Agnieszka Pietrosiuk

Antimitotic effect, G2/M accumulation, chromosomal and ultrastructure changes in meristematic cells of Allium cepa L. root tips treated with the extract from Rhodiola rosea roots

Abstract Rhodiola rosea is a Russian and Chinese medicinal plant used in medical practice as a stimulating, adaptogenic and antiarrhythmic agent. Many investigations have pointed to the anticancer and antimutagenic role of R. rosea. In spite of these therapeutic effects of R. rosea extract. little is known about the mechanism of its anticancer action, because few cytological investigations havebeen conducted on the mitotic activity, cellular cycle, structure, ultrastructure, and cytophysiology of both animal and plant cells treated with the extract. Our studies have shown that extract from R. rosea roots (containing rosavine and cinnamyl alcohol) cause the inhibition of a mitotic activity; however, cells resume their divisions after the postincubation period. During incubation in the extract, accumulation of G2/M cells was observed. Chromosomes inprophase and metaphase were shorter and thicker but the extract did not cause chromosomal aberrations or the formation of micronuclei. Ultrastructure changes after 24 h of incubation were connected with an increase in the electron density of ground cytoplasm, decrease in the number of mitochondria but with an increase of their surface. As early as after 6h postincubation, the number ofmitochondria doubled but their surface declined by over two times. After 24 h of incubation, the surface of plastids increased over twice in spite of their unchanged number. In postincubation (6h), their number doubled and the surface declined dramatically. The ultrastructure of the mitochondria was changed by the disappearance of cristae and a lowered electron density of the matrix, suggestingthat one of the mechanisms could be connected with changes in mitochondrial function.

Enhancement of taxane production in hairy root culture of Taxus x media var. Hicksii.

This study assessed the effect of two precursors (l-phenylalanine and p-amino benzoic acid) used alone or in combination with methyl jasmonate, on the growth and accumulation of paclitaxel, baccatin III and 10-deacetylbaccatin III in hairy root cultures of Taxus x media var. Hicksii. The greatest increase in dry biomass was observed after 4 weeks of culturing hairy roots in medium supplemented with 1microM of l-phenylalanine (6.2gL(-1)). Addition of 1microM of l-phenylalanine to the medium also resulted in the greatest 10-deacetylbaccatin III accumulation (422.7microg L(-1)), which was not detected in the untreated control culture. Supplementation with 100microM of l-phenylalanine together with 100microM of methyl jasmonate resulted in the enhancement of paclitaxel production from 40.3microg L(-1) (control untreated culture) to 568.2microg L(-1), the highest paclitaxel content detected in the study. The effect of p-amino benzoic acid on taxane production was less pronounced, and the highest yield of paclitaxel (221.8microg L(-1)) was observed when the medium was supplemented with 100microM of the precursor in combination with methyl jasmonate. Baccatin III was not detected under the conditions used in this experiment and the investigated taxanes were not excreted into the medium.

Biotechnological approaches to enhance salidroside, rosin and its derivatives production in selected Rhodiola spp. in vitro cultures

Rhodiola (Crassulaceae) an arctic-alpine plant, is extensively used in traditional folk medicine in Asian and European countries. A number of investigations have demonstrated that Rhodiola preparations exhibit adaptogenic, neuroprotective, anti-tumour, cardioprotective, and anti-depressant effects. The main compounds responsible for these activities are believed to be salidroside, rosin and its derivatives which became the target of biotechnological investigations. This review summarizes the results of the diverse biotechnological approaches undertaken to enhance the production of salidroside, rosin and its derivatives in callus, cell suspension and organ in vitro cultures of selected Rhodiola species.

Pyrrolizidine Alkaloids from Lithospermum canescens Lehm.

Seven pyrrolizidine alkaloids (PAs) have been isolated from Lithospermum canescens and their structures determined by spectroscopical methods. Besides the known lycopsamine, O7- acetyl-lycopsamine and O7-acetylintermedine four new PAs were found. Their structures are O7-(3-hydroxy-3-methyl-butanoyl)-O9-(+)-trachelanthoyl-heliotridine (= O7-(3-hydroxy-3- methyl-butanoyl)-rinderine = canescine), O7-(3-hydroxy-3-methyl-butanoyl)-O9-(D)-viridifloryl- heliotridine (= O7-(3-hydroxy-3-methyl-butanoyl)-echinatine = canescenine) and their O13-acetyl-derivatives (= acetylcanescine; acetylcanescenine).

