Pretreatment of partially delignified hybrid poplar for biofuels production: Characterization of organosolv hemicelluloses

In this study, extraction of hemicelluloses from the carbohydrate-enriched residues was successfully carried out with organic solvent and the residue was used for bio-based energy production. The chemical composition and physico-chemical properties of six hemicelluloses released were elucidated by a combination of sugar analysis, molecular determination, Fourier transform infrared, and ¹H, ¹³C and 2D-HSQC NMR spectroscopy. The results showed that the successful treatments resulted in a fractionation of the native hemicelluloses. The sugar analysis indicated that xylose (47.14-56.91%) was found to be the major sugar components and small amounts of glucose (14.1-19.06%) and mannose (12.41-18.09%) were also observed in these hemicellulosic fractions. Further studies by NMR spectroscopy exhibited that the acetylated hemicellulosic fraction had a main structure of (1 → 4)-linked β-d-xylopyranosyl backbone with 4-O-methyl-α-d-glucuronic acid as a side chain and a minor structure of linear β-(1 → 4)-linked glucomannans. Furthermore, these hemicelluloses possessed a low substituted degree which was beneficial for enzymatic saccharification.     

Triterpenoid saponins from corms of Crocus sativus: Localization, extraction and characterization

A mixture of highly glycosilated triterpenoid saponins (CS5) isolated from the corm of Crocus sativus or saffron showed cytotoxic activity against HeLa tumoral cells. The main reverse phase HPLC fraction of this mixture (CS5-1) contains two new oleanane-type saponins, denominated Azafrine 1 (1) and Azafrine 2 (2). The bidesmosidic saponins were respectively characterized as (1) 3-O-β-d-glucopyranosiduronic acid echinocystic acid 28-O-β-D-galactopyranosyl-(1 → 2)-α-l-arabinopyranosyl-(1 → 2)-[β-d-xylopyranosyl-(1 → 4)]-α-d-rhamnopyranosyl-(1 → 2)-[4-O-di-α-L-rhamnopyranosyl-3,16-dihydroxy-10-oxo-hexadecanoyl]-α-D-fucopyranoside and (2) 3-O-β-D-galactopyranosiduronic acid echinocystic acid 28-O-β-D-galactopyranosyl-(1 → 2)-α-L-arabinopyranosyl-(1 → 2)-[β-D-xylopyranosyl-(1 → 4)]-α-L-rhamnopyranosyl-(1 → 2)-[4-O-di-α-L-rhamnopyranosyl-3,16-dihydroxy-10-oxo-hexadecanoyl]-β-D-fucopyranoside. The surfactant properties of saponins, probably involved in the cytotoxic activity of CS5 and their exclusive localization in the external part or the corm, indicate their possible role as phytoprotectans. The similarity of their structural compositions to that of other triterpenoid saponins which are of special use in the pharmaceutical industry suggest a new application for C. sativus crops through the exploitation of corm for saponin extraction.     

Ethanol production from alkali- and ozone-treated cotton stalks using thermotolerant Pichia kudriavzevii HOP-1

In the present study, milled cotton stalks were subjected to alkali pretreatment with NaOH at 1-4% (w/v) concentrations at 121 °C for time ranging from 30 to 90 min. Ozone pretreatment was performed by passing 45 mg/L of ozone gas over 2 mm cotton stalks for 150 min at a flow rate of 0.37 L/min. The residual biomass from 4% alkali pretreatment for 60 min showed 46.6% lignin degradation accompanied by 83.2% increase in glucan content, compared with the untreated biomass. Hydrolysis of 4% alkali-treated and ozone-treated cotton stalks was conducted using enzyme combination of 20 filter paper cellulase units/gram dried substrate (FPU/g-ds), 45 IU/g-ds β-glucosidase and 15 IU/g-ds pectinase. Enzymatic hydrolysis of alkali-treated and ozone-treated biomass after 48 h resulted in 42.29 g/L glucose, 6.82 g/L xylose and 24.13 g/L glucose, 8.3 g/L xylose, respectively. About 99% of glucose was consumed in 24 h by Pichia kudriavzevii HOP-1 cells resulting in 19.82 g/L of ethanol from alkali-treated cotton stalks and 10.96 g/L of ethanol from ozone-treated cotton stalks. Simultaneous saccharification and fermentation of the alkali-treated cotton stalks after 12-h pre-hydrolysis resulted in ethanol concentration, ethanol yield on dry biomass basis and ethanol productivity of 19.48 g/L, 0.21 g/g and 0.41 g/L/h, respectively which holds promise for further scale-up studies. To the best of our knowledge, this is the first study employing SSF for ethanol production from cotton stalks.     

