Is there a role for oligosaccharides in seed longevity? An assessment of intracellular glass stability

We examined whether oligosaccharides extend seed longevity by increasing the intracellular glass stability. For that purpose, we used a spin probe technique to measure the molecular mobility and glass transition temperature of the cytoplasm of impatiens (Impatiens walleriana) and bell pepper (Capsicum annuum) seeds that were osmo-primed to change oligosaccharide content and longevity. Using saturation transfer electron paramagnetic resonance spectroscopy, we found that the rotational correlation time of the polar spin probe 3-carboxy-proxyl in the cytoplasm decreased, together with longevity, as a function of increasing seed water content, suggesting that longevity may indeed be regulated by cytoplasmic mobility. Osmo-priming of the seeds resulted in considerable decreases in longevity and oligosaccharide content, while the sucrose content increased. No difference in the glass transition temperature was found between control and primed impatiens seeds at the same temperature and water content. Similarly, there was no difference in the rotational motion of the spin probe in the cytoplasm between control and primed impatiens and bell pepper seeds. We therefore conclude that oligosaccharides in seeds do not affect the stability of the intracellular glassy state, and that the reduced longevity after priming is not the result of increased molecular mobility in the cytoplasm.     

Low-Moisture Food: A Physicochemical Approach to Investigate the Origin of Their Physical Instability versus Water or Sucrose

Low-moisture biopolymer-based systems are commonly encountered in food. Obviously, understanding the physical basis of their quality [texture, or performance over time or as a function of their composition (water or other added solutes)] is of primary importance. A polymer science approach using physical chemistry concepts based on physical state, phase transitions and molecular mobility can be applied to investigate the performances of food in particular versus moisture. Based on the example of starch-based samples and their texture property changes versus composition, the role of water and sucrose is considered through different aspects. The relations existing between the observed changes and physical state are investigated. While the motions associated with the glass transition were observed at high temperature, secondary relaxations are observed below Tg (at T _β): T _β decreased with water content and increased with increasing sucrose content. These local motions are suggested to contribute to the observed texture modifications versus water. Moreover, the stability of the glassy state was investigated by differential scanning calorimetry through the study of enthalpy relaxation (physical ageing). The amplitude of enthalpy relaxation decreased with both increasing sucrose and water content. All in all, this study strengthened the hypotheses that sub-Tg mobility could contribute to texture instability versus moisture or sugar content.     

Influence of Water Content and Temperature on Molecular Mobility and Intracellular Glasses in Seeds and Pollen

Although the occurrence of intracellular glasses in seeds and pollen has been established, physical properties such as rotational correlation times and viscosity have not been studied extensively. Using electron paramagnetic resonance spectroscopy, we examined changes in the molecular mobility of the hydrophilic nitroxide spin probe 3-carboxy-proxyl during melting of intracellular glasses in axes of pea (Pisum sativum L.) seeds and cattail (Typha latifolia L.) pollen. The rotational correlation time of the spin probe in intracellular glasses of both organisms was approximately 10⁻³ s. Using the distance between the outer extrema of the electron paramagnetic resonance spectrum (2A_zz) as a measure of molecular mobility, we found a sharp increase in mobility at a definite temperature during heating. This temperature increased with decreasing water content of the samples. Differential scanning calorimetry data on these samples indicated that this sharp increase corresponded to melting of the glassy matrix. Molecular mobility was found to be inversely correlated with storage stability. With decreasing water content, the molecular mobility reached a minimum, and increased again at very low water content. Minimum mobility and maximum storage stability occurred at a similar water content. This correlation suggests that storage stability might be at least partially controlled by molecular mobility. At low temperatures, when storage longevity cannot be determined on a realistic time scale, 2A_zz measurements can provide an estimate of the optimum storage conditions.     

