RESUMEN
Short open reading frames (sORFs) are a newly identified family of genes, and the functions of most sORF genes and their encoded peptides (SEPs) are still unknown. In this study, two ATP synthase subunits were identified in kuruma shrimp (Marsupenaeus japonicus) as SEPs, namely MjATP5I and MjATP5L. They were widely distributed in all of the tested tissues of shrimp and upregulated in hemocytes and intestines in response to WSSV challenge. The injection of recombinant proteins (rMjATP5I and rMjATP5L) increased the expression of Ie1 and Vp28, while the knockdown of MjATP5I and MjATP5L decreased the expression of Ie1 and Vp28. All of the results suggest that MjATP5I and MjATP5L were beneficial for WSSV replication. Further exploration found that MjATP5I and MjATP5L RNAi significantly improved the shrimp survival rates, reduced ATP production, and upregulated the expression of antimicrobial peptide genes post viral challenge, and the two ATPase subunits and Relish negatively regulated each other. These results reveal that MjATP5I and MjATP5L facilitated WSSV duplication by regulating the production of ATP contents and the expression of antimicrobial peptide genes in shrimp.
Asunto(s)
Penaeidae , Virus del Síndrome de la Mancha Blanca 1 , Animales , Virus del Síndrome de la Mancha Blanca 1/genética , Proteínas de Artrópodos/química , Sistemas de Lectura Abierta , Penaeidae/genética , Péptidos/genética , Adenosina TrifosfatoRESUMEN
Lysyl oxidases (LOXs) are copper-dependent monoamine oxidases, and they play critical roles in extracellular matrix (ECM) remodeling. The LOX and LOX-like (LOXL) proteins also have a variety of biological functions, such as development and growth regulation, tumor suppression, and cellular senescence. However, the functions of LOXLs containing repeated scavenger receptor cysteine-rich (SRCR) domains in immunity are rarely reported. In this study, we characterized the antiviral and antibacterial functions of a lysyl oxidase-like (LOXL) protein containing tandem SRCR domains in Marsupenaeus japonicus. The mRNA level of LoxL was significantly upregulated in the hemocytes and intestines of shrimp challenged using white spot syndrome virus (WSSV) or bacteria. After the knockdown of LoxL via RNA interference, WSSV replication and bacterial loads were apparently increased, and the survival rate of the shrimp decreased significantly, suggesting that LOXL functions against pathogen infection in shrimp. Mechanistically, LOXL interacted with the envelope proteins of WSSV or with lipopolysaccharide and peptidoglycan from bacteria in shrimp challenged using WSSV or bacteria, and it promoted the expression of a battery of antimicrobial peptides (AMPs) via the induction of Dorsal nuclear translocation against viral and bacterial infection. Moreover, LOXL expression was also positively regulated by Dorsal in the shrimp challenged by pathogens. These results indicate that, by acting as a pattern recognition receptor, LOXL plays vital roles in antiviral and antibacterial innate immunity by enhancing the expression of AMPs in shrimp.
Asunto(s)
Proteínas del Envoltorio Viral , Virus del Síndrome de la Mancha Blanca 1 , Animales , Antibacterianos , Péptidos Antimicrobianos , Antivirales , Cobre , Cisteína , Lipopolisacáridos , Peptidoglicano , Proteína-Lisina 6-Oxidasa/genética , Proteína-Lisina 6-Oxidasa/metabolismo , ARN Mensajero , Receptores de Reconocimiento de Patrones , Receptores Depuradores , Virus del Síndrome de la Mancha Blanca 1/genéticaRESUMEN
Maintaining eubiosis of the gastrointestinal (GI) microbiota is essential for animal health. White spot syndrome virus (WSSV) is the most lethal viral pathogen because it causes extremely high mortality in shrimp farming. However, it remains poorly understood how WSSV infection affects the microbiota in different regions of the GI tract of shrimp. In the present study, we established an experimental model of kuruma shrimp (Marsupenaeus japonicus) infection with WSSV and then investigated the effects of WSSV infection on the microbiota in the cardiac stomach, pyloric stomach, and intestines using metataxonomics. We identified 34 phyla and 576 genera of bacteria collectively. At the phylum level, Proteobacteria and Firmicutes were the most abundant in all the three GI segments. The WSSV infection decreased microbial diversity to a different extent in the stomachs and in a time-dependent manner. The infection with WSSV affected the microbiota composition in the two stomachs, but not the intestines. Firmicutes increased significantly, while Actinobacteria, Bacteroidetes, and Cyanobacteria decreased in the two stomachs of the WSSV-infected shrimp. At the genus level, Trichococcus and Vibrio increased, but Bradyrhizobium and Roseburia decreased in the cardiac stomach of the WSSV-infected shrimp. Trichococcus and Photobacterium increased in the pyloric stomach. Although Vibrio showed a slight downward trend, Aliivibrio (formerly Vibrio) increased in the pyloric stomach. Thiothrix, Fusibacter, and Shewanella decreased in the pyloric stomach, but no significant differences in these genera were detected in the cardiac stomach. Analysis of the predicted functions of the GI microbiota indicated that the WSSV infection resulted in losses of some microbiota functions. The new information from this study may help better understand the bacteria-virus interaction in the GI tract of shrimp and other crustacean species, and inform pathogen prevention/control and sustainable aquaculture production.
Asunto(s)
Microbioma Gastrointestinal , Penaeidae , Virus del Síndrome de la Mancha Blanca 1 , Animales , Intestinos , Penaeidae/microbiología , EstómagoRESUMEN
Substrate entrainment can greatly influence the mass movement process of a debris avalanche because it can enlarge the landslide volume and change the motion characteristics of the sliding masses. To study the interaction between debris avalanches and erodible substrate, physical modeling experiments varying in the mass of granular flow and substrate thickness were performed. The experimental results show that both the entrained materials and the maximum erosion depth are increased with increasing mass of the debris avalanche and decreasing substrate thickness. During the experiment, several tests were recorded using a high-speed digital camera with a frequency of 500 frames per second, so that the process of entrainment could be clearly observed. Combined with the experiment result and results of previous studies from predecessors, the entrainment mechanism during debris avalanches are analyzed and discussed. The entrainment effect of the sliding masses on the loose bed materials include basal abrasion and impact erosion of the avalanche front, the latter of which can contribute to the former by failing or yielding the erodible bed.