Morphological characteristics and composition of lipophilic extractives and lignin in Brazilian woods from different eucalypt hybrids

The morphological and chemical characteristics of the woods from several eucalypt hybrids from the Brazilian Genolyptus program were studied. The hybrids selected for this study were Eucalyptus grandis × E. urophylla (IP), E. urophylla × E. urophylla (U1 × U2), E. grandis × [E. urophylla × E. globulus] (G1 × UGL), and [E. dunnii × E. grandis] × E. urophylla (DG × U2). The analyses of the lipophilic extractives indicated a similar composition in all eucalypt hybrids, which were dominated by sitosterol, sitosterol esters and sitosteryl 3β-d-glucopyranoside. These compounds are responsible for pitch deposition during kraft pulping of eucalypt wood. Some quantitative differences were found in the abundances of different lipid classes, the wood from U1 × U2 having the lowest amounts of these pitch-forming compounds. The chemical composition and structure of lignins were characterized by Py-GC/MS and 2D-NMR that confirmed the predominance of syringyl over guaiacyl units and only showed traces of p-hydroxyphenyl units in all the woods, with the highest S/G ratio for G1 × UGL. The 2D-NMR spectra gave additional information about the inter-unit linkages in the lignin polymer. All the lignins showed a predominance of β-O-4′ ether linkages (75-79% of total side-chains), followed by β-β′ resinol-type linkages (9-11%) and lower amounts of β-5′ phenylcoumaran-type, β-1′ spirodienone-type linkages or β-1′ open substructures. The lignin from the hybrid G1 × UGL presented also the highest proportion of β-O-4′ linkages, and therefore, it is foreseen that the wood from this hybrid will be more easily delignifiable than the other selected Brazilian eucalypt hybrids. In complement to these chemical analyses, the morphological characterization of fibers, vessels and fines revealed that hybrid eucalypt clone DG × U2 presented the most interesting properties for the manufacture of paper pulps and biofuels.     

Bioethanol production from sweet sorghum bagasse by Mucor hiemalis

The present work deals with production of ethanol from sweet sorghum bagasse by a zygomycetes fungus Mucor hiemalis. The bagasse was treated with phosphoric acid and sodium hydroxide, with or without ultrasonication, prior to enzymatic hydrolysis by commercial cellulase and β-glucosidase enzymes. The phosphoric acid pretreatment was performed at 50 °C for 30 min, while the alkali treatment performed with 12% NaOH at 0 °C for 3 h. The pretreatments resulted in improving the subsequent enzymatic hydrolysis to 79-92% of the theoretical yield. The best hydrolysis performance was obtained after pretreatment by NaOH assisted with ultrasonication. The fungus showed promising results in fermentation of the hydrolyzates. In the best case, the hydrolyzate of NaOH-ultrasound pretreated bagasse followed by 24 h fermentation resulted in about 81% of the corresponding theoretical ethanol yield. Furthermore, the highest volumetric ethanol productivity was observed in the hydrolyzates of NaOH pretreated bagasse, especially after ultrasonication in pretreatment stage.     

Optimization of ethanol production by Saccharomyces cerevisiae UFPEDA 1238 in simultaneous saccharification and fermentation of delignified sugarcane bagasse

Ethanol production by Saccharomyces cerevisiae UFPEDA1238 was performed in simultaneous saccharification and fermentation of delignified sugarcane bagasse. Temperature (32 °C, 37 °C), agitation (80; 100 rpm), enzymatic load (20 FPU/g cellulose and 10%, v/v β-glucosidase or 10 FPU/g cellulose and 5% β-glucosidase) and composition of culture medium were evaluated. Ethanol concentration, enzymatic convertibility of cellulose and volumetric productivity were higher than 25 g/L, 72% and 0.70 g/L h, respectively, after 30 h, when the culture medium 1 and 20 FPU/g cellulose/10%, v/v β-glucosidase or the culture medium 2 and 10 FPU/g cellulose/5% β-glucosidase were used in SSF at 37 °C and 80 rpm. In the SSF with culture medium 2 (supplemented with ammonium, phosphate, potassium and magnesium), 150 L ethanol/t bagasse was achieved, with minimum enzyme loading (10 FPU/g cellulose and 5%, v/v β-glucosidase) for 8%, w/v of solids, which is often an important requirement to provide cost-efficient second generation ethanol processes.     