Enthalpy relaxation of freeze concentrated sucrose-water glass

The enthalpy relaxation of freeze concentrated sucrose-water glass was investigated using 40% sucrose, differential scanning calorimetry (DSC) with isothermal ageing for 1-6 days at various temperatures (-70, -65, -60, and -55 degrees C). The enthalpy relaxation was observed as an endothermic peak superimposed on the endothermic step-wise change due to the glass transition around -47 degrees C. The enthalpy relaxation was found to increase with ageing time and temperature. An 80% sucrose glass was also investigated at ageing temperatures of -60 and -65 degrees C, and this material exhibited a similar glass transition and enthalpy relaxation to that observed with the frozen 40% sucrose solution. The calculated activation energy of the enthalpy relaxation of the sucrose-water glass was smaller than that reported for pure sucrose. These results suggest that the freeze concentrated sucrose-water glass could have a higher molecular mobility and less stability than pure sucrose glass.     

Effects of thermal denaturation on the solid-state structure and molecular mobility of glycinin

The effects of moisture and thermal denaturation on the solid-state structure and molecular mobility of soy glycinin powder were investigated using multiple techniques that probe over a range of length and time scales. In native glycinin, increased moisture resulted in a decrease in both the glass transition temperature and the denaturation temperature. The sensitivity of the glass transition temperature to moisture is shown to follow the Gordon-Taylor equation, while the sensitivity of the denaturation temperature to moisture is modeled using Flory's melting point depression theory. While denaturation resulted in a loss of long-range order, the principal conformational structures as detected by infrared are maintained. The temperature range over which the glass to rubber transition occurred was extended on the high temperature side, leading to an increase in the midpoint glass transition temperature and suggesting that the amorphous regions of the newly disordered protein are less mobile. (13)C NMR results supported this hypothesis.     

Melting, glass transition, and apparent heat capacity of α-d-glucose by thermal analysis

The thermal behaviors of α-d-glucose in the melting and glass transition regions were examined utilizing the calorimetric methods of standard differential scanning calorimetry (DSC), standard temperature-modulated differential scanning calorimetry (TMDSC), quasi-isothermal temperature-modulated differential scanning calorimetry (quasi-TMDSC), and thermogravimetric analysis (TGA). The quantitative thermal analyses of experimental data of crystalline and amorphous α-d-glucose were performed based on heat capacities. The total, apparent and reversing heat capacities, and phase transitions were evaluated on heating and cooling. The melting temperature (T_m) of a crystalline carbohydrate such as α-d-glucose, shows a heating rate dependence, with the melting peak shifted to lower temperature for a lower heating rate, and with superheating of around 25 K. The superheating of crystalline α-d-glucose is observed as shifting the melting peak for higher heating rates, above the equilibrium melting temperature due to of the slow melting process. The equilibrium melting temperature and heat of fusion of crystalline α-d-glucose were estimated. Changes of reversing heat capacity evaluated by TMDSC at glass transition (T_g) of amorphous and melting process at T_m of fully crystalline α-d-glucose are similar. In both, the amorphous and crystalline phases, the same origin of heat capacity changes, in the T_g and T_m area, are attributable to molecular rotational motion. Degradation occurs simultaneously with the melting process of the crystalline phase. The stability of crystalline α-d-glucose was examined by TGA and TMDSC in the melting region, with the degradation shown to be resulting from changes of mass with temperature and time. The experimental heat capacities of fully crystalline and amorphous α-d-glucose were analyzed in reference to the solid, vibrational, and liquid heat capacities, which were approximated based on the ATHAS scheme and Data Bank.     

Influence of thermal history on the stability of gelatin gels

Gelatin gel properties have been studied using three techniques. Optical rotation measurements have shown that the lower the ageing temperature, the faster the helix content increases but the lower the helix stability. Rheological measurements show that a small increase in temperature leads to a melting of some junction zones followed by a build up of new ones. By ageing the gel at two successive temperatures one can show the existence of two populations of junction zones with different thermal stabilities. The same result is shown on melting thermograms obtained by differential scanning calorimetry. All these results are consistent with the hypothesis of the presence of junction zones of various lengths, the thermostability of which being proportional to their lengths.     

