Induction of betacyanin formation in cell cultures of Chenopodium album under UV-light irradiation

A cell culture of Chenopodium album was cultivated under differentlight conditions (UV, blue, white, red light, and darkness).Red-violet coloured betacyanin-containing cell groups were formedon the surface of the cell culture only under UV-light or UV-containinglight qualities (blue and white), respectively. A direct correlationexists between UV-portions of different light qualities andthe number of red-violet cell groups formed, whereas light conditionswithout UV-light do not induce betacyanin formation. Betacyaninsformed under UV-light were degraded after transfer to red lightand darkness. The phytohormone kinetin enhanced the betacyaninformation in white light, but was unable to induce betacyaninsin darkness. The phytohormone 2, 4-D totally inhibited the formationof betacyanins under all light conditions studied. All otherinvestigated parameters of primary and secondary metabolismof the cells (protein content, respiration activity, concentrationof carotenoids and flavonoids) only unspecifically increasedin reaction to continuous UV-light irradiation. A selected yellowcoloured cell strain autotroph to auxin was unable to form betacyaninunder continuous UV-light irradiation and under increased kinetinconcentration. Key words: Betacyanins, cell culture, phytohormones, secondary product formation, UV-light     

Light-stimulated Absorption of Nitrate by Wolffia arrhiza

The mechanism of the light-stimulated absorption of nitrate by Wolffia arrhiza was studied. The nitrate-absorption mechanism in ammonium-grown plants is stimulated by the presence of nitrate. In a manner similar to the absorption of many other ions, the absorption of nitrate follows a typical biphasic pattern in relation to external nitrate concentration. Mechanism 1 is effective at nitrate concentrations up to 0.5 to 0.75 mM and mechanism 2 becomes operative at higher nitrate levels. Light stimulates the absorption of nitrate independently of the effect of light on the reduction of nitrate. The effects of uncouplers, inhibitors, and light of wavelengths of 700 nm or longer indicate that nitrate absorption by Wolffia cells is reduced when non-cyclic electron transport is blocked. It is postulated that under this condition, ATP in the chloroplast (produced by cyclic photophosphorylation) may be less readily transported across the chloroplast envelope than when non-cyclic electron transport is proceeding.     

The effect of pH on the population growth of three species of duckweed: Spirodela oligorrhiza, Lemna minor and Wolffia arrhiza

Three species of duckweed, Spirodela oligorrhiza, Lemna minor and Wolffia arrhiza were grown under aseptic conditions on both buffered and unbuffered solutions of Jacob's media. Media with manually regulated pH levels were also used. Growth on unbuffered media is initially rapid but eventually inhibited, probably by increased pH levels. On buffered media growth is poor and effects of buffers cannot be separated out. These media give inadequate pictures of the species’ responses to changes in pH. Growth is most successful on media with regulated pH where sustained logarithmic population increases were achieved. Spirodela and Lemna rates are symmetrical about an almost neutral, optimal pH, declining fairly rapidly away from the optimum. Wolffia has an optimum at pH 5 and growth declined with increasing pH. All three species have optima at, or below, the neutral point. The range of tolerance of duckweeds is broader than has previously been suspected. Estimated lower limits, optimum and upper limits for each species are: Wolffia, pH 4·5–0·10, Lemna pH 4–6·2–10, Spirodela pH 3·7–0·10. Growth rate along a pH gradient is best described by means of polynomial equations: second-degree equations are sufficient for Spirodela and Lemna but a fifth-degree equation is required for Wolffia. Rates of population growth are similar for all species. In decreasing order they are: Wolffia, Lemna, Spirodela. However, in biomass units Lemna grew more than six and Spirodela seventeen times faster than Wolffia.     

Biosorption of cadmium and chromium in duckweed Wolffia globosa

The biosorption of cadmium (Cd) and chromium (Cr) by using dried Wolffia globosa biomass were investigated using batch technique. The effects of concentration and pH solution on the adsorption isotherm were measured by determining the adsorption isotherm at initial metal concentrations from 10 to 400 mg/L and pH 4 to 7 for Cd, and pH 1.5 to 6 for Cr. The adsorption equilibria were found to follow Langmuir models. The maximum adsorption capacity (Xm) at pH 7 in W. globosa-Cd system was estimated to be 80.7 mg/g, while the maximum removal achieved at pH 4, pH 5, and pH 6 were 35.1, 48.8, and 65.4 mg/g, respectively. The Xm at pH 1.5 in W. globosa--Cr system was estimated to be 73.5 mg/g, while the maximum removal achieved at pH 3, pH 5, and pH 6 were 47.4, 33.1, and 12.9 mg/g, respectively. The effects of contact times on Cd and Cr sorption indicated that they were absorbed rapidly and more efficiently at lower concentrations.     

