Cell Cultures Of Higher Plants Research Articles

Chlorella as the source of biologically active compounds for in vitro plant propagation

The use of nutrient media with an organic composition is a potentially commercially effective way to avoid the use of expensive components of nutrient media while maintaining and increasing the biosynthetic potential of cell cultures of higher plants in vitro. A method has been developed for the in vitro propagating of various taxonomic groups plants. It has been shown that the medium after cultivating chlorella at ¼ MS is optimal for the in vitro plant propagation. The method is intended for in vitro cultivation of seeds, microcuttings and microshoots of plants of different taxonomic groups and can be used for accelerated propagation of valuable plant varieties and hybrids.

Biotechnology based on cell cultures of higher plants

This paper reviews the role of plant cell culture as a biotechnological tool in preserving the botanical diversity of higher plants while meeting the growing demand of the commercial market for large volumes of plant raw material. The prospects of plant cell-based technology are discussed in the framework of creating an economy of sustainable development in the short and long term.

Получение и характеристика каллусных культур клеток Alhagi persarum Boiss. et Buhse - продуцентов изофлавоноидов

The physiological characteristics of the callus cell cultures of Alhagi persarum Boiss et Buhse, a member of the legume family, widely used in folk medicine, have been studied. It was shown that the source of the explant was an important factor in the initiation of callusogenesis: more intense callusogenesis (almost 100%) was observed for explants from various organs of sterile seedlings, rather than intact plants (less than 30%). As a result, more than 20 lines of morphologically different callus cell cultures were obtained, and the growth parameters for the 5 most intensively growing lines were determined. The composition of fatty acids (FA) of total lipids and secondary metabolites in the most physiologically stable callus line Aр-207 was analyzed. Using capillary gas-liquid chromatography with mass spectrometric detection (GLC-MS), 19 individual C12--C24 FAs were identified, the main fraction of which were palmitic (~ 23%), stearic (~ 22%), linoleic (~ 14%) and α-linolenic (~ 33%) acids. The established atypical ratio of FAs (a simultaneous high content of both saturated FAs and polyunsaturated α-linolenic acid) is possibly due to the adaptation of cells to in vitro growth conditions. Phytochemical analysis of the secondary metabolites was carried out using ultra-performance liquid chromatography with electrospray ionization mass spectrometric detection (UPLC MS). Compounds belonging to different structural groups of isoflavones were found. Aglycones (calycosin, formononetin and afrormosin isomer), glucosides (formononetin glucoside), as well as esters of glucosides (malonylglycosides of calicosin, formononetin, afrormosin isomers, glycitein and genistein) were detected. These secondary metabolites are widespread in plants of the Fabaceae family; however, isoflavones are rare in representatives of the Alhagi genus. The presence of malonylated isoflavone glycosides in Alhagi spp. was shown for the first time. endemic plant species, Alhagi, in vitro cell culture, callus cell culture, isoflavones, fatty acids All studies were carried out using the equipment of the "Experimental Biotechnological Facility" and the "All-Russian Collection of Cell Cultures of Higher Plants" of IРР RAS. This work was supported by the Russian Foundation for Basic Research (RFBR), contract no.18-54-06021 (Az_a), and the Government of the Russian Federation, Megagrant Project no. 075-15-2019-1882.

Evidence for 18O labeling of photorespiratory CO2 in photoautotrophic cell cultures of higher plants illuminated in the presence of 18O2

The 18O-enrichment of CO2 produced in the light or during the post-illumination burst was measured by mass spectrometry when a photoautotrophic cell suspension of Euphorbia characias L. was placed in photorespiratory conditions in the presence of molecular 18O2. The only 18O-labeled species produced was C18O16O; no C18O18O could be detected. Production of C18O16O ceased after addition of two inhibitors of the photosynthetic carbon-oxidation cycle, aminooxyacetate or aminoacetonitrile, and was inhibited by high levels of CO2. The average enrichment during the post-illumination burst was estimated to be 46 ± 15% of the enrichment of the O2 present during the preceding light period. Addition of exogenous carbonic anhydrase, by catalyzing the exchange between CO2 and H2O, drastically diminished the 18O-enrichment of the produced CO2. The very low carbonio-anhydrase level of the photoautotrophic cell suspension probably explains why the 18O labeling of photorespiratory CO2 could be observed for the first time. These data allow the establishment of a direct link between O2 consumption and CO2 production in the light, and the conclusion that CO2 produced in the light results, at least partially, from the mitochondrial decarboxylation of the glycine pool synthesized through the photosynthetic carbon-oxidation cycle. Analysis of the C18O16O and CO2 kinetics provides a direct and reliable way to assess in vivo the real contribution of photorespiratory metabolism to CO2 production in the light.