Characterization of molecular mobility in seed tissues: an electron paramagnetic resonance spin probe study.

The relationship between molecular mobility (tauR) of the polar spin probe 3-carboxy-proxyl and water content and temperature was established in pea axes by electron paramagnetic resonance (EPR) and saturation transfer EPR. At room temperature, tauR increased during drying from 10(-11) s at 2.0 g water/g dry weight to 10(-4) s in the dry state. At water contents below 0.07 g water/g dry weight, tauR remained constant upon further drying. At the glass transition temperature, tauR was constant at approximately 10(-4) s for all water contents studied. Above Tg, isomobility lines were found that were approximately parallel to the Tg curve. The temperature dependence of tauR at all water contents studied followed Arrhenius behavior, with a break at Tg. Above Tg the activation energy for rotational motion was approximately 25 kJ/mol compared to 10 kJ/mol below Tg. The temperature dependence of tauR could also be described by the WLF equation, using constants deviating considerably from the universal constants. The temperature effect on tauR above Tg was much smaller in pea axes, as found previously for sugar and polymer glasses. Thus, although glasses are present in seeds, the melting of the glass by raising the temperature will cause only a moderate increase in molecular mobility in the cytoplasm as compared to a huge increase in amorphous sugars.     

Melting, glass transition, and apparent heat capacity of α-D-glucose by thermal analysis

The thermal behaviors of α-D-glucose in the melting and glass transition regions were examined utilizing the calorimetric methods of standard differential scanning calorimetry (DSC), standard temperature-modulated differential scanning calorimetry (TMDSC), quasi-isothermal temperature-modulated differential scanning calorimetry (quasi-TMDSC), and thermogravimetric analysis (TGA). The quantitative thermal analyses of experimental data of crystalline and amorphous α-D-glucose were performed based on heat capacities. The total, apparent and reversingheat capacities, and phase transitions were evaluated on heating and cooling. The melting temperature (T(m)) of a crystalline carbohydrate such as α-D-glucose, shows a heating rate dependence, with the melting peak shifted to lower temperature for a lower heating rate, and with superheating of around 25K. The superheating of crystalline α-D-glucose is observed as shifting the melting peak for higher heating rates, above the equilibrium melting temperature due to of the slow melting process. The equilibrium melting temperature and heat of fusion of crystalline α-D-glucose were estimated. Changes of reversing heat capacity evaluated by TMDSC at glass transition (T(g)) of amorphous and melting process at T(m) of fully crystalline α-D-glucose are similar. In both, the amorphous and crystalline phases, the same origin of heat capacity changes, in the T(g) and T(m) area, are attributable to molecular rotational motion. Degradation occurs simultaneously with the melting process of the crystalline phase. The stability of crystalline α-D-glucose was examined by TGA and TMDSC in the melting region, with the degradation shown to be resulting from changes of mass with temperature and time. The experimental heat capacities of fully crystalline and amorphous α-D-glucose were analyzed in reference to the solid, vibrational, and liquid heat capacities, which were approximated based on the ATHAS scheme and Data Bank.     

Carbohydrate metabolism before and after dehiscence in the recalcitrant pollen of pumpkin (Cucurbita pepo L.)

