Effect of pile fermentation on the cells of Chinese Liupao tea: The first record of cell wall of Liupao tea on transmission electron microscope.

Fermentation often degrades the cell wall of dark tea, changes the carbohydrate components in the cell wall of tea, and thus affects the quality of tea. However, the lack of ultrastructural details limits our knowledge on the effect of fermentation on tea cell walls. Morphological studies of cell structures are important; thus, the cell wall of Liupao tea was analyzed under transmission electron microscopy for the first time, and the effects of different raw materials and fermentation methods on the cell wall and main carbohydrates of tea were compared. Overall, fermentation degrades the cell wall of Liupao tea under the action of microorganisms. Interestingly, the middle lamella degrades obviously, whereas the primary wall is complete. The decrease in hemicellulose and increase in water-soluble pectin (WSP) were remarkable, whereas the changes in cellulose and WSP were considerably correlated with the increase in tea polysaccharide (TPS). The results suggest that cell wall degradation might be related to the increase in TPS.

Tender leaf and fully-expanded leaf exhibited distinct cuticle structure and wax lipid composition in Camellia sinensis cv Fuyun 6.

The goal of the present study was to compare the structural and compositional differences of cuticle between tender leaf and fully-expanded leaf in Camellia sinensis, and provide metabolic base for the further characterization of wax biosynthesis in this economically important crop species. The tender second leaf and the fully-expanded fifth leaf from new twig were demonstrated to represent two different developmental stages, their cuticle thickness were measured by transmission electron microscopy. The thickness of the adaxial cuticle on the second and fifth leaf was 1.15 µm and 2.48 µm, respectively; the thickness of the abaxial cuticle on the second and fifth leaf was 0.47 µm and 1.05 µm, respectively. The thickness of the epicuticular wax layer from different leaf position or different sides of same leaf were similar. However, the intracuticular wax layer of the fifth leaf was much thicker than that of the second leaf. Total wax lipids were isolated from the second leaf and the fifth leaf, respectively. Gas chromatography-mass spectrometry analysis identified 51 wax constituents belonging to 13 chemical classes, including esters, glycols, terpenoids, fatty acids and their derivatives. Wax coverage on the second and fifth leaf was 4.76 µg/cm2 and 15.38 µg/cm2, respectively. Primary alcohols dominated in the tender second leaf. However, triterpenoids were the major components from the fully-expanded fifth leaf. The predominant carbon chains varied depending on chemical class. These data showed that the wax profiles of Camellia sinensis leaves are development stage dependent, suggesting distinct developmental dependent metabolic pathways and regulatory mechanisms.

Using magnetically responsive tea waste to remove lead in waters under environmentally relevant conditions.

We report the use of a simple yet highly effective magnetite-waste tea composite to remove lead(II) (Pb(2+)) ions from water. Magnetite-waste tea composites were dispersed in four different types of water-deionized (DI), artificial rainwater, artificial groundwater and artificial freshwater-that mimic actual environmental conditions. The water samples had varying initial concentrations (0.16-5.55 ppm) of Pb(2+) ions and were mixed with the magnetite-waste tea composite for at least 24 hours to allow adsorption of the Pb(2+) ions to reach equilibrium. The magnetite-waste tea composites were stable in all the water samples for at least 3 months and could be easily removed from the aqueous media via the use of permanent magnets. We detected no significant leaching of iron (Fe) ions into the water from the magnetite-waste tea composites. The percentage of Pb adsorbed onto the magnetite-waste tea composite ranged from ∼70% to 100%; the composites were as effective as activated carbon (AC) in removing the Pb(2+) ions from water, depending on the initial Pb concentration. Our prepared magnetite-waste tea composites show promise as a green, inexpensive and highly effective sorbent for removal of Pb in water under environmentally realistic conditions.

Microwaves and tea: new tools to process plant tissue for transmission electron microscopy.

