The role of lignin and cellulose in the carbon-cycling of degraded soils under semiarid climate and their relation to microbial biomass

A high level of biological degradation is usually observed in soils under semiarid climate where the low inputs of vegetal debris constraint the development of microbiota. Among vegetal inputs, cellulose and lignin are dominant substrates but their assimilation by the microbial community of semiarid soils is yet not understood. In the present study, ¹³C-labeled cellulose and ¹³C-labeled lignin (75 μg ¹³C g⁻¹ soil) were added to two semiarid soils with different properties and degradation level. Abanilla soil is a bare, highly degraded soil without plant cover growing on it and a total organic C content of 5.0 g kg⁻¹; Santomera soil is covered by plants (20% coverage) based on xerophytic shrubs and has a total organic C content of 12.0 g kg⁻¹. The fate of added carbon was evaluated by analysis of the carbon isotope signature of bulk soil-derived carbon and extractable carbon fractions (water and sodium-pyrophosphate extracts). At long-term (120 days), we observed that the stability of cellulose- and lignin-derived carbon was dependent on their chemical nature. The contribution of lignin-derived carbon to the pool of humic substances was higher than that of cellulose. However, at short-term (30 days), the mineralization of the added substrates was more related to the degradation level of soils (i.e. microbial biomass). Stable isotope probing (SIP) of phospholipid fatty acids (PLFA-SIP) analysis revealed that just a minor part of the microbial community assimilated the carbon derived from cellulose and lignin. Moreover, the relative contribution of each microbial group to the assimilation of lignin-derived carbon was different in each soil.     

Biochemical defense responses of black pepper (Piper nigrum L.) lines to Phytophthora capsici

A study on biochemical factors involved in black pepper defense response against Phytophthora capsici, was carried out in P. capsici susceptible (Sreekara) and resistant (04-P24, shows root resistance to the pathogen) black pepper lines. Seven important factors – change in membrane conductance, total phenols, orthodihydroxy (OD) phenols, lignin and defense related enzymes (peroxidase, β-1,3 glucanase and β-1,4 glucanase) – were studied under uninoculated and pathogen (P. capsici, isolate 06-04) inoculated condition to know the preformed and induced responses. The pathogen was inoculated (soil inoculation) and plants were observed for changes, at 24 h intervals for 10 days. On 8th day after inoculation symptoms started appearing on Sreekara and increased the severity till 10th day. Both root and stem samples were subjected for biochemical analysis. Of the factors analyzed, it was found that membrane conductance, OD phenol, lignin and peroxidase activity play significant role in root resistance to P. capsici in 04-P24. Light microscopy of the portion of root – where pathogen found attached – was also done.     

Soil aggregation and the stabilization of organic carbon as affected by erosion and deposition

The importance of soil aggregation in determining the dynamics of soil organic carbon (SOC) during erosion, transportation and deposition is poorly understood. Particularly, we do not know how aggregation contributes to the often-observed accumulation of SOC at depositional sites. Our objective was to assess how aggregation affects SOC stabilization in comparison to interactions of SOC with minerals. We determined and compared aggregate size distributions, SOC distribution in density fractions, and lignin-derived phenols from aggregated soil samples at both eroding and depositional sites. The stabilization effect of aggregation was quantified by comparing mineralization from intact and crushed macro-aggregates. Deposition of eroded soil material resulted in carbon (C) enrichment throughout the soil profile. Both macro-aggregate associated SOC and C associated with minerals (heavy fraction) increased in their importance from the eroding to the depositional site. In the uppermost topsoil (0–5 cm), SOC mineralization from intact aggregates was larger at the depositional site than at the eroding site, reflecting the large input of labile organic matter (plant residues) promoting aggregation. Contrastingly, in the subsoil, mineralization rates were lower at the depositional site because of effective stabilization by interactions with soil minerals. Aggregate crushing increased SOC mineralization by 10–80% at the eroding site, but not at the depositional site. The content of lignin-derived phenols did not differ between eroding and depositional sites in the topsoil (24.6–30.9 mg per g C) but was larger in the subsoil of the eroding site, which was accompanied by higher lignin oxidation. Lignin data indicated minor effects of soil erosion and deposition on the composition of SOC. We conclude that SOC is better protected in aggregates at the eroding than at the depositional site. During transport disaggregation and consequently SOC mineralization took place, while at the depositional site re-aggregation occurred mainly in the form of macro-aggregates. However, this macro-aggregation did not result in a direct stabilization of SOC. We propose that the occlusion of C inside aggregates serves as a pathway for the eroded C to be later stabilized by organo-mineral interaction.     

