scholarly journals Selection of functional EPHB2 genotypes from ENU mutated grass carp treated with GCRV

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Meher un Nissa ◽  
Zhu-Xiang Jiang ◽  
Guo-Dong Zheng ◽  
Shu-Ming Zou
Keyword(s):  
Grass Carp ◽  
Bulked Segregant Analysis ◽  
Grass Carp Reovirus ◽  
Synonymous Mutations ◽  
Resistant Group ◽  
Intron Region ◽  
Immune Related Genes ◽  
Haemorrhagic Disease ◽  
Functional Mutants ◽  
Selection Of

Abstract Background N-ethyl-N-nitrosourea (ENU) mutagenesis is a useful method for the genetic engineering of plants, and the production of functional mutants in animal models including mice and zebrafish. Grass carp reovirus (GCRV) is a haemorrhagic disease of grass carp which has caused noteworthy losses in fingerlings over the last few years. To overcome this problem, we used ENU mutant grass carp in an attempt to identify functional resistance genes for future hereditary rearing projects in grass carp. Results This study used ENU-mutated grass carp to identify genetic markers associated with resistance to the haemorrhagic disease caused by GCRV. Bulked segregant analysis (BSA) was performed on two homozygous gynogenetic ENU grass carp groups who were susceptible or resistant to GCRV. This analysis identified 466,162 SNPs and 197,644 InDels within the genomes of these mixed pools with a total of 170 genes annotated in the associated region, including 49 genes with non-synonymous mutations at SNP sites and 25 genes with frame shift mutations at InDel sites. Of these 170 mutated genes, 5 randomly selected immune-related genes were shown to be more strongly expressed in the resistant group as compared to the susceptible animals. In addition, we found that one immune-related gene, EPHB2, presented with two heterozygous SNP mutations which altered the animal’s responded to GCRV disease. These SNPs were found in the intron region of EPHB2 at positions 5859 (5859G > A) and 5968 (5968G > A) and were significantly (p = 0.002, 0.003) associated with resistance to GCRV. These SNP sites were also shown to correlate with the GCRV-resistant phenotype in these ENU grass carp. We also evaluated the mortality of the different ENU fish genotypes in response to GCRV and the SNPs in EPHB2. The outcomes of these evaluations will be useful in future selections of GCRV-resistant genes for genetic breeding in grass carp. Conclusion Our results provide a proof of concept for the application of BSA-sequence analysis in detecting genes responsible for specific functional genotypes and may help to develop better methods for marker-assisted selection, especially for disease resistance in response to GCRV.

scholarly journals Selection of functional EPHB2 genotype in ENU mutated grass carp response to GCRV by the method of BSA Sequence Analysis

2021 ◽  
Author(s):  
Meher un Nissa ◽  
ZhuXiang Jiang ◽  
GuoDong Zheng ◽  
ShuMing Zou
Keyword(s):  
Sequence Analysis ◽  
Grass Carp ◽  
Bulked Segregant Analysis ◽  
Hemorrhagic Disease ◽  
Grass Carp Reovirus ◽  
Synonymous Mutations ◽  
Genes Expression ◽  
Resistant Group ◽  
Intron Region ◽  
Immune Related Genes

Abstract Background N-ethyl-N-nitrosourea (ENU) mutagenesis is a useful method for genetic development of plants, as well as for inducing functional mutants in animal models including mice and zebrafish. GCRV (Grass carp Reovirus) is the hemorrhage disease of grass carp and causes noteworthy loss of fingerlings in the last few years. To overcome this problem, we have used ENU mutant grass carp might be a helpful to find out some resistant functional gene for future hereditary rearing projects in grass carp.Results In the present study, ENU-mutated grass carp were used to identify genetic markers correlated with anti hemorrhagic disease caused by grass carp reovirus (GCRV). The BSA (Bulked segregant analysis) technology were performed on two homozygous gynogenetic ENU grass carp groups with susceptible or resistant in response of GCRV. Our results showed that 466,162 SNPs and 197644 InDel were found out in the mixed pool. A total of 170 genes were annotated in the associated region, including 49 genes with non-synonymous mutations at SNP sites and 25 genes with frame shift mutations at InDel sites. Among these 170 mutated genes, 5 randomly selected immune-related genes expression was higher in resistant group as compared to susceptible. Furthermore, we found one immune-related gene EPHB2 has confirmed two heterozygous SNP mutations which responded to GCRV disease. The SNPs within gene EPHB2 were found out in the intron region with position 5859 (5859G>A) and 5968 (5968G>A) were significantly (p = 0.002, 0.003) associated with the resistance against GCRV. These SNP sites were confirmed to correlate with the GCRV-resistant phenotype of ENU grass carp. The SNPs associated with GCRV resistance are useful in selecting GCRV-resistant genes for genetic breeding in grass carp.Conclusion Our results proved the possibility of BSA-sequence analysis in detecting genes responsible for the exciting phenotypes and will help in carrying out the marker-assisted selection, especially for disease resistance in the response of GCRV.

