Oxidative cleavage of cellulose in the horse gut.

BACKGROUND: Lytic polysaccharide monooxygenases (LPMOs) belonging to the auxiliary activity 9 family (AA9) are widely found in aerobic fungi. These enzymes are O2-dependent copper oxidoreductases that catalyze the oxidative cleavage of cellulose. However, studies that have investigated AA9 LPMOs of aerobic fungi in the herbivore gut are scare. To date, whether oxidative cleavage of cellulose occurs in the herbivore gut is unknown. RESULTS: We report for the first time experimental evidence that AA9 LPMOs from aerobic thermophilic fungi catalyze the oxidative cleavage of cellulose present in the horse gut to C1-oxidized cellulose and C1- and C4-oxidized cello-oligosaccharides. We isolated and identified three thermophilic fungi and measured their growth and AA9 LPMO expression at 37 °C in vitro. We also assessed the expression and the presence of AA9 LPMOs from thermophilic fungi in situ. Finally, we used two recombinant AA9 LPMOs and a native AA9 LPMO from thermophilic fungi to cleave cellulose to yield C1-oxidized products at 37 °C in vitro. CONCLUSIONS: The oxidative cleavage of cellulose occurs in the horse gut. This finding will broaden the known the biological functions of the ubiquitous LPMOs and aid in determining biological significance of aerobic thermophilic fungi.

Cell wall modification and lignin biosynthesis involved in disease resistance against Diaporthe citri in harvested pummelo fruit elicited by carvacrol.

BACKGROUND: Phomopsis stem-end rot caused by Diaporthe citri, causes significant commercial postharvest losses of pummelo fruit during storage. Carvacrol (CVR) is a known generally recognized as safe and has the ability to prolong the preservation of harvested fruits. In the present study, the inhibitory effects of CVR treatment at the appropriate concentration on Phomopsis stem-end rot development of harvested pummelo fruit inoculated with D. citri were evaluated by the amounts of cell wall components, the activities and gene expressions of related enzymes involved in cell wall modification and lignin biosynthesis. RESULTS: Results indicated that CVR completely inhibited D. citri growth in vitro at 200 mg L-1 and significantly controlled Phomopsis stem-end rot development in harvested pummelo. The CVR treatment delayed peel softening and browning, and retarded electrolyte leakage, superoxide radical (O2 •- ) production, and malondialdehyde content. The CVR-treated fruit maintained higher amounts of cell wall material, protopectin, hemicelluloses, and cellulose, but exhibited lower water-soluble pectin amount. Moreover, in D. citri-inoculated fruit, CVR treatment suppressed the activities and gene expressions of cell wall disassembling-enzymes, including pectin methylesterase, polygalacturonase, cellulase, and ß-galactosidase, while the development of cell wall degradation was reduced. Meanwhile, the CVR treatment enhanced the lignin biosynthesis by increasing the activities and up-regulating the gene expressions of phenylalanine ammonialyase, cinnamic alcohol dehydrogenase, and peroxidase accompanied with elevated level of lignin in pummelo fruit. CONCLUSION: The disease resistance to D. citri in pummelo fruit elicited by CVR treatment is related to delaying cell wall degradation and enhancing lignin biosynthesis. © 2021 Society of Chemical Industry.

Alleviatory effects of salicylic acid on postharvest softening and cell wall degradation of 'Jinshayou' pummelo (Citrus maxima Merr.): A comparative physiological and transcriptomic analysis.

The regulatory mechanisms underlying the salicylic acid (SA)-mediated inhibition of senescence in pummelo fruit, the largest known citrus variety, remain unclear. Herein, postharvest 0.3% SA treatment was demonstrated to delay postharvest 'Jinshayou' pummelo senescence, as evidenced by the inhibitions in firmness loss, electrolyte leakage increase, and color change. Using comparative transcriptomic data, a total of 4367, 3769, and 1659 DEGs were identified between CK0 and CK60, CK0 and SA60, and CK60 and SA60, respectively. Further GO analysis revealed that DEGs were mainly implicated in the processes of cell wall modification and phenylpropanoid pathway during fruit senescence. More importantly, postharvest exogenous 0.3% SA treatment was observed to inhibit CWDEs activities and their encoding gene expression, retain higher protopectin, cellulose, and hemicelluloses contents, as well as reduce WSP content, thus maintaining cell wall structure. These findings collectively indicated that postharvest SA treatment was a green and useful preservative for alleviating fruit senescence and prolonging the storage life of harvested 'Jiashayou' pummelo fruit.

Comparison of cell wall metabolism in the pulp of three cultivars of 'Nanfeng' tangerine differing in mastication trait.

BACKGROUND: Like sweet orange (Citrus sinensis), tangerine (Citrus reticulata) is another citrus crop grown widely throughout the world. However, whether it shares a common mechanism with sweet orange in forming a given mastication trait is still unclear. In this study, three 'Nanfeng' tangerine cultivars, 'Yangxiao-26' ('YX-26') with inferior mastication trait, elite 'YX-26' with moderate mastication trait and 'Miguang' ('MG') with superior mastication trait, were selected to investigate the formation mechanism of mastication trait. RESULTS: 'MG' had the lowest contents of total pectin, protopectin and lignin and the highest gene expression levels of citrus polygalacturonase (PG) and pectin methylesterase (PME) at the end of fruit ripening, whereas 'YX-26' had the lowest water-soluble pectin (WSP) content, the highest lignin content and the lowest PG and PME expression levels. The contents of cellulose and hemicellulose were similar among the three tangerines. CONCLUSION: The fruit mastication trait of C. reticulata was determined by the proportions of WSP and protopectin as well as lignin content, not by cellulose and hemicellulose contents. Pectin content could be a major contribution to the feeling of mastication trait, while PG and PME exhibited an important role in forming a given mastication trait according to the present results as well as previous results for C. sinensis.

