Relationship between structural development of cell walls and degradation of tissues in maize stems

Sections of internodes of growing maize stems were used to study the behaviour of cell walls of different tissues during in-vitro degradation with rumen fluid. Tissues with primary cell walls-middle lamellae, at early stages of development, were degraded completely. In specific tissues, newly synthesised secondary walls were highly digestible whereas the primary walls-middle lamellae of these tissues were indigestible. These primary walls-middle lamellae stained positively with acid phloroglucinol but showed no fluorescence. At pollination, when secondary walls were of considerable thickness, these walls were still completely digestible even though they stained intensely with acid phloroglucinol and showed reduced fluorescence. However, at some distance from the cut end of sections, the secondary walls of the elongated tube-like cells of sclerenchyma tissue showed considerable reduction in digestibility. Cross-sectional area and dry weight measurements of different stem tissues revealed the importance of secondary wall digestion of sclerenchyma compared with the thin-walled parenchyma. Chemical treatment with KmnO₄ or NaOH resulted in colourless secondary walls after staining whereas primary walls still reacted positively. It was concluded that very small amounts of phenolic compounds (lignin) located in the primary wall-middle lamella that are not removed by KmnO₄ and NaOH treatment are responsible for the decrease in digestibility of tissues during plant development. Histochemical ‘lignin’ reactions and fluorescence just detect phenolic compounds and cannot be correlated with degradation.     

Metabolism and fate of dietary (U-14C)-labelled spinach cell walls in the rat

Using uniformly ¹⁴C-labelled spinach (Spinacia oleracea L) plant cell walls (PCW) the metabolism of PCW can be followed in a defined manner in the rat. Only 10% of the ¹⁴C was recoverable in the gut contents and faeces 18 h post-gavage. Two percent of the ¹⁴C was excreted in urine and 25% excreted as CO₂. ¹⁴C was found in all tissues of the body but was most concentrated in the adrenal glands, colon and caecum. The ¹⁴C was also present at moderately high concentration in the liver pelt, and owing to their larger size, these tissues accounted for a high percentage of the total ¹⁴C. In the liver the ¹⁴C was predominantly associated with phospholipid, whereas in the pelt it was present in protein and in fatty acid residues esterified to cholesterol. Dietary PCW material is extensively fermented in the caecum and colon. Also, the products of fermentation are a source of energy and important structural precursors of lipids and proteins for the animal.     

Fermentation characteristics of polysaccharide fractions extracted from the cell walls of maize endosperm

Cell walls were extracted from maize endosperm and separated into different polysaccharide fractions by sequential extraction with solutions of saturated Ba(OH)₂, demineralised water and 1 and 4 M KOH. Solubilised polysaccharides were collected after each extraction. Residues were collected following the extractions with demineralised water and 1 and 4 M KOH. The original cell wall (CW) material, extracts and residues were analysed for their fermentation characteristics using an in vitro cumulative gas production technique. The rate of fermentation of the alkali‐treated residues was faster than that of the original CW material, except for the 4 M KOH residue, which had a similar rate of degradation to the original CW material. The polysaccharides solubilised from the cell wall (extracts) were all rapidly fermented, more rapidly than both CW and residues. A division of the gas production profile into two phases using curve fitting was in good agreement with a division of the cell wall fermentation into the fermentation of arabinoxylans and cellulose. Therefore the likelihood of preferential degradation of arabinoxylans from the maize cell wall was discussed. The volatile fatty acid production pattern was fairly well explained by the fermentation rate and composition of the substrates. It was concluded that breaking the interactions of polysaccharides in the maize cell wall by mild alkali extraction increases the fermentability of maize cell walls in the gastrointestinal tract of farm animals. Contrarily, more severe alkali extractions will reduce the fermentability of maize cell walls. © 2002 Society of Chemical Industry     

Metabolism of dietary (Acetyl-1-14c)-plant cell walls in the rat intestine

To determine a fate of acetyl groups in intact dietary plant cell walls (PCW) spinach cells were labelled using [1-¹⁴C]acetate as a precursor and the PCW isolated. The (acetyl-¹⁴C)-PCW were fed to rats by garage and the ¹⁴C traced in the rat gut and in the body tissues and excreta of the rat. Some label was solubilised during passage through the upper intestine of the rat, and subsequently incorporated into the body tissues of the rat. A small amount of the radioactivity was excreted as CO₂ (4%) and in urine (<1%). (Acetyl-¹⁴C)-PCW which reached the caecum was solubilised and some [¹⁴C]acetate released. The solubilised ¹⁴C was incorporated in liver tissue mainly as amino acid present in protein.     

