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Carbonic anhydrases (CAs) are ubiquitous zinc metalloenzymes that catalyze the interconversion of carbon dioxide and bicarbonate. Higher plants mainly contain the three evolutionarily distinct CA families αCA, βCA, and γCA, with each represented by multiple isoforms. Alternative splicing (AS) of the CA transcripts is common. However, there is little information on the spliced variants of individual CA isoforms. In this study, we focused on the characterization of spliced variants of βCA1 from Arabidopsis. The expression patterns and subcellular localization of the individual spliced variants of βCA1 were examined. The results showed that the spliced variants of βCA1 possessed different subcellular and tissue distributions and responded differently to environmental stimuli. Additionally, we addressed the physiological role of βCA1 in heat stress response and its protein-protein interaction (PPI) network. Our results showed that βCA1 was regulated by heat stresses, and βca1 mutant was hypersensitive to heat stress, indicating a role for βCA1 in heat stress response. Furthermore, PPI network analysis revealed that βCA1 interacts with multiple proteins involved in several processes, including photosynthesis, metabolism, and the stress response, and these will provide new avenues for future investigations of βCA1.

White tail disease (WTD), a major disease prevailing in the larval stage of Macrobrachium rosenbergii, caused by Macrobrachium rosenbergii nodavirus (MrNV) associated with extra small virus (XSV), led to the economic loss of shrimp industry in China. In order to establish a convenient, sensitive and selective molecular diagnostic method to detect MrNV and XSV for the Chinese shrimp (MrNV/XSV-chin), a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay combined with a lateral flow dipstick (LFD) method were developed. A set of four specific primers and a labeled probe were designed according to the six conserved gene sequence regions encoding for the MrNV capsid protein CP43 and the XSV capsid protein CP17. The detection of MrNV and XSV simultaneously by RT-LAMP was performed at 61 °C in a single reaction for 60 min followed by hybridization with an FITC-labeled probe for 5 min and visualized by LFD. The RT-LAMP-LFD assay had a sensitivity of approximately 100-fold higher than conventional PCR. In addition, the assay could detect MrNV/XSV-chin from limited amount of RNA extracts as low as 1.0 pg extracted from Macrobrachium rosenbergii. This assay was simple to use, required little instrumentation, and exhibited excellent specificity for the MrNV/XSV-chin compared with other shrimp viruses. In conclusion, a convenient, sensitive and selective practical molecular diagnostic method was developed with the potential for diagnosis and prevention of WTD.

C-type lectins play an important role in innate immunity of invertebrates, especially in defending them against bacterial pathogens. In this study, the full-length genomic DNA of C-type lectin (designated as CTL gene) was isolated from the swimming crab Portunus trituberculatus. The genomic DNA of CTL consists of 1473 bp, containing four exons and three introns. Polymorphisms of the CTL gene were identified to explore their association with susceptibility/resistance to Vibrio alginolyticus infection. Four sites of single nucleotide polymorphisms (SNPs) and one site of insertion/deletion (ins/del) polymorphism were identified in CTL. Three of these SNPs were synonymous and one was non-synonymous. The distribution of these polymorphisms in the susceptible and resistant stocks was identified, according to the survival time after V. alginolyticus challenge. The non-synonymous SNP E4-205 C/T, a dimorphism caused from C to T transition that resulted in a Threonine to Isoleucine substitution at position 152 in the peptide of CTL protein, showed significant difference between the two stocks according to Chi-squared test (P

We report here a draft genome sequence of Aeromonas hydrophila strain BSK-10, belonging to serotype O97, isolated from crucian carp ( Carassius carassius ) with motile aeromonad septicemia in Zhejiang, China. The assembly resulted in 34 scaffolds totaling approximately 4.97 Mb, with an average G+C content of 60.97% and 4,594 predicted coding genes.

Ichthyophthirius is a severe disease of farmed freshwater fish caused by the parasitic ciliate Ichthyophthirius multifiliis (Ich). This disease can lead to considerable economic loss, but the protein profiles in different developmental stages of the parasite remain unknown. In the present study, proteins from trophonts and theronts of Ich were identified by isobaric tags for relative and absolute quantitation (iTRAQ). A total of 2300 proteins were identified in the two developmental stages, of which 1520 proteins were differentially expressed. Among them, 84 proteins were uniquely expressed in the theronts stage, while 656 proteins were expressed only in trophonts. The differentially expressed proteins were catalogued (assorted) to various functions of Ich life cycle, including biological process, cellular component, and molecular function that occur at distinct stages. Using a 1.5-fold change in expression as a physiologically significant benchmark, a lot of differentially expressed proteins were reliably quantified by iTRAQ analysis. Two hundred forty upregulated and 57 downregulated proteins in the trophonts stage were identified as compared with theronts. The identified proteins were involved in various functions of the I. multifiliis life cycle, including binding, catalytic activity, structural molecule activity, and transporter activity. Further investigation of the transcriptional levels of periplasmic immunogenic protein, transketolase, zinc finger, isocitrate dehydrogenase, etc., from the different protein profiles using quantitative RT-PCR showed identical results to the iTRAQ analysis. This work provides an effective resource to further our understanding of Ich biology, and lays the groundwork for the identification of potential drug targets and vaccines candidates for the control of this devastating fish pathogen.

A target sequence of 0. 9kb which codes the major capsid protein VP7 of grass carp reovirus (GCRV) was amplified by PCR. The target fragment was inserted into the pMD19-T vector. The positive clone was screened and sequenced. Sequencing result showed the nucleic acid sequence is right. Then the target gene was cloned into eukaryotic expression vector pCI and the positive clone was screened. The recombinant plasmid pCI-VP7 extracted from the positive clone were identified by PCR and digestion. Then the recombinant plasmid pCI-VP-GFP (fusion expression of GFP and partial VP7 gene) was constructed to testify the expression of VP7 gene. The recombinant plasmid pCI-VP7-GFP were transfected into COS-1 and CIK cells by lipofectamine 2000. Fluorescence microscope and RT-PCR was used to detect the transient expression. The results showed that the gene fragment was transfected and expressed in COS-1 and CIK cell successfully. The results indicated that the recombinant plasmid pCI-VP7 can also express successfully. It establishes foundation to research the gene vaccine of GCRV.