Impact of plant cell wall network on biodegradation in soil: Role of lignin composition and phenolic acids in roots from 16 maize genotypes

Residue quality is a key factor governing biodegradation and the fate of C in soil. Most investigations of relationships existing between crop residue quality and soil decomposition have been based on determining the relative proportions of soluble, cellulose, hemicellulose and lignin components. However, cell wall cohesion is increased by tight interconnections between polysaccharides and lignin that involve cross-linking agents (phenolic acids). The aim of this study was to determine the role of lignin composition and phenolic acids on short- to medium-term decomposition of maize roots in soil. Sixteen maize genotypes, presenting a range of chemical characteristics related to root lignin and phenolic acids, were used. The main components were characterized by Van Soest (VS) extraction and cell wall acid hydrolysis, and the non-condensed Syringyl and Guaicyl lignin monomers, esterified phenolic acids and etherified phenolic acids were determined. Maize roots were then incubated in soil under controlled conditions (15 °C, −80 kPa moisture) for 796 days. Results showed that VS extraction over-estimated the structural hemicellulose content and that VS lignin was more recalcitrant than Klason lignin. The tremendous effect of cell wall chemical characteristics was shown by marked variations (almost two-fold differences in C mineralization), between the 16 maize roots. Decomposition was controlled by soluble residue components in the short term whereas lignin and the interconnections between cell wall polymers were important in the long-term. Notably the cell wall domain rich in non-condensed lignin and esterified phenolic acids was prone to decomposition whereas the presence of etherified ferulic acids seemed to hamper cell wall decomposition.     

Chemical composition and enzymatic degradability of xylem and nonxylem walls isolated from alfalfa internodes

During plant maturation, degradability of alfalfa (Medicago sativa L.) stems declines due to accumulation of highly lignified xylary tissue. Xylem and nonxylem tissues dissected from lower alfalfa internodes were analyzed for cell wall constituents and degradability. Cell walls comprised 740 mg g(-1) of xylem and 533 mg g(-1) of nonxylem tissues. Xylem tissues contributed about 60% of the cell wall mass in internodes. Xylem walls contained 28% lignin, 4% pectin, 29% hemicellulose, and 39% cellulose as compared to 15% lignin, 25% pectin, 30% hemicellulose, and 30% cellulose in nonxylem walls. Fungal enzymes hydrolyzed 22 and 73% of the structural carbohydrates in xylem and nonxylem walls, respectively. In both cell wall fractions, the release of xylose was 56-90% lower than that of other sugars, indicating that lignin preferentially restricted xylan degradation in secondary walls and xyloglucan degradation in primary walls. Elucidation of lignin-xylose interactions may reveal strategies for improving fiber degradability of alfalfa.     

根系主要成分含量对根系固土效能的影响

选取中国北方两种常见树种油松(Pinus tabulaeformis)和元宝枫(Acer truncatum),对这两种树0.2~6mm直径的根系直径、根系抗拉强度、整根的根土复合体强度、根系3种主要成分含量(纤维素含量、半纤维素含量与木质素含量)进行测定。结果表明,随着根系直径的增加,植物根系半纤维素含量增高,纤维素含量与木质素含量降低,从而植物的抗拉强度降低。植物根系抗拉强度与纤维素含量、木质素含量成正相关关系,与半纤维素含量成负相关关系。因此,在排除根系结构影响的前提下,植物根系纤维素含量与木质素含量是导致不同植物种固土效果差异的直接原因。     

Degradation of hemicellulose, cellulose and lignin in decomposing spruce needle litter in relation to N

