Colocalising proteins and polysaccharides in plants for cell wall and trafficking studies.

Plant cell walls (PCWs) are intricate structures with complex polysaccharides delivered by distinct trafficking routes. Unravelling the intricate trafficking pathways of polysaccharides and proteins involved in PCW biosynthesis is a crucial first step towards understanding the complexities of plant growth and development. This study investigated the feasibility of employing a multi-modal approach that combines transmission electron microscopy (TEM) with molecular-genetic tagging and antibody labelling techniques to differentiate these pathways at the nanoscale. The genetically encoded electron microscopy (EM) tag APEX2 was fused to Arabidopsis thaliana cellulose synthase 6 (AtCESA6) and Nicotiana alata ARABINAN DEFICIENT LIKE 1 (NaARADL1), and these were transiently expressed in Nicotiana benthamiana leaves. APEX2 localization was then combined with immunolabeling using pectin-specific antibodies (JIM5 and JIM7). Our results demonstrate distinct trafficking patterns for AtCESA6 and NaARADL, with AtCESA6 localized primarily to the plasma membrane and vesicles, while NaARADL1 was found in the trans-Golgi network and cytoplasmic vesicles. Pectin epitopes were observed near the plasma membrane, in Golgi-associated vesicles, and in secretory vesicle clusters (SVCs) with both APEX2 constructs. Notably, JIM7 labelling was found in vesicles adjacent to APEX2-AtCESA6 vesicles, suggesting potential co-trafficking. This integrative approach offers a powerful tool for elucidating the dynamic interactions between PCW components at the nanoscale level. The methodology presented here facilitates the precise mapping of protein and polysaccharide trafficking pathways, advancing our understanding of PCW biosynthesis and providing avenues for future research aimed at engineering plant cell walls for various applications.

Unlocking the Potential of Food Waste: A Review of Multifunctional Pectins.

This review comprehensively explores the multifunctional applications of pectins derived from food waste and by-products, emphasizing their role as versatile biomaterials in the medical-related sectors. Pectins, known for their polyelectrolytic nature and ability to form hydrogels, influence the chemical composition, sensory properties, and overall acceptability of food and pharmaceutical products. The study presents an in-depth analysis of molecular parameters and structural features of pectins, such as the degree of esterification (DE), monosaccharide composition, galacturonic acid (GalA) content, and relative amounts of homogalacturonan (HG) and rhamnogalacturonan I (RG-I), which are critical for their technofunctional properties and biological activity. Emphasis is placed on pectins obtained from various waste sources, including fruits, vegetables, herbs, and nuts. The review also highlights the importance of structure-function relationships, especially with respect to the interfacial properties and rheological behavior of pectin solutions and gels. Biological applications, including antioxidant, immunomodulatory, anticancer, and antimicrobial activities, are also discussed, positioning pectins as promising biomaterials for various functional and therapeutic applications. Recalled pectins can also support the growth of probiotic bacteria, thus increasing the health benefits of the final product. This detailed review highlights the potential of using pectins from food waste to develop advanced and sustainable biopolymer-based products.

Aerial and terrestrial root habits influence the composition of the cell walls of Vanilla phaeantha (Orchidaceae).

In response to the restrictions imposed by their epiphytic habit, orchids have developed structural traits that allow greater efficiency in water uptake and use, such as a complex adventitious root system with velamen. The composition of cell wall of this specialized epidermis can be altered according to the substrate to which it is fixed, influencing wall permeability, absorption, and storage of water in roots. The current study aimed to evaluate the cell wall composition of adventitious roots of Vanilla phaeantha (Orchidaceae) that grow attached to the phorophyte, fixed in the soil, or hung free. Immunocytochemical analyses were used to determine the protein, hemicellulose, and pectin composition of the cell walls of aerial and terrestrial roots. We observed that pectins are present in the different tissues of the aerial roots, while in the terrestrial roots, they are concentrated in the cortical parenchyma. The deposition of xyloglucans, extensins, and arabinogalactans was greater in the epidermis of the free side of the roots attached to the phorophyte. The strong labeling of pectins in aerial roots may be related to the influx of water and nutrients, which are generally scarce in this environment. The arrangement of hemicelluloses and proteins with the pectins may be associated with increased cell rigidity and sustainability, a feature of interest for the aerial roots. In summary, the habit of roots can interfere with the non-cellulosic composition of the cell walls of V. phaeantha, possibly related to changes in cell functionality.

