Comparative biochemical analysis of lectin and nuclease from Chelidonium majus L

It has been recently recognized that lectins exhibit other activities besides hemagglutination. Previously we have found that purified lectin from Chelidonium majus showed DNase activity (Fik, Go?dzicka-Józefiak & Kedzia, 1995, Herba Polon. 41, 84-95). Comparison of lectin and DNase from the sap from leaves and roots of Chelidonium majus proved that both these compounds are composed of 24 kDa monomer subunits which have an identical N-terminal sequence but differ in amino-acid composition and degree of glycosylation. Possible interrelationship between lectin and DNase is discussed.     

Propagation of Chelidonium majus L. by Somatic Embryogenesis

Direct somatic embryogenesis in celandine (Chelidonium majus L.) was achieved in epicotyl explants of seedlings after prolonged cultivation on Murashige and Skoog medium with or without plant growth regulators. Somatic embryos developed into plantlets which entered additional cycles of somatic embryogenesis. Cultures consisting of plantlets with prolonged embryogenic potential were maintained for five years on plant growth regulator free medium. Embryos which developed into rooted plantlets could be acclimated in a glasshouse enabling thus a continuous propagation scheme to be established.     

Nucleases isolated from Chelidonium majus L. milky sap can induce apoptosis in human cervical carcinoma HeLa cells but not in Chinese Hamster Ovary CHO cells

Milky sap isolated from Chelidonium majus L. (Greater Celandine) serves as a rich source of various biologically active substances such as alkaloids, flavonoids and phenolic acids. Previous research showed that the activity of Ch. majus milky sap may depend also on the presence of biologically active proteins. The goal of this study was to evaluate the biological effect of two nucleases isolated from Ch. majus milk sap, CMN1 of 20 kDa and CMN2 of 36 kDa, on HeLa and CHO tumour cell lines. Both studied nucleases together with other proteins in the sap of the plant are involved in stress and defence reactions against different pathogens. After 48 h incubation of CMN1 and CMN2 only with HeLa cells, the dependence between the number of apoptotic lesions and the concentration of applied nuclease was observed. The highest proapoptotic activity was induced by 13.3 ng/ml concentration of CMN2 collected in May (62 +/- 3% HeLa cells were apoptotic). Moreover, the proportion of necrotic cells in all concentrations of the nucleases and both cell lines was relatively low (1-8 +/- 0.5%). In summary, results of this study show that purified nucleases CMN1 and CMN2 isolated from Ch. majus milky sap exhibit apoptotic activity in HeLa tumour cell line, but not in CHO cells, without inflammatory reaction.     

Multiplication of Antirrhinum majus L. by shoot-tip culture

Nine varieties of Antirrhinum majus L. have been used in a study of in vitro multiplication of plants using shoot-tip culture. Acceptable multiplication rates were obtained in several media with only variety Victory showing significantly lower rates of shoot production. Wounded shoots of this variety produced callus in the absence of added auxin and some of this callus produced prolific roots.     

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In vitro and in vivo investigations on the antitumour activity of Chelidonium majus

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Modified alkaloids from Chelidonium majus L. induce G2/M arrest, caspase-3 activation, and apoptosis in human acute lymphoblastic leukemia MT-4 cells

The novel anticancer drug amitosine representing the mixture of thiophosphamide-modified alkaloids from Chelidonium majus L. has been reported to inhibit growth of various solid tumors in vivo. However, its antileukemic activity as well as the mechanisms of anticancer action have not been yet extensively examined. In this study, amitosine treatment at a dose of 100-250 microg/mL for 24 h resulted in dose-dependent inhibition of MT-4 cell proliferation in vitro with apoptosis induction in the setting of the significant G2/M phase arrest (up to 70% of cells). While amitosine induced caspase-3 activation in MT-4 cells, the increase in the number of cells containing the active caspase-3 did not correlate with the increase of apoptotic cell percentage. Western blotting data revealed the accumulation of cytochrome c in cytosolic fraction of MT-4 cells within 6 h after treatment with 100 microg/mL amitosine. To sum up, amitosine has been shown to possess strong antiproliferative and apoptosis-inducing activities in MT-4 cells in vitro, which seem to be mediated partially through caspase-dependent mitochondrial death pathways.     

High frequency plant regeneration from immature ovule-derived embryogenic cell suspension cultures of Chelidonium majus var. asiaticum

Culture conditions for high frequency plant regeneration via somatic embryogenesis in cell suspension cultures of Chelidonium majus var. asiaticum are described. Immature ovules formed embryogenic calluses at a frequency of 40% when cultured on Murashige and Skoog (MS) medium supplemented with 4.52 μM 2,4-dichlorophenoxyacetic acid (2,4-D). The optimum ovule size for embryogenic callus formation ranged from 1 to 1.5 mm in length. Cell suspension cultures were established from embryogenic calluses using MS liquid medium containing 4.52 μM 2,4-D. Upon plating onto MS basal medium, cell aggregates from cell suspension cultures produced somatic embryos which then developed into plantlets. Regenerated plantlets were transplanted to potting soil and grown to maturity in a growth chamber. This revised version was published online in June 2006 with corrections to the Cover Date.     