Lignin composition is more important than content for maize stem cell wall degradation

BACKGROUND

The relationship between the chemical and molecular properties – in particular the (acid detergent) lignin (ADL) content and composition expressed as the ratio between syringyl and guaiacyl compounds (S:G ratio) – of maize stems and in vitro gas production was studied in order to determine which is more important in the degradability of maize stem cell walls in the rumen of ruminants. Different internodes from two contrasting maize cultivars (Ambrosini and Aastar) were harvested during the growing season.

RESULTS

The ADL content decreased with greater internode number within the stem, whereas the ADL content fluctuated during the season for both cultivars. The S:G ratio was lower in younger tissue (greater internode number or earlier harvest date) in both cultivars. For the gas produced between 3 and 20 h, representing the fermentation of cell walls in rumen fluid, a stronger correlation (R² = 0.80) was found with the S:G ratio than with the ADL content (R² = 0.68). The relationship between ADL content or S:G ratio and 72‐h gas production, representing total organic matter degradation, was weaker than that with gas produced between 3 and 20 h.

CONCLUSION

The S:G ratio plays a more dominant role than ADL content in maize stem cell wall degradation. © 2017 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Forage quality variation among maize inbreds: Relationships of cell-wall composition and in-vitro degradability for stem internodes

Forty-five inbred maize (Zea mays L) lines were evaluated for genetic variation in stem cell-wall concentration, composition and degradability, and for relationships among cell-wall components and polysaccharide degradability. Cell-wall neutral sugars, uronic acids, Klason lignin, and ester- and ether-linked phenolic acids were measured on lower stem internode samples collected at the time of silking in 2 years. Twenty-four and 96 h in-vitro ruminal fermentations were used to determine the rapidly and potentially degradable cell-wall polysaccharide fractions, respectively. Genetic variation (P < 0.05) was found for all measures of cell-wall composition and many estimates of rapidly and potentially degradable cell-wall polysaccharide components. Inbred line means varied by 50–300% for most traits. Three brown midrib mutant inbred lines included in the study were not the lowest in lignin content nor did they exhibit the greatest cell-wall degradabilities in this population of inbred maize. Year of growth (environment) influenced (P < 0.05) cell-wall traits even though reproductive physiological maturity at sampling was similar in both years. Degradability of the cell-wall polysaccharide components were intercorrelated (P < 0.05) within the rapidly and potentially degradable fractions, but rate and extent of degradation of the cell-wall components were not correlated (P > 0.05), except for uronic acids. A multiple regression model of principal components (R² = 0.41, P < 0.05) indicated that cell-wall lignification and substitution of wall polymers with phenolic and uronic acids were negatively associated, and pectic substances were positively related with rapid polysaccharide degradation. Very little of the variation (R² = 0.15, P < 0.05) in potential cell-wall polysaccharide degradation could be explained by this multiple regression analysis. There is a large degree of genetic variation among current inbred maize lines for stem cell-wall quality traits, which should allow improvement of maize as a forage crop. Because of the complex matrix interactions in cell-wall organization, however, no single cell-wall component, or simple combination, can accurately predict degradability of maize cell walls.     

