Discrepancy in photosynthetic responses of the red alga Pyropia yezoensis to dehydration stresses under exposure to desiccation, high salinity, and high mannitol concentration

Macroalgae that inhabit intertidal zones are exposed to the air for several hours during low tide and must endure desiccation and high variations in temperature, light intensity, and salinity. Pyropia yezoensis (Rhodophyta, Bangiales), a typical intertidal red macroalga that is commercially cultivated in the northwestern Pacific Ocean, was investigated under different dehydration stresses of desiccation, high salinity, and high mannitol concentration. Using chlorophyll fluorescence imaging, photosynthetic activities of P. yezoensis thalli were analyzed using six parameters derived from quenching curves and rapid light curves. A distinct discrepancy was revealed in photosynthetic responses to different dehydration stresses. Dehydration caused by exposure to air resulted in rapid decreases in photosynthetic activities, which were always lower than two other stresses at the same water loss (WL) level. High salinity only reduced photosynthesis significantly at its maximum WL of 40% but maintained a relatively stable maximum quantum yield of photosystem II (PSII) (Fv/Fm). High mannitol concentration induced maximum WL of 20% for a longer time (60 min) than the other two treatments and caused no adverse influences on the six parameters at different WL except for a significant decrease in non-photochemical quenching (NPQ) at 20% WL. Illustrated by chlorophyll fluorescence images, severe spatial heterogeneities were induced by desiccation with lower values in the upper parts than the middle or basal parts of the thalli. The NPQ and rETRmax (maximum relative electron transport rate) demonstrated clear distinctions for evaluating photosynthetic responses, indicating their sensitivity and applicability. The findings of this study indicated that the natural dehydration of exposure to air results in stronger and more heterogeneous effects than those of high salinity or high mannitol concentration.

Efficient isolation and purification of tissue-specific protoplasts from tea plants (Camellia sinensis (L.) O. Kuntze)

Background Plant protoplasts constitute unique single-cell systems that can be subjected to genomic, proteomic, and metabolomic analysis. An effective and sustainable method for preparing protoplasts from tea plants has yet to be established. The protoplasts were osmotically isolated, and the isolation and purification procedures were optimized. Various potential factors affecting protoplast preparation, including enzymatic composition and type, enzymatic hydrolysis duration, mannitol concentration in the enzyme solution, and iodixanol concentration, were evaluated. Results The optimal conditions were 1.5% (w/v) cellulase and 0.4–0.6% (w/v) macerozyme in a solution containing 0.4 M mannitol, enzymatic hydrolysis over 10 h, and an iodixanol concentration of 65%. The highest protoplast yield was 3.27 × 106 protoplasts g−1 fresh weight. As determined through fluorescein diacetate staining, maximal cell viability was 92.94%. The isolated protoplasts were round and regularly shaped without agglomeration, and they were less than 20 μm in diameter. Differences in preparation, with regard to yield and viability in the tissues (roots, branches, and leaves), cultivars, and cultivation method, were also observed. Conclusions In summary, we reported on a simple, efficient method for preparing protoplasts of whole-organ tissue from tea plant. The findings are expected to contribute to the rapid development of tea plant biology.

Morphology Regulation Mechanism and Enhancement of Photocatalytic Performance of BiOX (X=Cl, Br, I) via Mannitol-Assisted Synthesis

BiOX (X=Cl, Br, I) photocatalysts with dominant (110) facets were synthesized via a mannitol-assisted solvothermal method. This is the first report on the exposed (110) facets-, size-, and defects-controlled synthesis of BiOX achieved by solvothermal synthesis with mannitol. This polyol alcohol acted simultaneously as a solvent, capping agent, and/or soft template. The mannitol concentration on the new photocatalysts morphology and surface properties was investigated in detail. At the lowest concentration tested, mannitol acted as a structure-directing agent, causing unification of nanoparticles, while at higher concentrations, it functioned as a solvent and soft template. The effect of exposed (110) facet and surface defects (Bi(3−x)+, Bi4+, Bi5+) of BiOX on the photocatalytic activity of nanomaterials under the UV–Vis irradiation were evaluated by oxidation of Rhodamine B (RhB) and 5-fluorouracil (5-FU), an anticancer drug, and by reduction of Cr(VI). Additionally, the influence of crucial factors on the formation of BiOX in the synthesis with mannitol was discussed extensively, and the mechanism of BiOX formation was proposed. These studies presented a new simple method for synthesizing BiOX without any additional surfactants or shape control agents with good photocatalytic activity. The study also provided a better understanding of the effects of solvothermal conditions on the BiOX crystal growth.

