Lipids in plant tissue cultures. III. Very long-chain fatty acids in the lipids of callus cultures and suspension cultures

The lipids in plant tissue cultures contain in addition to the common saturated and unsaturated fatty acids even- and odd-numbered fatty acids having chain-lengths up to 26 carbon atoms.     

Growth models of cultures with two liquid phases. III. Continuous cultures

Some mathematical models, which have been used to describe batch growth in fermentations with two liquid phases present, are used to predict the behavior of continuous fermentations in a chemostat. Two types of dispersed systems are considered in this investigation. In the first, type, it is assumed that the composition of the dispersed phase is such that, increased substrate utilization results in a decreased substrate concentration with no change in the interfacial area. In the second type of system, the dispersed phase is assumed to be pure substrate; therefore, the substrate concentration in the dispersed phase remains constant but the interfacial area is affected by changes in dilution rate. Three special cases are examined for each type of system in order to examine the effect of the interfacial area, the phase equilibrium constant, and the mass transfer coefficient on system performance. Comparison of two of the models with available experimental data shows fair agreement, between model and data.     

Lipids in plant tissue cultures IV. The characteristic patterns of lipid classes in callus cultures and suspension cultures

Lipids from callus cultures and suspension cultures of higher plants constitute 5 to 8% of the dry tissue's weight.The predominant lipid classes are the sterols, steryl esters, steryl glycosides and esterified steryl glycosides. Considerable amounts of a variety of sterylglycolipids, whose structures are not completely elucidated, are also present. Triglycerides and phospholipids occur in small proportions, whereas monogalactosyl diglycerides, digalactosyl diglycerides and sulfoquinovosyl diglycerides are present only in traces, if at all.β-Sitosterol is the predominant constituent sterol, stigmasterol and campesterol as well as a variety of as yet unidentified sterols occur in smaller proportions. The major constituent fatty acids are palmitic, oleic, linoleic and linolenic acids. Saturated very long-chain fatty acids are found in smaller proportions. Unusual fatty acids, such as epoxy acids, which occur in the seed lipids of certain plants, are not found in tissue cultures derived from these plants. Clucose and traces of galactose are the only sugars obtained by acid hydrolysis of the glycolipids occurring in plant tissue cultures.     

An analysis of extended and exponentially-fed-batch cultures.

Under certain conditions it is shown that an extended culture is equivalent to an exponentially-fed-batch culture, that an exponentially-fed-batch culture (and an extended culture) can be maintained at a steady state and that an exponentially-fed-batch culture may be mimicked by a continuous-flow culture with a constant dilution rate. Operational conditions required to maintain steady states are specified.     

Lipids in plant tissue cultures. VII: Heterotrophic and mixotrophic cell cultures of Chenopodium rubrum

Mixotrophic cell cultures of Chenopodium rubrum were found to synthesize 5 to 33 times more monogalactosyldiacylglycerols and 5 to 16 times more digalactosyldiacylglycerols than heterotrophic ones. The monogalactosyldiacylglycerols and digalactosyldiacylglycerols from mixotrophic cultures contained higher levels of linolenic acid as compared to heterotrophic cultures. It is concluded that the active synthesis of these galactolipids with high levels of constituent linolenic acid is associated with the onset of photosynthesis in plant cell cultures, as is the case in intact plants.     

Lipids in plant tissue cultures. V. Effect of environmental conditions on the lipids of Glycine soja and Brassica napus cultures.

The environmental conditions prevailing during growth of plant tissue cultures affect the concentration of certain lipid classes and the fatty acid patterns of the total lipids. Irrespective of whether the cultures are grown under continuous illumination or in the dark, aerated cultures contain larger proportions of sterols, steryl esters, steryl glycosides and various sterylglycolipids and lower concentrations of squalene than non-aerated cultures. The lipids of the latter contain larger proportions of saturated very long-chain fatty acids than those of the former cultures.     

Ammonia assimilation by rhizobium cultures and bacteroids.