Diferulate cross-links impede the enzymatic degradation of non-lignified maize walls

We assessed the effect of ferulate substitution and diferulate cross-linking of xylans on the degradation of cell walls by two fungal enzyme mixtures, one of which contained feruloyl esterase and high xylanase activities. Non-lignified cell suspensions of maize (Zea mays) were grown with 0 or 40 μM 2-aminoindan-2-phosphonic acid to produce walls with normal (17·2 mg g⁻¹) or reduced (5·1 mg g⁻¹) ferulate concentrations. Walls were incubated with mercaptoethanol to inhibit diferulate formation or with hydrogen peroxide to stimulate diferulate formation by wall bound peroxidases. Varying the ferulate substitution of xylans did not affect cell wall hydrolysis. In contrast, increasing ferulate dimerisation from 18 to 40% reduced carbohydrate release by 94–122 mg g⁻¹ after 3 h and by 0–48 mg g⁻¹ after 54 h of enzymatic hydrolysis. Diferulate cross-links impeded the release of xylans, cellulose and pectins from walls. These results provide compelling evidence that diferulate cross-links reduce the rate and, to a lesser degree, the extent of wall hydrolysis by fungal enzymes. Our results also suggest that enzyme mixtures containing high xylanase activity but not feruloyl esterase activity can partially overcome the inhibitory effects of diferulate cross-linking on wall hydrolysis. © 1998 SCI.     

The newly extended maize internode: A model for the study of secondary cell wall formation and consequences for digestibility

The upper five internodes were collected from maize (Zea mays L) inbred cell lines Co 125 and W401 harvested at the same developmental stage, 5 days after silking. Each internode was dissected into ten equal lengths labelled A (top) to J (base). The youngest cells were found in section J, which contained the intercalary meristem, and the oldest in section A. Internodes 1, 3 and 5 provided material for chemical analysis and internodes 2 and 4 for degradability measurements. Cell wall material accounted for one-third of dry matter in section J, doubling to two-thirds in the upper half of each internode. Only section J exhibited a polysaccharide profile typical of primary cell walls. In all other sections, 1,4-linked glucose (± 46% of cell wall) and xylan largely free from side chains (± 25% of cell wall) predominated. Net accretion of cell wall polysaccharide reached a maximum by segment G and thereafter little additional carbohydrate was deposited. Lignification appeared to be separated from the biogenesis of structural carbohydrate and continued over much of each internode reaching a maximum in section C. Degradability measurements, made using a modified neutral-detergent cellulase digestibility method, showed substantial differences between sections. In line Co 125, cell wall degradability fell from over 95% in the youngest section (J) to approximately 24% in section B. Internode 4 of line W401 failed to show the same pattern of degradabilities, probably because of a sequential rather than simultaneous pattern of internode elongation. Saponifiable p-coumaric acid appeared to provide a more sensitive marker than lignin of the extent of secondary wall development. The inverse relationship between extent of lignification in each section and its degradability confirmed the value of the internode model for the study of secondary wall formation and its biological consequences.     

Lax leaf maize: cell wall composition and nutritional value

Forage nutritive value, which comprises traits such as digestibility, fibre, lignin and protein content, is an important criterion for maize (Zea mays L) harvested as silage. Lines with a characteristic phenotype (‘lax leaf’) could be useful sources of genes for improved nutritive value in maize. A study was conducted to characterise the cell wall composition of the lax leaf line. Lax leaf inbreds and inbreds representing ‘normal’ maize were evaluated for cell wall neutral sugars, uronic acids, Klason lignin and phenolic acids in five tissues from the ear node and the internode above it. Acid detergent fibre (ADF) and neutral detergent fibre (NDF) and 48 h in vitro true digestibility (IVTD) were predicted using near‐infrared reflectance spectrophotometry (NIRS) calibrated with a subset of the scanned samples. Lax leaf inbred tissues had lower levels of ADF, NDF, lignin and xylose and were more digestible than tissues from the inbreds representing ‘normal’ maize. It was not known whether the lax leaf phenotype resulted from alterations in nutritive value traits or whether laxness and nutritive value traits are independent from one another. A second study was conducted to determine the nature of genetic control of the lax leaf character and to determine the genotypic relation between the lax leaf character and nutritive value. A recombinant inbred mapping population was developed from a cross between the lax leaf line and an inbred line with stiff upright leaves. Whole‐plant samples from each recombinant inbred line were evaluated for ADF, NDF, acid detergent lignin (ADL) and IVTD of dry matter using NIRS. Laxness, measured by number of broken leaves, was associated with lower nutritive value in this population (genetic correlations 0.16–0.34), which was contrary to expectation. Amplified restriction fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers were used to identify linkage groups associated with the lax leaf character, digestibility and fibre content. Several linkage groups were associated with both the lax leaf character and nutritive value. Where these characters were associated with the same linkage group, the lax leaf parent allele was associated with greater laxness but reduced nutritive value. The lax leaf parent allele was associated with increased nutritive value in linkage groups unassociated with the lax leaf character. While the lax leaf line may be a good source for alleles for improved nutritive value, selection for laxness will not likely be accompanied by improvement in forage quality. © 2000 Society of Chemical Industry     