Effect of Cadmium on the Level of Isoprenoid-Derived Phytohormones in Duckweed Wolffia arrhiza

Journal of Plant Growth Regulation - Wolffia arrhiza (L.) Horkel ex Wimm. is an aquatic plant belonging to the Lemnaceae family. It does not have leaves, stems, and roots, flowers rarely occur,...     

Increases of secondary metabolite production in various plant cell cultures by co-cultivation with cork

Cork tissues increased secondary metabolite production of various plant cell cultures in a different manner from those of conventional elicitors. In Sophora flavescens and Glycyrrhiza glabra cultured cells, cork tissues increased the amounts of both lipophilic and hydrophilic flavonoids without affecting the cell growth, although elicitors such as copper ion and yeast extracts showed a clear inhibition of cell growth with the increasing amount of these lipophilic ones. The validity of this effect of cork tissues covered a wide range of aromatic compounds produced by suspension cell cultures derived from diverse plant species. Woody tissues of Japanese cypress had a very similar effect to that of cork. Partial purification of cork tissues suggested that the production-stimulating factor was present in the hemicellulose B fraction that was not included in the dedifferentiated cultured tissues.     

Correlation of betacyanin synthesis with cell division in cell suspension cultures of Phytolacca americana

In suspension cultures of Phytolacca americana , betacyanin accumulation was reduced when cell division was inhibited by treatment with various inhibitors of DNA synthesis or anti-microtubule drugs. Aphidicolin (APC), an inhibitor of DNA synthesis, reduced the incorporation of radioactivity from labeled tyrosine into betacyanin, but the incorporation of radioactivity from labeled 3,4-dihydroxyphenylalanine (DOPA) into betacyanin was not affected by similar treatments. Propyzamide, another anti-microtubule drug, reduced incorporation of radioactivity from tyrosine and DOPA into betacyanin. However, the rate of incorporation from DOPA was higher than that from tyrosine. The results suggest that inhibition of betacyanin accumulation in Phytolacca americana cells by APC and propyzamide is due to suppression of the reaction converting tyrosine to DOPA, which may be closely related to cell division.     

Phytoaccumulation and phytotoxicity of cadmium and chromium in duckweed Wolffia globosa

The phytoaccumulation and phytotoxicity of heavy metals, cadmium (Cd), and chromium (Cr) on a common duckweed, Wolffia globosa, were studied. W. globosa were cultured in 3% Hoagland's nutrient medium, which was supplemented with 1, 2, 4, and 8 mg/L of Cd and Cr and were separately harvested after 3, 6, 9, and 12 days. The accumulation of Cd and Cr in W. globosa showed significant increases when the exposure time and metal concentration were increased. The effects of Cd and Cr on the biomass productivity and total chlorophyll content in W. globosa indicated that there were significant decreases in the biomass productivity and total chlorophyll content when the exposure time and metal concentration were increased.     

Biosynthesis of podophyllotoxin in Linum album cell cultures

Cell cultures of Linum album Kotschy ex Boiss. (Linaceae) showing high accumulation of the lignan podophyllotoxin (PTOX) were established. Enzymological studies revealed highest activities of phenylalanine ammonia-lyase, cinnamyl alcohol dehydrogenase, 4-hydroxycinnamate:CoA ligase and cinnamoyl-CoA:NADP oxidoreductase immediately prior to PTOX accumulation. To investigate PTOX biosynthesis, feeding experiments were performed with [2-¹³C]3′,4′-dimethoxycinnamic acid, [2-¹³C]3′,4′-methylenedioxycinnamic acid (MDCA), [2-¹³C]3′,4′,5′-trimethoxycinnamic acid, [2-¹³C]sinapic acid, [2-¹³C]- and [2,3-¹³C₂]ferulic acid. Analysis of the metabolites by HPLC coupled to tandem mass spectrometry revealed incorporation of label from ferulic acid into PTOX and deoxypodophyllotoxin (DOP). In addition, MDCA was also unambiguously incorporated intact into PTOX. These observations suggest that in L. album both ferulic acid and methylenedioxy-substituted cinnamic acid can be incorporated into lignans. Furthermore, it appears that, in this species, the hydroxylation of DOP is a rate-limiting point in the pathway leading to PTOX. Electronic supplementary material to this article is available at and is accessible for authorized users. Electronic Publication     