Pumpkin (Cucurbita pepo L.) pollen is starchy, sucrose‐poor and recalcitrant, features opposite to those of several model species; therefore, some differences in carbohydrate metabolism could be expected in this species. By studying pumpkin recalcitrant pollen, the objective was to provide new biochemical evidence to improve understanding of how carbohydrate metabolism might be involved in pollen functioning in advanced stages. Four stages were analysed: immature pollen from 1 day before anthesis, mature pollen, mature pollen exposed to the environment for 7 h, and pollen rehydrated in a culture medium. Pollen viability, water and carbohydrate content and activity of enzymes involved in carbohydrate metabolism were quantified in each stage. Pollen viability and water content dropped quickly after dehiscence, as expected. The slight changes in carbohydrate concentration and enzyme activity during pollen maturation contrast with major changes recorded with ageing and rehydration. Pumpkin pollen seems highly active and closely related to its surrounding environment in all the stages analysed; the latter is particularly evident among insoluble sucrolytic enzymes, mainly wall‐bound acid invertase, which would be the most relevant for sucrose cleavage. Each stage was characterised by a particular metabolic/enzymatic profile; some particular features, such as the minor changes during maturation, fast sucrolysis upon rehydration or sharp decrease in insoluble sucrolytic activity with ageing seem to be related to the lack of dormancy and recalcitrant nature of pumpkin pollen.     

Naked Plasmid DNA Formulation: Effect of Different Disaccharides on Stability after Lyophilisation

Since plasmid DNA (pDNA) is unstable in solution, lyophilisation can be used to increase product shelf life. To prevent stress on pDNA molecules during lyophilisation, cryo- and lyoprotectants have to be added to the formulation. This study assessed the effect of disaccharides on naked pDNA stability after lyophilisation using accelerated stability studies. Naked pDNA was lyophilised with sucrose, trehalose, maltose or lactose in an excipient/DNA w/w ratio of 20. To one part of the vials extra residual moisture was introduced by placing the vials half opened in a 25°C/60% RH climate chamber, before placing all vials in climate chambers (25°C/60% RH and 40°C/75% RH) for stability studies. An ex vivo human skin model was used to assess the effect of disaccharides on transfection efficiency. Lyophilisation resulted in amorphous cakes for all disaccharides with a residual water content of 0.8% w/w. Storage at 40°C/75% RH resulted in decreasing supercoiled (SC) purity levels (sucrose and trehalose maintained approximately 80% SC purity), but not in physical collapse. The addition of residual moisture (values between 7.5% and 10% w/w) resulted in rapid collapse except for trehalose and decreasing SC purity for all formulations. In a separate experiment disaccharide formulation solutions show a slight but significant reduction (<3% with sucrose and maltose) in transfection efficiency when compared to pDNA dissolved in water. We demonstrate that disaccharides, like sucrose and trehalose, are effective lyoprotectants for naked pDNA.     

Occurrence of mono- or disaccharides and polysaccharide reserves in mature pollen grains

 Pollen from 13 species of gymnosperms and angiosperms was studied for soluble and insoluble carbohydrates at dispersal. Starch reserves stored during pollen development give rise to carbohydrates at maturity. Combinations of different types of carbohydrates in mature pollen may depend on the extent of starch hydrolysis. An inverse relationship was found between the extent of starch hydrolysis and sucrose content. If the starch was scarcely de-polymerized, the cytoplasm had very low levels of soluble sugars and none of the periodic acid-Schiff (PAS)-positive material as found in pollen not subject to high dehydration (Cucurbita pepo L., Zea mays L.). After total or partial starch hydrolysis, insoluble PAS-positive oligo/polysaccharides were found in the cytoplasm associated with much soluble sugar, and the pollen grains were dehydrated at dispersal as in Typha latifolia L., Chamaerops humilis L., Trachycarpus excelsa Wendl., and other specimens. Intermediate levels of starch and soluble sugars, together with cytoplasmic PAS-positive material, characterized species with dehydrated pollen such as Pinus halepensis Miller. Carbohydrates may be related to pollen longevity, which largely depends on the abundance of sucrose, which is known to protect membrane integrity. The relationship between PAS-positive material and pollen viability is unclear at present. Received: 30 July 1996 / Revision accepted: 18 December 1996     

Glass transitions in soybean seed : relevance to anhydrous biology

We have investigated the mechanism by which anhydrobiotic organisms can survive severe dehydration. The method used was measurement of the rotational diffusion coefficient of a hydrophilic spin probe, inserted in the cytoplasm of soybean (Glycine max L.) axes, as a function of temperature and sample water content. Results indicate the existence of a hydration-dependent glass-like transition at physiological temperatures. No glass transitions have been observed in desiccation-intolerant samples, suggesting that the ability to withstand dehydration is associated with glass formation.     