Differential effects of enzymatically modified Ougan (Citrus Suavissima Hort. ex Tanaka) peel pectins extracted with different methods on inhibiting the proliferation of Hela cells.

Previous studies have shown that modified citrus pectin (MCP) is an anti-tumor material of food grade. In this study, two enzymatically modified Ougan (Citrus Suavissima Hort. ex Tanaka) peel pectins (EMP1 and EMP2, the ones extracted by alkali and enzymatic methods) were used to investigate their differential effects on viability and physiology of Hela cells. The results showed that EMP1 and EMP2 had 88.00 % and 81.01 % galacturonic acid, 21.31 % and 20.25 % esterification degree, 10,417 g/mol and 6416 g/mol molecular weight (Mw), 82.86 % and 50.62 % RG-I, and 8.91 % and 15.70 % HG, respectively. EMP2 had higher intensities of absorption peaks than EMP1. They were irregularly shaped, with more holes on EMP1 but more wrinkles on EMP2. Both could inhibit the growth, proliferation, migration, and invasion of HeLa cells in a concentration-dependent manner, with better efficiency in EMP2. Meanwhile, EMP2 was more efficient than EMP1 in blocking the cell cycle in S phase, resulting in apoptosis. In conclusion, the variations caused by extraction resulted in differences in anti-tumor activity of MCP and EMP2 with lower Mw and higher HG exhibited better anti-tumor effects. This study would provide an experimental basis and reference for the research and development of anti-tumor supplements from citrus pectin.

A time-course analysis of Aspergillus terreus secretomes reveals the importance of pectin-degrading enzymes to increase the digestibility of soybean meal.

Considering an ever-growing global population, which hit 8 billion people in the fall of 2022, it is essential to find solutions to avoid croplands competition between human food and animal feed. Agricultural co-products such as soybean meals have become important components of the circular economy thanks to their use in animal feed. Their implementation was made possible by the addition of exogenous enzymes in the diet of monogastric animals, especially fungal carbohydrate-active enzymes (CAZymes). Here, we describe a time-course production and analysis of Aspergillus terreus secretomes for the identification of CAZymes able to enhance the digestibility of soybean meals. Functional assays revealed that the release of nutrients and the degradation of pectins in soybean meals can be tightly interconnected. Using a comparative proteomics approach, we identified several fungal pectin-degrading enzymes leading to increased assimilable nutrients in the soluble fraction of soybean meals. Our results reinforce the importance of deconstructing pectic polysaccharides in feedstuffs and contribute to sharpen our understanding of the fungal enzymatic interplays involved in pectin hydrolysis.IMPORTANCEIn the present study, we developed a strategy to identify the key fungal enzymatic activities involved in the improvement of soybean meal (SBM) digestibility. Our data unravel the importance of pectin degradation for the release of nutrients from SBM and provide some insights regarding the degradation of rhamnogalacturonan-I (RG-I) by ascomycetes. Indeed, the hydrolysis of pectins and RG-I by human microbiota is well documented in the literature, but our knowledge of the fungal CAZymes at play for the degradation of soybean pectins remains hitherto underexplored. Due to its wide use in animal feed, improving the digestibility of SBM by enzymatic treatments is a current challenge for feed additive suppliers. Since non-starch polysaccharides and pectins have often been reported for their anti-nutritional role in SBM, we believe this study will provide new avenues toward the improvement of enzymatic cocktails for animal nutrition and health.

Immunocytochemical evaluation of aquaporins and cell wall components and their influence on foliar water uptake in Andean Melastomataceae.

Andean ecosystems are characterized by high humidity, mainly from rain and fog events. Because of differences in altitude two Andean ecosystems - sub-Andean forest and Páramo -face different environmental pressures that affect leaf anatomy and cell wall composition and, consequently, species foliar water uptake (FWU) capacity. Here, FWU capacity of eight species in the Melastomataceae was evaluated and found to be related to proportions of cell wall components and aquaporins in the two ecosystems. Cellulose was labelled with Calcofluor white, and aquaporin and pectins were labelled with monoclonal antibodies. There were differences in plant FWU capacity in both ecosystems, with higher FWU capacity in sub-Andean forest species than in Páramo forest species. Cell wall components were positively related to FWU, with increased FWU related to pectin and aquaporin content of the plasma membrane. Differences in water availability in the two analysed environments led to differences in FWU capacity that are associated with leaf anatomical traits and cell wall composition. In these two environments, plants with similar traits are selected to respond to given environmental pressures. Traits that favour FWU in sub-Andean forest species may lead to further advances of these species in this environments.

