbona??) exposed to 60 ??M Cu over 7 days were studied. Genome size was 10.6 and 5.5 pg DNA/2C and chromosome number 2n?=?36 and 2n?=?18 in tetraploids and diploid, respectively. Root tissue water content and dry biomass were more reduced in diploid cultivar. Soluble proteins were depressed by Cu excess in all cultivars without respect to ploidy. Lignin accumulation and cinnamylalcohol dehydrogenase activity was the highest in Novbona. Phenylalanine ammonia-lyase activity was stimulated in tetraploid but reduced in diploid roots. Diploid contained higher amount of Cu and soluble phenols in both shoots and roots. Sulphur-containing amino acids were stimulated in Cu-exposed shoots independently on ploidy level. Potassium content was more reduced in the diploid cultivar. Our data indicate that induction of phenolic metabolism is faster in diploid cultivar which also contained higher amount of Cu. In this view, polyploidisation is a good tool for the reduction of shoot metal accumulation and growth tolerance to Cu excess. Identity of Czech tetraploid cultivar ??Unknown?? is also discussed.     

Inhibition and reactivation of chicken kynurenine formamidase: In vitro studies with organophosphates, N-alkylcarbamates,and phenylmethanesulfonyl fluoride

In studies on the mechanism of avian teratogenesis induced by organophosphorus and methylcarbamate insecticides, chicken liver kynurenine formamidase (KFase) was used as a convenient model for the more difficultly accessible but teratogenically relevant KFase of the yolk sac membrane. The parameters of in vitro KFase inhibition kinetics do not correlate with the teratogenic potency of various organophosphates and N-alkylcarbamates. The phosphorylated, carbamoylated, and sulfonylated enzymes are relatively resistant to reactivation of enzyme activity either spontaneously or by aldoximes or hydroxylamine. This refractory property may contribute to the prolonged in ovo KFase inhibition necessary for the NAD-deficiency type of teratogenesis.     

Field efficacy of phoxim 50% (ByeMite) against the poultry red mite Dermanyssus gallinae in battery cages stocked with laying hens

Infestations with the poultry red mite Dermanyssus gallinae represent a major ectoparasite problem in poultry and can affect egg layers worldwide. There is presently a lack of an ectoparasiticide in Europe for poultry which can assure a 0-day withholding period for eggs. In this study, ByeMite (phoxim 50%, Bayer HealthCare, Animal Health Division) was administered to treat a D. gallinae infestation in a poultry house stocked with egg-laying hens kept in a cage system. A layer house was sprayed twice within a 7-day interval using a solution containing 2000 ppm phoxim and a similar layer house was used as an untreated control unit. Specially developed D. gallinae traps made of cardboard were used to assess the mite density in both layer houses during a 49-day period after the treatment. In order to collect mites, the traps were placed on days--1, 2, 6, 9, 13, 20, 34 and 48 and always removed after 24 h. The collected mites were counted and differentiated according to their developmental stage (mite eggs, larvae, nymphs, adults). Three days after the first spray treatment, the efficacy against all mite stages (larvae, nymphs, adults) was 96.1%, and from day 7 post-treatment until the end of the trial (day 49) the efficacy exceeded 99%. In contrast, in the untreated layer house (negative control group) the mite population showed a 400% increase. No treatment-related side effects in chickens were detectable. It is concluded that two administrations of ByeMite within a 7-day interval are highly effective against D. gallinae infestations in a stocked poultry house.