RESISTANCE OF SIGCRV TRANSGENIC RARE MINNOW(GOBIOCYPRIS RARUS)TO GRASS CARP REOVIRUS

2010 ◽  
Vol 36 (4) ◽  
pp. 837-842 ◽  
Author(s):  
Sha LIAO ◽  
Yun CHEN ◽  
Fu-Kuan DU ◽  
Ya-Ping WANG ◽  
Lan-Jie LIAO ◽  
...  
Keyword(s):  
Grass Carp ◽  
Gobiocypris Rarus ◽  
Grass Carp Reovirus ◽  
Rare Minnow

Establishment of multiplex PCR for detection of grass carp reovirus and its application

2013 ◽  
Vol 20 (2) ◽  
pp. 419-426 ◽  
Author(s):  
Weiwei ZENG ◽  
Qing WANG ◽  
Yingying WANG ◽  
Lesheng ZHANG ◽  
Baoqin LIU ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Grass Carp ◽  
Grass Carp Reovirus

Isolation and identification of grass carp reovirus strain JX-0902

2013 ◽  
Vol 18 (5) ◽  
pp. 1077-1083 ◽  
Author(s):  
Yongkui LIU ◽  
Qing WANG ◽  
Weiwei ZENG ◽  
Cunbin SHI ◽  
Chao ZHANG ◽  
...  
Keyword(s):  
Grass Carp ◽  
Isolation And Identification ◽  
Grass Carp Reovirus

Preparation and identification of monoclonal antibody against VP4 protein of grass carp reovirus HZ08 strain

2013 ◽  
Vol 37 (3) ◽  
pp. 450 ◽  
Author(s):  
Weiwei ZENG ◽  
Qing WANG ◽  
Yingying WANG ◽  
Cunbin SHI ◽  
Shuqin WU
Keyword(s):  
Monoclonal Antibody ◽  
Grass Carp ◽  
Grass Carp Reovirus

Development and efficacy of a grass carp reovirus (GCRV) outer capsid protein VP7 subunit vaccine

2012 ◽  
Vol 37 (6) ◽  
pp. 659-664 ◽  
Author(s):  
Shi-ying XU ◽  
Jing-hui LI ◽  
Yong ZOU ◽  
Lin LIU ◽  
Cheng-liang GONG ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Grass Carp ◽  
Subunit Vaccine ◽  
Outer Capsid Protein ◽  
Grass Carp Reovirus ◽  
Outer Capsid

scholarly journals Oral Vaccination of Grass Carp (Ctenopharyngodon idella) with Baculovirus-Expressed Grass Carp Reovirus (GCRV) Proteins Induces Protective Immunity against GCRV Infection

Vaccines ◽  
2021 ◽  
Vol 9 (1) ◽  
pp. 41
Author(s):  
Changyong Mu ◽  
Qiwang Zhong ◽  
Yan Meng ◽  
Yong Zhou ◽  
Nan Jiang ◽  
...  
Keyword(s):  
Survival Rate ◽  
Grass Carp ◽  
Oral Vaccine ◽  
Serum Antibody ◽  
Ctenopharyngodon Idella ◽  
Control Group ◽  
Oral Vaccination ◽  
Grass Carp Reovirus ◽  
Silkworm Pupae ◽  
Grass Carp Ctenopharyngodon Idella