Optimizing sample processing, reducing the duration of the preparation of specimen, and adjusting procedures to adhere to new health and safety regulations, are the current challenges of plant electron microscopists. To address these issues, plant processing protocols for TEM, combining the use of polyphenolic compounds as substitute for uranyl acetate with microwave technology are being developed. In the present work, we optimized microwave-assisted processing of different types of plant tissue for ultrastuctural and immunocytochemical studies. We also explored Oolong tea extract as alternative for uranyl acetate for the staining of plant samples. We obtained excellent preservation of cell ultrastructure when samples were embedded in epoxy resin, and of cell antigenicity, when embedded in LR-White resin. Furthermore, Oolong tea extract successfully replaced uranyl acetate as a counterstain on ultrathin sections, and for in block staining. These novel protocols reduce the time spent at the bench, and improve safety conditions for the investigator. The preservation of the cell components when following these approaches is of high quality. Altogether, they offer significant simplification of the procedures required for electron microscopy of plant ultrastructure.

Investigation of morphological change of green tea polysaccharides by SEM and AFM.

The objective of this study is to investigate the morphological structure and its change of green tea polysaccharides (GTPS) before and after enzyme reaction by scanning electron microscope (SEM) and atomic force microscope (AFM). Before enzyme reaction, with the novel sample preparation method SEM images of GTPS have obtained many branches and network structures. After enzyme reaction, the morphological structure of GTPS changed, and surface roughness increased. The microstructure of GTPS from SEM with the novel sample preparation method was in accordance with the results from AFM with the tapping mode. The results indicate that the novel sample preparation of GTPS for SEM is a simple, feasible, and reliable method for observing the surface morphology.

Removal of Ni(II) ions from aqueous solutions using waste of tea factory: adsorption on a fixed-bed column.

The removal efficiency of waste tea from nickel containing aqueous solutions was investigated. All experiments were conducted fixed-bed columns. Experiments were carried out as a function of liquid flow rate (5-20 mL/min), initial Ni(II) concentration (50-200 mg/L), bed height (10-30 cm), pH of feed solution (2.0-5.0) and particle size (0.15-0.25 to 1.0-3.0 mm) of adsorbent. The total adsorbed quantities, equilibrium uptakes and total removal percents of Ni(II) related to the effluent volumes were determined by evaluating the breakthrough curves obtained at different flow rates, different inlet Ni(II) concentrations, different pH value, different bed height and different particle size for waste tea. The longest breakthrough time and maximum of Ni(II) adsorption is obtained at pH 4.0. Decrease in the particle size from 1.0-3.0 to 0.15-0.25 mm resulted in significant increase in the treated volume, breakthrough time and bed capacity. The results show that the column performed well at lowest flow rate. Also, column bed capacity and exhaustion time increased with increasing bed height. When the initial Ni(II) concentration is increased from 50 to 200 mg/L, the corresponding adsorption bed capacity appears to increase from 7.31 to 11.17 mg/g. The bed depth service time (BDST) model and the Thomas model were used to analyze the experimental data and the model parameters were evaluated. Good agreement of the experimental breakthrough curves with the model predictions was observed.

Gravitropic reaction in the growth of tea roots.

In Japan, tea (Camellia sinenis (L.) Kuntze) seedlings are propagated by cutting. A root system of clonal plants by cutting consists of adventitious roots and lateral roots. Most of the roots grow horizontally, which results in a shallow distribution of the root system. Such a shallow root system could be one of the factors contributing to the deterioration of nutrient uptake and resistance to water stress. Gravitropism of the roots is considered to be a decisive factor that controls the depth of a root system. The authors have investigated changes in the growth direction of roots to gravitative stimulus, using several kinds of roots (seminal roots, lateral roots and adventitious roots). Furthermore, amyloplasts in the root-cap cells, which are considered to be an equipment sensing gravistimulus, were observed. Seminal roots prominently showed orthogravitropism and contained many amyloplast particles in their root cap cells. Most lateral and adventitious roots showed plagiogravitropism, growing in an angle to gravistimulus, and lacked observable amyloplast particles in their root cap cells. The results suggest that the shallowing of root systems of elonal tea plants could be attributed to a gravitropic reaction of the adventitious and lateral roots composing the root system. There could also be a close relationship between the growth direction of roots and the presence of amyloplasts in root-cap cells.