High throughput RNA sequencing reveals temperature tolerance mechanisms in Scylla serrata
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The effects of increasing temperature from global climate change threaten the sustainability and production of mud crabs from farms and wild populations in mangroves. Adaptation of mud crab populations to temperature stress is difficult to evaluate until now, with the emergence of RNA-Seq, a method which evaluates total mRNA expression under different conditions. In this study, 10 individuals each of S. serrata from Buguey, Cagayan were exposed to 26~’C and 32~’C for two weeks and the mRNA profiles were compared based on 186 million high quality pair-end reads which were aligned to a S. serrata reference transcriptome assembled de novo from 24,350 contigs with an average N50 of 1564 bp. Temperature related differences in gene expression were not significantly detected between the control and treatment groups and this was mostly due to the highly expressed genes such as the low and high molecular weight heat shock proteins. However, variations were greater among genes involved in the process of cell cycle regulation, the dissimilation processes such as oxidative phosphorylation, reproduction and transport across membranes. Greater differences were observed between immature or mature males and females.
Ablan-Lagman, M. C., & Meyer, E. (2017). High throughput RNA sequencing reveals temperature tolerance mechanisms in Scylla serrata. In E. T. Quinitio, F. D. Parado-Estepa, & R. M. Coloso (Eds.), Philippines : In the forefront of the mud crab industry development : proceedings of the 1st National Mud Crab Congress, 16-18 November 2015, Iloilo City, Philippines (p. 140). Tigbauan, Iloilo, Philippines: Aquaculture Department, Southeast Asian Fisheries Development Center. http://hdl.handle.net/10862/3174
PublisherAquaculture Department, Southeast Asian Fisheries Development Center
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Targeting essential gene utilizing RNA interference to protect the ailing shrimp/prawn industry against WSSV JMS Lazarte & MBB Maningas - In MRR Romana-Eguia, FD Parado-Estepa, ND Salayo & MJH Lebata-Ramos (Eds.), Resource Enhancement and Sustainable Aquaculture Practices in Southeast Asia: Challenges in Responsible Production … International Workshop on Resource Enhancement and Sustainable Aquaculture Practices in Southeast Asia 2014 (RESA), 2015 - Aquaculture Department, Southeast Asian Fisheries Development CenterThe white spot syndrome virus (WSSV) remains to be the most widespread and devastating infectious agent that has hit particularly the marine shrimp aquaculture industry worldwide. To date, there are no known effective strategies that can combat WSSV infection. This study aimed to elucidate host-pathogen interaction through the functional study of host - gene. Utilizing RNA Interference, the function of contig23 (c23) in the shrimp genome, identified to have high homology with WSSVORF-325, was determined. Three set-ups were prepared for treatment of c23-, GFP-dsRNA, and PBS using Macrobrachium rosenbergii freshwater prawns. Each treatment group was challenged with WSSV and survival rate was recorded. C23-, and GFP-dsRNA injected prawns showed a significant survival rate of 100%, in contrast to 20% of the PBS injected prawns at 10 days post-infection (dpi). Results showed that injection of c23- and GFP-dsRNA prior to challenge with WSSV, delayed and reduced mortality in contrast to PBS-treated prawns, which showed high mortality. Gene expression analysis showed silencing of both WSSV and c23 at day 3 post-WSSV challenge. This study proved that c23-dsRNA has a protective effect on WSSVchallenged prawns and highlights its involvement in the infectivity of WSSV in M. rosenbergii.
Shrimp metabolism: The roles of lactate dehydrogenase (c31), glycogen phosphorylase (c34) and protein kinase (PK) as revealed by RNA interference MVR Tare, H Kondo, I Hirono & MBB Maningas - In MRR Romana-Eguia, FD Parado-Estepa, ND Salayo & MJH Lebata-Ramos (Eds.), Resource Enhancement and Sustainable Aquaculture Practices in Southeast Asia: Challenges in Responsible Production … International Workshop on Resource Enhancement and Sustainable Aquaculture Practices in Southeast Asia 2014 (RESA), 2015 - Aquaculture Department, Southeast Asian Fisheries Development CenterEnergy metabolism is well-studied in vertebrate systems, providing insights on the genes and mechanisms involved in different pathways necessary for the survival of an organism. Yet, such studies are still lacking in invertebrate systems much more in shrimp. An earlier study has showed several contigs from the black tiger shrimp to be homologous to white spot syndrome virus (WSSV), a devastating pathogen in shrimp, including contig 31-WSSVORF82 (c31) and contig 34-WSSVORF21 (c34). This study aims to unveil the roles of three genes: c31, c34 and protein kinase (PK) in the shrimp system and its possible role in WSSV-infection. Rapid amplification of cDNA ends-polymerase chain reaction or RACE-PCR was used to obtain the full-length sequence of c31 and c34, followed by in vivo gene silencing using RNAi technology, and intramuscularly injecting dsRNA to WSSVchallenged Macrobrachium rosenbergii and Penaeus (Marsupenaeus) japonicus. Gene expression followed for healthy shrimps and dsRNA-treated shrimps. Mrc31 was revealed to be the enzyme lactase dehydrogenase (LDH), commonly released during tissue damage and is a marker for disease. The most parsimonious tree pictured Mrc31 to be sister clades to LDH of other shrimp species, Penaeus monodon and P. vannamei, supported with 100% and 72% bootstrap values, respectively. Mrc34 was highly homologous to the glycogen phosphorylase (GP) enzymes of other organisms including that of another shrimp, M. japonicus, bearing a bootstrap value of 99%. For PK, phylogenetic analysis revealed that the three open reading frames (ORFs) from P. monodon, M. rosenbergii and P. japonicus have 30% homology to WSSV-PK supported by a 98% bootstrap value. Mortality data from dsRNA-treated and WSSV-infected shrimps showed that treatment with dsRNA-LDH, GP and PK had significantly higher survival rates compared to that of the controls, Phosphate Buffered Saline (PBS) and Green Fluorescent Protein (GFP). Silencing the three genes in the shrimp has rendered some protective effect against the virus. Gene expression showed that all three genes are present in immune-related organs such as the gills, hepatopancreas and hemocyte. This study is the first to report the possible identities and functions of contigs 31, 34 and PK providing valuable data on the shrimp's genome.
Milkfish (Chanos chanos) growth hormone cDNA cloning and mRNA expression in embryos and early larval stages. EGT de Jesus, FG Ayson, Y Amemiya, S Moriyama, S Hyodo, T Hirano & H Kawauchi -
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