Genetic Aspects of the Relationship between Isoquinoline Alkaloids and Mineral Elements in Greater Celandine (Chelidonium majus L.)

Interrelations between the total content of isoquinoline alkaloids, the concentrations of quaternary protoberberines and benzophenanthridines, and the amount of K, Cu, Co, Al, Ba, and Zn in aerial parts of individual celandine plants were revealed, within a single cenopopulation, using correlation analysis and regression analysis. Mathematical models describing the regulation of isoquinoline metabolism by some of the mineral elements were obtained in analytical form. The results suggest that this process is genetically determined.     

Genetic aspects of the relationship between isoquinoline alkaloids and mineral elements in greater celandine (Chelidonium majus L.)

Interrelations between the total content of isoquinoline alkaloids, concentrations of quaternary protoberberines and benzophenanthridines, and the amount of K, Cu, Co, Al, Ba, and Zn in aerial parts of individual celandine plants were revealed, within a single cenopopulation, using correlation analysis and regression analysis. Mathematical models describing the regulation of isoquinoline metabolism by some of the mineral elements were obtained in the analytical form. The results suggest that this process is genetically determined.     

Regeneration of plants from Antirrhinum majus L. callus

Somaclone production in Antirrhinum majus plants by regeneration of plants from callus cultures has been achieved using three types of explant tissue. Regeneration from mature stem internode-derived callus was extremely poor. Callus derived from seedling shoot tips could be induced to form new shoots in six of seven cultivars tested. Regeneration was achieved in all seven cultivars when callus was produced from segments of hypocotyl and was most effective using agar-solidified medium containing 0.25 mgl^-1 naphthoxyacetic acid + 10% coconut milk. In this case, five of the cultivars produced shoots directly, one produced leaves from the petioles of which new shoots emerged, and one regenerated plants chiefly through the production of embryoids.     

Differential temperature sensitivity of normal and cancer cells in culture

Serially-propagated growing heteroploid and growing diploid cell cultures do not survive incubation at 42° for 24 hours, whereas contact-inhibited diploid monolayers are still viable after at least nine days at this elevated temperature. Heat-treated heteroploid HeLa cells and growing diploid cells exhibit a variety of morphologic abnormalities, but contact-inhibited cells are only minimally affected. A similar differential temperature sensitivity exists in the synthesis of cellular macromolecular components such as DNA, RNA, and protein: incorporation of radioactive precursors is drastically reduced in growing diploid and heteroploid cells after 24 hours at 42°, but not in contact-inhibited cells. Incorporation of labelled glucose, choline, or linolenic acid is actually enhanced in heat-treated contact-inhibited cells.     

Antifungal Activities of Chelidonium majus Extract on Botrytis cinerea in vitro and Ultrastructural Changes in its Conidia

The extract from the aboveground parts of Chelidonium majus reduced the mycelial growth of Botrytis cinerea on Czapek agar medium. In minimum fungicidal concentration (60  μ l/ml), the extract induced irreversible ultrastructural changes in B. cinerea conidia.     

Micro-Nutrient Deficiency and Transpiration in Tropaeolum majus L.

The transpiration of Tropaeolum plants, grown in water culture,on full nutrient solutions and on solutions deficient in oneof the following, i.e. boron, copper, manganese, and zinc, wasstudied. The transpiration rate of the control plants grownon the full nutrient solution was significantly higher thanthat of the deficient ones. The more acute the deficiency, thegreater was the disparity. Under the condition of the experiment, the control plants weresmaller in size, having fewer and smaller leaves than the deficientones, but the foliage was much greener and healthier in appearance.     

Isolation and Partial Characterization of a New Lectin from Seeds of the Greater Celandine (Chelidonium majus)

Seeds of the greater celandine (Chelidonium majus L.) contain a lectin which could be isolated using a combination of affinity chromatography on chitin and ion exchange chromatography on sulphopropyl-Sephadex. The purified lectin was partially characterized with respect to its biochemical and physicochemical properties. It is a small dimeric protein composed of two different subunits of M_r 9,500 and 11,500, respectively. Its amino acid composition is typified by high contents of glycine and cysteine. No covalently bound carbohydrate could be detected. Hapten inhibition experiments indicated that the lectin exhibits specificity towards oligomers of N-acetylglucosamine, the potency of inhibition increasing with chain length up to four residues. The greater celandine lectin is the first lectin to be isolated from a species belonging to the plant family Papaveraceae (poppy family). Although it represents a new type of plant lectin, resemblances to phytohemaglutinins from diverse taxonomic origin are obvious.