Organization of lipid reserves in cotyledons of primed and aged sunflower seeds

Imbibing sunflower (Helianthus annuus L., cv. Briosol) seeds at water potentials between –2 MPa and –5 MPa leads to faster (priming) or slower (accelerated ageing) germination depending on the temperature and duration of treatment. Mobilization of food reserves may be associated with the changes in seed vigor. To study this, morphological, biochemical and phase properties of lipid, the major food reserve in sunflower, were compared in freshly harvested (i.e., control), primed and aged sunflower cotyledons using electron microscopy, biochemical analyses and differential scanning calorimetry, respectively. Lipid bodies became smaller and more dispersed throughout the cytoplasm during priming and ageing. Despite ultrastructural changes, there were few measured changes in biochemistry of the neutral lipid component; lipid content, proportion of saturated and unsaturated fatty acids and level of free fatty acids were unchanged in primed and slightly aged seeds, with only severely aged seeds showing a net decrease in polyunsaturated fatty acids and an increase in free fatty acids. Subtle changes in the calorimetric behavior of lipids within sunflower cotyledons were observed. Sunflower lipids exhibited polymorphic crystalline and amorphous solid phases when cooled to <–100°C, but priming decreased the rate of crystallization in vivo and ageing increased the rate of crystallization, but decreased percentage crystallinity. The observed changes in thermal behavior in vivo are consistent with losses and gains, respectively, of interacting non-lipid moieties in the triacylglycerol matrix.     

土壤湿度对东北3种主要树种凋落物分解的影响

结合丰林国家自然保护区原始阔叶红松林1998-2017年表层(0～10 cm)土壤湿度监测数据,评估了全球变化背景下土壤湿度变化对东北主要森林树种红松、臭冷杉和白桦凋落物分解的影响.结果表明: 在同一土壤湿度水平下,凋落物分解速率随着凋落物质量的增加而增大,即表现为白桦>臭冷杉>红松.凋落物的分解速率随着土壤湿度的降低而减小.白桦、臭冷杉和红松3种凋落物的土壤湿度敏感性指数(M₁₀)分别为0.782、0.789和0.827,土壤湿度水平每降低10%,初始分解速率分别减小21.8%、21.1%和17.3%.高质量凋落物(高氮含量、低碳氮比、低木质素含量)的分解速率对土壤湿度变化的响应更敏感.凋落物分解速率在不同凋落物类型间的差异随着土壤湿度的降低而缩小.近20年间,原始阔叶红松林土壤湿度呈显著减少趋势,对凋落物分解表现为抑制作用.在全球变化背景下,随着气温的升高,土壤湿度将继续降低,对凋落物的抑制作用会进一步增强,并将部分抵消因温度升高所带来的凋落物分解速率增大的压力.     

