Structure and functional properties of taro starch modified by dry heat treatment.

Taro starch (TS) was modified by dry heat treatment (DHT) for different periods (1, 3, 5, and 7 h at 130 °C) and temperatures (90, 110, 130, and 150 °C for 5 h) to expand its applications in food and other industries. The structure and functional properties of DHT-modified TS were characterized. It was found that TS granules became agglomerated after DHT, and the particle size, amylose content, solubility, and retrogradation enthalpy change of TS increased with increasing dry heating time and temperature, whereas the relative crystallinity, molecular weight, swelling power, gelatinization temperature, and enthalpy change decreased. The absorbance ratio of 1047 cm-1/1022 cm-1 for DHT-modified TS (except at 7 h) was higher than that of native TS. DHT increased the contact angle of TS in a time- and temperature-dependent manner. At a moderate strength, DHT increased the pasting viscosity, relative setback value, and storage modulus but decreased the relative breakdown value. Moreover, DHT (except at 150 °C) caused a decrease in the rapid digestive starch content and estimated glycemic index of TS. These results suggested that DHT-modified TS could be used in foods with high viscosity requirements, gel foods, and low-glycemic index starch-based foods.

Mechanism of synergistic stabilization of emulsions by amorphous taro starch and protein and emulsion stability.

Amorphous taro starch (TS)/whey protein isolate (WPI) mixtures were prepared using pasting treatment. The TS/WPI mixtures and their stabilized emulsions were characterized to determine the emulsion stability and the mechanism of synergistic stabilization of emulsions. As WPI content increased from 0% to 13%, the paste final viscosity and retrogradation ratio of the TS/WPI mixture gradually decreased from 3683 cP to 2532 cP and from 80.65% to 30.51%, respectively. As the WPI content increased from 0% to 10%, the emulsion droplet size decreased gradually from 96.81 µm to 10.32 µm, and the storage modulus G' and stabilities of freeze-thaw, centrifugal, and storage increased gradually. Confocal laser scanning microscopy revealed that WPI and TS were mainly distributed at the oil-water interface and droplet interstice, respectively. Thermal treatment, pH, and ionic strength had little influence on the appearance but had different influences on the droplet size and G', and the rates of droplet size and G' increase under storage varied with different environmental factors.

Ligninolytic characteristics of Pleurotus ostreatus cultivated in cotton stalk media.

Biodelignification is widely regarded as a low-efficiency process because it is usually slow and difficult to control. To improve its efficiency and understand its mechanism, the present study analyzed the delignification characteristics of Pleurotus ostreatus grown on a cotton stalk medium. The results demonstrated that all strains of P. ostreatus can selectively degrade the cotton stalk lignin. When cultured in a cotton stalk medium for 60 days, P. ostreatus degraded lignin primarily during its mycelium growth with up to 54.04% lignin degradation and produced laccase and manganese dependent peroxidase with high activity levels at the peaks of 70.17 U/ml and 62.39 U/ml, respectively, but no detectable lignin peroxidase. The results of nuclear magnetic resonance spectroscopy and Fourier transform infrared spectroscopy analyses of significant changes in lignin structure revealed that syringyl (S) lignin units were more degraded than guaiacyl (G) lignin units, with a significantly elevated G/S ratio. The Gas Chromatography-Mass Spectrometer analysis of low-molecular-weight compounds revealed that the delignification resulted in the formation of alcohols, organic acids, benzodiazepines, and alkanes. Identified benzodiazepines implied the degradation of G and S units of lignin. These findings will help to improve the efficiency of biodelignification and expand our understanding of its mechanism.

Functional Characterization of Laccase Isozyme (PoLcc1) from the Edible Mushroom Pleurotus ostreatus Involved in Lignin Degradation in Cotton Straw.

Fungal laccases play important roles in the degradation of lignocellulose. In this study, the laccase producing cotton straw medium for Pleurotus ostreatus was optimized by single-factor and orthogonal experiments, and to investigate the role of Lacc1 gene, one of the laccase-encoding genes, in the degradation of cotton straw lignin, an overexpression strain of Lacc1 gene was constructed, which was analyzed for the characteristics of lignin degradation. The results demonstrated that the culture conditions with the highest lignin degradation efficiency of the P. ostreatus were the cotton straw particle size of 0.75 mm, a solid-liquid ratio of 1:3 and containing 0.25 g/L of Tween in the medium, as well as an incubation temperature of 26 °C. Two overexpression strains (OE L1-1 and OE L1-4) of Lacc1 gene were obtained, and the gene expression increased 12.08- and 33.04-fold, respectively. The results of 1H-NMR and FTIR analyses of significant changes in lignin structure revealed that Lacc1 gene accelerated the degradation of lignin G-units and involved in the cleavage of ß-O-4 linkages and the demethylation of lignin units. These findings will help to improve the efficiency of biodelignification and expand our understanding of its mechanism.