Approaches of Rhodiola kirilowii and Rhodiola rosea field cultivation in Poland and their potential health benefits.

Numerous researches have been carried out on plants of the Rhodiola species, especially Rhodiola kirilowii (Regel) Maxim. and Rhodiola rosea. Various compounds have been reported to be isolated from R. kirilowii and R. rosea, including cyanogenic glycosides, monoterpene alcohols and their glycosides, aryl glycosides, phenylethanoids, phenylpropanoids and their glycosides (salidroside and rosavins respectively), as well as flavonoids, flavonlignans, proanthocyanidins and gallic acid derivatives and the latter have free radical scavenging capacity. The benefits claimed for Rhodiola include adapogenic, neuroprotective, anti-depresive anti-tumour and cardioprotective activities. Currently, the adaptogenic activity of Rhodiola compounds are properties evaluated mainly in human clinical trials. The mechanism of the action of Rhodiola extracts include affecting the levels of cortisol and NO by interactions with glucocorticoid receptors directly or via the c-Jun N-terminal protein kinase (JNK) pathway. However, the natural populations of R. rosea in Poland are threatened; therefore, the cultivation of R. rosea and alternative species R. kirilowii might be a possible solution for producing these kinds of plants in Poland in sufficient quantities and quality for pharmaceutical purposes. Lack of proven interaction with other drugs and no confirmed adverse effects during clinical trials encourages further investigation. These herb preparations ought to be studied extensively to establish their position as potential drugs for a variety of diseases.

The chemical profile of pyrrolizidine alkaloids from selected greek endemic boraginaceae plants determined by gas chromatography/mass spectrometry.

Four Greek endemic Boraginaceae plants, Onosma erecta Sibth. & Sm., Onosma kaheirei Teppner, Onosma leptantha Heldr., and Cynoglossum columnae L. (aerial parts), were screened for their content of pyrrolizidine alkaloids (PAs). TLC with the Mattocks-Molyneux visualization reagent was used as a preliminary qualitative test for PA or PA N-oxide detection. The extracts of the species found to contain PAs and their N-oxides were further analyzed by GC/MS, so as to identify their structures by means of the mass spectra and retention index values of known PAs already published in the literature. Twenty-three PAs were identified. For additional peaks, recognized as possible PAs by their MS pattern, no exact structures were tentatively suggested, as a result of lack of matching literature data. Furthermore, a quantitative PA profile of the species was obtained.

Hairy root cultures for the production of anti-cancer naphthoquinone compounds

BACKGROUND Recent years have brought the dynamic development in studies of naphthoquinones obtained from plants, in vitro cultures and semi- or total synthesis. This review presents the hairy root cultures approach for producing naphthoquinones and summarizes their most recent anti-cancer investigations. OBJECTIVE This review aimed to define biotechnological strategies impacted on naphthoquinones production in hairy root cultures. Up to now the major source of shikonin/alkannin derivatives, rhinacanthins and ramentaceone is isolation from plant material, also derived via biotechnological methods. Moreover, the most recent anti-cancer activity studies on naphthoquinones which could be produced in hairy root cultures were outlined. METHODS For databases survey two selection criteria were used: (i) naphthoquinone could be produced in hairy roots, and (ii) it exhibits anti-cancer properties. RESULTS Ninety two papers were included in the review, thirty described biotechnological approaches enhancing naphthoquinones production, among them twenty seven were dedicated to hairy root cultures. Forty papers outlined the anti-cancer activity of targeted naphthoquinones including the type of cancer and bioassays description. The synergistic effect of natural naphthoquinones and other anti-cancer therapies was reviewed and toxicity of natural naphthoquinones and plant extracts was discussed. The review highlights tendencies in hairy root investigations and indicates the possible future research directions for improving biotechnological production efficacy. CONCLUSION This review demonstrates a great potential of hairy root cultures for naphthoquinones production, which could be furtherly developed for future medical purposes, especially as anti-cancer agents. This area of plant biotechnology will be surely still developed with traditional and new strategies.