SUMMARYRice (Oryza sativa L.) accumulates prolamines and glutelins as its major storage proteins. Glutelins are synthesized on rough endoplasmic reticulum as 57-kDa precursors; they are then sorted into protein storage vacuoles where they are processed into acidic and basic subunits. We report a novel rice glutelin mutant, W379, which accumulates higher levels of the 57-kDa glutelin precursor. Genetic analysis revealed that the W379 phenotype is controlled by a single recessive nuclear gene. Using a map-based cloning strategy, we identified this gene, OsVPE1, which is a homolog of the Arabidopsis bVPE gene. OsVPE1 encodes a 497-aminoacid polypeptide. Nucleotide sequence analysis revealed a missense mutation in W379 that changes Cys269 to Gly. Like the wild-type protein, the mutant protein is sorted into vacuoles; however, the enzymatic activity of the mutant OsVPE1 is almost completely eliminated. Further, we show that OsVPE1 is incorrectly cleaved, resulting in a mature protein that is smaller than the wild-type mature protein. Taken together, these results demonstrate that OsVPE1 is a cysteine protease that plays a crucial role in the maturation of rice glutelins. Further, OsVPE1 Cys269 is a key residue for maintaining the Asn-specific cleavage activity of OsVPE1.

Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ac83 is a baculovirus core gene whose function in the AcMNPV life cycle is unknown. In the present study, an ac83-knockout AcMNPV (vAc83KO) was constructed to investigate the function of ac83 through homologous recombination in Escherichia coli. No budded virions were produced in vAc83KO-transfected Sf9 cells, although viral DNA replication was unaffected. Electron microscopy revealed that nucleocapsid assembly was aborted due to the ac83 deletion. Domain-mapping studies revealed that the expression of Ac83 amino acid residues 451 to 600 partially rescued the ability of AcMNPV to produce infectious budded virions. Bioassays indicated that deletion of the chitinbinding domain of Ac83 resulted in the failure of oral infection of Trichoplusia ni larvae by AcMNPV, but AcMNPV remained infectious following intrahemocoelic injection, suggesting that the domain is involved in the binding of occlusion-derived virions to the peritrophic membrane and/or to other chitin-containing insect tissues. It has been demonstrated that Ac83 is the only component with a chitin-binding domain in the per os infectivity factor complex on the occlusion-derived virion envelope. Interestingly, a functional inner nuclear membrane sorting motif, which may facilitate the localization of Ac83 to the envelopes of occlusion-derived virions, was identified by immunofluorescence analysis. Taken together, these results demonstrate that Ac83 plays an important role in nucleocapsid assembly and the establishment of oral infection.

Rabies is an acute, fatal, neurological disease that affects almost all kinds of mammals. Vaccination (using an inactivated rabies vaccine), combined with administration of rabies immune globulin, is the only approved, effective method for post-exposure prophylaxis against rabies in humans. In the search for novel rabies control and treatment strategies, live-attenuated viruses have recently emerged as a practical and promising approach for immunizing and controlling rabies. Unlike the conventional, inactivated rabies vaccine, live-attenuated viruses are genetically modified viruses that are able to replicate in an inoculated recipient without causing adverse effects, while still eliciting robust and effective immune responses against rabies virus infection. A number of viruses with an intrinsic capacity that could be used as putative candidates for live-attenuated rabies vaccine have been intensively evaluated for therapeutic purposes. Additional novel strategies, such as a monoclonal antibody-based approach, nucleic acid-based vaccines, or small interfering RNAs (siRNAs) interfering with virus replication, could further add to the arena of strategies to combat rabies. In this review, we highlight current advances in rabies therapy and discuss the role that they might have in the future of rabies treatment. Given the pronounced and complex impact of rabies on a patient, a combination of these novel modalities has the potential to achieve maximal anti-rabies efficacy, or may even have promising curative effects in the future. However, several hurdles regarding clinical safety considerations and public awareness should be overcome before these approaches can ultimately become clinically relevant therapies.