Analytical methodologies for quantification of ferulic acid and its oligomers

Ferulic acid (4‐hydroxy‐3‐methoxycinnamic acid) is the most widespread hydroxycinnamic acid in the plant world, where it is a key molecule in cell wall architecture. Owing to its high antioxidant properties, ferulic acid shows large potential applications in food industry as well as in the health and cosmetic markets. There is thus a high interest in extracting this high‐value compound from waste materials of the agricultural industry, which requires the selection of an appropriate quantification method. This paper therefore gives an overview of analytical methodologies developed over past decades for quantification of ferulic acid and its oligomers. Copyright © 2008 Society of Chemical Industry     

Calcium effect on mechanical properties of model cell walls and apple tissue

Bacterial cellulose alone, with pectin added and with both pectin and xyloglucan added were produced as models for plant cell walls. The artificial cell wall materials and natural apple tissue were treated with calcium and then subjected to tests of mechanical properties; a tension test for the artificial cell wall materials and a compression test for the apple tissue. It was found that pectin and xyloglucan had a significant effect on the mechanical properties of artificial cell wall materials, making them more extensible. A high concentration of calcium increased the failure strain and decreased the failure stress of these materials. Treatment of apple tissue with calcium had similar effects on the failure stress and the secant modulus, whereas the differences of failure strain and work to failure resulted in behaviour different from that of the artificial material.     

Microspectrophotometry of bermudagrass (Cynodon dactylon) cell walls in relation to lignification and wall biodegradability

The walls of the major plant cell types, sclerenchyma and parenchyma, of the second (upper) and fifth (lower) internodes from the apex of bermudagrass (Cynodon dactylon L) stem have been examined by ultraviolet absorption microspectrophotometry and the results related to wall digestibility (measured in vitro with rumen liquor) and to histochemical reactions for ‘lignin’ and phenolics. Sclerenchyma walls from the lower internode gave high values of absorbance in the ultraviolet region of the spectrum equivalent to 109 mg trans-ferulic acid g^?1 dry walls; similar walls from the upper internode also gave high values but these were lower than values from the lower internode. Histochemical examination showed that the sclerenchyma walls, which were indigestible to rumen microorganisms, gave positive tests with acid phloroglucinol reagent for lignin. Parenchyma walls, which were either digested or partially digested, gave much lower absorbance values in the ultraviolet region and negative tests with acid phloroglucinol but positive tests with diazotised sulphanilic acid (upper and lower internodes) and chlorine-sulphite (lower internode) reagents. Ultraviolet absorption microspectrophotometry is a useful technique for examining phenolics within individual cell walls that vary in biodegradability.     

Effects of conventional boiling on the polyphenols and cell walls of pears

Pears of the cultivar Gieser Wildeman were cooked for up to 24 h and changes in polyphenol and cell wall components were monitored. The main polyphenols were flavan‐3‐ols (epicatechin and its procyanidin oligomers), with an average degree of polymerisation of 6, and caffeoylquinic acid. Upon cooking, flavan‐3‐ols were retained in the pear tissue while the hydroxycinnamic acids were partially leached into the cooking water. After 1 h of cooking, 65% of the original flavan‐3‐ols and 40% of the original caffeoylquinic acid were still detectable in the pear tissue; the cooking water contained only 2% of the flavan‐3‐ols but 24% of the caffeoylquinic acid. Cell walls represented 23 g kg^?1 of the fresh pear and were composed of cellulose, pectins and xylans. The pectic fractions was degraded during cooking while xylans and cellulose were not affected. Copyright © 2004 Society of Chemical Industry     