The study of the consequences of preserving berry crops in vitro on the processes of subsequent clonal micropropagation

As a result of studies, it was found that collection samples of berry crops in vitro can be maintained in viable condition on specially selected nutrient media for 12 months. The effect of mannitol concentration (0.45, 0.75 and 1.05%; sucrose control -3%) on the regenerative potential of microprobe of the studied berry crop varieties was estimated. When studying the effect of the reproduction cycle on the morphogenesis of meristemic cultures, it was found that the dynamics of changes in shoot length, number of leaves and shoots depended on the concentration of the active substance. In general, the majority of berry crops that were viable after storage of explants retained the ability to grow again, in many cases reaching or exceeding the control level.

Microencapsulation of Bifidobacterium animalis subsp. lactis BB-12 with mannitol

Bifidobacterium animalis subsp. lactis BB-12 (BB-12) was microencapsulated using co-extrusion technology with chitosan coating and the incorporation of mannitol as prebiotic. Optimization of coating material chitosan concentration (0–0.5% w/v) and mannitol concentration (0–5% w/v) as prebiotic were performed to determine the formulation that produces beads with desired properties. The microencapsulation efficiency (MEE) of free and microencapsulated BB-12 (with and without mannitol) were determined. All forms of BB-12 further subjected to sequential digestion in simulated gastric juice (SGJ, pH 2.0) for 2 hours and simulated intestinal juice (SIJ, pH 7.5) for 3 hours. The results indicated that 0.4% (w/v) of chitosan coating and 3% (w/v) of mannitol were the optimum concentrations to produce microencapsulated BB-12 with the highest MEE of 89.15% and the average bead size of 805 µm. The BB-12 beads produced through co-extrusion were spherical with a smooth surface. Throughout the five hours sequential gastrointestinal digestion, both microencapsulated BB-12 with and without mannitol were able to maintain their viable cell count at least 106 CFU/g at the end of the incubation. The presence of prebiotic mannitol showed a significant protective effect on the microencapsulated BB-12 during gastrointestinal transit.

In vitro response of three contrasting cassava (Manihot esculenta Crantz) varieties to mannitol-induced drought stress

In vitro selection of drought-tolerant cassava varieties is essential for rapid breeding for drought tolerance. The objectives of this study were to determine the response of three contrasting cassava varieties to mannitol-induced drought stress to establish its suitability for in vitro screening and examine relationships among growth parameters. Plantlets were raised from nodal segments on Murashige and Skoog (MS) medium containing 0 (control), 5, 10, 15, 20, 25 and 30 g/l mannitol. Variety CH 140 had the highest survival of explants and frequency of root formation, while MV 99/0395 recorded the highest number of chlorotic leaves and the lowest survival of explants. The lowest numbers of leaves were produced at 25 and 30 g/l mannitol by the three varieties. In CH 140, the highest number of leaves was produced in medium free of mannitol, while the highest number of leaves was produced at 5 and 10 g/l mannitol in MV 99/0395 and TMS 01/1206, respectively. In TMS 01/1206, number of roots produced decreased as the concentration of mannitol in culture media increased, whereas in CH140, number of roots increased as the concentration of mannitol increased before decreasing; while in MV 99/0395, number of roots was not affected by an increase in mannitol concentration. As the concentration of mannitol in the culture media increased shoot height of plantlets decreased with a sharp decline at 20 mg/l mannitol. Concentration of mannitol and survival of explants had significant negative correlation with all parameters. However, frequency of root formation only had significant positive correlation with shoot length. The study concluded that differential responses were expressed by the three varieties to mannitol-induced drought stress and mannitol at 20 g/l concentration was a suitable in vitro drought inducing-agent for screening cassava varieties for drought tolerance.