Conversion of starch from dry common beans (Phaseolus vulgaris L.) to ethanol

Dry common beans (Phaseolus vulgaris L.) were evaluated for potential conversion of starch to ethanol. Eight varieties of beans with average starch content of 46% (db) were assayed in a laboratory-scaled process based upon the commercial corn dry grind fermentation process. Ethanol yield was 0.43-0.51 g ethanol/g glucose (0.19-0.23 g ethanol/g beans). The average ethanol yield for the eight bean types was 92% of maximum theoretical yield, demonstrating that starch from beans could be efficiently converted to ethanol. Ethanol concentration obtained from 20% (w/w) solids loading was 3.5-4.4% (w/v). The residual fermentation solids contained, on a dry basis, 37.1-43.6% crude protein, 10.8-15.1% acid detergent fiber and 19.1-31.3% neutral detergent fiber.     

Optimisation of a solvent for the complete extraction of prolamins from heated foods

Wheat bread was lyophilised, ground, extracted and centrifuged. The supernatants were analysed for gluten content by RP-HPLC and a commercial sandwich ELISA. Prolamin extraction solvents contained tris(2-carboxyethyl)phosphine (TCEP; 1, 2, 5, 10, 20, 50 mmol/L), guanidine hydrochloride (GUA; 0 or 2 mol/L) and a buffered salt solution. A commercial cocktail solution (250 mmol/L mercaptoethanol (ME), 2 mol/L GUA, buffered salt solution) as well as 60% (v/v) ethanol were used as control solvents. Wheat flour was the control for the extractability of the native prolamins. 60% ethanol only extracted 37% of the prolamins from wheat bread (cocktail = 100%). When ME was replaced by TCEP the protein yield increased from 35% at the lowest TCEP-level to 95% when 20–50 mmol/L TCEP were used. The use of GUA was essential to extract prolamins quantitatively. Comparative protein analysis using RP-HPLC and ELISA showed that both methods provided comparable prolamin (gliadin) concentrations of the wheat flour (40.3–45.7 mg/g), when 60% ethanol was used as extraction solvent. The extraction yields from bread were considerably lower (16.7–24.7 mg/g). Cocktail and TCEP extracted almost the same amount of protein from flour and bread with TCEP showing slightly lower yields. Total extractable protein (gliadin + glutenin) as determined by RP-HPLC was 70.5–75.3 mg/g, and total gliadin as determined by ELISA was 42.7–44.2 mg/g. Thus, the study has shown that TCEP in combination with GUA extracts proteins from heated, gluten-containing foods as effective as the commercial cocktail solution.     

Analysis of starch swelling power in Australian breeding lines of hexaploid wheat (Triticum aestivum L.)

Starch is the major component of wheat (Triticum aestivum L.) grain and is composed of two large glucan molecules, amylose and amylopectin. The ratio between the two polymers types influences the water absorbing properties of starch upon heating, and thus affects the end-use of grain and purified starch. In this study, we evaluated the starch swelling power (SSP) values in seven wheat populations developed from crosses involving low-SSP lines. Analysis of starch produced by the F₂ generation plants showed that the largest SSP variation (11.4–16.2) and lowest SSP mean (13.9) was obtained for a population derived from doubled haploid lines SM1028 (SSP = 14.5) and VK306 (SSP = 13.6). The population of 360 lines was advanced by single seed descent to the following generations for further studies. Starch analysis of grain produced by F₄ generation lines in two field locations during 2006 and in a greenhouse environment during 2005 showed that SSP values were relatively stably inherited. The average broad-sense heritability was 73% and significant (P < 0.001) genotype × genotype and genotype × environment interactions were seen. Starches with the highest and lowest SSP values were inversely related to amylose concentration determined by high pressure liquid chromatography (HPLC)–size exclusion chromatography (SEC) of debranched starch. Developed lines with the lowest SSP values surpassed 40% in apparent amylose concentration. The study illustrates that screening for SSP in early generations can be used to develop wheat lines with desired starch swelling characteristics.     

