RNA extraction involves several main stages, regardless of the method of extraction: homogenization, effective denaturation of proteins from RNA, inactivation of ribonuclease and removal of any DNA, protein, and some residual contamination. Isolation of undamaged intact RNA is challenging when the related tissue contains high levels of polysaccharides and phenols. Several efforts have been made towards the comparison and optimization of extraction and purification methods for RNA from plant tissues. This is dictated by the necessity of obtaining RNA of a good quality and in a sufficient quantity for further molecular analyzes. Plant storage organs (such as bulbs or seeds) rich in polysaccharide and polyphenolic compounds present distinct challenges for total RNA isolation. Such components, considered in this case as contamination, may bind and co-precipitate with nucleic acids and negatively affect later assays. Since standard routine protocols yield unacceptable results in bulbs, we have designed a new method for RNA extraction. We used two modified procedures (based on CTAB and sarkosyl reagents) of RNA extraction from so called “difficult plant material” and compared them to a popular RNA isolation base on the column isolation kit and TriPure reagent. Our modified protocols dealt with problems of both RNA degradation and low yield caused by co-purification with polysaccharides present in plant bulbs. In this study we have shown that improvement of the CTAB and sarkosyl method with a lyophilization step of plant tissues leads to isolation of high quality RNA from difficult material like storage organs of bulbous plants. The main changes in the procedure compared to the previously described methods concerned the different order of lithium chloride and sodium acetate addition, lithium chloride concentration increase and modification of centrifugation conditions. Gel electrophoresis and spectrophotometer analysis confirmed the high quality and integrity of the obtained RNA. The modified procedures allowed for obtaining a satisfying amount of RNA concentration in the range from 280 to 950 ng/μl depending on the plant species. Thus, the demonstrated RNA isolation methods are efficient and can be used for plant material rich in polysaccharides, such as bulbs.

The aim of the research conducted in a 2-year pot experiment in an unheated plastic tunnel was to determine suitability of Miscanthus × giganteus for phytoextraction of nickel from soil as well as to assess tolerance of this species on increasing concentrations of this metal in soil. Pots were filled with mineral soil (sand) and a mixture of soil with high-moor peat and three levels of nickel were introduced, i.e. 75 mg dm-3, 150 mg dm-3 and 600 mg dm-3 and the control combinations used substrates without the addition of nickel. Nickel was introduced only in the first year of the experiment in the form of nickel sulfate (NiSO4 · 6H2O). Miscanthus × giganteus accumulated a considerable amount of nickel in biomass. Miscanthus × giganteus growing in contaminated mineral soil turned out to be a species tolerant to high nickel concentrations

The phytoextraction is a process that uses living plants for cleaning up the heavy metals from contaminated soil. The cadmium and lead contamination of soils results from the application of sludge or urban composts, fertilizers, pesticides, motorization, metallurgy, and different technological processes. In industrial terrain the content of cadmium and lead in soils has increased in the recent years. This study was undertaken to evaluate the potential of Amaranthus caudatus L. ‘Atropurpureus’ and Ricinus communis L. ‘Sanguineus Apache’ for phytoextraction of cadmium and lead. Two species of ornament plants, i.e. Amaranthus caudatus L. ‘Atropurpureus’ and Ricinus communis L. ‘Sanguineus Apache’, were planted in drainless containers in a substrate artificially polluted with cadmium and lead in order to evaluate their suitability for phytoremediation of soils or substrates contaminated with these metals. Cadmium was applied at increasing rates of 0, 1, 5 and 10 mg Cd∙dm-3 in the form of cadmium sulfate 3CdSO4∙8H2O, while lead was used at 0, 100, 500 and 1000 mg Pb∙dm-3 in the form of lead acetate (CH3COO)2Pb∙3H2O. The applied doses of cadmium and lead in the experiment reflected different degrees of soil pollution. After five months of growth it was found that Amaranthus caudatus L. accumulated the biggest concentrations of cadmium and lead in leaves and the lowest concentrations in inflorescences. Ricinus communis L. accumulated the highest concentrations of cadmium in stems, while the lowest concentrations in inflorescences, whereas the biggest concentration of lead was accumulated in inflorescences and the least lead was accumulated in leaves. The biggest reduction of cadmium and lead concentrations after the completion of the experiment was found in substrates, in which Amaranthus caudatus L. was grown. The tested species of ornamental plants may be used in the phytoextraction of cadmium and lead from soils contaminated.

Chrysanthemum stunt viroid (CSVd) is a serious pathogen infecting chrysanthemum worldwide. To improve and enhance the detection procedure, a colorimetric loop-mediated isothermal amplification (LAMP) technique was developed. Six LAMP primers were newly designed and tested to determine the optimal conditions using a recombinant plasmid of CSVd as a DNA template. The optimal conditions for colorimetric LAMP were incubation at 65°C for 45 min. Under these conditions, a ladder-like pattern of LAMP products was detected along with a change of color from pink to yellow in the positive reactions. Limits of the detection (LOD) of colorimetric LAMP were up to 1 fg ∙ μl–1 of plasmid DNA concentration which was 104 times greater than that of polymerase chain reaction (PCR). The developed colorimetric LAMP was not cross reacted to other viruses and viroids. From detection of actual samples and chrysanthemum plantlets which were obtained from meristem tip culture, the colorimetric LAMP showed effective potential in detecting CSVd. Therefore, the colorimetric LAMP can be used as a main technique to detect CSVd and ensure CSVd-free chrysanthemum plantlet production due to its accuracy, rapidness and sensitivity.

The aim of presented investigation was to determine the composition of scale insects species and intensity of their occurrence on some greenhouse’s ornamental plants. The investigations were carried out in the greenhouse of Maria Curie Skłodowska Botanical Garden in Lublin in years 2002–2004. Eight species belonging to seven botanical families were observed: Abutilon striatum cv. Thomsoni, Cyrtomium falcatum Presl., Dizygotheca elegantissima (Veitch), Hedera helix L., Hypoestes phyllostachya Presl., Nerium oleander L., Passiflora guadrangularia L., Ruscus aculeatus L. The quantitative analysis of the studied material was performed making use of the following ecological indicators: number and density. Identification of the scale insects species was performed on the basis of microscope slides. Three species of the scale insects belonging to three families were observed on ornamental plants: Pseudococcidae [Pseudococcus maritimus (Ehrh.)], Coccidae [Saissetia coffeae (Walker)] and Diaspididae (Aspidiotus nerii Bouchè). The scale insects were noted on all species of studied plants. Observed scale insects are typical polyphagous and all of them are considered as harmful pests in greenhouses. Among scale insects inhabiting this group of plants distinctly numerous on particular host plants were S. coffea and A. nerii. On the studied plants scale insects were stated at four degree of density.