Artur Zdunek

Sensing the structural differences in cellulose from apple and bacterial cell wall materials by Raman and FT-IR spectroscopy.

Raman and Fourier Transform Infrared (FT-IR) spectroscopy was used for assessment of structural differences of celluloses of various origins. Investigated celluloses were: bacterial celluloses cultured in presence of pectin and/or xyloglucan, as well as commercial celluloses and cellulose extracted from apple parenchyma. FT-IR spectra were used to estimate of the Iβ content, whereas Raman spectra were used to evaluate the degree of crystallinity of the cellulose. The crystallinity index (XCRAMAN%) varied from −25% for apple cellulose to 53% for microcrystalline commercial cellulose. Considering bacterial cellulose, addition of xyloglucan has an impact on the percentage content of cellulose Iβ. However, addition of only xyloglucan or only pectins to pure bacterial cellulose both resulted in a slight decrease of crystallinity. However, culturing bacterial cellulose in the presence of mixtures of xyloglucan and pectins results in an increase of crystallinity. The results confirmed that the higher degree of crystallinity, the broader the peak around 913 cm−1. Among all bacterial celluloses the bacterial cellulose cultured in presence of xyloglucan and pectin (BCPX) has the most similar structure to those observed in natural primary cell walls.

The relation of apple texture with cell wall nanostructure studied using an atomic force microscope

In this study, the relation of the nanostructure of cell walls with their texture was investigated for six different apple cultivars. Cell wall material (CWM) and cellulose microfibrils were imaged by atomic force microscope (AFM). The mean diameter of cellulose microfibrils for each cultivar was estimated based on the AFM height topographs obtained using the tapping mode of dried specimens. Additionally, crystallinity of cellulose microfibrils and pectin content was determined. Texture of apple cultivars was evaluated by sensory and instrumental analysis. Differences in cellulose diameter as determined from the AFM height topographs of the nanostructure of cell walls of the apple cultivars are found to relate to the degree of crystallinity and pectin content. Cultivars with thicker cellulose microfibrils also revealed crisper, harder and juicier texture, and greater acoustic emission. The data suggest that microfibril thickness affects the mechanical strength of cell walls which has consequences for sensory and instrumental texture.

Imaging of polysaccharides in the tomato cell wall with Raman microspectroscopy

BackgroundThe primary cell wall of fruits and vegetables is a structure mainly composed of polysaccharides (pectins, hemicelluloses, cellulose). Polysaccharides are assembled into a network and linked together. It is thought that the percentage of components and of plant cell wall has an important influence on mechanical properties of fruits and vegetables.ResultsIn this study the Raman microspectroscopy technique was introduced to the visualization of the distribution of polysaccharides in cell wall of fruit. The methodology of the sample preparation, the measurement using Raman microscope and multivariate image analysis are discussed. Single band imaging (for preliminary analysis) and multivariate image analysis methods (principal component analysis and multivariate curve resolution) were used for the identification and localization of the components in the primary cell wall.ConclusionsRaman microspectroscopy supported by multivariate image analysis methods is useful in distinguishing cellulose and pectins in the cell wall in tomatoes. It presents how the localization of biopolymers was possible with minimally prepared samples.

Relation of Biospeckle Activity with Quality Attributes of Apples

Biospeckle is nondestructive optical technique based on the analysis of variations of laser light scattered from biological samples. Biospeckle activity reflects the state of the investigated object. In this study the relation of biospeckle activity (BA) with firmness, soluble solids content (SSC), titratable acidity (TA) and starch content (SC) during the shelf life of seven apple cultivars was studied. The results showed that the quality attributes change significantly during storage. Significant and pronounced positive correlation between BA and SC was found. This result shows that degradation of starch granules, which could be stimulated to vibration by intracellular cyclosis, causes a lesser number of laser light scattering centers and results in smaller apparent biospeckle activity.

Determination of the Elastic Properties of Tomato Fruit Cells with an Atomic Force Microscope

Since the mechanical properties of single cells together with the intercellular adhesive properties determine the macro-mechanical properties of plants, a method for evaluation of the cell elastic properties is needed to help explanation of the behavior of fruits and vegetables in handling and food processing. For this purpose, indentation of tomato mesocarp cells with an atomic force microscope was used. The Youngs modulus of a cell using the Hertz and Sneddon models, and stiffness were calculated from force-indentation curves. Use of two probes of distinct radius of curvature (20 nm and 10,000nm) showed that the measured elastic properties were significantly affected by tip geometry. The Youngs modulus was about 100 kPa ± 35 kPa and 20 kPa ± 14 kPa for the sharper tip and a bead tip, respectively. Moreover, large variability regarding elastic properties (>100%) among cells sampled from the same region in the fruit was observed. We showed that AFM provides the possibility of combining nano-mechanical properties with topography imaging, which could be very useful for the study of structure-related properties of fruits and vegetables at the cellular and sub-cellular scale.

