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Previously, VpPR-10.1 was isolated and characterized from a cDNA library of a fungus-resistant accession of Chinese wild grape (Vitis pseudoreticulata). We found that expression of VpPR-10.1 is affected by the fungal pathogen Erysiphe necator. To investigate the biochemical basis of the nuclease activity of VpPR-10.1 and its role in antifungal resistance, we generated recombinant VpPR-10.1 as well as site-directed mutations targeting three conserved amino acid residues among plant PR-10 s: Lys55, Glu149, and Tyr151. We showed that wild-type recombinant VpPR-10.1 exhibits both RNase and DNase activities. Mutant VpPR10.1-Y151H essentially retained all these activities. In contrast, VpPR10.1-K55N, where Lys55 in the P-loop region is mutated to Asn, and VpPR10.1-E149G, where Glu149 is mutated to Gly, lost their nuclease activity, indicating that both residues play a critical role in catalyzing RNA and DNA degradation. Furthermore, VpPR10.1 and VpPR10.1-Y151H inhibited the growth of the cultured fungal pathogen Alternaria alternate. Through transient expression in grapevine, we also demonstrated that VpPR10.1-K55N and VpPR10.1-E149G compromised resistance to E. necator. Finally, we further found that VpPR-10.1 can lead to programmed cell death and DNA degradation when incubated with tobacco BY-2 suspension cells. We show here that Lys55 and Glu149, but not Tyr151, are required for the RNase, DNase and antifungal activities of VpPR-10.1. The strong correlation between the level of VpPR-10.1 nuclease activity and its antifungal property indicates that the former is the biochemical basis for the latter. Taken together, our experiments revealed that VpPR-10.1 is critical in mediating fungal resistance in grape, potentially playing a dual role by degrading pathogen RNA and inducing programmed death of host cells. [ABSTRACT FROM AUTHOR]

Torrefaction of sawdust has the advantages of increasing its heating value, lowering its O/C ratio, and making it more convenient to transport and crush. Torrefied sawdust has characteristics that are more similar to coal than to sawdust. The initial pyrolysis temperature and residue content of torrefied sawdust are higher than those of sawdust. With the increase of coalification degree of three different coals, the initial pyrolysis temperature and residue contents are raised. The initial pyrolysis temperatures and remaining contents of lignite, bituminous coal, and anthracite are 330, 380, and 500 °C and 61.3, 75.1, and 89.5%, respectively. The torrefied sawdust also has a synergistic effect on the conversion of anthracite and bituminous coal, but it has an inhibitory effect on lignite. The composition of gaseous products was also measured after pyrolysis. The results demonstrate that with the addition of torrefied sawdust to anthracite and bituminous coal, the gaseous products contain more combustible components, such as H2, CO, and CH4, which increase the heating value. Moreover, the effect is more obvious with the co-pyrolysis of torrefied sawdust and anthracite. However, the co-pyrolysis of torrefied sawdust and lignite leads to decreasing CO and light hydrocarbons (CnHm (n=1, 2)) in the resulting gaseous products, which has a negative effect on the quality of the gaseous products. The co-pyrolysis characteristics of torrefied sawdust with different rank coals are discussed in this paper. [ABSTRACT FROM AUTHOR]