Selecting in Higher Plant Cell Cultures. A Discussion

Summary Procedures of somatic cell genetics provide additional methods for manipulation in plant breeding which must, however, be standardized to be used effectively and successfully. The expectations connected with the employment of selection in higher plant cell cultures have not been fulfilled yet. Undoubtedly, this is mainly due to the lack of adapted selection procedures. Cell cultures of higher plants possess qualities that have to be taken into account when elaborating selection procedures: (1) cell suspensions mostly contain cell colonies; (2) higher plant cell cultures are characterized by genetic instability; (3) cell cultures do not form expected traits, but exhibit traits in another form or develop new ones; (4) useful individual plants have to be regenerated from single cells in most cases; (5) plating efficiency is reduced; (6) spontaneous selection processes may diminish the range of traits selectable from cell cultures. These problems as well as advantages and drawbacks of selection procedures in current use are discussed.

Heat shock proteins from cell cultures of higher plants and their somatic hybrids

The species specificity of heat shock proteins of callus cultures of Nicotiana chinensis, Nicotiana glauca, Nicotiana tabacum, Atropa belladonna, Lycopersicon peruvianum, as well as some somatic hybrids of A. belladonna + N. chinensis, was investigated by means of polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate. Despite the general similarity of electrophoretic mobility of heat shock proteins from different plants, a number of species-specific differences between the proteins of distantly related genera were found. Heat shock proteins may thus serve as genetic markers in somatic hybridization.

A Comparison of 2,4-Dichlorophenoxyacetic Acid Metabolism in Cultured Soybean Cells and in Embryogenic Carrot Cells

The removal or reduction in concentration of auxin is often a successful method for obtaining morphogenesis in cell cultures of higher plants, such as carrot, but not for soybean. For this reason, the metabolism of one auxin, 2,4-dichlorophenoxyacetic acid (2,4-D), was compared in both carrot and soybean cells. Whereas soybean cells conjugated a high percentage of their 2,4-D to amino acids, carrot cells contained primarily free 2,4-D. Moreover, after long-term exposure to 2,4-D, carrot cells released much more 2,4-D upon transfer to 2,4-D-free (embryogenic) medium than did soybean cells. It appears that the retention of 2,4-D by soybean cells might interfere with subsequent morphogenesis. Because no impairment of 2,4-D efflux was found with short-term exposure to radiolabeled 2,4-D, it was concluded that 2,4-D retention in soybean cells might be due to a time-dependent, metabolic process. The conjugation of 2,4-D to amino acids was shown to be one such time-dependent process. Additionally, the release of 2,4-D from the cells was shown to be due primarily to a loss of free 2,4-D and not 2,4-D-amino acid conjugates. It seems that the greater retention of 2,4-D by soybean cells upon transfer to 2,4-D-free medium is due to greater formation of 2,4-D-amino acid conjugates.

Localisation and characterization of the fatty acid synthesizing system in cells of Glycine max (soybean) suspension cultures

In course of a study of fatty acid synthetase in higher plants, non-green cell suspension cultures of Glycine max (soybean) served as model tissues. For the first time, a fatty acid synthesizing system was characterized in cell cultures of higher plants and was found to be solely located in proplastids of the cells. Optimum activity of the fatty acid synthesizing system in proplastids was observed between pH 8.0 and 8.2; with [1- 14C]acetate as substrate, cofactors required were CoA, ATP, Mn 2+, Mg 2+, HCO 3 −, NADH and NADPH. The system was more sensitive towards NADH than NADHP. [1- 14C]Acetate, [2- 14C]-malonate and [3- 14C]pyruvate served as precursors for fatty acids, indicating the presence of pyruvate dehydrogenase activity in proplastids. In disrupted proplastids, [2- 14C]malonylCoA was a better precursor than [1- 14C]acetylCoA. After incubation of proplastids with [2- 14C]malonate, a small shift, from palmitic acid to higher homologs, of label incorporated was observed, as compared to incorporation of label from [1- 14C]acetate and [3- 14C]pyruvate. Under the conditions of the experiment, only small amounts of polyunsaturated fatty acids, the main fatty acid components of this organelle, were synthesized. In respect to fatty acid synthesis, the non-green cell suspension culture resembles photosynthetic leaf tissue.

Extracellular polysaccharide from cell suspension cultures of bush bean (Phaseolus vulgaris cv. Contender)

High yields of extracellular polysaccharide were obtained from cell suspension cultures of root, hypocotyl, and cotyledon of bush bean. Hydrolysates of the three polysaccharide samples contained the same sugars: galacturonic acid, galactose, glucose, mannose, arabinose, and xylose. The relative amounts of the six sugars were not the same in the hydrolysates from the three sources. The extracellular polysaccharide was produced at all times during the culture cycle. Semilogarithmic plots of increase in cell number, and production of extracellular polysaccharide, indicate that production per cell decreased during the logarithmic phase, and increased at the onset of the stationary phase. Production of extracellular polysaccharide, per culture and per cell, was much higher than that reported for other cell cultures of higher plants.