Enzymatic Degradation of Chlorophyll in Chenopodium album

The breakdown of chlorophyll (Chi) in crude extracts of Chenopodiumalbum (white goose foot) in the dark was examined. Derivativesof pheophorbide were formed when Chi or chlorophyllide wasincubated with depigmented crude extracts. The formation ofpheophorbide was completely prevented by heat treatment of extracts,indicating that the reaction was enzymatic, and the presenceof a Mg-releasing enzyme, the so called Mg-dechelatase, waspostulated. This hypothesis was strongly supported by the observationthat the formation of pheophorbide was inhibited by 51% by 10mM MgCl₂. Analysis by high-performance thin-layer chromatography(HPTLC) and liquid chromatography (HPLC) showed that the appearanceof chlorophyllide , pheophorbide 13²-hydroxychlorophyllide and pyropheophorbide was accompanied by a concomitant decreasein levels of Chi The formation of 13²-hydroxychloro-phyllide was not clearly an enzymatic reaction and requires furtherexamination. It appears that Chl is degraded in a crude extractof C. album via the following enzymatically catalyzed reactions (Received September 10, 1990; Accepted November 15, 1990)     

Induction of shikonin formation by agar in Lithospermum erythrorhizon cell suspension cultures

Cultured cells of Lithospermum erythrorhizon capable of producing red naphthoquinone (shikonin) derivatives on Linsmaier-Skoog agar medium ceased s     

Jasmonic acid as modulator of lead toxicity in aquatic plant Wolffia arrhiza (Lemnaceae)

The present study was undertaken to test the influence of exogenously applied jasmonic acid (JA) upon the growth and metabolism of Wolffia arrhiza (Lemnaceae), the smallest vessel aquatic plant exposed to lead (Pb) stress. It was found, that JA acted in a concentration-dependent manner. Treatment with JA at the highest concentration 100 μM resulted in the enhancement of heavy metal toxicity leading to increase in metal biosorption and formation of lipid peroxides as well as decrease in fresh weight, chlorophyll a, carotenoid, monosaccharide and soluble protein content. In contrast, this phytohormone applied at 0.1 μM protected W. arrhiza fronds against Pb stress inhibiting heavy metal accumulation, restoring plant growth and primary metabolite level. Moreover, JA at 0.1 μM activated enzymatic (catalase, ascorbate peroxidase, NADH peroxidase) and non-enzymatic antioxidant (ascorbate, glutathione) system in W. arrhiza, and therefore, suppressed oxidative destruction of cellular components induced by heavy metal. The data suggest that JA plays an important role in the growth and metabolism of W. arrhiza exposed to abiotic stressor and its high adaptation ability to metal contamination of aquatic environment.     

Induction of benzoquinone formation by activated carbon in Lithospermum erythrorhizon cell suspension cultures

Cultured cells of Lithospermum erythrorhizon which were capable of producing red naphthoquinone (shikonin) derivatives on Linsmaier-Skoog's agar medium stopped synthesizing these compounds when grown in liquid medium without agar. However, when the liquid medium was supplemented with a small amount of activated carbon, the cells produced a new orange benzoquinone derivative, echinofuran B, which may be considered an abnormal metabolite of geranylquinol, the key intermediate in the biosynthesis of shikonin. A similar effect of activated carbon was also observed with a variant cell line incapable of producing shikonin derivatives even on the agar medium. By contrast, the callus cultures grown on the agar medium as well as the dried roots of the intact plant were found to contain a small amount of echinofuran C, another new benzoquinone related to echinofuran B, in addition to shikonin derivatives.     

Purification and Some Properties of Pheophorbidase in Chenopodium album

A novel enzyme, pheophorbidase, which catalyzes the conversionof pheophorbide a to C-13²-carboxylpyropheophorbide a, was purifiedfrom Chenopodium album leaves. The purified enzyme showed twobands of 28 kDa and 29 kDa on SDS-PAGE. The molecular mass ofthe native pheophorbidase was 105 kDa. The N-terminal aminoacid sequence for the 28-kDa protein could be determined, whereasthe N-terminus of the 29-kDa protein was blocked. Immunochemicaland enzyme activity analyses revealed that pheophorbidase islocated in an extra-plastidic part of the cell. (Received September 7, 1998; Accepted October 26, 1998)     