Fast pollen tube growth in Conospermum species

BACKGROUND AND AIMS: An unusual form of pollen tube growth was observed for several Conospermum species (family Proteaceae). The rate of pollen tube growth, the number of tubes to emerge and the ultrastructure of these tubes are given here. METHODS: Pollen was germinated in vitro in different sucrose concentrations and in the presence of calcium channel blockers, and tube emergence and growth were recorded on a VCR. Measurements were taken of the number of tubes to emerge and rate of tube emergence. Pollen behaviour in vivo was also observed. The ultrastructure of germinated and ungerminated pollen was observed using TEM. RESULTS: After 10 s to 3 min in germination medium, up to three pollen tubes emerged and grew at rates of up to 55 micro m s(-1); the rate then slowed to around 2 micro m s(-1), 30 s after the initial growth spurt. Tubes were observed to grow in pulses, and the pulsed growth continued in the presence of calcium channel blockers. Optimal sugar concentration for pollen germination was 300 g L(-1), in which up to 81 % of pollen grains showed fast germination. Germination and emergence of multiple tubes were observed in sucrose concentrations of 100-800 g L(-1). The vegetative and generative nuclei moved into one of the tubes. Multiple tubes from a single grain were observed on the stigma. Under light microscopy, the cytoplasm in the tube showed a clear region at the tip. The ultrastructure of C. amoenum pollen showed a bilayered exine, with the intine being very thick at the pores, and elsewhere having large intrusions into the plasma membrane. The cytoplasm was dense with vesicles packed with inner tube cell wall material. Golgi apparatus producing secretory vesicles, and mitochondria were found throughout the tube. The tube wall was bilayered; both layers being fibrous and loosely packed. CONCLUSIONS: It is proposed that, for Conospermum, initial pollen tube wall constituents are manufactured and stored prior to pollen germination, and that tube extension occurs as described in the literature for other species, but at an exceptionally fast rate.     

Ripe pollen carbohydrate changes in Trachycarpus fortunei: the effect of relative humidity

Pollen of the palm Trachycarpus fortunei was kept at 25°C and relative humidities (RH) of 20, 55 and 98%. Changes in viability, water content and carbohydrates were measured over 2–17 days. Water content remained almost constant at 20 and 50% RH and increased dramatically at 98%. Pollen viability and germination rate remained almost constant over 14 days at 20% RH and decreased to about 2% after 7–9 days at 55% and to even less at 98% RH. Although the three experimental conditions were constant, qualitative and quantitative variations in pollen carbohydrates were recorded, even after pollen had lost its viability. The quantities of mono-, di- and polysaccharides varied with the period of pollen storage at the various RH. The greatest changes in glucose, fructose and sucrose content were recorded at 55 and 98% RH. At these relative humidities, maximum glucose and fructose content and minimum sucrose content occurred at maximum water content. Starch was not present in mature pollen but appeared and peaked after 7–9 days of pollen storage at 55 and 98%. Appearance of starch coincided with an increase in pectin content. PAS-positive cytoplasmic polysaccharides showed an increasing trend at 20% RH. A relation was found between pollen viability, water content and monosaccharide content. Pollen viability and germination capacity remained high at 20% RH for 14 days. At this relative humidity, pollen water, glucose and fructose contents remained almost constant, while sucrose reached its maximum value. The fluctuations of more complex carbohydrates (starch, pectins and PAS-positive cytoplasmic polysaccharides) were less easy to interpret. Changes observed under experimental conditions could simulate processes occurring in nature during pollen presentation and dispersal.     

Citrate increases glass transition temperature of vitrified sucrose preparations

The aim of this study was to investigate the effect of sodium citrate on the properties of dried amorphous sucrose glasses. Addition of sodium citrate to a sucrose solution followed by freeze-drying or convective drying resulted in a glass transition temperature (Tg) that was higher than the well-studied sucrose Tg. This result was obtained either at reduced water content of the analysed sample or by removal of water during Modulated DSC analysis. After removal of the remaining water ( < 3.5% w/w), a Tg of approximately 105 degrees C was obtained at a mass ratio of sodium citrate to sucrose of 0.3. FTIR analysis showed a similar increase in Tg as was found with Modulated DSC analysis. The Tg values were derived from breaks in the vibrational frequency vs. temperature plots in the OH stretching and bending regions. Elevated average strength of hydrogen bonding in the sucrose/citrate glass was concluded from the downshift of the OH stretching band of 25 cm(-1) and from the reduced wavenumber temperature coefficient (WTC). The antisymmetric carboxylate stretch of citrate sensed the glass transition of the mixture, from which we conclude that citrate interacts with the sucrose OH via its carboxylate groups.     

