We isolated an MgATG1 gene encoding a serine/threonine protein kinase from the rice blast fungus Magnaporthe grisea. In the ⌬Mgatg1 mutant, in which the MgATG1 gene had been deleted, autophagy was blocked; the mutant also showed fewer lipid droplets in its conidia, lower turgor pressure of the appressorium, and such defects in morphogenesis as delayed initiation and slower germination of conidia. As a result of lower turgor pressure of the appressorium, the ⌬Mgatg1 mutant lost its ability to penetrate and infect the two host plants, namely, rice and barley. However, normal values of the parameters and infective abilities were restored on reintroducing an intact copy of the MgATG1 gene into the mutant. Autophagy is thus necessary for turnover of organic matter during the formation of conidia and appressoria and for normal development and pathogenicity in M. grisea.Magnaporthe grisea, a filamentous ascomycete fungus, is best known as the causal agent of rice blast, the most serious disease of cultivated rice throughout the world (15), and has been developed as a model organism for investigating fungus-host interactions (4, 27). The appressorium, a specialized cell necessary for infection by the rice blast fungus, generates tremendous intracellular turgor pressure (as much as 8.0 MPa) to penetrate the leaf cuticle (5, 26). Such enormous turgor in the appressorium is a consequence of accumulation of very large quantities of glycerol in the cell, and potential sources of glycerol biosynthesis are lipid, glycogen, and two sugars, trehalose and mannitol, in the conidium (21).The process of development from the conidium to the appressorium, and then from the appressorium to the penetration peg or infectious hypha, requires a cell to undergo significant phenotypic changes accompanied by the breakdown and recycling of old cellular components in about 30 h (21). In M. grisea, appressorium formation involves autophagy, but nuclei in conidia of the ⌬Mgatg8 mutant were not degraded during appressorium formation, and the mutant failed to infect the plant through the appressoria (29). Also, the mutant produced far fewer conidia. Autophagy is a common and evolutionarily preserved process that degrades and recycles old proteins and organelles in all eukaryotic cells (8,9,18,31). For many years, autophagy was believed to be involved in changes in cellular architecture during differentiation and development, presumably through its role in the turnover of organelles and proteins (9). Our study sought to find out whether autophagy has a role in the turnover of organic matter that enables a hypha to form a conidium, which then goes on to develop an appressorium, and in generating the turgor pressure in the appressorium required for successful infection.An expressed sequence tag (clone s197; GenBank accession no. CK828251) for MgATG1 (autophagy-related gene 1), homologous to ATG1 of yeast in its protein sequence, was found in the appressorium of the rice blast fungus (10). Because the fungus is used as a primary model for host-pathogen...

Rab GTPases are required for vesicle-vacuolar fusion during vacuolar biogenesis in fungi. To date, little is known about the biological functions of the Rab small GTPase components in Magnaporthe oryzae. In this study, we investigated MoYpt7 of M. oryzae, a homologue of the small Ras-like GTPase Ypt7 in Saccharomyces cerevisiae. Cellular localization assays showed that MoYpt7 was predominantly localized to vacuolar membranes. Using a targeted gene disruption strategy, a ΔMoYPT7 mutant was generated that exhibited defects in mycelial growth and production of conidia. The conidia of the ΔMoYPT7 mutant were malformed and defective in the formation of appressoria. Consequently, the ΔMoYPT7 mutant failed to cause disease in rice and barley. Furthermore, the ΔMoYPT7 mutant showed impairment in autophagy, breached cell wall integrity, and higher sensitivity to both calcium and heavy metal stress. Transformants constitutively expressing an active MoYPT7 allele (MoYPT7-CA, Gln67Leu) exhibited distinct phenotypes from the ΔMoYPT7 mutant. Expression of MoYPT7-CA in MoYpt7 reduced pathogenicity and produced more appressoria-forming single-septum conidia. These results indicate that MoYPT7 is required for fungal morphogenesis, vacuole fusion, autophagy, stress resistance and pathogenicity in M. oryzae.

Chromosome 17 is unusual among the human chromosomes in many respects. It is the largest human autosome with orthology to only a single mouse chromosome1, mapping entirely to the distal half of mouse chromosome 11. Chromosome 17 is rich in protein-coding genes, having the second highest gene density in the genome2,3. It is also enriched in segmental duplications, ranking third in density among the autosomes4. Here we report a finished sequence for human chromosome 17, as well as a structural comparison with the finished sequence for mouse chromosome 11, the first finished mouse chromosome. Comparison of the orthologous regions reveals striking differences. In contrast to the typical pattern seen in mammalian evolution5,6, the human sequence has undergone extensive intrachromosomal rearrangement, whereas the mouse sequence has been remarkably stable. Moreover, although the human sequence has a high density of segmental duplication, the mouse sequence has a very low density. Notably, these segmental duplications correspond closely to the sites of structural rearrangement, demonstrating a link between duplication and rearrangement. Examination of the main classes of duplicated segments provides insight into the dynamics underlying expansion of chromosome-specific, low-copy repeats in the human genome.

BackgroundThere was a large scale outbreak of the highly pathogenic porcine reproductive and respiratory syndrome (PRRS) in China and Vietnam during 2006 and 2007 that resulted in unusually high morbidity and mortality among pigs of all ages. The mechanisms underlying the molecular pathogenesis of the highly virulent PRRS virus (H-PRRSV) remains unknown. Therefore, the relationship between pulmonary gene expression profiles after H-PRRSV infection and infection pathology were analyzed in this study using high-throughput deep sequencing and histopathology.ResultsH-PRRSV infection resulted in severe lung pathology. The results indicate that aberrant host innate immune responses to H-PRRSV and induction of an anti-apoptotic state could be responsible for the aggressive replication and dissemination of H-PRRSV. Prolific rapid replication of H-PRRSV could have triggered aberrant sustained expression of pro-inflammatory cytokines and chemokines leading to a markedly robust inflammatory response compounded by significant cell death and increased oxidative damage. The end result was severe tissue damage and high pathogenicity.ConclusionsThe systems analysis utilized in this study provides a comprehensive basis for better understanding the pathogenesis of H-PRRSV. Furthermore, it allows the genetic components involved in H-PRRSV resistance/susceptibility in swine populations to be identified.