Biological control of Chenopodium album L. in Europe

Ascochyta caulina (P. Karst) v.d. Aa and v. Kest is aplant pathogenic fungus which is specific to Chenopodium albumL. It has been suggested as a potential mycoherbicide to this weed,which is important and wide spread in arable crops throughout Europe. Toinvestigate its potential as a biocontrol agent, the fungus has beentested in glasshouse and field experiments. Formulations containingdifferent combinations of A. caulina conidia, the phytotoxinsfrom the fungus and low doses of herbicides have been tested.Significant improvement in the efficacy of the fungus was achieved inglasshouse trials with an aqueous formulation containing PVA(0.1% v/v), Psyllium (0.4% w/v), Sylgard 309(0.1% v/v), nutrients and conidia (5 ×10⁶/ml). The extracellular, hydrophilic phytotoxinsproduced by A. caulina were purified and their structuresdetermined. The main toxin, named ascaulitoxin, was characterised as theN²--D-glucopyranoside of the unusual bis-aminoacid2,4,7-triamino-5-hydroxyoctandioic acid. Two other toxins proved to betrans-4-amino-D-proline and the aglycone of ascaulitoxin. Thesetoxins have shown promising herbicidal properties. Field trials haveinvestigated the performance of A. caulina conidia applied atdifferent developmental stages of C. album either as a singletreatment or combined with sub-lethal doses of herbicides or with thefungal phytotoxins. With the available formulation, favourable weatherconditions are needed to obtain infection in the field. The efficacy ofthe strain of A. caulina used so far has proved to beinadequate to justify its development as a bioherbicide. This isprobably due to its low virulence.     

Suppression of Wolffia arrhiza growth by brassinazole, an inhibitor of brassinosteroid biosynthesis and its restoration by endogenous 24-epibrassinolide

The effect of the brassinosteroid (BR) 24-epibrassinolide (epiBL; 10(-13)-10(-6)M) on growth and levels of chlorophylls, carotenoids, sugars and protein in Wolffia arrhiza after 7 days of cultivation is reported. Application of epiBL to W. arrhiza cultures stimulates the growth and increases the content of photosynthetic pigments, sugar and protein. The greatest effect of epiBL is observed at a concentration of 10(-9)M. We tested the action of Brz2001, a specific BR biosynthesis inhibitor, in the range of 10(-6)-10(-4)M. Addition of Brz2001 to W. arrhiza cultures inhibits their growth after 7 days of cultivation. The inhibition of growth could be reversed by the addition of epiBL. Moreover, there was not complete recovery to the level of control, especially at 5 x 10(-5)-10(-4)M Brz2001. The effects of treatment with 10(-9)M epiBL mixed with a mevalonate pathway inhibitor (mevinolin), or a 2-methylerythritol 4-phosphate pathway inhibitor (clomazone), were also investigated. Mevinolin did not inhibit growth of W. arrhiza after 7 days of cultivation. However, clomazone did. Addition of epiBL overcame this inhibition. These results suggest that the mevalonate pathway may not function well in W. arrhiza and that biosynthesis of BRs through the non-mevalonate pathway in W. arrhiza could be possible.     

Production of polysaccharides by callus cultures of common duckweed

Callus lines of common duckweed produced acid arabinogalactan and pectin in an amount varying from 1 to 3% of dry weight. The arabinogalactan specimens from the cell lines studied displayed a similar monosaccharide composition. The duckweed callus lines whose arabinogalactans contained apiose residues (1–2%) were found. All pectin specimens had a similar qualitative monosaccharide composition but differed in the quantitative content of monosaccharide residues. The lines with high contents of galactose, arabinose, and apiose in pectin specimens were obtained. The total content of neutral monosaccharide residues in pectins varied from 26 to 50%.     

Uptake and Effect of Abscisic Acid during Induction and Progress of Radicle Growth in Seeds of Chenopodium album

In the seeds of Chenopodium album L. visible phenomena preceding the final protrusion of the radicle enable a clear distinction between the induction and the progress of growth inside the covering structures. The light-dependent induction of radicle growth is not inhibited by exogenously applied abscisic acid (ABA). Experiments with 1-¹⁴C-ABA ruled out a lack of penetration of the hormone. However, ABA does inhibit the growth of the radicle before final protrusion. This inhibition and the uptake of 1-¹⁴C-ABA are enhanced at lower pH values, indicating absorption of the undissociated molecule. The uptake of labeled hormone strongly increases during the growth of the radicle. This increase is not merely a reflection of extra water uptake. Seeds of different degrees of dormancy contain equallly low levels of endogenous ABA. Much higher levels of ABA in the seeds were obtained by exogenous application of the hormone but these levels stills do not prevent the breaking the dormancy by light. It is concluded that ABA has no function in the regulation of dormancy in C. album seeds.     

Production of polysaccharides by callus cultures of common duckweed

Callus lines of common duckweed produced acid arabinogalactan and pectin in an amount varying from 1 to 3% of dry weight. The arabinogalactan specimens from the cell lines studied displayed a similar monosaccharide composition. The duckweed callus lines whose arabinogalactans contained apiose residues (1-2%) were found. All pectin specimens had a similar qualitative monosaccharide composition but differed in the quantitative content of monosaccharide residues. The lines with high contents of galactose, arabinose, and apiose in pectin specimens were obtained. The total content of neutral monosaccharide residues in pectins varied from 26 to 50%.