Carbon sources and their osmotic potential in plant tissue culture: does it matter (original) (raw)
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Review: role of carbon sources for in vitro plant growth and development
Molecular Biology Reports, 2012
In vitro plant cells, tissues and organ cultures are not fully autotrophic establishing a need for carbohydrates in culture media to maintain the osmotic potential, as well as to serve as energy and carbon sources for developmental processes including shoot proliferation, root induction as well as emission, embryogenesis and organogenesis, which are highly energy demanding developmental processes in plant biology. A variety of carbon sources (both reducing and non-reducing) are used in culture media depending upon genotypes and specific stages of growth. However, sucrose is most widely used as a major transport-sugar in the phloem sap of many plants. In micropropagation systems, morphogenetic potential of plant tissues can greatly be manipulated by varying type and concentration of carbon sources. The present article reviews the past and current findings on carbon sources and their sustainable utilization for in vitro plant tissue culture to achieve better growth rate and development.
Scientia Agricola, 2001
Cell suspension cultures of Bauhinia forficata Link, Curcuma zedoaria Roscoe and Phaseolus vulgaris L. were used to test plant ability to utilize an alternative to sucrose as carbon source and energy for growth. Glycerol, sorbitol and galactose were the alternative carbon sources tested. Cell suspension cultures established on liquid medium containing sucrose were transferred to culture medium supplemented with sucrose or glycerol, or sorbitol, or galactose as the sole carbon source. Fresh and dry weight increasing and protein content showed marked differences among the different carbon sources used. Sucrose was the best carbon source for all the three plant species tested. Galactose and glycerol promoted slow or no growth of the three studied species. Sucrose in liquid medium promoted initiation of meristemoid formation. Sorbitol, which was ineffective on promoting significant growth, was the only alternative carbon source tested that also promoted this effect.
The dynamic fluctuations of CO2 concentration in the tissue culture growth chamber after transplantation of petunia, chrysanthemum and tomato plantlets were recorded with a real-time control system to determine the critical CO2 concentration levels of 35 ll l 1 at which CO2 enrichment is needed. The experimental data showed that the tissue-cultured plantlets of petunia, chrysanthemum and tomato had the same CO2 concentration dynamics. The results indicated that CO2 enrichment was proper on the second day after transplantation. Petunia plantlets were used to conduct experiments under PPFD of 80 lmol m 2 s 1, and CO2 concentrations of 350 ± 50 ll l 1, 650 ± 50 ll l 1 and 950 ± 50 ll l 1 as well as medium moisture contents of 60%, 70% and 80%, with the result that plantlets grew better under CO2 concentration of 650 ± 50 ll l 1 than under the other two concentrations with all the different media water contents. Three media water contents under the same CO2 concentration produced plantlets with the same quality. The impacts of CO2 concentrations on plantlets are more important than those of the media water contents. Sugar-free tissue culture, as compared with the conventional culture, showed that CO2 enrichment to 350 ± 50 ll l 1 can promote the growth of the cultured plantlets. Sugar-free tissue culture produced healthy plantlets with thick roots, almost equivalent to the common plantlets.
Plant Production Science, 2000
The effects of various carbon sources, sucrose, glucose and fructose alone or in combination on the in vitro growth of banana plantlets were studied. Banana plants were cultured on the media supplemented with these carbon sources at 0.08 M for 13 weeks. The water potential of the medium was the highest in the medium supplemented with sucrose+ glucose (-0.3 MPa) , and was significantly lower in the medium supplemented with fructose alone or in combination with other carbon sources (-0.7 to-1.0 MPa) than in the other media. The leaf water potential was also the highest in the plants cultured on the medium supplemented with sucrose+ glucose, and lowest in the plants cultured on that with fructose. The leaf water potential of plants cultured on sucrose+ glucose, sucrose and glucose correlated well with their growth and photosynthetic activity, but the correlation was not observed in the plants cultured on fructose alone or in combination with other carbon sources. Plants cultured on fructose had a lower chlorophyll content (400 Jlg dm-2) and lower photosynthetic rate (3 Jlmol0 2 m-2 s-1) than those cultured on sucrose+glucose (15,950 Jlg dm-2 for chlorophyll and 8.5 Jlmol0 2 m-2 s-1 for photosynthesis), and these differences were statistically significant. Both chlorophyll content and photosynthetic oxygen evolution were the highest in the plants cultured on sucrose+ glucose, and the superior growth of plants on this medium was attributed to their high photosynthetic efficiency.
Culture-induced changes in osmolality of tobacco cell suspensions using four exogenous sugars
Plant Cell Tissue and Organ Culture, 1998
Suspension cultures of tobacco cells were grown in B5 media supplemented with sucrose, glucose, mannitol and sorbitol as exogenous sugars to examine culture-induced changes in the osmolality of the medium. Osmolality decreases were greatest in sucrose and glucose media during the 14 days in culture, and in glucose media were essentially linear, presumably reflecting the use of this sugar as
Plant Cell Reports, 2020
Key message Poplar callus maintained a specific difference in osmotic potential with respect to media when supplemented with different carbohydrate concentrations. This balance in osmotic potential guaranteed the growth capacity. Abstract Osmotic stress is caused by several abiotic factors such as drought, salinity, or freezing. However, the threshold of osmotic potential that allows the growth under stress conditions has not been thoroughly studied. In this study, different levels of osmotic stress in Populus alba (L.) callus have been induced with the addition of mannitol or sorbitol in the medium (from 0 to 500 mM). The key factor for preserving the growth was observed to be the restoration of a constant difference in osmotic potential between callus and medium for all the tested conditions. The osmotic adjustments were primarily achieved with the uptake of mannitol or sorbitol from the media considering their chemical properties instead of their biological functions. The decrease in water content (from − 1 to − 10% after 21 days) and mineral elements, such as potassium, calcium, and magnesium, together with the alterations in cell morphology, did not show negative effects on growth. The activity of sorbitol dehydrogenase was detected for the first time in poplar (+ 4.7 U l −1 in callus treated with sorbitol compared to control callus). This finding suggested the importance of choosing carefully the molecules used to exert osmotic stress for separating the dual function of carbohydrates in osmotic adjustments and cell metabolism.
