Expression of a fungal laccase fused with a bacterial cellulose-binding module improves the enzymatic saccharification efficiency of lignocellulose biomass in transgenic Arabidopsis thaliana

Delignification is effective for improving the saccharification efficiency of lignocellulosic biomass materials. We previously identified that the expression of a fungal laccase (Lac) fused with a bacterial cellulose-binding module domain (CBD) improved the enzymatic saccharification efficiency of rice plants. In this work, to evaluate the ability of the Lac-CBD fused chimeric enzyme to improve saccharification efficiency in a dicot plant, we introduced the chimeric gene into a dicot model plant, Arabidopsis thaliana. Transgenic plants expressing the Lac-CBD chimeric gene showed normal morphology and growth, and showed a significant increase of enzymatic saccharification efficiency compared to control plants. The transgenic plants with the largest improvement of enzymatic saccharification efficiency also showed an increase of crystalline cellulose in their cell wall fractions. These results indicated that expression of the Lac-CBD chimeric protein in dicotyledonous plants improved the enzymatic saccharification of plant biomass by increasing the crystallinity of cellulose in the cell wall

Comparative Assessment of Biochar Stability Using Multiple Indicators

Biochar application is one strategy proposed to improve carbon sequestration in soil. Maintaining high carbon content in soil for a long period requires stable biochar. In this work, we assessed biochar stability by two methodologies, i.e., laboratory incubation and chemical oxidation. Biochar was produced at four different temperatures (400 °C, 500 °C, 600 °C, and 800 °C) from rice (Oryza sativa L.) straw and husk, applewood branch (Malus pumila), and oak (Quercus serrata Murray) residues. Results showed that the high-temperature biochars were more stable in both abiotic and biotic incubations, whereas the low-temperature biochars had reduced longevity. In addition, we showed biochars originated from woody material have higher stable carbon than those produced from rice residues. Finally, the oxidative assessment method provided a more reliable estimation of stability than the biotic incubation method and showed a strong correlation with other stability indicators

The efficiency of a low dose of biochar in enhancing the aromaticity of humic-like substance extracted from poultry manure compost

Using biochar as a bulking agent in composting is gradually becoming popular for the minimization of nitrogen losses during the process and the improvement in compost quality. While a wide range of different biochar doses is applied, not much clear information was available about the optimum ratio. This study presents the impact of adding a low dose (2% v/v) of slow-pyrolysis oak biochar (Quercus serrate Murray), into poultry manure on the recalcitrant characteristic of humified organic matter. The influence in the chemical composition of humic-like substance was evaluated in poultry manure compost prepared with (PM+B) and without biochar (PM). The shift to slightly more stable chemical composition was shown in humic acid-like (HA) and fulvic acid-like (FA) extracted from PM+B compost, by increasing the proportion of aromatic carbon groups and thermal stability measured by thermogravimetry. We conclude that the addition of 2% biochar moderately enhances the recalcitrance of humified organic carbon and this could be feasible for the implementation of the biochar use in composting since only a small amount is required.</p

Methodological interference of biochar in the determination of extracellular enzyme activities in composting samples

Biochar application has received increasing attention as a means to trap recalcitrant carbon and enhance soil fertility. Hydrolytic enzymatic assays, such as β-glucosidase and phosphatase activities, are used for the assessment of soil quality and composting process, which are based on use of p-nitrophenol (PNP) derivatives as substrate. However, sorption capacity of biochar can interfere with colorimetric determination of the hydrolysed PNP, either by the sorption of the substrate or the reaction product of hydrolysis into biochar surface. The aim of the present work is to study the biochar sorption capacity for PNP in biochar-blended composting mixtures in order to assess its impact on the estimation of the colorimetric-based enzymatic assays. A retention test was conducted by adding a solution of known amounts of PNP in universal buffer solution (pH = 5, 6.5 and 11, corresponding to the β-glucosidase, acid and alkaline phosphatase activity assays, respectively), in samples taken at the initial stage and after maturation stage from four different composting piles (two manure composting piles; PM: poultry manure, CM: cow manure and two other similar piles containing 10% of additional biochar (PM + B, CM + B)). The results show that biochar-blended composts (PM + B, CM + B) generally exhibited low enzymatic activities, compared to manure compost without biochar (PM, CM). In terms of the difference between the initial and maturation stage of composting process, the PNP retention in biochar was shown higher at maturation stage, caused most probably by an enlarged proportion of biochar inside compost mixture after the selective degradation of easily decomposable organic matter. TThe retention of PNP on biochar was influenced by pH dependency of sorption capacity of biochar and/or PNP solubility, since PNP was more efficiently retained by biochar at low pH values (5 and 6.5) than at high pH values (11).We are very grateful for financial support by the Japan Society of the Promotion of science as well as CSIC programmes of bilateral project. This research was financed by a grant from the Spanish Ministry of Science and Innovation, research project AGL2012-40143-C02-01.Peer reviewe

Physical and chemical characterization of biochars derived from different agricultural residues

Biochar is widely recognized as an efficient tool for carbon sequestration and soil fertility. The understanding of its chemical and physical properties, which are strongly related to the type of the initial material used and pyrolysis conditions, is crucial to identify the most suitable application of biochar in soil. A selection of organic wastes with different characteristics (e.g., rice husk (RH), rice straw (RS), wood chips of apple tree (Malus pumila) (AB), and oak tree (Quercus serrata) (OB)) were pyrolyzed at different temperatures (400, 500, 600, 700, and 800 °C) in order to optimize the physicochemical properties of biochar as a soil amendment. Low-temperature pyrolysis produced high biochar yields; in contrast, high-temperature pyrolysis led to biochars with a high C content, large surface area, and high adsorption characteristics. Biochar obtained at 600 °C leads to a high recalcitrant character, whereas that obtained at 400 °C retains volatile and easily labile compounds. The biochar obtained from rice materials (RH and RS) showed a high yield and unique chemical properties because of the incorporation of silica elements into its chemical structure. The biochar obtained from wood materials (AB and OB) showed high carbon content and a high absorption character.This work was partly supported by the bilateral project of the Japan Society for the Promotion of Science (JSPS) and the Spanish National Research Council (CSIC).Peer reviewe

Influence of biochar addition on the humic substances of composting manures

Application of biochar (10% v/v) to a manure composting matrix was investigated to evaluate its effect on the chemical composition of humic substances during the composting process. The characteristics of the humic acid (HA) and fulvic acid (FA) fractions were analyzed in compost mixtures originating from two different manures (poultry manure (PM) and cow manure (CM)). The C contents of HA and FA from the manure compost/biochar blends (PM+B and CM+B) were higher than those from PM and CM, with an enhanced recalcitrant fraction, as determined by thermogravimetric analysis. Spectroscopic analysis showed that enrichment of aromatic-C and carboxylic-C occurred in the FA fractions of PM+B and CM+B to a greater extent than in PM and CM. Biochar addition into the composting mixture improved the final compost quality, especially for the light humified fraction (FA).We are very grateful to the Japan Society for the Promotion of Science for their financial support as well as to the CSIC (Consejo Superior de Investigaciones Científicas) programs for supporting this bilateral project. The financial support from the Fundación Séneca (Murcia, Spain), the Spanish Ministry of Economy and Competitiveness (research project AGL2012-40143-C02-01) and the government agency Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) (Program “Atração de Jovens Talentos”) are also appreciated.Peer reviewe