Transcriptome Analysis of Moso Bamboo (Phyllostachys edulis) Reveals Candidate Genes Involved in Response to Dehydration and Cold Stresses.

Bamboo (Bambusoideae) belongs to the grass family (Poaceae) and has been utilized as one of the most important nontimber forest resources in the world. Moso bamboo (Phyllostachys edulis) is a large woody bamboo with high ecological and economic values. Global climate change brings potential challenges to the normal growth of moso bamboo, and hence its production. Despite the release of moso bamboo genome sequence, the knowledge on genome-wide responses to abiotic stress is still limited. In this study, we generated a transcriptome data set with respect to dehydration and cold responses of moso bamboo using RNA-seq technology. The differentially expressed genes (DEGs) under treatments of dehydration and cold stresses were identified. By combining comprehensive gene ontology (GO) analysis, time-series analysis, and co-expression analysis, candidate genes involved in dehydration and cold responses were identified, which encode abscisic acid (ABA)/water deficit stress (WDS)-induced protein, late embryogenesis abundant (LEA) protein, 9-cis-epoxycarotenoid dioxygenase (NCED), anti-oxidation enzymes, transcription factors, etc. Additionally, we used PeLEA14, a dehydration-induced gene encoding an "atypical" LEA protein, as an example to validate the function of the identified stress-related gene in tolerance to abiotic stresses, such as drought and salt. In this study, we provided a valuable genomic resource for future excavation of key genes involved in abiotic stress responses and genetic improvement of moso bamboo to meet the requirement for environmental resilience and sustainable production.

Overexpressing PhytochromeInteractingFactor 8 of Myrothamnus flabellifolia Enhanced Drought and Salt Tolerance in Arabidopsis.

Myrothamnus flabellifolia is the only woody resurrection plant found in the world and can survive from long-term desiccation. Therefore, M. flabellifolia could be considered as a valuable resource for study of plant adaptation to abiotic stress. However, few genes related to its drought tolerance have been functionally characterized and the molecular mechanisms underlying the stress tolerance of M. flabellifolia are largely unknown. The phytochrome interacting factor (PIF) family is a group of basic helix-loop-helix (bHLH) transcription factors and functions as the core regulator in plant growth and development. However, less is known of its participation in abiotic stress response. In this study, we isolated and characterized a dehydration-inducible PIF gene MfPIF8 from M. flabellifolia. Heterologous expression of MfPIF8 in Arabidopsis enhanced tolerance to drought and salinity stresses at seedling and adult stages. It significantly increased primary root length and stomatal aperture (ration of length/width) under stress treatments and decreased water loss rate. Compared with WT, the transgenic lines overexpressing MfPIF8 exhibited higher chlorophyll content and lower malondialdehyde accumulation. The abilities of osmotic adjustment and reactive oxygen species scavenging were also enhanced in MfPIF8 transgenic lines. These results suggest that MfPIF8 may participate in the positive regulation of abiotic stress responses. Additional investigation of its mechanism is needed in the future.

Genome-wide identification and expression analyses of C2H2 zinc finger transcription factors in Pleurotus ostreatus.

The C2H2-type zinc finger proteins (C2H2-ZFPs) regulate various developmental processes and abiotic stress responses in eukaryotes. Yet, a comprehensive analysis of these transcription factors which could be used to find candidate genes related to the control the development and abiotic stress tolerance has not been performed in Pleurotus ostreatus. To fill this knowledge gap, 18 C2H2-ZFs were identified in the P. ostreatus genome. Phylogenetic analysis indicated that these proteins have dissimilar amino acid sequences. In addition, these proteins had variable protein characteristics, gene intron-exon structures, and motif compositions. The expression patterns of PoC2H2-ZFs in mycelia, primordia, and young and mature fruiting bodies were investigated using qRT-PCR. The expression of some PoC2H2-ZFs is regulated by auxin and cytokinin. Moreover, members of PoC2H2-ZFs expression levels are changed dramatically under heat and cold stress, suggesting that these genes may participate in abiotic stress responses. These findings could be used to study the role of P. ostreatus-derived C2H2-ZFs in development and stress tolerance.

Heterologous Expression of Dehydration-Inducible MfWRKY17 of Myrothamnus Flabellifolia Confers Drought and Salt Tolerance in Arabidopsis.