Dyeing properties and colour fastness of wool dyed with indicaxanthin natural dye

This research work involves the dyeing of wool with indicaxanthin, a natural dye extracted from fruits of Opuntia ficus-indica. The optimal conditions for dye extraction were to mix 50 g of Juice from cactus pears with 100 mL of 80% aqueous ethanol as solvent for dye extraction. Liquid chromatography was applied for the separation. Two main dyes were obtained, which were identified as indicaxanthin (75 mg per 50 g) and betanin (5 mg per 50 g). The effect of dye bath pH, salt concentration, dyeing time and temperature were studied. The optimal conditions for wool dyeing with indicaxanthin dye were carried out at 70 °C for 90 min with the pre-treatment of various metal salts as mordant. The colour yields of the dye on the wool were found to be highly dependent of the pH, optimum results being obtained at pH 4. The K/S of wool increased in the order of the dyeing using KAl (SO₄)₂ > MnSO₄ > CoSO₄ > FeSO₄ > none > ZnSO₄ > CuSO₄. Un-mordanted samples have good properties of water and washing fastness. Mordants KAl (SO₄)₂ and CoSO₄ were found to give good light fastness (rating 5).     

Fractionation of Oat (1→3), (1→4)-β-D-Glucans and Characterisation of the Fractions

A commercial sample of oat β-glucan was fractionated through stepwise precipitation with ammonium sulphate to yield four fractions. These were analysed for apparent viscosity values, relative molecular weights using high-performance size-exclusion chromatography, and chemical structure by methylation analysis and analysis of water-soluble fragments released from the fractions by lichenase. Fractions exhibiting lower molecular size and decreased limiting viscosity values were precipitated at higher concentrations of ammonium sulphate. Methylation analysis indicated that in all fractions β-(1→3) and β-(1→4) linkages represented more than 90% of all linkages present. The β-(1→3) to β-(1→4) ratio in the fractions decreased with molecular size. β-glucan fractions with distinct physical and structural characteristics were obtained with the ammonium sulphate fractional precipitation technique.     

Impacts of weather,shade cover and elevation on coffee leafminer Leucoptera coffeella (Lepidoptera: Lyonetiidae) population dynamics and natural enemies

We assessed the influences of ambient temperature, rainfall, shade cover and elevation on seasonal abundance of coffee leafminer Leucoptera coffeella (Guérin-Ménèville) and its natural enemies in coffee farms in the Soconusco region of Chiapas, Mexico. Mined coffee leaves were most abundant during the rainy season (i.e. historical average rainfall >200 mm/mo, April–November) compared to the dry season (<100 mm/mo, December–March), and at low (<600 m asl) relative to high (>900 m asl) elevations. The abundance of mined leaves increased with rainfall, and decreased with maximum daily temperatures. Coffee leafminer survivorship was highest during the dry season (>40%), when predation was lowest (<10%). Predation was the main source of coffee leafminer mortality, and was greatest during the rainy season (>25%) when coffee leafminer incidence was highest (>30% mined leaves per plant). None of the weather variables that were evaluated (viz. maximum and minimum temperatures, and rainfall) significantly impacted parasitism ratios. Shade cover moderated on-farm temperatures, by reducing maximum daily temperatures and any potential, direct impacts of rainfall on coffee leafminer, by providing partial shelter from rainfall, but did not significantly affect coffee leafminer incidence. In 48 h laboratory trials, coffee leafminer oviposition was highest at 28 °C (∼15 eggs/female), minimal at 25 °C (∼3 eggs) and nil at 20 °C, and higher during night-time hours (>8 eggs/female/day) compared to day-time hours (<1 egg). Historical average temperatures were higher at low elevation (yearly average ca. 25 °C; range = 18.0–32.0 °C) than at high elevation (ca. 21 °C; 13.5–28.5 °C), and we predicted that physical environmental conditions (i.e. night-time hours with temperatures > 20 °C) were permissive of coffee leafminer oviposition during twice as many hours each year at low elevation (4060 h) compared to high elevation (2081 h). Overall, our results suggested that evident differences in the abundance of coffee leafminer between elevations may be due in considerable part to differences in ambient temperatures, particularly night-time temperatures, rather than rainfall, shade cover, or elevation per se.     