Decomposing needles from a Norway spruce forest in southern Sweden were studied for 559 days under laboratory conditions. Falling needles were collected in control (Co) plots and plots that had received 100 kg N ha⁻¹ yr⁻¹ as (NH₄)₂SO₄ for 9 years under field conditions. One of the aims was to determine whether the previously documented low decomposition rate of the N fertilized (NS) needles could be explained by a lower degradation degree of lignin. The lignin content was studied using the alkaline CuO oxidation method, the Klason lignin method and CPMAS ¹³C NMR spectroscopy. The amounts of cellulose and hemicellulose were also determined.The fertilized needle litters initially decomposed faster than the unfertilized, but later this reaction reversed, so that at the end the mass loss was 45% initial C in the control and 35% initial C in NS. Klason lignin decreased with time in both treatments and overall, the change of Klason lignin mirrored the litter mass loss. No major difference as regards the decomposition of hemicellulose occurred between the treatments, whereas significantly lower concentrations of cellulose were found in NS needles throughout the incubation. The CuO derived compounds (VSC) were somewhat lower in NS needles throughout the decomposition time. Initially, VSC increased slightly in both treatments, which contradicts the Klason lignin data. There was a weak positive relationship (p>0.05) between VSC and Klason lignin. Both vanillyls compounds (V) and cinnamyl compounds (Ci) increased slightly during decomposition, whereas syringyl compounds (S) vanished entirely. The lignin degradation degree, i.e. the acid-to-aldehyde ratio of the vanillyl compounds expressed as (Ac/Al)_v, showed no significant effect of treatment. The ¹³C NMR analyses of the combined samples showed increased content of aromatic C with increasing decomposition time. The carbohydrate content (O-alkyl C) was lower in the fertilized needle litter throughout the incubation time. The alkyl C content tended to increase with decomposition time and N fertilization. The alkyl C/O-alkyl C ratios increased in both treatments during the incubation. The NMR results were not tested statistically.In conclusion, no major difference concerning lignin degradation could be found between the unfertilized and N fertilized needle litter. Thus, the study contradicts the hypothesis that higher amounts of N reduce lignin degradation. The reduced biological activity is probably due to direct N effects on the microorganisms and their decomposing ability.     

甜菜细胞壁界面特征显著影响纤维素酶解效率

生物质不同器官、组织理化性质各异,这种异质性对纤维素酶解效率的影响有待深入研究。该研究以甜菜为研究对象,系统分析不同器官理化性质与酶解效率间的关系,探究细胞壁与纤维素酶的吸附特点及在酶解过程中的形态变化。结果表明,甜菜根、茎、叶酶解还原糖得率不同,根和茎的还原糖得率相近且高于叶,分别为14.64%和14.26%,叶还原糖得率较低为10.15%;酶解还原糖得率与木质素（P<0.01）、半纤维素（P<0.01）呈极显著负相关,而与比表面积（P<0.01）呈极显著正相关;甜菜根、茎、叶的薄壁组织更容易与纤维素酶结合,并且更易被降解。甜菜细胞壁界面理化特征是影响酶与底物有效吸附,进而影响纤维素酶解效率的关键因素。研究结果可为甜菜农业废弃物的利用提供参考。     

Effect of chemical composition on the release of nitrogen from agricultural plant materials decomposing in soil under field conditions

Summary In two field experiments, plant materials labelled with ¹⁵N were buried separately within mesh bags in soil, which was subsequently sown with barley. In the first experiment, different parts of white clover (Trifolium repens), red clover (T. pratense), subterranean clover (T. subterraneum), field bean (Vicia faba), and timothy (Phleum pratense) were used, and in the second, parts of subterranean clover of different maturity. The plant materials were analysed for their initial concentrations of total N, ¹⁵N, C, ethanol-soluble compounds, starch, hemicellulose, cellulose, lignin, and ash. After the barley had been harvested, the bags were collected and analysed for their total N and ¹⁵N. In the first experiment the release of N was highest from white clover stems + petioles (86%) and lowest from field bean roots (20%). In stepwise regression analysis, the release of N was explained best by the initial concentrations of lignin, cellulose, hemicellulose, and N (listed according to decreasing partial correlations). Although the C/N ratio of the plant materials varied widely (11–46), statistically the release of N was not significantly correlated with this variable. The results of the second experiment using subterranean clover of different maturity confirmed those of the first experiment.     

铝胁迫下植物根尖细胞壁组成物质变化抑制根伸长的生理机制研究

根伸长受抑制是植物受铝毒害的主要症状,铝诱导的细胞壁组成物质的变化是其主要原因。本文主要对铝胁迫下植物根尖细胞壁组成物质如木质素、 胼胝质、 纤维素、 半纤维素、 果胶、 细胞壁多糖蛋白及相关代谢酶类在铝胁迫下的变化对根伸长的影响及生理机理的研究进展进行了综述,明确了铝胁迫诱导的植物根尖细胞壁组成物质含量、 比例及结构的变化导致细胞壁刚性降低,从而抑制细胞伸长,最终抑制根伸长。本文还指出,鉴于缺乏对同一植物甚至同一个种类的植物根尖细胞壁各主要组成物质铝胁迫下变化的系统研究,不能对造成该植物根伸长受抑制的原因做出全面合理的解释,所以今后应侧重于铝胁迫下各细胞壁组分变化在抑制根伸长中的贡献率的研究,尤其要针对主要粮食作物进行系统研究,以有效解决铝胁迫造成的产质量降低。     