: As the only woody resurrection plant, Myrothamnus flabellifolia has a strong tolerance to drought and can survive long-term in a desiccated environment. However, the molecular mechanisms related to the stress tolerance of M. flabellifolia are largely unknown, and few tolerance-related genes previously identified had been functionally characterized. WRKYs are a group of unique and complex plant transcription factors, and have reported functions in diverse biological processes, especially in the regulation of abiotic stress tolerances, in various species. However, little is known about their roles in response to abiotic stresses in M. flabellifolia. In this study, we characterized a dehydration-inducible WRKY transcription factor gene, MfWRKY17, from M. flabellifolia. MfWRKY17 shows high degree of homology with genes from Vitis vinifera and Vitis pseudoreticulata, belonging to group II of the WRKY family. Unlike known WRKY17s in other organisms acting as negative regulators in biotic or abiotic stress responses, overexpression of MfWRKY17 in Arabidopsis significantly increased drought and salt tolerance. Further investigations indicated that MfWRKY17 participated in increasing water retention, maintaining chlorophyll content, and regulating ABA biosynthesis and stress-related gene expression. These results suggest that MfWRKY17 possibly acts as a positive regulator of stress tolerance in the resurrection plant M. flabellifolia.

Purification, characterization and immunomodulatory activity of polysaccharides from stem lettuce.

Stem lettuce has a long history of cultivation in China and possesses high nutritional and medicinal value. In our previous studies, extraction optimization, characterization, and bioactivities of stem lettuce polysaccharides (SLP) were investigated. In this study, SLP were further separated into two purified polysaccharides, SLP-1 and SLP-2, by anion exchange chromatography followed by size exclusion chromatography. SLP-1, with a molecular weight of 90 KDa, was mainly composed of galacturonic acid, galactose and arabinose in a molar ratio of 17.6:41.7:33.9. SLP-2, with a molecular weight of 44 KDa, was mainly composed of mannose, galacturonic acid, galactose and arabinose in a molar ratio of 11.5:69.5:9.3:8.2. In addition, both purified polysaccharides contain sulphate radicals, have triple helical structures and can promote macrophage proliferation without cytotoxicity. SLP-2 was better able to stimulate phagocytic and nitric oxide production than SLP-1. The results suggest that polysaccharides from stem lettuce could be explored as immunomodulatory agents in the field of pharmaceuticals and functional foods.

Purification, characterization and immunomodulatory activity of water extractable polysaccharides from the swollen culms of Zizania latifolia.

The swollen culms of Zizania latifolia is a popular vegetable and traditional herbal medicine mainly consumed in some Asian countries. In our previous study, water extractable polysaccharide (ZLPs-W) and alkali extractable polysaccharide (ZLPs-A) was sequentially prepared from the swollen culms of Zizania latifolia. In vitro cell assay revealed that ZLPs-W without cytotoxicity had higher immunomodulatory activity than ZLPs-A. Therefore, in this study, ZLPs-W was further separated to three purified polysaccharides of ZLPs-W1, ZLPs-W2, and ZLPs-W3 by anion exchange chromatography and size exclusive chromatography in sequence. They were found to be homogeneous polysaccharides in ß-type glycosidic linkage. ZLPs-W1 and ZLPs-W2 without triple helix conformation mainly contained Glc in molar percentage of 96.47% and 85.1%, respectively. ZLPs-W3 with triple helix conformation mainly consisted of Man, Rha, GalA, Glc, Gal and Ara in the molar ratio of 10.23:14.14:37.44:17.14:16.54:4.51. In addition, all the three purified polysaccharides could effectively enhanced the proliferation, phagocytosis, and nitric oxide production of murine macrophage RAW 264.7, suggesting that they possessed a potent immunostimulatory activity and could be developed as immunomodulator in medicine or functional food areas.

Optimization of water-soluble polysaccharides from stem lettuce by response surface methodology and study on its characterization and bioactivities.

Stem lettuce is widely consumed as a vegetable in China. It is also used as a traditional Chinese medicine for a long time. Up to now, no information is available for stem lettuce polysaccharide (SLP). In this study, extraction optimization, characterization, and antioxidant activity of SLP was investigated.The maximum SLP yield of 20.89% was obtained under the optimal extraction conditions as follows: extraction temperature 95°C, extraction time 3.3h and ratio of water to material 25mL/g. The sulfate content of SLP was 5.82% and the main monosaccharides were GalA, Gal, and Ara with a mole ratio of 49.2: 23.3: 22.9. FT-IR spectrum and HPGPC result further indicated that SLP is a sulfated polydisperse heterpolysaccharide. Congo-red test and AFM scan indicated that SLP might be branching and intertwining structure with triple helix conformation. Moreover, SLP exhibited potent antioxidant activity and α-amylase inhibitory activity, hence it could be used as a potential natural antioxidant and hypoglycemic agent in medicine or functional food fields.