Persimmon Fiber-Rich Ingredients Promote Anti-Inflammatory Responses and the Growth of Beneficial Anti-Inflammatory Firmicutes Species from the Human Colon.

Persimmon fruit processing-derived waste and by-products, such as peels and pomace, are important sources of dietary fiber and phytochemicals. Revalorizing these by-products could help promote circular nutrition and agricultural sustainability while tackling dietary deficiencies and chronic diseases. In this study, fiber-rich fractions were prepared from the by-products of Sharoni and Brilliant Red persimmon varieties. These fractions were quantified for their phenolic composition and assessed for their ability to promote the growth of beneficial human colonic Firmicutes species and for their in vitro anti-inflammatory potential. Gallic and protocatechuic acids, delphinidin, and cyanidin were the main phenolics identified. Faecalibacterium prausnitzii strains showed significantly higher growth rates in the presence of the Brilliant Red fraction, generating more than double butyrate as a proportion of the total short-chain fatty acids (39.5% vs. 17.8%) when compared to glucose. The fiber-rich fractions significantly decreased the inflammatory effect of interleukin-1ß in Caco-2 cells, and the fermented fractions (both from Sharoni and Brilliant Red) significantly decreased the inflammatory effect of interleukin-6 and tumor necrosis factor-α in the RAW 264.7 cells. Therefore, fiber-rich fractions from persimmon by-products could be part of nutritional therapies as they reduce systemic inflammation, promote the growth of beneficial human gut bacteria, and increase the production of beneficial microbial metabolites such as butyrate.

Evaluation of Different Pectic Materials Coming from Citrus Residues in the Production of Films.

This article explores the use of citrus residues as a source of different pectic materials for packaging film production: a water-soluble orange residue extract (WSE) (~5% pectin), semi-pure pectins extracted in citric acid (SP) (~50% pectin), and commercial pure citrus pectins (CP). First, these materials were characterized in terms of chemical composition. Then, films were produced using them pure or mixed with chitosan or glycerol through solvent-casting. Finally, antioxidant activity, functional properties (e.g., mechanical and gas barrier properties), and visual appearance of the films were assessed. WSE films showed the highest antioxidant activity but the lowest mechanical strength with the highest elongation at break (EB) (54%); incorporating chitosan increased the films' strength (Young's modulus 35.5 times higher). SP films showed intermediate mechanical properties, reinforced by chitosan addition (Young's modulus 4.7 times higher); they showed an outstanding dry O2 barrier. CP films showed a similar O2 barrier to SP films and had the highest Young's modulus (~29 MPa), but their brittleness required glycerol for improved pliability, and chitosan addition compromised their surface regularity. Overall, the type of pectic material determined the film's properties, with less-refined pectins offering just as many benefits as pure commercial ones.

Structural and biochemical characterization of SmoPG1, an exo-polygalacturonase from Selaginella moellendorffii.

Polygalacturonases (PGs) can modulate chemistry and mechanical properties of the plant cell wall through the degradation of pectins, one of its major constituents. PGs are largely used in food, beverage, textile, and paper industries to increase processes' performances. To improve the use of PGs, knowledge of their biochemical, structural and functional features is of prime importance. Our study aims at characterizing SmoPG1, a polygalacturonase from Selaginella moellendorffii, that belongs to the lycophytes. Transcription data showed that SmoPG1 was mainly expressed in S. moellendorffii shoots while phylogenetic analyses suggested that SmoPG1 is an exo-PG, which was confirmed by the biochemical characterization following its expression in heterologous system. Indeed, LC-MS/MS oligoprofiling using various pectic substrates identified galacturonic acid (GalA) as the main hydrolysis product. We found that SmoPG1 was most active on polygalacturonic acid (PGA) at pH 5, and that its activity could be modulated by different cations (Ca2+, Cu2+, Fe2+, Mg2+, Mn2+, Na2+, Zn2+). In addition, SmoPG1 was inhibited by green tea catechins, including (-)-epigallocatechin-3-gallate (EGCG). Docking analyses and MD simulations showed in detail amino acids responsible for the SmoPG1-EGCG interaction. Considering its expression yield and activity, SmoPG1 appears as a prime candidate for the industrial production of GalA.

