This study examined genetic variation in the major histocompatibility complex(MHC) Class II B gene in turbot(Scophthalmus maximus) by virulent bacterial pathogen challenge.One hundred fry from each of six families wer...This study examined genetic variation in the major histocompatibility complex(MHC) Class II B gene in turbot(Scophthalmus maximus) by virulent bacterial pathogen challenge.One hundred fry from each of six families were infected with Edwardsiella tarda by intraperitoneal injection.Family mortality ranged from 28.0% to 83.3%.Complete exon 2 and intron 1 sequences of MHC Class II B genes were amplified from five survivor and five non-survivor individuals per family using the clone-sequence method.Thirty-seven sequences from 60 individuals revealed 37 different alleles,25 of which were unique to this study.The 25 unique alleles belonged to 16 major allele types.Nine alleles were used to examine the association between alleles and resistance/susceptibility to disease.Five alleles were present in an individual,suggesting a minimum of three loci or copies of the turbot MHC Class II B gene.The rate of non-synonymous substitution(d N) was 2.30 and 1.58 times higher than synonymous substitution(d S) in the peptide-binding regions(PBR) and non-PBR in whole families,respectively,which suggested balancing selection on exon 2 of the MHC Class II B gene in turbot.One allele,Scma-DBB1*02,was significantly more prevalent in survivor stock than in non-survivor stock(P=0.001).Therefore,this allele might be associated with resistance to bacteria.A second allele,Scma-DBB1*10,was significantly more prevalent in non-survivor stock(P=0.021),and is likely associated with susceptibility to bacteria.展开更多
The worldwide declines in amphibian populations have largely been caused by infectious fungi and bacteria. Given that vertebrate immunity against these extracellular pathogens is primarily functioned by the major hist...The worldwide declines in amphibian populations have largely been caused by infectious fungi and bacteria. Given that vertebrate immunity against these extracellular pathogens is primarily functioned by the major histocompatibility complex(MHC) class Ⅱ molecules, the characterization and the evolution of amphibian MHC class Ⅱ genes have attracted increasing attention. The polymorphism of MHC class Ⅱ genes was found to be correlated with susceptibility to fungal pathogens in many amphibian species, suggesting the importance of studies on MHC class Ⅱ genes for amphibians. However, such studies on MHC class Ⅱ gene evolution have rarely been conducted on amphibians in China. In this study, we chose Omei treefrog(Rhacophorus omeimontis), which lived moist environments easy for breeding bacteria, to study the polymorphism of its MHC class Ⅱ genes and the underlying evolutionary mechanisms. We amplified the entire MHC class ⅡB exon 2 sequence in the R. omeimontis using newly designed primers. We detected 102 putative alleles in 146 individuals. The number of alleles per individual ranged from one to seven, indicating that there are at least four loci containing MHC class ⅡB genes in R. omeimontis. The allelic polymorphism estimated from the 102 alleles in R. omeimontis was not high compared to that estimated in other anuran species. No significant gene recombination was detected in the 102 MHC class ⅡB exon 2 sequences. In contrast, both gene duplication and balancing selection greatly contributed to the variability in MHC class ⅡB exon 2 sequences of R. omeimontis. This study lays the groundwork for the future researches to comprehensively analyze the evolution of amphibian MHC genes and to assess the role of MHC gene polymorphisms in resistance against extracellular pathogens for amphibians in China.展开更多
基金supported by the National Key Basic Research Program of China (2010CB126303)the Tai Shan Scholar Project of Shandong Province,China
文摘This study examined genetic variation in the major histocompatibility complex(MHC) Class II B gene in turbot(Scophthalmus maximus) by virulent bacterial pathogen challenge.One hundred fry from each of six families were infected with Edwardsiella tarda by intraperitoneal injection.Family mortality ranged from 28.0% to 83.3%.Complete exon 2 and intron 1 sequences of MHC Class II B genes were amplified from five survivor and five non-survivor individuals per family using the clone-sequence method.Thirty-seven sequences from 60 individuals revealed 37 different alleles,25 of which were unique to this study.The 25 unique alleles belonged to 16 major allele types.Nine alleles were used to examine the association between alleles and resistance/susceptibility to disease.Five alleles were present in an individual,suggesting a minimum of three loci or copies of the turbot MHC Class II B gene.The rate of non-synonymous substitution(d N) was 2.30 and 1.58 times higher than synonymous substitution(d S) in the peptide-binding regions(PBR) and non-PBR in whole families,respectively,which suggested balancing selection on exon 2 of the MHC Class II B gene in turbot.One allele,Scma-DBB1*02,was significantly more prevalent in survivor stock than in non-survivor stock(P=0.001).Therefore,this allele might be associated with resistance to bacteria.A second allele,Scma-DBB1*10,was significantly more prevalent in non-survivor stock(P=0.021),and is likely associated with susceptibility to bacteria.
基金supported by the National Natural Science Foundation of China(No.31201713,No.31270425 and No.31470442)
文摘The worldwide declines in amphibian populations have largely been caused by infectious fungi and bacteria. Given that vertebrate immunity against these extracellular pathogens is primarily functioned by the major histocompatibility complex(MHC) class Ⅱ molecules, the characterization and the evolution of amphibian MHC class Ⅱ genes have attracted increasing attention. The polymorphism of MHC class Ⅱ genes was found to be correlated with susceptibility to fungal pathogens in many amphibian species, suggesting the importance of studies on MHC class Ⅱ genes for amphibians. However, such studies on MHC class Ⅱ gene evolution have rarely been conducted on amphibians in China. In this study, we chose Omei treefrog(Rhacophorus omeimontis), which lived moist environments easy for breeding bacteria, to study the polymorphism of its MHC class Ⅱ genes and the underlying evolutionary mechanisms. We amplified the entire MHC class ⅡB exon 2 sequence in the R. omeimontis using newly designed primers. We detected 102 putative alleles in 146 individuals. The number of alleles per individual ranged from one to seven, indicating that there are at least four loci containing MHC class ⅡB genes in R. omeimontis. The allelic polymorphism estimated from the 102 alleles in R. omeimontis was not high compared to that estimated in other anuran species. No significant gene recombination was detected in the 102 MHC class ⅡB exon 2 sequences. In contrast, both gene duplication and balancing selection greatly contributed to the variability in MHC class ⅡB exon 2 sequences of R. omeimontis. This study lays the groundwork for the future researches to comprehensively analyze the evolution of amphibian MHC genes and to assess the role of MHC gene polymorphisms in resistance against extracellular pathogens for amphibians in China.