Human Enterovirus 68 Interferes with the Host Cell Cycle to Facilitate Viral Production.

Enterovirus D68 (EV-D68) is an emerging pathogen that recently caused a large outbreak of severe respiratory disease in the United States and other countries. Little is known about the relationship between EV-D68 virus and host cells. In this study, we assessed the effect of the host cell cycle on EV-D68 viral production, as well as the ability of EV-D68 to manipulate host cell cycle progression. The results suggest that synchronization in G0/G1 phase, but not S phase, promotes viral production, while synchronization in G2/M inhibits viral production. Both an early EV-D68 isolate and currently circulating strains of EV-D68 can manipulate the host cell cycle to arrest cells in the G0/G1 phase, thus providing favorable conditions for virus production. Cell cycle regulation by EV-D68 was associated with corresponding effects on the expression of cyclins and CDKs, which were observed at the level of the protein and/or mRNA. Furthermore, the viral non-structural protein 3D of EV-D68 prevents progression from G0/G1 to S. Interestingly, another member of the Picornaviridae family, EV-A71, differs from EV-D68 in that G0/G1 synchronization inhibits, rather than promotes, EV-A71 viral replication. However, these viruses are similar in that G2/M synchronization inhibits the production and activity of both viruses, which is suggestive of a common therapeutic target for both types of enterovirus. These results further clarify the pathogenic mechanisms of enteroviruses and provide a potential strategy for the treatment and prevention of EV-D68-related disease.

Plastidial glutathione reductase from Haynaldia villosa is an enhancer of powdery mildew resistance in wheat (Triticum aestivum).

A full-length cDNA (Hv-GR) whose transcript accumulation increased in response to infection by Blumeria graminis DC.f.sp. tritici (Bgt) was isolated from Haynaldia villosa. Southern analysis revealed a single copy of Hv-GR present in H. villosa. This gene encodes a glutathione reductase (GR) with high similarity to chloroplastic GRs from other plant species. Chloroplastic localization of Hv-GR was confirmed by targeting of the green fluorescent protein (GFP)-Hv-GR fusion protein to chloroplasts of epidermal guard cells. Following inoculation with Bgt, transcript accumulation of Hv-GR increased in a resistant line of wheat, but no significant change was observed in a susceptible line. In vivo function of Hv-GR in converting oxidized glutathione (GSSG) to the reduced form (GSH) was verified through heterologous expression of Hv-GR in a yeast GR-deficient mutant. As expected, overexpression of this gene resulted in increased resistance of the mutant to H(2)O(2), indicating a critical role for Hv-GR in protecting cells against oxidative stress. Moreover, overexpression of Hv-GR in a susceptible wheat variety, Triticum aestivum cv. Yangmai 158, enhanced resistance to powdery mildew and induced transcript accumulation of other pathogenesis-related genes, PR-1a and PR-5, through increasing the foliar GSH/GSSG ratio. Therefore, we concluded that a high ratio of GSH to GSSG is required for wheat defense against Bgt, and that chloroplastic GR enzymes might serve as a redox mediator for NPR1 activation.

[Transformation of powdery mildew resistance-related genes of wheat].

Anthocyanin synthesis regulation gene C1-Lc was used as the reporter gene to optimize the parameters of gene-gun transformation protocol through counting of red spots on wheat calli after transient expression. Wheat Beclin1 like gene TaTBL and thiosulfate sulfutransferase gene TaTST proved to have an increased expression level after induction of wheat powdery mildew fungus (Erysiphe graminis f.sp. tritici Em. Marchal.). These two resistance-related genes were constructed into expression vectors driven by the strong ubi promoter and used to perform genetic transformation on wheat cv Yangmai158 immature embryo-derived calli through particle bombardment. After two rounds of herbicide bialaphos selection and regeneration, herbicide-resistance plants were obtained, which were subsequently subjected to PCR analysis. Five TaTBL transgenic plants and six TaTST transgenic plants were identified. Pathogen inoculation of detached leaves showed that the introduction of exogenous gene increased wheat resistance level by delaying the development of powdery mildew symptoms.