Lignin Biosynthesis Pathway and Redox Balance Act Synergistically in Conferring Resistance against Penicillium italicum Infection in 7-Demethoxytylophorine-Treated Navel Orange.

7-Demethoxytylophorine (DEM), a natural water-soluble phenanthroindolizidine alkaloid, has a great potential for in vitro suppression of Penicillium italicum growth. In the present study, we investigated the ability of DEM to confer resistance against P. italicum in harvested "Newhall" navel orange and the underlying mechanism. Results from the in vivo experiment showed that DEM treatment delayed blue mold development. The water-soaked lesion diameter in 40 mg L-1 DEM-treated fruit was 35.2% lower than that in the control after 96 h. Moreover, the decrease in peel firmness loss and increase in electrolyte leakage, superoxide anion (O2•-) production, and malondialdehyde (MDA) content were significantly inhibited by DEM treatment. Hydrogen peroxide (H2O2) burst in DEM-treated fruit at the early stage of P. italicum infection contributed to the conferred resistance by increasing the activities of lignin biosynthesis-related enzymes, along with the expressions of their encoding genes, resulting in lignin accumulation. The DEM-treated fruit maintained an elevated antioxidant capacity, as evidenced by high levels of ascorbic acid and glutathione content, and enhanced or upregulated the activities and gene expression levels of APX, GR, MDHAR, DHAR, GPX, and GST, thereby maintaining ROS homeostasis and reducing postharvest blue mold. Collectively, the results in the present study revealed a control mechanism in which DEM treatment conferred the resistance against P. italicum infection in harvested "Newhall" navel orange fruit by activating lignin biosynthesis and maintaining the redox balance.

Suppression on postharvest juice sac granulation and cell wall modification by chitosan treatment in harvested pummelo (Citrus grandis L. Osbeck) stored at room temperature.

Deposition of both lignin and cellulose accompanied by juice sac granulation is widespread in harvested citrus fruit. Hence, measures to suppress postharvest granulation of 'Majiayou' pummelo is of great importance. The fruit was treated with 1.5% chitosan and then stored at room temperature (20 ± 2 °C) for 150 d. As compared to the control fruits, chitosan coating significantly suppressed granulation index and maintained good quality. Chitosan coating inhibited lignification by suppressing the activities and expression levels of lignin synthesis-related enzymes (PAL, CAD and POD). By contrast, chitosan treatment enhanced the activities and expression levels of cell wall degrading enzymes, including PME, PG, Cx, XTH and ß-Gal, which might contribute to the decrease in cellulose. In a nutshell, chitosan coating can effectively suppress juice sac granulation and fruit senescence of pummelo fruits, and play a crucial role in maintaining the cell wall modification.

Mitigating effects of chitosan coating on postharvest senescence and energy depletion of harvested pummelo fruit response to granulation stress.

The effect of chitosan coating exposure on juice sac granulation and energy metabolism in harvested pummelo fruit was investigated. Pummelo fruits were exposed to 1.5% chitosan coating, and then stored at 20 ± 2 °C for about 150 days. Postharvest chitosan coating treatment apparently alleviated the development of juice sac granulation as well as the increases in weight loss, pulp firmness, cell membrane permeability and cellulose content. The levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP) and energy charge (EC) in the chitosan-coated fruit showed significantly higher levels than those of the respective controls. Meanwhile, the enzymses actively engaged in energy metabolism such as H+-ATPase, Ca2+-ATPase, Mg2+-ATPase, cytochrome C oxidase (CCO), succinate dehydrogenase (SDH) and malate dehydrogenase (MDH) were markedly maintained by chitosan coating. Besides, notably high contents of acetyl-CoA, cis-aconitate, succinate, fumarate and oxaloacetate were observed in the chitosan-coated fruit. The results highlighted that chitosan coating could delay postharvest senescence of pummelo fruit by reducing the rate of energy depletion while maintaining higher levels of key metabolites taking part in tricarboxylic acid (TCA) cycle at room temperature storage.

Pharmacological Properties and Health Benefits of Eugenol: A Comprehensive Review.

The biologically active phytochemicals are sourced from edible and medicinally important plants and are important molecules being used for the formulation of thousands of drugs. These phytochemicals have great benefits against many ailments particularly the inflammatory diseases or oxidative stress-mediated chronic diseases. Eugenol (EUG) is a versatile naturally occurring molecule as phenolic monoterpenoid and frequently found in essential oils in a wide range of plant species. EUG bears huge industrial applications particularly in pharmaceutics, dentistry, flavoring of foods, agriculture, and cosmeceutics. It is being focused recently due to its great potential in preventing several chronic conditions. The World Health Organization (WHO) has declared EUG as a nonmutant and generally recognized as safe (GRAS) molecule. The available literature about pharmacological activities of EUG shows remarkable anti-inflammatory, antioxidant, analgesic, and antimicrobial properties and has a significant effect on human health. The current manuscript summarizes the pharmacological characteristics of EUG and its potential health benefits.