Role of acemannan and pectic polysaccharides in saline-water stress tolerance of Aloe vera (Aloe barbadensis Miller) plant.

This study investigates the impact of water and salinity stress on Aloe vera, focusing on the role of Aloe vera polysaccharides in mitigating these stresses. Pectins and acemannan were the most affected polymers. Low soil moisture and high salinity (NaCl 80 mM) increased pectic substances, altering rhamnogalacturonan type I in Aloe vera gel. Aloe vera pectins maintained a consistent 60 % methyl-esterification regardless of conditions. Interestingly, acemannan content rose with salinity, particularly under low moisture, accompanied by 90 to 150 % acetylation increase. These changes improved the functionality of Aloe vera polysaccharides: pectins increased cell wall reinforcement and interactions, while highly acetylated acemannan retained water for sustained plant functions. This study highlights the crucial role of Aloe vera polysaccharides in enhancing plant resilience to water and salinity stress, leading to improved functional properties.

Immunodetection of Cell Wall Components in Studies on Cell Wall Rebuilding in Fagopyrum esculentum and Fagopyrum tataricum.

Immunocytochemical studies of the cell wall are used to visualize specific epitopes of pectins, arabinogalactan proteins, hemicelluloses, extensins, and other wall components using specific primary antibodies. This reaction, combined with calcofluor staining, allows to comprehend how the cell wall is rebuilt during the protoplast culture. In this protocol, the method of immunostaining using antibodies against cell wall components based on Fagopyrum esculentum and Fagopyrum tataricum protoplasts is described.

Immunohistochemical Detection of the Wall Components on the Example of Shoot Apical Meristem of Fagopyrum esculentum and Fagopyrum tataricum.

Immunohistochemistry is a method that allows the detection of individual components of cell walls in an extremely precise way at the level of a single cell and wall domains. The cell wall antibodies detect specific epitopes of pectins, arabinogalactan proteins (AGP), hemicelluloses, and extensins. The presented method visualization of the selected pectic and AGP epitopes using antibodies directed to wall components is described. The method of the analysis of the chemical composition of the wall is present on the example of the shoot apical meristems of Fagopurum esculentum and Fagopyrum tataricum. Recommended protocols for immunostaining and examination on fluorescence microscopy level are presented.

Implications of cell wall immunocytochemical profiles on the structural and functional traits of root and stem galls induced by Eriosoma lanigerum on Malus domestica.

Alterations in cell wall composition imply in new structural and functional traits in gall developmental sites, even when the inducer is a sucking exophytophagous insect with strict feeding sites as the aphid associated to Malus domestica Borkh. This host plant is an economically important, fruit-bearing species, susceptible to gall induction by the sucking aphid Eriosoma lanigerum Hausmann, 1802. Herein, the immunocytochemical detection of arabinogalactan-proteins (AGPs), pectins, and hemicelluloses using monoclonal antibodies was performed in samples of non-galled roots and stems, and of root and stem galls on M. domestica. The dynamics of these cell wall components was discussed under the structural and functional traits of the galls proximal, median, and distal regions, according to the proximity of E. lanigerum colony feeding site. In the proximal region, the epitopes of AGPs and homogalacturonans (HGs) are related to cell growth and divisions, which result in the overproduction of parenchyma cells both in root and stem galls. In the proximal and median regions, the co-occurrence of HGs and arabinans in the cell walls of parenchyma and secondary tissues favors the nutrient flow and water-holding capacity, while the xylogalacturonans and hemicelluloses may function as additional carbohydrate resources to E. lanigerum. The immunocytochemical profile of the cell walls support the feeding activity of E. lanigerum mainly in the gall proximal region. The similarity of the cell wall components of the gall distal region and the non-galled portions, both in roots and stems, relates to the decrease of the cecidogenetic field the more distant the E. lanigerum colony is.

Chromatography based profiling of feruloylated arabinans and galactans.

Profiling of pectic arabinans and galactans by analysis of the released oligosaccharides after backbone cleavage provides information on the complexity of the polymer structure. In plants of the family Amaranthaceae, arabinan and galactan substitution with ferulates extends the polysaccharide complexity, changing its chemical properties. Knowledge of the ferulate environment is crucial to understand structure-function-relationships of feruloylated pectins. Here, we present an approach to separate enzymatically generated feruloylated and non-feruloylated arabino- and galactooligosaccharides, followed by deesterification and semiquantitative analysis by HPAEC-PAD using previously reported relative response factors. Application of this approach to sugar beet pectins and insoluble and soluble dietary fiber preparations of amaranth and quinoa suggests that ferulates are preferably incorporated into more complex structures, as nicely demonstrated for feruloylated galactans. Also, ferulate substitution appears to negatively affect enzymatic cleavage by using endo-enzymes. As a consequence, we were able to tentatively identify new feruloylated tri- and tetrasaccharides of galactans isolated from sugar beet pectins.