Proper chloroplast development and chlorophyll biosynthesis are essential for the photoautotrophic plants. The insertion of magnesium (Mg 2+ ) into protoporphyrin IX (Proto), catalyzed by magnesium chelatase (Mgchelatase), is the first committed step of chlorophyll biosynthesis. In dicot plants, a proposed model revealed that Mg-chelatase I subunit (CHLI) and Mg-chelatase D subunit (CHLD) can interact directly; however, their relation remains elusive in rice, a monocot model plant. In this study, we characterized a chlorophyll-deficiency mutant, etiolated leaf and lethal (ell), which displayed a yellow leaf in young seedlings and became lethal after three-leaf stage. Chlorophyll content in homozygous ell mutant was approximately 1 % of that in the wild type. Besides, chloroplast development in the mutant was entirely arrested and no thylakoid structure was observed. By map-based cloning, the ell locus was delimited to a 3.9-Mb interval in chromosome 3. A single-base mutation (G529C) inOsCHLI was identified, leading to an amino acid substitution (G177R) in a highly conserved region. Compared with the wild type, more Proto but less magnesium protoporphyrin IX (Mg-Proto) was measured in the ell mutant. Using protoplast transfection and callus transformation, we found that exogenous OsCHLI could consistently recover the lesion of chloroplast in the ell mutant. By subcellar localization analysis, OsCHLI was detected in the chloroplast. Despite the secondary structure of OsCHLI that was predicted to be altered in the mutant, the point mutation did not affect subcellular localization. Real-time PCR demonstrated that the ell mutation induced significantly transcriptional downregulation of the photosynthesis-associated nuclear and plastid genes. Additionally, yeast-two-hybrid experiments indicated that the single amino acid substitution blocked the intrinsic interaction between OsCHLI and OsCHLD. Moreover, OsCHLI showed physical interactions with some thioredoxins (TRXs), suggesting a similar regulatory mechanism of Mg-chelatase activity in both monocot and dicot plants. Keywords Rice . Map-based cloning . Mg-chelatase . OsCHLI subunit . OsCHLD subunit . Thioredoxin Abbreviations Proto Protoporphyrin IX Mg-chelatase Magnesium chelatase ell Etiolated leaf and lethal Mg-Proto Magnesium protoporphyrin IX TRX Thioredoxin EMS Ethyl methyl sulfonate CHLH Mg-chelatase H subunit Electronic supplementary material The online version of this article (

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Producing sufficient food with finite resources to feed the growing global population while having a smaller impact on the environment has always been a great challenge. Here, we review the concept and practices of Green Super Rice (GSR) that have led to a paradigm shift in goals for crop genetic improvement and models of food production for promoting sustainable agriculture. The momentous achievements and global deliveries of GSR have been fueled by the integration of abundant genetic resources, functional gene discoveries, and innovative breeding techniques with precise gene and whole-genome selection and efficient agronomic management to promote resource-saving, environmentally friendly crop production systems. We also provide perspectives on new horizons in genomic breeding technologies geared toward delivering green and nutritious crop varieties to further enhance the development of green agriculture and better nourish the world population.

To study the prevalence and isoforms of the pathogenicity island ETT2 among pathogenic Escherichia coli, as well as to determine the relationship between the ETT2 locus and other virulence factors, PCR amplifications target to the 35 ETT2-associated genes were established and used to investigate the presence of the ETT2 locus in 168 E. coli isolates from weaned piglets with edema and/or diarrhea or dairy cows with mastitis. The results showed that the ETT2 locus could be identified in the pathogenic E. coli isolates from colibacillosis in pigs and in the ones from mastitis in cows, but the presence of ETT2 among the isolates of porcine origin were significantly higher (85.87%) than that (47.37%) of bovine origin. Furthermore, 11 ETT2 isoforms were found in this research, including an intact form and 10 deletion types. The intact ETT2 was the prevalent form among the pathogenic E. coli isolates of porcine origin, and highly associated with the presence of shigatoxin type 2e (Stx2e), while the great majority isolates of bovine origin just carried various deletion types, and no distinct association with other virulence factors, e.g., the presence/absence of LT1, ST2, Cnf2, Tra, HPI, Hly, and F17a fimbriae.

Starch synthesized and stored in amyloplasts serves as the major energy storage molecule in cereal endosperm. To elucidate the molecular mechanisms underlying amyloplast development and starch synthesis, we isolated a series of floury endosperm mutants in rice (). We identified the rice mutant (), which exhibited obvious defects in the development of compound starch grains, decreased starch content, and altered starch physicochemical features. Map-based cloning showed that encodes a phospholipase-like protein homologous to phosphatidic acid-preferring phospholipase A was expressed ubiquitously with abundant levels observed in developing seeds and roots. FSE1 was localized to both the cytosol and intracellular membranes. Lipid profiling indicated that total extra-plastidic lipids and phosphatidic acid were increased in plants, suggesting that FSE1 may exhibit in vivo phospholipase A activity on phosphatidylcholine, phosphatidylinositol, phosphatidyl-Ser, phosphatidylethanolamine, and, in particular, phosphatidic acid. Additionally, the total galactolipid content in developing endosperm was significantly reduced, which may cause abnormal amyloplast development. Our results identify FSE1 as a phospholipase-like protein that controls the synthesis of galactolipids in rice endosperm and provide a novel connection between lipid metabolism and starch synthesis in rice grains during endosperm development.