Characterization, antioxidant activity and immunomodulatory activity of polysaccharides from the swollen culms of Zizania latifolia.

The swollen culms of Zizania latifolia have been used as a vegetable and traditional herbal medicine in China, Japan, Korea and Southeast Asia countries. Up to date, there is little information about the polysaccharides from the swollen culms of Zizania latifolia and their potential bioactivities. In the present study, water extractable polysaccharide (ZLPs-W) and alkali extractable polysaccharide (ZLPs-A) was sequentially prepared from the swollen culms of Zizania latifolia. Both of ZLPs-W and ZLPs-A was found to be non-starch polydisperse heterpolysaccharide with ß-type glycosidic linkage. ZLPs-W with triple helix conformation mainly composed of GalA, Glc and Gal. ZLPs-A without triple helix conformation mainly composed of Glc, Gal, Xyl and Ara. In in vitro antioxidant assay, ZLPs-W and ZLPs-A exhibited good scavenging activities. The EC50 of DPPH radical, superoxide radical and hydroxy radical scavenging activities for ZLPs-A is 1.87, 1.13 and 0.38mg/mL compared with that for ZLPs-W is 2.95, 3.99 and 0.5mg/mL, respectively. Moreover, in vitro cell assay revealed that ZLPs-W without cytotoxicity has higher immunomodulatory activity than ZLPs-A in terms of stimulation of phagocytic ability and NO production in murine macrophage RAW 264.7. At the treated concentration of 400µg/mL and 100µg/mL, ZLPs-W induced a highest phagocytosis index (1.76) and NO product (29.12µmol/L), respectively. The results suggest that polysaccharide from the swollen culms of Zizania latifolia could be explored as potential natural antioxidant and immunomodulatory agents in medicine or functional food fields.

[Identification of hexavalent chromium reducing bacteria Cr4-1 and optimization of its reduction conditions].

OBJECTIVE: To identify the hexavalent chromium reduction Cr4-1, and to study the better conditions of the bacterial growth and its Cr(VI) reduction. METHODS: The physiological and biochemical methods and 16s rDNA sequencing were used for identification of bacteria Cr4-1. The influence of temperature, pH, initial Cr (VI) concentration and shaking speed on bacterial growth and Cr (VI) reduction were studied. Mass balance analysis was used to analyze the end products of the reduction reaction. RESULTS: A Cr(VI) reducing bacteria Cr4-1, screened from acclimated activated sludge, was identified as Bacillus cereus. The appropriate conditions of bacterial growth was 25 °C, pH 7 to 8, shaking speed 150 r/min, while the suitable conditions for Cr(VI) reduction was 35 °C, pH 8 to 9. When the initial Cr(VI) concentration increased from 20 mg/L to 60 mg/L, the reduction rate decreased gradually. Under the suitable reducing conditions, when the initial concentration of Cr (VI) was 30 mg/L, the reduction rate could up to 100% in 9 h. The end product was soluble trivalent chromium. CONCLUSION: Strain Cr4-1 had a good effect on Cr (VI) reduction and the final product was soluble trivalent chromium.

Preliminary characterization, antioxidant activity in vitro and hepatoprotective effect on acute alcohol-induced liver injury in mice of polysaccharides from the peduncles of Hovenia dulcis.

The fresh fleshy peduncles of Hovenia dulcis have been used as a food supplement and traditional herbal medicine for the treatment of liver diseases and alcoholic poisoning for more than a millennium. The objectives of the present study, therefore, were to determine the antioxidant activity of polysaccharides from the peduncles of H. dulcis (HDPS) and to evaluate its hepatoprotective effect on acute alcohol-induced liver injury in mice. HDPS, prepared by hot water extraction, ethanol precipitation and treatment of macroporous resin, was found to be non-starch polysaccharide and mainly composed of galactose, arabinose, rhamnose and galacturonic acid. In in vitro antioxidant assay, HDPS exhibited high superoxide radical scavenging activity, strong inhibition effect on lipid peroxidation and a medium ferrous ion-chelating activity. For hepatoprotective activity in vivo, the administration of HDPS significantly decreased the serum levels of alanine aminotransferase and aspartate aminotransferase, significantly decreased the liver level of malondialdehyde and remarkably restored the liver activities of superoxide dismutase and glutathione peroxidase in alcohol-induced liver injury mice. The results suggested that HDPS had a significant protective effect against acute alcohol-induced liver injury possibly via its antioxidant activity to protect biological systems against the oxidative stress.