Revista Brasileira de Botânica, 2000
This review considers the importance of compartmentation in the regulation of carbohydrate metabolism in leaves. We draw particular attention to the role of the vacuole as a site for storage of soluble sugars based on sucrose, and discuss briefly their characteristic metabolism. We also point out inconsistencies between the observed properties of vacuoles and the behaviour in vitro of the enzymes of fructan biosynthesis that do not support the hypothesis that the vacuole is the site of synthesis as well as of storage. We also consider compartmentation of carbohydrate metabolism between different cell types, using mainly our studies on leaves of temperate C3 gramineae. Here we present evidence of significant differences in carbon metabolism between epidermis, mesophyll, bundle sheath and vasculature based upon both single-cell sampling and immunolocalisation. The implications of these differences for the control of metabolism in leaves are discussed. RESUMO-(Compartimentação intracelular e intercelular do metabolismo de carboidratos em folhas de gramíneas de regiões temperadas). Esta revisão considera a importância da compartimentação para a regulação do metabolismo de carboidratos nas folhas, com destaque especial para o papel do vacúolo como local de armazenamento de carboidratos solúveis derivados da sacarose e seu característico metabolismo. Inconsistências entre as propriedades dos vacúolos e o comportamento in vitro das enzimas de biossíntese de frutanos são também destacadas, uma vez que não dão suporte à hipótese de que o vacúolo, é o local de síntese e armazenamento desses compostos. A compartimentação do metabolismo de carboidratos em diferentes tipos de células também é considerada em folhas de gramíneas C3 de regiões temperadas. Através de estudos com células isoladas e de imunolocalização são apresentadas evidências de diferenças significativas entre o metabolismo de carbono em epiderme, mesófilo, bainha de feixes e tecidos vasculares; são discutidas as implicações dessas diferenças no controle do metabolismo foliar.
Bioprocess and Biosystems Engineering, 2012
The kinetic behavior of the sugar uptake phenomena of a suspension of Arabidopsis cells was investigated by mid-infrared spectroscopy using Fourier transform infrared spectrometers and attenuated total reflection techniques. The kinetic behavior of the cell growth was also studied and the growth and the sugar uptake behaviors were discussed for three typical plant cells (Arabidopsis, TBY-2, and rice cells). The cell growth rate and the lag period were influenced by not only the types of the plant cells, but also the sugar species used as the carbon source. The characteristics of the sugar uptake behavior were clarified based on the difference in the three types of plant cells. The cell growth and the sugar uptake progressed at approximately the same time in the TBY-2 cells. In the rice cells, the sugar uptake rate was relatively lower than that of the others. On the other hand, the sugar uptake of the Arabidopsis cells started before the cell growth. Furthermore, glucose as the carbon source of the Arabidopsis cell cultivation seems to significantly influence the sugar metabolism. Glucose had a significant influence on the sugar metabolism of the other sugar under the conditions for the mixture of glucose and the other sugar. The characteristics of the sugar uptake phenomena based on the cell growth stage was typical for each plant cell except for some sugars, such as galactose and trehalose, and the behavior of the total sugar uptake had not changed. These results suggested that the cell growth and the sugar uptake in the plant cell cultivation processes may be controlled by the combined supply of the sugar species as the carbon source. The detailed data for plant cell cultivation using each sugar obtained in this study would be useful for bioscience research and for cultivation process control using various sugars, for example, purified or sugar mixtures formed from biomass materials.
Mixotrophic in vitro cultivations: the way to go astray in plant physiology
Physiologia Plantarum, 2018
Rate of photosynthesis and related plant carbohydrate status are crucial factors affecting plant vigour. Sugars providing carbon and energy sources serve also as important signalling molecules governing plant growth and development through a complex regulatory network. These facts are often neglected when mixotrophic cultivation of plants in vitro is used, where artificial exogenous sugar supply hinders studies of metabolism as well as sugar-driven developmental processes. We compared the growth, selected gasexchange parameters and sugar metabolism characteristics in four model plants, potato (Solanum tuberosum 'Lada'), tobacco (Nicotiana tabacum 'Samsun'), rapeseed (Brassica napus 'Asgard') and strawberry (Fragaria vesca), under both photomixotrophic (PM) and photoautotrophic (PA) conditions. To ensure PA conditions we used our improved sun caps that serve as gas and light permeable covers for cultivation vessels. We found bigger biomass accumulation, larger leaf areas, higher stomatal conductance and higher instantaneous water use efficiency and lower root sugar contents in PA plants compared to PM ones. However, for other characteristics (root biomass, root/shoot ratio, pigment contents, leaf sugar and starch levels and transpiration rates), a strong species-dependent reactions to the exogenous sugar supply was noted, which does not allow to create a general view on the overall impact of photomixotrophic nutrition under in vitro conditions.