The Quality of Stemwood of Pinus sylvestris in an Alkalised Environment

The impact of long-term dust pollution emitted from a cement plant on soil chemistry, and the concentrations of plant nutrients, lignin, cellulose and hemicellulose in the stemwood of 80–85-year-old Pinus sylvestris was investigated at different distances from the emission source. It was found that alkaline cement dust (pH 12.3–12.6) emissions for over 40 years resulted in an alkalisation (pH 6.7–7.9) of the polluted soil compared to a pH value of 3.8 in unpolluted soil. There were also nutrient imbalances in the soil, as well as certain disturbances in mineral nutrition processes and accumulation of nutrients in the tree stems. The average concentrations of K, Ca and Mg in stems were higher and those of N and P lower than in the unpolluted area. The lignin (L) content in stemwood increased, hemicellulose (Hc) decreased, while cellulose (Ce) did not change. A variation in the partitioning of L, Ce, Hc and nutrients between different sections of stems and between trees from different sample plots was found. L, Ce and Hc were not related to the internal K, Ca and Mg concentrations. Correlations were established between L, Ce, or Hc content and C content, and between L and Hc content in stem tissues. The contents of wood components were not related to N or P in the alkalised areas, but seemed to be more associated with P than with N. Alterations in the arbitrarily chosen ratio L/(Ce + Hc) indicated changes in wood quality, and a negative correlation with N/P was found in stem tissue in the polluted area, while positive correlations with N/Mg and Ca/Mg were found in the control area.     

Fractionation of flax shives by water and aqueous ammonia treatment in a pressurized low-polarity water extractor

Fractionation of flax shives into cellulose, hemicellulose, and lignin with a two-stage extraction process using water and aqueous ammonia was carried out in a pressurized low-polarity water extractor operated at different temperatures, flow rates, and ammonia concentrations. During the first stage with water, 84% of hemicellulose and 32% of lignin were removed at 190 degrees C at a flow rate of 1.5 mL/min for 30 min. During the second stage with aqueous ammonia, more than 77% of the lignin was removed, and hemicellulose removal reached 95% at 200 degrees C at a flow rate of 0.5 mL/min and with a solvent/feed ratio of 40 mL/g. The temperature and flow rate had a significant effect on lignin removal. The impact of additives (anthraquinone and hydrogen peroxide) and modifications (overnight soaking, reduced particle size, and elevated temperature) on lignin extraction was also studied. The combination of higher temperatures and reduced particle sizes resulted in enhanced lignin extraction. The extraction profiles of free phenolics (vanillin, acetovanillone, and vanillic acid) during the two-stage processing were monitored and compared with those of lignin.     

Hydrothermal Treatment of Date Palm Lignocellulose Residue for Organic Fertilizer Conversion: Effect on Cell Wall and Aerobic Degradation Rate

Date palm (Phoenix roebelenii) woodchips, a residue of palm tree plantations, was subjected to hydrothermal treatment (HTT) at mild reaction conditions (160°C < T < 220°C, 0.6 MPa < P < 2.4 MPa) for 30 min, and the effect of treatments on the cell wall (CW) solubilization and subsequent aerobic degradation rate (as CO production) was tested under controlled composting conditions during 63 days of incubation (38°C). The HTT at 160 and 180°C reaction temperatures notably solubilized hemicellulose, decreasing the fraction of this CW polymer from 34.1% in the untreated material, to 9.5 and 4.6% in the respective residues. However, treatment at 200 and 220°C reaction temperatures rapidly liquefied the lignin, which apparently went into solution with hemicellulose and appeared to stabilize a portion of this polysaccharides against hydrolysis. Consequently, the fraction of hemicellulose in 200 and 220°C – treated residues gradually increased; the respective values were 5.8 and 9.4%. The treatment temperature of 180°C was the most effective HTT temperature for subsequent aerobic degradation by solubilizing the largest portion of hemicellulose within the CW, which had two consequences: 1) it supplied additional readily bioavailable form of carbon, which in turn promoted rapid microbial activities in the early stage of decomposition; and 2) it created pores and cavities within the CW, which permitted rapid bacterial penetration and CW degradation. As a consequence, biodegradation of the residue treated under this reaction temperature proceeded rapidly and stability was reached within 21 days, compared to 63 days of continued degradation for the untreated CW. The enhanced biodegradability was also partially linked to the effect of 180°C treatment temperature on solubilization of amorphous cellulose and partial hydrolysis of lignin. Based on the results, the HTT system can successfully be used as a pretreatment step to accelerate the aerobic digestion rate of date palm residues for the production of organic fertilizers.     