The enzymes involved in the assimilation of ammonia by free-living cultures of Rhizobium spp. are glutamine synthetase (EC. 6.o.I.2), glutamate synthase (L-glutamine:2-oxoglutarate amino transferase) and glutamate dehydrogenase (ED I.4.I.4). Under conditions of ammonia or nitrate limitation in a chemostat the assimilation of ammonia by cultures of R. leguminosarum, R. trifolii and R. japonicum proceeded via glutamine synthetase and glutamate synthase. Under glucose limitation and with an excess of inorganic nitrogen, ammonia was assimilated via glutamate dehydrogenase, neither glutamine synthetase nor glutamate synthase activities being detected in extracts. The coenzyme specificity of glutamate synthase varied according to species, being linked to NADP for the fast-growing R. leguminosarum, R. melitoti, R. phaseoli and R. trifolii but to NAD for the slow-growing R. japonicum and R. lupini. Glutamine synthetase, glutamate synthase and glutamate dehydrogenase activities were assayed in sonicated bacteroid preparations and in the nodule supernatants of Glycine max, Vicia faba, Pisum sativum, Lupinus luteus, Medicago sativa, Phaseolus coccineus and P. vulgaris nodules. All bacteroid preparations, except those from M. sativa and P. coccineus, contained glutamate synthase but substantial activities were found only in Glycine max and Lupinus luteus. The glutamine synthetase activities of bacteroids were low, although high activities were found in all the nodule supernatants. Glutamate dehydrogenase activity was present in all bacteroid samples examined. There was no evidence for the operation of the glutamine synthetase/glutamate synthase system in ammonia assimilation in root nodules, suggesting that ammonia produced by nitrogen fixation in the bacteroid is assimilated by enzymes of the plant system.     

Lipids in plant tissue cultures. VI. Effect of temperature on the lipids of Brassica napus and Tropaeolum majus cultures

The lipid contents of callus cultures of rape (Brassica napus) and nasturtium (Tropaeolum majus) increase in response to decreasing the temperature, though to different degree. Irrespective of the incubation temperature the lipids in cultures of both plants contain as predominant classes steryl glycosides, esterified steryl glycosides, sterols, steryl esters and fatty acids and, as minor constituents, various proportions of triacylglycerols, phospholipids and several unidentified fractions.The ratio of phospholipids to triacylglycerols as well as the ratio of diacylglycerophosphorylethanolamines to dicylglycerophosphorylcholines increase with time both in rape cultures incubated at 30°C and in those kept at 5°C.The lipids in rape and nasturtium cultures grown at 30°C contain smaller proportions of polyunsaturated fatty acids than the lipids in cultures incubated at 5°C. Erucic acid, the major constituent fatty acid of the seed lipids, in both plants, occurs only in trace amounts in the lipids of callus cultures. In contrast, linoleic and linolenic acids, which occur only in traces in the seed lipids of nasturtium, are major constituent fatty acids in the lipids of callus cultures derived from seedlings of this plant.The levels of constituent polyunsaturated fatty acids in the diacylglycerophosphorylethanolamines and the diacylglycerophosphorylcholines increase with time whereas in the triacylglycerols only linolenic acid is slightly increased.     

Bacterial breakdown of benomyl. II. Mixed cultures

Experiments with mixed bacterial cultures grown in liquid media which contained the benzimidazole fungicide benomyl, with or without Na-lactate, as source of carbon provided circumstantial evidence for cleavage of the benzimidazole heterocyclic ring. Yet, neither 2-aminobenzimidazole (2-AB) nor benzimidazole, as sole source of carbon, supported any bacterial growth. Total ¹⁴C-balance analysis experiments conclusively showed production of ¹⁴CO₂ from [2-¹⁴C] methyl benzimidazol-2-yl carbamate (MBC), and thus cleavage of the benzimidazole nucleus; bioassays, however, showed that the actual rate of benomyl and MBC breakdown was only small, the parent compound benomyl being still recovered in substantial quantities after up to 80 days of incubation. Therefore, cleavage of the benzimidazole ring is probably a matter of cometabolism, n-butylamine which originates from the butylcarbamoyl side chain serving as the proper source of carbon.Besides radiolabelled 2-AB and CO₂, an unknown metabolite was isolated which showed characteristics of a 2-AB-nucleotide. Probably, 2-AB was incorporated into bacterial DNA, which upon lysis of the bacterial cells gave rise to the nucleotide in question. Therefore, 2-AB might exert its inhibitory action by interfering with the normal functioning of DNA.