The grass carp reovirus (GCRV) causes severe hemorrhagic disease with high mortality and leads to serious economic losses in the grass carp (Ctenopharyngodon idella) industry in China. Oral vaccine has been proven to be an effective method to provide protection against fish viruses. In this study, a recombinant baculovirus BmNPV-VP35-VP4 was generated to express VP35 and VP4 proteins from GCRV type Ⅱ via Bac-to-Bac baculovirus expression system. The expression of recombinant VP35-VP4 protein (rVP35-VP4) in Bombyx mori embryo cells (BmE) and silkworm pupae was confirmed by Western blotting and immunofluorescence assay (IFA) after infection with BmNPV-VP35-VP4. To vaccinate the grass carp by oral route, the silkworm pupae expressing the rVP35-VP4 proteins were converted into a powder after freeze-drying, added to artificial feed at 5% and fed to grass carp (18 ± 1.5 g) for six weeks, and the immune response and protective efficacy in grass carp after oral vaccination trial was thoroughly investigated. This included blood cell counting and classification, serum antibody titer detection, immune-related gene expression and the relative percent survival rate in immunized grass carp. The results of blood cell counts show that the number of white blood cells in the peripheral blood of immunized grass carp increased significantly from 14 to 28 days post-immunization (dpi). The differential leukocyte count of neutrophils and monocytes were significantly higher than those in the control group at 14 dpi. Additionally, the number of lymphocytes increased significantly and reached a peak at 28 dpi. The serum antibody levels were significantly increased at Day 14 and continued until 42 days post-vaccination. The mRNA expression levels of immune-related genes (IFN-1, TLR22, IL-1β, MHC I, Mx and IgM) were significantly upregulated in liver, spleen, kidney and hindgut after immunization. Four weeks post-immunization, fish were challenged with virulent GCRV by intraperitoneal injection. The results of this challenge study show that orally immunized group exhibited a survival rate of 60% and relative percent survival (RPS) of 56%, whereas the control group had a survival rate of 13% and RPS of 4%. Taken together, our results demonstrate that the silkworm pupae powder containing baculovirus-expressed VP35-VP4 proteins could induce both non-specific and specific immune responses and protect grass carp against GCRV infection, suggesting it could be used as an oral vaccine.

scholarly journals Grass Carp Reovirus (GCRV) Giving Its All to Suppress IFN Production by Countering MAVS Signaling Transduction

2020 ◽  
Vol 11 ◽  
Author(s):  
Long-Feng Lu ◽  
Zhuo-Cong Li ◽  
Can Zhang ◽  
Xiao-Yu Zhou ◽  
Yu Zhou ◽  
...  
Keyword(s):  
Grass Carp ◽  
Signaling Transduction ◽  
Grass Carp Reovirus

scholarly journals Isolation, Identification, and Genomic Analysis of a Novel Reovirus from Healthy Grass Carp and Its Dynamic Proliferation In Vitro and In Vivo

Viruses ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 690
Author(s):  
Ke Zhang ◽  
Wenzhi Liu ◽  
Yiqun Li ◽  
Yong Zhou ◽  
Yan Meng ◽  
...  
Keyword(s):  
Cell Lines ◽  
Grass Carp ◽  
Amino Acid Sequences ◽  
Gibel Carp ◽  
Fish Cell ◽  
Fish Cell Lines ◽  
Total Size ◽  
Sequence Comparisons ◽  
Grass Carp Reovirus

A new grass carp reovirus (GCRV), healthy grass carp reovirus (HGCRV), was isolated from grass carp in 2019. Its complete genome sequence was determined and contained 11 dsRNAs with a total size of 23,688 bp and 57.2 mol% G+C content, encoding 12 proteins. All segments had conserved 5' and 3' termini. Sequence comparisons showed that HGCRV was closely related to GCRV-873 (GCRV-I; 69.57–96.71% protein sequence identity) but shared only 22.65–45.85% and 23.37–43.39% identities with GCRV-HZ08 and Hubei grass carp disease reovirus (HGDRV), respectively. RNA-dependent RNA-polymerase (RdRp) protein-based phylogenetic analysis showed that HGCRV clustered with Aquareovirus-C (AqRV-C) prior to joining a branch common with other aquareoviruses. Further analysis using VP6 amino acid sequences from Chinese GCRV strains showed that HGCRV was in the same evolutionary cluster as GCRV-I. Thus, HGCRV could be a new GCRV isolate of GCRV-I but is distantly related to other known GCRVs. Grass carp infected with HGCRV did not exhibit signs of hemorrhage. Interestingly, the isolate induced a typical cytopathic effect in fish cell lines, such as infected cell shrank, apoptosis, and plague-like syncytia. Further analysis showed that HGCRV could proliferate in grass carp liver (L28824), gibel carp brain (GiCB), and other fish cell lines, reaching a titer of up to 7.5 × 104 copies/μL.

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