Studies on the metabolism in roots of lowland rice

In the previous studies we investigated some characteristics of paddy root in comparison with shoot by analysis of inorganic elements¹⁾ and metabolizable forms of organic compounds²⁾ of shoot and root at successive growth stages, and found that the contents of the above-mentioned components in root were generally lower than those in shoot with few exceptions. From this it is concluded that in root the contents of the residual components, mainly cell wall substances, must be high. To ascertain this surmise, we carried out analysis of shoot, root and inflorescences of rice plant with respect to hemicellulose, cellulose and lignin which construct cell walls.     

乙二醇-氯化铁预处理对棉秆酶水解效率的影响

为提高棉秆的纤维素酶水解效率,该研究以乙二醇为预处理溶剂,氯化铁为催化剂对棉秆进行预处理,实现了棉秆木质素和半纤维素的有效去除,提高了酶水解效率。以木质素和半纤维素的去除率为指标,运用正交试验方法优化乙二醇-氯化铁预处理条件。结果表明,棉秆在90%乙二醇水溶液,0.1 mol/L氯化铁,固液比1∶15,160 ℃条件下处理20 min,木质素和半纤维素去除率分别为85.7%和88.9%。相较原料,预处理后棉秆酶解率提高了7.6倍,葡萄糖产率达到100%（基质浓度5%,酶载量8.3 FPU/g,水解72 h条件下）。通过结构表征发现乙二醇-氯化铁预处理使棉秆的比表面积增大,致密结构被破坏,有效提高了棉秆的纤维素酶可及性。     

C and N Turnover and Lignocellulose Degradation During Composting of Miscanthus Straw And Liquid Pig Manure

To obtain a peat substitute for pot plants, Miscanthus straw and liquid pig manure were composted in two different systems (open box and closed reactor) and examined for changes in pH, water content and chemical composition (nitrogen, carbon, hemicellulose, cellulose and lignin). Temperature maxima of 65-70°C were achieved within a few days in both systems. Composting and sampling were continued for 190 days in the box system and 150 days in the reactor. Major loss of nitrogen was found in a period of eight days after temperature maximum followed by stabilization. Accumulation of nitrate was observed during final weeks of composting in both systems. A marked decrease in content of hemicellulose was seen just after temperature maximum followed by slow, constant decrease throughout remaining composting period. The same pattern was observed for degradation of cellulose, though degradation was initiated later in the experiment. Degradation of hemicellulose was nearly 100% in both systems, whereas the cellulose decrease was 64 and 70% in the box and reactor, respectively. Lignin was not degraded during the experiments. Though initial C/N ratio was different in the two systems (25 in box and 16 in reactor), final ratios were the same (C/N=13). In spite of the similarities, the more complicated closed system is preferred if removal of ammonia from exit gas is considered.     

Distribution of Accumulated Aluminum and Changes in Cell Wall Polysaccharides in Eucalyptus camaldulensis and Melaleuca cajuputi under Aluminum Stress

Distribution of aluminum (Al) within plant components and Al-induced changes in cell wall polysaccharides in root tips of Eucalyptus camaldulensis Dehnh. seedlings were compared with those of Melaleuca cajuputi Powell. In E. camaldulensis , 0.5 mM Al (pH 4.2 for 40 d) reduced plant dry weight by 50%, increased callose concentration in the root tips and induced leaf necrosis. In comparison with M. cajuputi , Al concentrations were higher in roots and leaves of E. camaldulensis on both a fresh weight basis and in the cell sap, but were lower in the cell wall. Al increased pectin, hemicellulose and cellulose concentration in the cell walls of E. camaldulensis root tips. Al-induced leaf necrosis and growth reduction in E. camaldulensis is discussed in the context of potentially toxic concentrations of Al in plant tissue and changes in polysaccharide content which could reduce water and nutrient uptake and cell wall extensibility in roots.     