Utilization of pretreated bagasse for the sustainable bioproduction of cellulase by Aspergillus nidulans MTCC344 using response surface methodology

Response surface methodology (RSM) was used to optimize the conditions for the production of endo β-1,4 glucanase, a component of cellulase by Aspergillus nidulans MTCC344 under solid state fermentation, using bagasse as the chief substrate. A four-factor-five-level central composite design was employed for experimental design and analysis of the results. Maximum cellulase activity (CMCase was 28.96 U g⁻¹) can be attained at the optimum conditions, 16.8 mm bagasse bed height, 60% moisture content, pH 4.25 and temperature 40 °C in the solid state fermenter. These data were rather close to the experimental results obtained (CMCase was 28.84 U g⁻¹). A. nidulans MTCC344 was able to hydrolyze pretreated bagasse completely after 8 days of incubation with significant endo β-1,4 glucanase activities. The results of Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and scanning electron microscopy (SEM) of bagasse showed structural changes through pretreatment, in favor of enzymatic hydrolysis. Bagasse with alkali pretreatment using sodium hydroxide is a source of lignocelluloses able to improve the yield of endo β-1,4 glucanase by the strain of A. nidulans. The endo β-1,4 glucanase produced during the bioconversion of cellulose to glucose by A. nidulans MTCC344 is strongly dependent on the pretreatment given before hydrolysis.     

Enzyme inhibitory and antioxidant activity of Melia azedarach L. naturalized in Anatolia and its phenolic acid and fatty acid composition

In the present study, acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and tyrosinase inhibitory and antioxidant activities of the fruit and leaf extracts of Melia azedarach L. (Meliaceae) of Turkish origin were evaluated. Enzyme inhibitory activity of the extracts was tested in vitro using ELISA microplate reader. Antioxidant activity of the extracts was tested using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferrous ion-chelation, and ferric-reducing antioxidant power (FRAP) assays. Phenolic composition of the extracts was elucidated by high performance liquid chromatography (HPLC) and fatty acid compositions of the fatty oils of the fruits and leaves were elucidated by gas chromatography-mass spectrometry (GC-MS). The ethyl acetate extract from the leaves showed the highest inhibition against AChE (33.63 ± 1.40%) and BChE (92.89 ± 3.05%). The methanol extract from the leaves exerted the best antioxidant activity in DPPH radical scavenging and FRAP assays, while the ethyl acetate extracts of the fruits and leaves had the most notable effect in metal-chelation assay.     

Lavender and hyssop productivity, oil content, and bioactivity as a function of harvest time and drying

Field and laboratory experiments were conducted to evaluate the productivity and essential oil composition of lavender (Lavandula angustifolia Mill.) and hyssop (Hyssopus officinalis L.) as functions of year, harvest time, and drying. Lavender essential oil content ranged from 0.71 to 1.3% (overall average of 0.89%) and hyssop oil content ranged from 0.13 to 0.26% (overall average of 0.19%). Lavender and hyssop essential oil yields increased with time. Hyssop oil yields varied from 7.3 kg ha⁻¹ to 19.6 kg ha⁻¹, and lavender oil yields varied from 7.8 kg ha⁻¹ to 55.5 kg ha⁻¹. The major constituents of lavender oil were linalool (23.3-43.4%) and linalylacetate (20.2-39.6%), while the major constituents of hyssop oil were pinocamphene + isopinocamphene (57-75%) and β-pinene (5-15%). Lavender oil extracted from dry material had higher concentrations of linalyl acetate and caryophyllene but lower concentrations of myrcene than the oil from the fresh material. Delayed harvest of hyssop increased the concentrations of β-pinene, myrcene, and limonene + cineole but reduced pinocamphone + isopinocamphone. The chemical composition of the lavender and hyssop oil produced in Mississippi was similar to commercial oils from Bulgaria, Canada, France, and US. Lavender and hyssop can be established as essential oil crops in areas of the southeastern United States. Lavender and hyssop essential oils did not show significant antimicrobial, antileishmanial, antimalarial activity, and did not alter ruminal fermentation. However, commercial oil from L. latifolia reduced methane production in an in vitro digestibility study. The antioxidant activity of hyssop essential oil was 2039 μmol of TE L⁻¹, whereas the antioxidant activity of lavender essential oil was 328 μmol of TE L⁻¹.     