An insight into tissue culture-induced variation origin shared between anther culture-derived triticale regenerants.

BACKGROUND: The development of the plant in vitro techniques has brought about the variation identified in regenerants known as somaclonal or tissue culture-induced variation (TCIV). S-adenosyl-L-methionine (SAM), glutathione (GSH), low methylated pectins (LMP), and Cu(II) ions may be implicated in green plant regeneration efficiency (GPRE) and TCIV, according to studies in barley (Hordeum vulgare L.) and partially in triticale (× Triticosecale spp. Wittmack ex A. Camus 1927). Using structural equation models (SEM), these metabolites have been connected to the metabolic pathways (Krebs and Yang cycles, glycolysis, transsulfuration), but not for triticale. Using metabolomic and (epi)genetic data, the study sought to develop a triticale regeneration efficiency statistical model. The culture's induction medium was supplemented with various quantities of Cu(II) and Ag(I) ions for regeneration. The period of plant regeneration has also changed. The donor plant, anther-derived regenerants, and metAFLP were utilized to analyze TCIV concerning DNA in symmetric (CG, CHG) and asymmetric (CHH) sequence contexts. Attenuated Total Reflectance-Fourier Transfer Infrared (ATR-FTIR) spectroscopy was used to gather the metabolomic information on LMP, SAM, and GSH. To frame the data, a structural equation model was employed. RESULTS: According to metAFLP analysis, the average sequence change in the CHH context was 8.65%, and 0.58% was de novo methylation. Absorbances of FTIR spectra in regions specific for LMP, SAM, and GSH were used as variables values introduced to the SEM model. The average number of green regenerants per 100 plated anthers was 2.55. CONCLUSIONS: The amounts of pectin demethylation, SAM, de novo methylation, and GSH are connected in the model to explain GPRE. By altering the concentration of Cu(II) ions in the medium, which influences the amount of pectin, triticale's GPRE can be increased.

Extraction of pectins from renewable grapefruit (Citrus paradisi) peels using deep eutectic solvents and analysis of their structural and physicochemical properties.

This study presents an innovative attempt to extract high-quality pectins from grapefruit (Citrus paradisi) peels by using deep eutectic solvents (DESs) as extraction agents. The maximum yield of betaine-citric acid (BC)-extracted pectin (BC-P) reached 36.47 % under the optimum process conditions: an L/S ratio of 25 mL/g, a pH of 2.0, and a temperature of 85 °C for 120 min. The yield of BC-P was significantly higher than HCl-extracted pectin (HCl-P, 8.76 %) under a pH of 2.0. In addition, the structural, physicochemical, and emulsifying properties of the purified pectins (BC-P and HCl-P) and commercial pectin (CP) were comparatively analyzed. Results showed that BC-P exhibited higher RG-I value, more arabinan side-chains, bigger Mw and Mn value than HCl-P. Moreover, the viscosity, G' and G'' of BC-P were significantly higher than those of HCl-P and CP. More importantly, BC-P demonstrated better emulsifying activity and stability compared to HCl-P and CP. When the concentration of BC-P was increased to 1.50 %, a stable emulsion containing a 50 % soybean oil fraction could be obtained. Our results confirmed that DESs can be considered as high-effective agents for pectin extraction. Pectins extracted from grapefruit peels can be as a promising natural emulsifiers that can be used in the food industry.

Foliar Pectins and Physiology of Diploid and Autotetraploid Mango Genotypes under Water Stress.