Role of wood macromolecules on selective sorption of phenolic compounds by wood

Wood is a complex structure of various macromolecules, mainly cellulose, hemicellulose, and lignin. Although the sorption process of some organic compounds by wood has been elucidated, the relative contribution of its different fractions in the sorption mechanism is not clearly determined. Certain works predict the amount of organic compounds sorbed on wood as a direct relationship to its lignin fraction. All wood macromolecules, however, seem to have the capacity to sorb organic compounds. Sorption of phenolic compounds on individual wood macromolecules has been studied and compared to that on wood. Wood-water partition coefficients ( K wood) for phenolic volatiles and their sorption rates in the presence of lignin display a linear relationship. Results show that cellulose and hemicellulose sorb all phenolic compounds without apparent distinction, whereas lignin is a selective sorbent of these compounds. Sorbant availability and sorbate chemical structure seem to be the key factors governing the sorption mechanism. Sorption kinetics study gives apparent diffusion coefficient values of aroma compounds, bringing new kinetic data for understanding the ternary system of wood, hydroalcoholic solution, and phenolic compounds.     

Synthesis and characterization of dehydrogenation polymers in Gluconacetobacter xylinus cellulose and cellulose/pectin composite

To mimic the lignin polymerization process, mats of bacterial cellulose and of a pectin/cellulose composite were used as a host matrix for in vitro polymerization of coniferyl alcohol. A diffusion cell was used to allow the diffusion of both hydrogen peroxide and coniferyl alcohol into the peroxidase impregnated cellulose mats through dialysis membranes. The results indicate that significant polymerization occurs within the mats. The resulting binary and ternary blends were imaged by scanning electron microscopy (SEM) and characterized by chemical means. The presence of pectin induces a better dispersion of the synthetic lignin in the cellulose network and enhances the proportion of alkyl-aryl-ether in the polymer.     

城市污泥与调理剂混合堆肥过程中有机质组分的变化

【目的】研究城市污泥堆肥过程中各项有机质组分及碳、 氮在堆肥过程中的形成与转化,以期改善堆肥的生物有效性,促进其土地利用。【方法】在工厂规模化下,以城市污泥、 蘑菇渣锯末以及返混料按照6∶3∶1的质量比混合形成堆肥物料,辅以强制通风措施和翻抛,进行为期18 d的高温堆肥试验。堆肥期间定期采样,测定指标包括温度、 C/N值、 pH、 含水率、 有机质降解率、 水溶性组分、 半纤维素、 纤维素和木质素,研究堆肥期间不同阶段堆肥物料中有机质组分的动态变化。【结果】堆体温度随着发酵时间的延长呈现先升高后降低的趋势,最高温度达到71.3℃; 含水率由60.7%降低到51.4%,pH呈现先升高后降低的趋势,总体处于6.0~7.5之间; 总有机碳含量持续下降,氮素含量表现为高温期持续下降随后呈上升的趋势; 初始阶段,堆肥物料中四种成分含量分布为: 水溶性组分纤维素半纤维素木质素,至堆肥结束变化为: 纤维素水溶性组分木质素半纤维素,经过堆肥之后水溶性组分及半纤维素含量分别由39.5%和20.1%下降为27.9%和14.4%,纤维素含量由初始的21.8%上升至29.5%,木质素含量相对稳定不变。物料经过堆肥化处理后达到腐熟标准,水溶性组分和半纤维素含量分别降低了38.6%和38.8%,纤维素和木质素含量在高温期分别降解了11.7%和18.5%; 物料总量降低了9.8%。水溶性组分的主要降解阶段为高温期,期间降解部分占总降解量的65.5%; 半纤维素的主要降解阶段为稳定期,稳定期降解部分占总降解量的69.1%,且有继续降解的趋势; 纤维素和木质素仅在高温期有少量降解; 氮素则表现为高温期铵态氮的损失及稳定期硝态氮的积累。【结论】堆肥化处理在实现污泥减量化基础上,污泥中有机质得到了稳定化,有利于城市污泥的土地利用。     