甘蓝型油菜茎秆菌核病抗性与木质素及其单体比例的相关性分析及QTL定位

菌核病是一类非专一性的植物真菌病原菌，寄主范围广泛，严重危害农作物的生产。对高世代重组自交系群体(RIL)及F₂群体终花期茎秆进行菌核病抗性接种鉴定，根据构建的近红外模型对接种鉴定的茎秆木质素含量、单体组分比例进行测定，并进行相关性分析和QTL定位。结果表明在2013年和2014年RIL群体茎秆菌斑大小与木质素含量呈极显著负相关，相关系数分别为–0.348和–0.286，与单体G/S呈显著正相关，相关系数分别为0.198和0.167。2014年F₂群体菌斑大小与木质素含量呈极显著负相关，相关系数为–0.306，与单体G/S相关性为0.142。F_2:3家系抗(感)植株茎部切片间苯三酚染色观察表明抗性较强的材料木质素含量高于抗性较弱的材料。根据已构建的重组自交系高密度SNP遗传图谱，利用复合区间作图法对上述性状进行QTL分析，共检测到18个QTL，其中9个菌核病抗性相关QTL分布于A05、A06、C04和C06染色体，单个QTL可解释的表型变异为2.38%~12.05%；3个木质素含量QTL分别位于A04、A05和C01染色体，单个QTL可解释表型变异的2.03%~13.75%。6个木质素单体G/S QTL分布于A08、C03和C07染色体，单个QTL可解释表型变异的2.06%~8.66%。本文研究结果为油菜菌核病抗性育种提供了新的思路和理论基础。     

棉花纤维中木质素的相对分子量

分子量是聚合物的重要特性之一,木质素的分子量及其分布是研究苯丙烷类结构的反应、物理化学特性和评价其改性产物质量的内容之一。本研究以陆地棉TM-1成熟纤维为材料,分别利用酶解-温和酸解木质素法和二氧六环法提取棉纤维中木质素,结合凝胶渗透色谱法(gel permeation chromatography,GPC)调查和评价2种方法获得的棉纤维中木质素的相对分子量。结果表明,经二氧六环处理提取的棉花纤维中的木质素(dioxane lignin,DL)的重均分子量为2924g mol~(–1)、数均分子量2403 g mol~(–1),略高于由酶解-温和酸解处理提取的木质素(enzymatic hydrolysis-mild acidolysis lignin,EMAL)的重均分子量(2169 g mol~(–1))和数均分子量(1970 g mol~(–1)),EMAL的多分散系数稍低,说明木质素的均一性比DL高。表明EMAL法提取的木质素更适用于分析棉纤维中木质素的相对分子量。利用EMAL法分析棉纤维中木质素相对分子量表明,不同棉花品种的木质素重均分子量分布范围为938~2169 g mol~(–1),数均分子量分布范围为857~1970 g mol~(–1),多分散性系数在1.09~1.74间,均小于2。重均分子量与纤维马克隆值呈显著负相关,数均分子量与纤维长度呈显著负相关,与纤维马克隆值呈极显著负相关。     

Variation in the chemical composition of green leaves and leaf litters from three deciduous tree species growing on different soil types

