Changes in chromatin of Daucus carota cells during embryogenesis

Cells of carrot (Daucus carota var. Rote Riesen) were cultured on media inductive and non-inductive for embryogenesis and analyzed for differences in their chromosomal proteins and chromatin template activity. Non-histone proteins were prepared from dehistonized chromatin and their properties were investigated. Non-histone proteins proved to be acidic and associated easily with calf thymus histone. Non-histone proteins were able to counteract the inhibitory effect of histone on DNA-directed RNA synthesis in vitro. Almost the same rate of restoration occurred regardless of the interaction between DNA and protein, when sufficient amounts of non-histone proteins were added. However, once the histone-DNA complex was established, the restoration by non-histone proteins at comparably lower concentration was poor. Another acidic protein, bovine serum albumin, had no effect on histone inhibited RNA synthesis. Also non-histone proteins enhanced the chromatin directed RNA synthesis more than 100%. The template activity of chromatin changed after the inductive treatment of embryo formation and induced cells showed higher template activity than non-indiiced controls after embryo cells were formed. Histone components were the same in inductive and non-inductive cells. On the other hand, there was a correlation between template activity and the stimulation by non-histone proteins of histone-inhibited RNA synthesis.     

Lipid composition of Daucus carota roots

Lipids extracted from Daucus carota roots were analyzed and the fatty acid composition of the triglycerides and phospholipids determined. Compariso     

Location and density labelling of acid invertase in aging discs of Daucus carota storage tissue

Cell walls prepared from aged discs by extraction in 0·1 M acetate buffer, pH 4·8, possess ionically bound acid invertase which can be removed from the wall by incubation in 1 M sodium chloride in 0·1 M acetate buffer, pH 4·8, and more firmly attached enzyme which is not removed. Cell walls prepared in 0·195 M phosphate-0·003 M citrate-buffer, pH 8·0, do not possess ionically bound enzyme. Ionically bound invertase is density labelled when discs are aged in 90% deuterium oxide suggesting that at least part of the increase in activity observed during aging is due to de novo protein synthesis.     

Abnormal metabolites produced by Daucus carota roots stored under conditions of stress

The formation of dihydroisocoumarins (I and II), the chromones (III and IV) and scopoletin (V) is induced in carrot roots by storage in the presence of low concentrations of ethylene and by inoculation with various fungi. There has been concern over possible toxic effects of dihydroisocoumarins and some preliminary toxicological data on (I), (II) and (IV) are included.     

A Unique pattern of anthocyanins in Daucus carota and other umbelliferae

Three cyanidin glycosides have been identified in the black carrot: the known 3-lathyroside and two new pigments, a 3-xylosylglucosylgalactoside and its ferulyl derivative. The same pigments, together with the sinapyl derivative of the triglycoside, occur variously in other tissues of Daucus carota. Ferulyl and sinapyl derivatives of cyanidin 3-glucosylgalactoside occur exceptionally in stem of one subspecies, maritimus. One or other of the same pigments have been found to occur variously in 20 of 22 other umbellifer species surveyed. Both ferulyl and sinapyl derivatives occur in stem of Conium maculatum and Foeniculum vulgare. A further novel acylated pigment based on p-coumaric acid was found in wild celery, Apiurn graveolens. The systematic significance of these various findings is discussed.     

Guaiane type sesquiterpenes and other constituents from Daucus carota L.

In a chemosystematic investigation of the fruits of Daucus carota L. we have isolated eleven compounds including four guaiane type sesquiterpenes (1–4), one eudesmane type sesquiterpene (5) and six other constituents (6–11). Among them, the five compounds 1–5 all are new for Daucus carota L. and all have been reported in Torilis japonica. The results show that Daucus carota L. is chemically similar with Torilis japonica providing chemical evidence for both species used as the same traditional Chinese medicine “nanheshi”.     

