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.
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Cloning of mangrove red snapper (Lutjanus argentimaculatus) growth hormone cDNA and mRNA expression during early development LP Samentar, FG Ayson, EGT de Jesus-Ayson & MJ Formacion -
Philippine Journal of Natural Sciences, 2013 - University of the Philippines VisayasGrowth hormone regulates growth and development in vertebrates. As a first step to understand the role of growth hormone in the regulation of growth and development of the mangrove red snapper Lutjanus argentimaculatus, the red snapper growth hormone (sGH) cDNA was cloned using reverse transcription - polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. The expression of sGH mRNA in embryos and larvae was examined also by RT-PCR. Excluding the poly-A tail, the full-length red snapper GH cDNA is 945 base pairs (bp) long. It contains untranslated regions of 99 bp and 234 bp in the 5’ and 3’ ends, respectively. It has an open reading frame of 612 bp coding for a signal peptide of 17 amino acids and a mature hormone of 187 amino acid residues. Red snapper GH contains 4 cysteine residues and the typical polyadenylation site 16 bp upstream of the poly-A tail. Based on the amino acid sequence of the mature hormone, sGH shows higher sequence identity (>75%) to GHs of perciforms like grouper, seabass, tilapia and rabbitfish than to GHs of salmonids and carps. Semi-quantitative RT-PCR showed that expression of sGH mRNA commenced two days after hatching.