The chemical composition of green leaves and leaf litters of sweet chestnut (Castanea sativa), oak (Quercus robur) and beech (Fagus sylvatica) were determined for 26 sites grouped into high fertility (HF) and low fertility (LF) soils according to base saturation and N-mineralization potentials. Measurements were made of total carbon, acid detergent fibre (ADF), Klason lignin, holo-cellulose, sugar constituents of hemicellulose and phenylpropanoid derivatives of lignin, and nutrient concentrations (N, Ca, P, Mg, K and Mn). Leaf and litter constituents varied within and between species according to soil groups, but beech showed contrasting responses to oak and chestnut. Beech leaves had lower ADF, lignin and cellulose on HF soils than LF soils, whereas oak and chestnut leaves had higher ADF, lignin and cellulose on HF than the LF soils. Conversely, the same constituents in beech leaf litter were higher on HF soils than LF soils, but lower in oak and chestnut leaf litter on HF soils than LF soils. The phenylpropanoid derivatives of lignin and sugar constituents of hemicellulose also showed similar variations in relation to soil groups with contrasting patterns for in leaves and litters. Re-absorption of N from leaves before litter fall was negatively correlated with soil N mineralization potential for beech (highest on LF soils) but showed an unexpected, positive relationship for oak and chestnut (highest on HF soils). These intra-specific differences of leaf and litter chemistry in relation to soil fertility status are unprecedented and largely unexplained. The observed patterns reflect phenotypic responses to soil type that result in continuum of litter quality, within and between tree species, that have been shown in related studies to significantly influence litter decomposition rates.     

Infrared spectroscopic study of alkaline oxygen treatment of lignin with ATR technique in aqueous state 1: method for determining quantitative spectra of oxygen-degraded lignin

For the fundamental study of oxygen delignification of kraft pulp, structural changes of kraft lignin during alkaline oxygen treatment were investigated with the use of infrared measurement with attenuated total reflectance (ATR) technique. In the neutralized reaction mixture of alkaline oxygen-treated kraft lignin, there is a significant amount of NaCl, so that the spectral changes of water due to the coexistence of NaCl was investigated, and how to remove the huge absorption of NaCl solution is discussed. Sodium vanillate–NaCl solutions were employed as model solutions for the reaction mixture. Partial least square (PLS) regression was applied for the prediction of NaCl concentration, and the spectrum of NaCl solution was subtracted from the spectrum of sodium vanillate–NaCl solution as background measurement. This allowed us to obtain the vanillate spectra free from the absorption of NaCl solution. In addition, the mathematical method for reconstructing the spectrum of NaCl solution is discussed. The spectrum of NaCl solution is reconstructed as the linear combination of basic spectra calculated by singular value decomposition (SVD), and it was subtracted from that of the sodium vanillate–NaCl solution. By this procedure, the vanillate spectra were also obtained quantitatively, as has been demonstrated in PLS regression study. It was also confirmed that the quantitative spectra of high molecular weight fraction of alkaline oxygen-treated kraft lignin were obtained by the use of this reconstruction technique.Parts of this report were presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, Japan, April 2002 and the 12th International Symposium on Wood and Pulping Chemistry, Madison, USA, June 2003     

Semi-quantitative method to evaluate the α-carbonyl content in lignin

A method to estimate the content of -carbonyl structures in lignin was developed. This method consists of two successive treatments: NaBD₄ treatment of pulp to reduce an -carbonyl structure in lignin, and nitrobenzene oxidation. NaBD₄ was used to convert an -carbonyl structure to a deuterium-labeled hydroxymethine structure. The ratio of D-vanillin [(HO)(H₃CO)C₆H₃CDO] to H-vanillin [(HO)(H₃CO)C₆H₃CHO] or that of their syringyl analogues obtained by nitrobenzene oxidation was used as the measure of the content of -carbonyl structure. Model experiments demonstrated that when sodium hydroxide was used as alkali for the nitrobenzene oxidation, the retention of deuterium at the side chain -position was very low due to the displacement of deuterium with hydrogen by an unknown reaction mechanism. In order to depress this unexpected displacement, the reaction conditions of the nitrobenzene oxidation were modified. The modified nitrobenzene oxidation employs 0.5mol/l of lithium hydroxide as a reaction medium instead of 2.0mol/l sodium hydroxide. By this modification, this method could successfully trace the formation and the degradation of the -carbonyl structure in milled wood lignins.This paper was presented in part at the 11th International Symposium on Wood and Pulping Chemistry, Nice, France, June 2001 and at the 46th Lignin Symposium, Kyoto, Japan, November 2001     