Genetic variation for volatile terpenoids in roots of carrot, Daucus carota, backcrosses and F2 generations

Volatile terpenoid levels were measured in segregating carrot populations. Genetic analysis suggested dominance and multigenic control for low total volatile terpenoids, and low terpinolene quantities. The inheritance of caryophyllene quantities differed in different populations. Percent (E)-γ-bisabolene exhibited simple genetic control in several carrot populations with dominance for high levels. Compensatory shifts between the major volatile terpenes indicates an inter-relationship between the mono- and sesquiterpenoid biosynthetic pools.     

Genetic variation for volatile terpenoids in roots of carrot, Daucus carota, inbreds and F1 hybrids

Volatile terpenoid levels were determined in eight carrot inbreds and ten F₁ hybrids. Genetic variation between inbreds spanned a 5-10-fold range f     

Effect of 2,4-dichlorophenoxyacetic acid on the activity of o-methyltransferase in carrot cell culture

The activity of caffeic acid-O-methyltransferase (OMT) in carrot cells was greatly affected by the amount of 2,4-dichlorophenoxyacetic acid (2,4-D) supplemented to the culture medium. The OMT fraction was purified by (NH₄)₂SO₄ followed by ultrafiltration and gel filtration or DEAE-Sephadex chromatography after cells were cultured in the medium containing [2-¹⁴C]-2,4-D. Thus, this purified fraction revealed high OMT activity and was still radioactive. The OMT activity was about eight-fold higher (or more) in cells cultured at 0.05 ppm 2,4-D than in those at 1.0 ppm 2,4-D. The ratio of radioactivity to OMT activity was about four-fold higher in cells cultured at 1.0 ppm 2,4-D than those at 0.05 ppm 2,4-D. On the other hand, the OMT fraction was separated into two radioactive protein fractions by DEAE-Sephadex chromatography. The radioactive fractions became Et₂O-soluble after HCl hydrolysis, but not after salt-urea treatment. From these results, it was concluded that 2,4-D is covalently bound to proteins in the OMT fraction. Such 2,4-D protein conjugates may play a role in the regulation of OMT activity.     

Behaviour of phenylalanine ammonia-lyase in carrot cells in suspension cultures

The high PAL activity in carrot cells in suspension culture was found at the linear and early stationary phases, with concomitant increases in phenylal     

Effects of adenyl cytokinins on the solutes of cultured cells

When zeatin acts upon cultured carrot explants in a basal medium lacking IAA its most significant effect is to increase the Na⁺/K⁺ ratio. In the presence of IAA, zeatin greatly stirnulates cell multiplication and the Na⁺/K⁺ ratio is then low. The effect of a series of N 6-substituted adenines with side chains (-CH₂)_nH varying from 1 to 10 have been tested for their effect upon cell multiplication and cell size and upon the total osmotic value and the relative concentrations of Na⁺, K⁺ and Cl_? in the cells. Whereas the maximum response in terms of cell multiplication occurred with the compound having a ?(CH₂)₅ · H sidechain, the maximum effect on the Na⁺/K⁺ ratio of the cultured cells as so grown was caused when the side chain was ?(CH₂₄H.     

Differentiation of metabolic pathways in the umbel of Daucus carota

The extractable activities of PAL, chalcone synthase and chalcone isomerase of white and coloured petals of the inflorescence of the umbel of wild carr     

Development of aminoacyl tRNA synthetases in cultured Nicotiana tabacum cells

Changes in the activity of aminoacyl tRNA synthetases during growth of tobacco XD cells in suspension culture have been determined by the pyrophosphate exchange assay. Alanyl, arginyl, glutamyl, glutaminyl and seryl tRNA synthetases showed the lowest activity, whilst lysyl, histidyl, leucyl, isoleucyl, phenylalanyl threonyl and valyl tRNA synthetases were most active. Most synthetases, and total protein, increased to a maximum, at around 7 days, just before mid-exponential phase, and then fell.     