In vitro response of three contrasting cassava (Manihot esculenta Crantz) varieties to mannitol-induced drought stress

In vitro selection of drought-tolerant cassava varieties is essential for rapid breeding for drought tolerance. The objectives of this study were to determine the response of three contrasting cassava varieties to mannitol-induced drought stress to establish its suitability for in vitro screening and examine relationships among growth parameters. Plantlets were raised from nodal segments on Murashige and Skoog (MS) medium containing 0 (control), 5, 10, 15, 20, 25 and 30 g/l mannitol. Variety CH 140 had the highest survival of explants and frequency of root formation, while MV 99/0395 recorded the highest number of chlorotic leaves and the lowest survival of explants. The lowest numbers of leaves were produced at 25 and 30 g/l mannitol by the three varieties. In CH 140, the highest number of leaves was produced in medium free of mannitol, while the highest number of leaves was produced at 5 and 10 g/l mannitol in MV 99/0395 and TMS 01/1206, respectively. In TMS 01/1206, number of roots produced decreased as the concentration of mannitol in culture media increased, whereas in CH140, number of roots increased as the concentration of mannitol increased before decreasing; while in MV 99/0395, number of roots was not affected by an increase in mannitol concentration. As the concentration of mannitol in the culture media increased shoot height of plantlets decreased with a sharp decline at 20 mg/l mannitol. Concentration of mannitol and survival of explants had significant negative correlation with all parameters. However, frequency of root formation only had significant positive correlation with shoot length. The study concluded that differential responses were expressed by the three varieties to mannitol-induced drought stress and mannitol at 20 g/l concentration was a suitable in vitro drought inducing-agent for screening cassava varieties for drought tolerance.

Carbohydrate Content and Root Growth in Seeds Germinated Under Salt Stress

Sugars and sugar alcohols have well-documented roles in salt tolerance of whole plants and maturing seeds. Less is known, however, about possible effects of these compounds during germination. Seeds from mannitol-accumulating salt-tolerant celery [Apium graveolens L. var. dulce (P. Mill.) DC], non-mannitol-accumulating salt-tolerant cabbage (Brassica oleracea L. var. capitata L. ‘Golden Acre’), and salt-sensitive non-mannitol-accumulating tobacco (Nicotiana tabacum L.) and arabidopsis [Arabidopsis thaliana (L.) Heynh.] were placed on vertical Phytagel plates containing 0 to 300 mm NaCl. Germination percentage, root elongation, and carbohydrate content of seeds and seedlings were assessed. With the exception of cabbage, there was no positive relationship between ability to germinate in NaCl and the reported species salt tolerance of the mature plant. For instance, while cabbage seeds germinated in 300 mm NaCl, germination of two celery cultivars was inhibited completely by 150 mm NaCl. In contrast, seeds from salt-sensitive tobacco and arabidopsis germinated in 200 mm NaCl. There was also no obvious relationship between the observed salt tolerance and total soluble carbohydrates in either non-imbibed seeds or in seedlings germinated in salt. For example, the most-salt tolerant species in these studies, cabbage, had the third highest seed and seedling carbohydrate concentration, while the next most tolerant, arabidopsis, had the lowest. However, both species contained significant amounts of the osmoprotective oligosaccharides raffinose or stachyose. In addition, although celery seedling mannitol concentration initially increased at low NaCl concentrations (50 mm), germination and mannitol concentration decreased at higher NaCl concentrations (100 mm). Finally, the broadest response observed was a large increase in seedling sucrose at the lowest salt concentration that significantly inhibited germination. Although most seeds, with the notable exception of cabbage, did not germinate at 150 mm NaCl, they were still metabolically active because the sucrose content was two to eight times higher than in non-imbibed seeds, suggesting a possible role for sucrose in salt-stressed germinating seeds. These results not only suggest that mechanisms providing salt tolerance in mature plants are different from those in germinating seeds, but also that, even when the same mechanisms are employed, they may be less effective in seeds.