Differential desiccation sensitivity of corn and Pennisetum pollen linked to their sucrose contents

Abstract. Two pollen species from the Gramineae family were investigated as to their differential sensitivity to desiccation. Germination in vivo ceased to occur after previous drying to 7–8% moisture content in Zea mays and 3% in Pennisetum typhoides. The reduced vitality coincided with extensive membrane damage, as evidenced by the considerable leakage of fluorescein and K⁺ from the prehumidified grains into liquid media. Further experiments were undertaken to investigate the basis for this interspecific difference: (1) phospholipid analyses revealed little difference in composition and content both before and after drying; (2) free fatty acid contents increased in both species upon drying but levels were generally low; and (3) sucrose was the sole soluble carbohydrate found in both species. Fresh maize pollen contained 5% sucrose, as compared to 14% in Pennisetum pollen. During slow drying (8h) this level went up to 12% and 17%, respectively. Drying corn pollen in the cold (2°C), or at high rate, limited sucrose levels and affected the resistance to drying. The authors conclude that survival of dehydration is correlated with the presence of sucrose. Finally, they tested the ability of sucrose to preserve dry liposomes prepared from phospholipids purified from the two species. When liposomes were dried in the presence of sucrose, fusion and leakage could largely be prevented at mass ratio's of sugar to lipid of > 4. Trehalose was also effective but myo-inositol was not. No species differences were observed. The authors suggest that the presence of sucrose is a key factor in preserving membranes in dry pollen.     

Change in sugar, sterol and fatty acid composition in banana meristems caused by sucrose-induced acclimation and its effects on cryopreservation

To understand the mechanisms of sucrose‐induced acclimation in relation to plant cryopreservation, sugars, sterols, fatty acids of different lipid fractions (neutral lipids, glycolipids and sphingolipids and phospholipids), as well as free fatty acids were analyzed in proliferating meristem cultures of different banana varieties. The four banana varieties that were selected show different post‐thaw shoot regeneration rates (0–53.4%). All mentioned parameters were analyzed using (1) control meristems that were cultured on a normal sucrose concentration (0.09 M), which resulted in low survival after cryopreservation; and (2) 2‐week sucrose precultured meristems (0.4 M). This sucrose preculture, essential for regeneration after cryopreservation, resulted in a significant increase of each of seven sugars detected. The ratio of stigmasterol/sitosterol (St/Si) in sucrose‐pretreated meristems significantly increased. The sucrose pretreatment also resulted in a significant increase of total fatty acid content of the neutral lipid fraction and of the glycolipid and sphingolipid fraction, as well as the total free fatty acid content. The individual fatty acid content of the phospholipids was differently changed by the sucrose pretreatment for the given varieties studied. In most cases, sucrose pretreatment resulted in an increase of the double bond index (DBI) in the neutral lipids and a decrease of DBI in the glycolipids and sphingolipids, in phospholipids as well as in free fatty acids. Principal component analysis of all collected data revealed that (1) for the control material, sucrose and total sugar contents were closely linked to the post‐thaw shoot regeneration, suggesting that sucrose and total sugar may be main limiting factors to survive cryopreservation; (2) accumulation of large quantities of sugars (glucose, fructose, sucrose and total sugar) in sucrose‐pretreated material cannot explain the differences in survival after cryopreservation of the four banana varieties. We assume that a minimal amount of sugars is needed in meristem cultures to survive cryopreservation. Still, other limiting factors do influence the survival following the sucrose pretreatment. We observed that the parameters which are closely linked to the post‐thaw shoot regeneration are a minimal change in the ratios of St/Si, the minimal change of the DBI of phospholipids and free fatty acids, as well as linoleic acid content (C18:2); and (3) inositol, raffinose, myristic acid (C14:0) and oleic acid (C18:1) were present in small quantities; however, they could be correlated to survival after cryopreservation, suggesting that they may be also involved in cryopreservation process.