植物细胞壁中伸展蛋白的研究

首先测定8种蔬菜细胞壁中的羟脯氨酸含量,研究结果表明:豆皮、豆荚、花粉、胡萝卜根、油菜叶柄、芹菜叶柄、青菜和洋葱细胞壁中羟脯氨酸的含量分别为6.02μg/mg、3.30μg/mg、1.74μg/mg、1.53μg/mg、0.64μg/mg、0.52μg/mg、0.42μg/mg和0.31μg/mg。然后通过CM-纤维素柱和Sephadex G-75柱色谱纯化豆渣细胞壁中的伸展蛋白。用CM-纤维素柱色谱纯化豆渣细胞壁蛋白时,得到3个组分,即S1、S2和S3;对S2组分进一步采用Sephadex G-75柱色谱纯化,得到S2a、S2b和S2c,并对S2c组分进行单糖组分分析,S2c中的单糖以阿拉伯糖和半乳糖为主,它们的摩尔之比为21∶。     

Impact of accessibility and chemical composition on cell wall polysaccharide degradability of maize and lucerne stems

Although lignification of forages is generally accepted as limiting cell wall degradability, prediction of degradation from cell wall composition is often difficult when forages are of similar maturity. It has been proposed that rumen microbe accessibility to potentially degradable cell walls is limited by the presence of non‐disrupted cells in forage particles with lignified middle lamella/primary walls acting as barriers to microbial access. We tested this accessibility hypothesis by evaluating the impact of reducing particle size of maize and lucerne stems to the level of individual cells by ball‐milling, in order to eliminate accessibility as a limiting factor. While cell wall concentration and composition were not influenced by ball‐milling compared with grinding to pass a 1 mm screen in a cyclone‐type mill, degradability of total cell wall polysaccharides was dramatically increased. However, only those polysaccharides (cellulose and xylan) which are most abundant in cell types with lignified middle lamella/primary and secondary walls increased in degradability owing to particle size reduction. Degradability of pectins, which are abundant in non‐lignified tissues in lucerne, did not respond to ball‐milling. Contrary to our expectations, ball‐milled forages showed fewer correlations for cell wall composition with degradability than observed for the larger‐particle‐size grinding treatment. Many components of the cell wall were correlated with polysaccharide degradation for the cyclone‐ground samples; however, the results were inconsistent as to which cell wall components were correlated with degradation among and within forages. This observation does not clarify the role of cell wall chemical structure as a limiting agent to wall degradation in the absence of accessibility barriers, but this study does provide support for the hypothesis that lignified middle lamella/primary walls act as barriers to microbial access for degradation. © 2000 Society of Chemical Industry     

Effect of heat treatments on the phytic acid content of maize products

Processing of maize (Zea mays L fresh and dry) for the production of various traditional products results in the loss of phytic acid. Fresh mature corn contains less phytic acid (1·71 g kg^?1) than dry corn (7·15–7.60 g kg^?1). The loss of phytic acid varies from 18·1 to 46·7% for fresh maize and from 11·5 to 52·6% for dry maize respectively among the heat treatments given. From a nutritional viewpoint, consumption of maize as chapati and after roasting in a sand bath or on charcoal will improve the availability of minerals.     

Relationship of cell wall composition to in vitro cell wall digestibility of maize inbred line stems

The phenolic equipment of maize stem tissues was investigated in relation to the feeding value of the detergent fibre components. Sixteen maize inbred lines, including three brown‐midrib 3 mutants and their normal counterparts, were selected for highly divergent in vitro cell wall digestibility. These lines were grown during two years. Maize stems were analysed for detergent fibre concentration, esterified and etherified p‐hydroxycinnamic acids, lignin content and structure and in vitro digestibility. A large genotypic variation was found for neutral detergent fibre, cell wall phenolic composition and cell wall digestibility. Within the normal maize lines the in vitro neutral detergent fibre digestibility (IVNDFD) of stem fractions was negatively correlated with their Klason lignin content. A multiple regression model based on esterified p‐coumaric acid and lignin composition as two explanatory variates accounted for 58% of the IVNDFD variation. In this study, three normal maize inbred lines displaying a lignin content and a cell wall digestibility level close to those observed in the three bm3 lines could be detected, which opens up new breeding avenues. © 2000 Society of Chemical Industry