Influence of non-cellulose structural carbohydrate composition on plant material decomposition in soil

The C mineralisation pattern during the early stage of decomposition of plant materials is largely determined by their content of different carbohydrates. This study investigated whether detailed plant analysis could provide a better prediction of C mineralisation during decomposition than proximate analysis [neutral detergent solution (NDF)/acid detergent solution (ADF)]. The detailed analysis included sugars, fructans, starch, pectin, cellulose, lignin and organic N. To determine whether differences in decomposition rate were related to differences in hemicellulose composition, the analysis particularly emphasised the concentrations of arabinose and xylose in hemicelluloses. Carbon dioxide evolution was monitored hourly in soil amended with ten different plant materials. Principal component and regression analysis showed that C mineralisation during day 1 was closely related to free sugars, fructans and soluble organic N components (R ² = 0.83). The sum of non-cellulose structural carbohydrates (intermediate NDF/ADF fraction) was not related to C mineralisation between days 1 and 9. In contrast, a model including starch and protein in addition to the non-cellulose structural carbohydrates, with the hemicelluloses replaced by arabinose and xylose, showed a strong relationship with evolved CO₂ (R ² = 0.87). Carbon mineralisation between days 9 and 34 was better explained by xylan, cellulose and lignin (R ² = 0.72) than by lignocellulose in the ADF fraction. Our results indicated that proximate analyses were not sufficient to explain differences in decomposition. To predict C mineralisation from the range of plant materials studied, we propose a minimum set of analyses comprising total N, free sugars, starch, arabinose, xylan, cellulose and lignin.     

Biosynthesis, molecular structure, and domain architecture of potato suberin: a (13)C NMR study using isotopically labeled precursors

Although suberin in potato wound periderm is known to be a polyester containing long-chain fatty acids and phenolics embedded within the cell wall, many aspects of its molecular structure and polymer-polymer connectivities remain elusive. The present work combines biosynthetic incorporation of site-specifically (13)C-enriched acetates and phenylalanines with one- and two-dimensional solid-state (13)C NMR spectroscopic methods to monitor the developing suberin polymer. Exogenous acetate is found to be incorporated preferentially at the carboxyl end of the aliphatic carbon chains, suggesting addition during the later elongation steps of fatty acid synthesis. Carboxyl-labeled phenylalanine precursors provide evidence for the concurrent development of phenolic esters and of monolignols typical of lignin. Experiments with ring-labeled phenylalanine precursors demonstrate a predominance of sinapyl and guaiacyl structures among suberin's phenolic moieties. Finally, the analysis of spin-exchange (solid-state NOESY) NMR experiments in ring-labeled suberin indicates distances of no more than 0.5 nm between pairs of phenolic and oxymethine carbons, which are attributed to the aromatic-aliphatic polyester and the cell wall polysaccharide matrix, respectively. These results offer direct and detailed molecular information regarding the insoluble intermediates of suberin biosynthesis, indicate probable covalent linkages between moieties of its polyester and polysaccharide domains, and yield a clearer overall picture of this agriculturally important protective material.     

Tissue-specific patterns of lignification are disturbed in the brown midrib2 mutant of maize (Zea mays L.)

Despite recent progress, several aspects of lignin biosynthesis, including variation in lignin composition between species and between tissues within a given species, are still poorly understood. The analysis of mutants affected in cell wall biosynthesis may help increase the understanding of these processes. We have analyzed the maize brown midrib2 (bm2) mutant, one of the four bm mutants of maize, using pyrolysis-mass spectrometry (Py-MS) and pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS). Vascular tissues from the leaf blade and leaf sheath from different parts of the plant were investigated and compared to the corresponding samples from a wild-type plant of the same genetic background (inbred line A619). Multivariate analysis revealed that the bm2 mutant had reduced amounts of di- and trimeric lignin derivatives, notably species with m/z 272 and m/z 330, and that the ratio of guaiacyl residues to polysaccharides was reduced in the bm2 mutant. In addition, differences in cell wall composition between different parts of the plant (blade versus sheath, young versus old tissue) were much less pronounced in the bm2 mutant. These changes suggest that the functional Bm2 gene is important for the establishment of tissue-specific cell wall composition.