Ethanol metabolism in suspension cultured carrot cells

Ethanol production in plant tissues deprived of oxygen is a well known process. Nevertheless, little information is available on the toxic effects of ethanol on plant cells and tissues, or on the possible role of acetaldehyde, the first oxidative product of ethanol, in inducing toxic effects in plants. Data on the metabolism of ethanol in suspension cultured cells of carrot ( Daucus carola L. cv. S. Valery, cell line T22), a system highly sensitive to the presence of ethanol in the culture medium, indicate that carrot cells oxidize only small amounts of ethanol to CO₂. Instead, they convert ethanol mainly to acetaldehyde, which accumulates in the culture medium. This suggests a possible role of acetaldehyde in causing ethanol-induced injury to carrot cells.     

Phytochelatins govern zinc/copper homeostasis and cadmium detoxification in Cuscuta campestris parasitizing Daucus carota

Cuscuta sp., known with the common name of “dodder”, is an obligate parasite capable of invading stems and leaves of a wide range of host plants. Dodder stem usually coils counterclockwise around the host and, within a few days, develops haustorial structures at each point of contact. As soon as dodder haustoria reach host vascular bundles, they start tapping water, photosynthates and minerals. Metal ions such as zinc (Zn) and copper (Cu) are essential for dodder growth and metabolism, although an exceedingly high (over-homeostatic) supply of these micronutrients can result in growth inhibition and cellular toxicity. Even more so, non-essential metals such as cadmium (Cd), if transferred from the host to the parasite, need to be neutralized by timely detoxification mechanisms. In this work, we showed that Cuscuta campestris Yuncker establishes effective haustorial connections with leaf petioles and blades of Daucus carota L. (carrot), with the consequent transfer of Cd and essential metals (such as Zn and Cu) from the host vascular bundles to the parasite. Following up to this point, we detected the presence in the parasite of significant amounts of glutathione and phytochelatins, even in the absence of Cd exposure. This suggests that thiol peptides in dodder might be particularly important for Zn and Cu homeostasis as well as for Cd detoxification. Finally, we demonstrated that dodder is capable of synthesizing phytochelatins on its own, rather than massively importing them from the host, and also provided evidence for the existence of an endogenous, constitutively expressed, dodder's phytochelatin synthase.     

Increases in enzyme levels during the formation of phenolic acids in carrot cell cultures

The levels of glutamic acid dehydrogenase (GDH), phenylalanine ammonia-lyase (PAL), cinnamic acid 4-hydroxylase (CAH) and O-methyltransferase (OMT) were measured during the formation of phenolic acids in carrot cells in suspension culture. Caffeic, ferulic and p-hydroxybenzoic acids were always present as the culture proceded. Total content of these acids increased at the early logarithmic and linear phases. GDH showed high activity at the early logarithmic and stationary phases. PAL activity was much enhanced at the linear and stationary phases. CAH activity was found in actively growing cells, especially at the early and late logarithmic phases OMT behaved similarly to PAL. The increases in GDH and CAH might be responsible for the rapid synthesis of phenolic acid at the early logarithmic phase. The increase in phenolic acid at the linear phase would certainly be due to enhancements of both PAL and OMT. On the other hand, the accumulation of vanillic acid was observed in cells which were transferred and cultured on an agar medium, but not in cells in suspension culture. This accumulation is related to increases in OMT levels and also to changes in the degree of β-oxidation.     

Sucrose metabolism during tobacco callus growth

Activities of soluble and insoluble invertases and sucrose synthetase in tobacco callus increased significantly within the first 3 days of culture. After this period soluble invertase activity declined, while the activities of the insoluble invertase and the sucrose synthetase were relatively unchanged.     

Phytoalexin production in cultured carrot cells treated with pectinolytic enzymes

6-Methoxymellein, a phytoalexin of carrot, was produced in cultured cells upon addition of partial hydrolysates of carrot cells obtained by treatment with purified endo-polygalacturonase or endo-pectin lyase. Direct addition of these enzymes to the cell culture also stimulated the accumulation of this 6-methoxymellein. When the hydrolysates obtained by these enzymes were subsequently treated within pectin esterase, the activity for the elicitation of 6-methoxymellein production decreased appreciably. These results suggest that pectinolytic enzymes release elicitor-active cell wall fragments from carrot cells and that a certain degree of esterification of the galacturonosyl moiety in these pectic polysaccharides is required for elicitor activity.