Characteristics and decomposition rates of pruning residues from a shaded coffee system in Southeastern Brazil

In the Zona da Mata Mineira of Southeastern Brazil the development of sustainable land requires the integration of crops with trees. The objectives of this study then were to (i) characterize prunings from the main tree species in an agroforestry system; (ii) determine the effects of the physical and chemical characteristics of the prunings on their decomposition patterns in the laboratory; (iii) assess the effect of mixing leaves of different species on decomposition rates; and (iv) propose a decomposition index for the residues studied. The study was carried out with pruning residues from Cajanus cajan, Solanum variable, Cassia ferruginea, Piptadenia gonoacantha, Croton urucurana, and Melinis multiflora. The materials were characterized for total C, N, P, Ca, Mg and K contents; lignin, cellulose, hemicellulose and soluble polyphenols contents. The pruning residues had high polyphenols and lignin contents, high C:N and C:P ratios, and low contents of Ca, Mg, and K. The low decomposition rates of the prunings were related to the P, K, hemicellulose and polyphenol contents. The rates of N mineralization from most of the residues indicate that there is a potential to supply the needs of a crop of maize. The residues of some species, if decomposed alone, would not supply sufficient nutrients, and need to be mixed with leaves of other species.This revised version was published online in November 2005 with corrections to the Cover Date.     

Reaction behavior of lignin in supercritical methanol as studied with lignin model compounds

 The reaction behavior and kinetics of lignin model compounds were studied in supercritical methanol with a batch-type supercritical biomass conversion system. Guaiacol, veratrole, 2,6-dimethoxyphenol, and 1,2,3-trimethoxybenzene were used as model compounds for aromatic rings in lignin. In addition, 5-5, β-1, β-O-4, and α-O-4 types of dimeric lignin model compounds were used as representatives of linkages in lignin. As a result, aromatic rings and 5-5 (biphenyl)-type structures were stable in supercritical methanol, and the β-1 linkage was not cleaved in the β-1-type structure but converted rapidly to stilbene. On the other hand, β-ether and α-ether linkages of β-O-4 and α-O-4 lignin model compounds were cleaved rapidly, and these compounds decomposed to some monomeric compounds. Phenolic compounds were found to be more reactive than nonphenolic compounds. These results indicate that cleavages of ether linkages mainly contribute to the depolymerization of lignin, whereas condensed linkages such as the 5-5 and β-1 types are not cleaved in supercritical methanol. Therefore, it is suggested that the supercritical methanol treatment effectively depolymerizes lignin into the lower-molecular-weight products as a methanol-soluble portion mainly by cleavage of the β-ether structure, which is the dominant linkage in lignin. Received: December 19, 2001 / Accepted: April 30, 2002 Acknowledgments This research has been done under the research program for the development of technologies for establishing an ecosystem based on recycling in rural villages for the twenty-first century from the Ministry of Agriculture, Forestry and Fisheries, Japan; by a Grant-in-Aid for Scientific Research (B)(2) (no.12460144, 2001.4–2003.3) from the Ministry of Education, Culture, Sports, Science and Technology, Japan; and under the research program from Kansai Research Foundation for Technology Promotion, Japan. The authors thank them for their financial support. This study was presented in part at the 45th Lignin Symposium, Ehime, Japan, October 2000 and the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, Japan, April 2002 Correspondence to:S. Saka