The cultivation of mango in Mediterranean-type climates is challenged by the depletion of freshwater. Polyploids are alternative genotypes with potential greater water use efficiency, but field evaluations of the anatomy and physiology of conspecific adult polyploid trees under water stress remain poorly explored. We combined field anatomical evaluations with measurements of leaf water potential (Ψl) and stomatal conductance (Gs) comparing one diploid and one autotetraploid tree per treatment with and without irrigation during dry summers (when fruits develop). Autotetraploid leaves displayed lower Ψl and Gs in both treatments, but the lack of irrigation only affected Gs. Foliar cells of the adaxial epidermis and the spongy mesophyll contained linear pectin epitopes, whereas branched pectins were localized in the abaxial epidermis, the chloroplast membrane, and the sieve tube elements of the phloem. Cell and fruit organ size was larger in autotetraploid than in diploid mango trees, but the sugar content in the fruits was similar between both cytotypes. Specific cell wall hygroscopic pectins correlate with more stable Ψl of autotetraploid leaves under soil water shortage, keeping lower Gs compared with diploids. These preliminary results point to diploids as more susceptible to water deficits than tetraploids.

Evaluation of the Carbohydrate Composition of Crabapple Fruit Tissues Native to Northern Asia.

A comprehensive comparative analysis of the carbohydrate composition (soluble sugars and pectins) of fruit tissues of Malus baccata, Malus mandshurica, Malus chamardabanica, and Malus sachalinensis, characteristic of the vast territory of Eastern Siberia and the Far East, was carried out. It was shown that a large part of the soluble carbohydrates of the studied species were represented by transport sugars-sorbitol and sucrose. These compounds also provided the main variability in the carbohydrate composition of fruits in the studied material. The polymers pectins and protopectins isolated from the studied fruits were highly methoxylated (up to 60-70%), and their content averaged about 6% of dry weight. The greatest length of pectin polymers was found in the fruit tissues of M. chamardabanica and M. sachalinensis. Data on elemental analysis of fractions of pectins and protopectins of all studied species showed the absence of potentially toxic concentrations of heavy metals. Of note is the rather high content of calcium in both polymer fractions of the four studied species, while its content in protopectin is significantly higher. In addition, in all cases, the presence of low-molecular-weight oligosaccharide molecules with a low-dispersed linear structure was revealed in the tissues of the fruits. It is worth noting that the high content of ascorbic acid was observed in the fruits of all studied species. In addition to being of fundamental interest, information about the carbohydrate composition of the wild Malus species can be useful for apple breeding when choosing sources of genes underlying useful traits.

Apoplastic and Symplasmic Markers of Somatic Embryogenesis.

Somatic embryogenesis (SE) is a process that scientists have been trying to understand for many years because, on the one hand, it is a manifestation of the totipotency of plant cells, so it enables the study of the mechanisms regulating this process, and, on the other hand, it is an important method of plant propagation. Using SE in basic research and in practice is invaluable. This article describes the latest, but also historical, information on changes in the chemical composition of the cell wall during the transition of cells from the somatic to embryogenic state, and the importance of symplasmic communication during SE. Among wall chemical components, different pectic, AGP, extensin epitopes, and lipid transfer proteins have been discussed as potential apoplastic markers of explant cells during the acquisition of embryogenic competence. The role of symplasmic communication/isolation during SE has also been discussed, paying particular attention to the formation of symplasmic domains within and between cells that carry out different developmental processes. Information about the number and functionality of plasmodesmata (PD) and callose deposition as the main player in symplasmic isolation has also been presented.

Contrasting effects of polysaccharide components on the cooking properties of roots, tubers and bananas.

BACKGROUND: Consumer preferences for boiled or fried pieces of roots, tubers and bananas (RTBs) are mainly related to their texture. Different raw and cooked RTBs were physiochemically characterized to determine the effect of biochemical components on their cooking properties. RESULTS: Firmness in boiled sweetpotato increases with sugar and amylose contents but no significant correlation was observed between other physicochemical characteristics and cooking behaviour. Hardness of boiled yam can be predicted by dry matter (DM) and galacturonic acid (GalA) levels. For cassava, no significant correlation was found between textural properties of boiled roots and DM, but amylose and Ca2+ content were correlated with firmness, negatively and positively, respectively. Water absorption of cassava root pieces boiled in calcium chloride solutions was much lower, providing indirect evidence that pectins are involved in determining cooking quality. A highly positive correlation between textural attributes and DM was observed for fried plantain, but no significant correlation was found with GalA, although frying slightly reduced GalA. CONCLUSION: The effect of main components on texture after cooking differs for the various RTBs. The effect of global DM and major components (i.e. starch, amylose) is prominent for yam, plantain and sweetpotato. Pectins also play an important role on the texture of boiled yam and play a prominent role for cassava through interaction with Ca2+ . © 2023 Bill and Melinda Gates Foundation. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.