Combining FT-IR spectroscopy and multivariate analysis for qualitative and quantitative analysis of the cell wall composition changes during apples development

The aim of this work was to quantitatively and qualitatively determine the composition of the cell wall material from apples during development by means of Fourier transform infrared (FT-IR) spectroscopy. The FT-IR region of 1500-800 cm(-1), containing characteristic bands for galacturonic acid, hemicellulose and cellulose, was examined using principal component analysis (PCA), k-means clustering and partial least squares (PLS). The samples were differentiated by development stage and cultivar using PCA and k-means clustering. PLS calibration models for galacturonic acid, hemicellulose and cellulose content from FT-IR spectra were developed and validated with the reference data. PLS models were tested using the root-mean-square errors of cross-validation for contents of galacturonic acid, hemicellulose and cellulose which was 8.30 mg/g, 4.08% and 1.74%, respectively. It was proven that FT-IR spectroscopy combined with chemometric methods has potential for fast and reliable determination of the main constituents of fruit cell walls.

Study on dietary fibre by Fourier transform-infrared spectroscopy and chemometric methods.

Fresh fruit is an important part of the diet of people all over the world as a significant source of water, vitamins and natural sugars. Nowadays it is also one of the main sources of dietary fibre. In fruit the dietary fibre is simply cell wall consisting essentially of polysaccharides. The aim of present study was to predict the contents of pectins, cellulose and hemicelluloses by partial least squares regression (PLS) analysis on the basis of Fourier transform-infrared (FT-IR) spectra of fruit cell wall residue. The second purpose was to analyse the composition of dietary fibre from fruit based on FT-IR spectral information in combination with chemometric methods (principle components analysis (PCA) and hierarchical cluster analysis (HCA)). Additionally the contents of polysaccharides in studied fruits were determined by analytical methods. It has been shown that the analysis of infrared spectra and the use of multivariate statistical methods can be useful for studying the composition of dietary fibre.

Application of the Biospeckle Method for Monitoring Bull’s Eye Rot Development and Quality Changes of Apples Subjected to Various Storage Methods—Preliminary Studies

In this study, the biospeckle technique was evaluated for monitoring of apple bull’s eye rot development and product quality in general, during storage under various conditions and during subsequent shelf life. This non-destructive optical method is based on the analysis of laser light variations scattered from the sample. Apples of the cultivars ‘Pinova’ and ‘Topaz’, susceptible to bull’s eye rot, were used in two independent experiments. In the first, apples were non-destructively monitored for five months during cold storage. After that time, 34% of ‘Pinova’ and 21% of ‘Topaz’ apples displayed visible surface lesions. The increase of biospeckle activity was observed during the development of fungal disease. In the second experiment various storage conditions were used and apples were tested during their shelf life by non-destructive and destructive methods. This study showed that biospeckle activity decreased during shelf life, irrespective of storage conditions.

Determination of the Optimum Harvest Window for Apples Using the Non-Destructive Biospeckle Method

Determination of the optimum harvest window plays a key role in the agro-food chain as the quality of fruit depends on the right harvesting time and appropriate storage conditions during the postharvest period. Usually, indices based on destructive measurements are used for this purpose, like the De Jager Index (PFW-1), FARS index and the most popular Streif Index. In this study, we proposed a biospeckle method for the evaluation of the optimum harvest window (OHW) of the “Ligol” and “Szampion” apple cultivars. The experiment involved eight different maturity stages, of which four were followed by long cold storage and shelf life to assist the determination of the optimum harvest window. The biospeckle activity was studied in relation to standard quality attributes (firmness, acidity, starch, soluble solids content, Streif Index) and physiological parameters (respiration and ethylene emission) of both apple cultivars. Changes of biospeckle activity (BA) over time showed moderate relationships with biochemical changes during apple maturation and ripening. The harvest date suggested by the Streif Index and postharvest quality indicators matched with characteristic decrease in BA. The ability of biospeckle method to characterize the biological state of apples was confirmed by significant correlations of BA with firmness, starch index, total soluble solids and Streif Index, as well as good match with changes in carbon dioxide and ethylene emission. However, it should be noted that correlations between variables changing over time are not as meaningful as independent observations. Also, it is a well-known property of the Pearson’s correlation that its value is highly susceptible to outlier data. Due to its non-selective nature the BA reflected only the current biological state of the fruit and could be affected by many other factors. The investigations showed that the optimum harvest window for apples was indicated by the characteristic drop of BA during pre-harvest development. Despite this, at the current state of development the BA method cannot be used as an indicator alone. Due to rather poor results for prediction in OHW the BA measurements should be supported by other destructive methods to compensate its low selectivity.

Changes of pectin nanostructure and cell wall stiffness induced in vitro by pectinase

Structural modifications of fruit cell-wall pectins are controlled by various enzymes. In this in vitro study, the cell wall material (CWM) from pear fruit (Pyrus communis L.) was treated using pectinases in two concentrations. Water soluble (WSP), chelator soluble (CSP) and sodium carbonate soluble (DASP) pectin fractions were extracted from CWM. By visualization of enzymatic-induced changes of structure and CWM stiffness using an atomic force microscopy (AFM), the role of pectins in the mechanical properties of cell walls was shown. Galacturonic acid (GalA) content in pectin fractions was assayed as well. This experiment unveiled evidence of the structural degradation of molecules in pectin fractions extracted from CWM caused by in vitro pectinase action and softening of CWM due to pectin removal that might be related to the creation of empty spaces in the cellulose-hemicellulose network.