Intra-specific competition in maize: Early established hierarchies differ in plant growth and biomass partitioning to the ear around silking

Early interferences among plants within a maize stand determine the establishment of extreme plant types (i.e. dominant and dominated individuals). The development of these hierarchies takes place well before [from the seventh leaf stage (V₇) onwards] the start of the critical period for kernel set (i.e. a 30-day period centered in silking). Kernel number per plant (KNP) is significantly related to plant growth rate around silking (PGR_S) and biomass partitioning to the ear during this period. Previous evidence has demonstrated that at high stand densities, extreme plant types may exhibit similar PGR_S values but set different KNP. We tested the hypothesis that early established plant hierarchies differ in biomass allocation to the ears during the period around silking. Two hybrids of contrasting tolerance to crowding (DK752 and DK765 as the tolerant and the intolerant hybrid, respectively) were cropped at different interplant competition intensities (6, 12, 12 pl m⁻² thinned to 6 pl m⁻² at V₉ and 6 pl m⁻² shading from V₉ onwards) during 2003/2004 and 2004/2005 in Argentina. For all treatments, the coefficient of variation (CV) of plant biomass increased from V₃ (ca. 1.2%) to V_9-10 (ca. 22%). From V₇ onwards, plant growth rate of dominant individuals was higher (P < 0.05) than that of the dominated plants. Hence, dominant plants exhibited higher (P < 0.05) PGR_S (ca. 4.5 g pl day⁻¹) than dominated individuals (ca. 3.7 g pl day⁻¹). As PGR_S declined in response to increased plant population density (ca. 5.1 and 2.8 for 6 and 12 pl m⁻², respectively), biomass partitioning to the ear was reduced (ca. 0.44 and 0.33 for 6 and 12 pl m⁻², respectively). For all treatments, however, dominant plants exhibited a greater biomass partitioning to the ear (ca. 0.41) than the dominated individuals (ca. 0.36). Consequently, the former were the individuals with the highest ear growth rate (ca. 1.9 and 1.4 g per ear per day for the dominant and dominated plant, respectively) and KNP (ca. 623 and 490 kernels per plant for the dominant and dominated plant, respectively) of the stand. We identified three traits on DK765 related to the low tolerance to high-density stress of this genotype: (i) a higher plant-to-plant variability (CV ca. 26% and 19%, for DK765 and DK752, respectively), (ii) a lower biomass partitioning to the ear around silking (ca. 0.26 and 0.39 for DK765 and DK752, respectively), and (iii) a higher response rate of KNP to ear growth rate around silking (ca. 370–738 and 360–414 kernels per g, for DK765 and DK752, respectively). Hence, as stand density was increased, KNP of DK765 was sharply reduced, especially in the dominated individuals of the stand.     

Processing of Lespedeza stalks by pretreatment with low severity steam and post-treatment with alkaline peroxide

The steam pre-treatment with low severity preserves valuable biomass components, and further delignification with alkaline peroxide could improve hydrolysis. A combination of low severity steam pretreatment and alkaline peroxide post-treatment of Lespedeza stalks was investigated. The post-treatment of steam-pretreated Lespedeza stalks with alkaline peroxide significantly increased the cellulose content and changed the structure of the cellulose-rich fractions. A glucose yield of 503.5 mg g⁻¹ raw material from enzyme hydrolysis was obtained when the steam-pretreated material (184 °C for 4 min) was post-treated with 2% hydrogen peroxide at 60 °C for 24 h with a substrate concentration of 3.3%. Its hydrolysis yield is 88.8%, which is higher than that of samples processed by steam pretreatment alone (63.7%). The samples obtained by post-treatment with alkaline peroxide were found to have a smoother surface and looser structure in scanning electron microscopy images. The isolated lignin preparations had a yield range from 10.9 to 14.7 (% dry matter). The lignin was characterized by thermogravimetric analysis/differential thermal analysis, Fourier transform infrared spectroscopy, and gel permeation chromatography. Alkaline peroxide treatment increased the thermal stability of lignin, and decreased the amounts of all functional groups. Depolymerization and repolymerization occurred during the alkaline peroxide treatment.     

Microwave-enhanced alkali treatment of Pinus yunnanensis: Physiochemical characterization of the dissolved lignins

Pinus yunnanensis was subjected to water-bath and microwave treatments in 1% NaOH aqueous solutions at 100 °C with various ratios of bath heating time to microwave heating time (0/120, 20/100, 40/80, 60/60, 80/40, 100/20 and 120/0 min). The lignins dissolved in the alkali liquors were separated and purified, and their physicochemical features were comparatively characterized by sugar analysis, GPC, FT-IR, ¹³C and HSQC NMR, as well as thermogravimetric analysis (TGA). The results showed that the lignin fractions extracted with microwave heating (20-120 min) had high molecular weights and polydispersities (M_w 3150-5710 g/mol, M_n 2130-3020 g/mol, M_w/M_n 1.48-2.00) as compared to those prepared without microwave heating (M_w 3080 g/mol, M_n 2080 g/mol, M_w/M_n 1.48). The most striking characteristic of all lignin fractions was the almost absence of associated sugars (0.16-3.25%). The TGA results indicated that the thermal stability of the lignin fraction increased with the increment of the molecular weight. FT-IR and NMR spectra suggested that the lignin fractions showed similar structures which were mainly composed of guaiacyl (G) and minor amounts of p-hydroxyphenyl (H) units. Moreover, HSQC NMR spectrum of a typical lignin fraction (prepared with microwave heating for 120 min) revealed that it contained dominant amounts of β-O-4′ linkages (64.6%) and phenylcoumaran (β-5′) substructures (25.8%) together with small amounts of resinol (β-β′) substructures (6.7%) and coniferyl alcohol end groups (2.9%).     

Synthesis of withanolide A depends on carbon source and medium pH in hairy root cultures of Withania somnifera

Withania somnifera (L.) Dunal. (Indian ginseng) is an important medicinal plant which yields pharmaceutically active compounds called withanolides. The present work deals with optimization of parameters of hairy root culture of W. somnifera for the production of biomass and withanolide A. We also investigated the effects of carbon source [sucrose, glucose, fructose, maltose, glucose + fructose (1:1), fructose + sucrose (1:1) and sucrose + glucose (1:1)], sucrose concentration (1%, 2%, 3%, 4%, 6% and 8%) and the initial medium pH (4.0, 4.5, 5.0, 5.5, 5.8, 6.0 and 6.5) on growth and production of withanolide A in hairy root cultures of W. somnifera. We found that biomass accumulation and production of withanolide A was highest when sucrose was used as the carbon source (11.92 g l⁻¹ DW and 11.96 mg g⁻¹ DW of withanolide A). Further 3% sucrose concentration was found to be optimal for biomass accumulation (11.92 g l⁻¹ DW) and 4% sucrose favoured the production of withanolide A (13.28 mg g⁻¹ DW) in the tested range of concentrations (1-8%). The biomass of hairy roots was optimal when the initial medium pH was 5.8 (12.1 g l⁻¹ DW) and the withanolide A production was highest in the medium pH set at 6.0 (13.84 mg g⁻¹ DW).     

Effect of boron on the development of brown rot (Monilinia laxa) on peaches

Requirements of consumers for products with low residues of pesticides have increased the need for alternative disease management practices. The concentration of boron in fruit affects its quality, shelf life and the development of physiological disorders. However, the effect of boron on the susceptibility of peach to fruit rots has not been reported. This study investigated the effect of boron (Power B and Borax) on the development of Monilinia laxa on peaches (cv Andross). Mycelial growth of M. laxa was inhibited on potato dextrose agar supplemented with 750 μg ml⁻¹ of Borax or 1000 μg ml⁻¹ of Power B. The EC 50 values were 107.9 and 522.4 for Borax and Power B respectively. Field investigations showed that the incidence of peach infections by M. laxa was negatively correlated with the content of Boron in the leaves. Post-harvest dipping of peaches in Power B or Borax solution, at concentrations recommended by manufacturer (2 μg ml⁻¹ for Power B and 1 mg ml⁻¹ for Borax), significantly reduced the development of M. laxa. Power B, at rates of 6 μg ml⁻¹, and Borax at rates of 3 mg ml⁻¹ were the most effective in reducing infections by M. laxa. Finally, post-harvest dipping of fruit in Power B or Borax reduced losses of fruit weight and improved fruit firmness one month after storage, showing that boron increased the maintainability of peaches in cold storage. Peaches treated with 6 μg ml⁻¹ Power B or 3 mg ml⁻¹ Borax had the highest flesh firmness and the lowest water losses, while untreated control peaches were the least firm. Generally, Borax was significantly less effective than Power B, but better than the control treatment.