Background： Porcine reproductive and respiratory syndrome virus （PRRSV）, and particularly its highly pathogenic genotype （HP-PRRSV）, have caused massive economic losses to the global swine industry. Results： To ...Background： Porcine reproductive and respiratory syndrome virus （PRRSV）, and particularly its highly pathogenic genotype （HP-PRRSV）, have caused massive economic losses to the global swine industry. Results： To rapidly identify HP-PRRSV, we developed a direct reaL-time reverse transcription polymerase chain reaction method （dRT-PCR） that could detect the virus from serum specimen without the need of RNA purification Our dRT-PCR assay can be completed in 1.5 h from when a sample is received to obtaining a result. Additionally, the sensitivity of dRT-PCR matched that of conventional reverse transcription PCR （cRT-PCR） that used purified RNA The lowest detection limit of HP-PRRSV was 6.3 TCIDs0 using dRT-PCR. We applied dRT-PCR assay to 144 field samples and the results showed strong consistency with those obtained by cRT-PCR. Moreover, the dRT-PCR method was able to tolerate 5-20% （v/v） serum. Conclusions： Our dRT-PCR assay allows for easier, faster, more cost-effective and higher throughput detection of HP-PRRSV compared with cRT-PCR methods. To the best of our knowledge, this is the first report to describe a real-time RT-PCR assay capable of detecting PRRSV in crude serum samples without the requirement for purifying RNA. We believe our approach has a great potential for application to other RNA viruses.展开更多
[Objective] To isolate three strains of avian leukosis virus subgroup J（ALV -J） , and then amplify and sequence the gp85 gene. [ Method] Three strains of ALV- J were isolated from Hubei Province, which were identifi...[Objective] To isolate three strains of avian leukosis virus subgroup J（ALV -J） , and then amplify and sequence the gp85 gene. [ Method] Three strains of ALV- J were isolated from Hubei Province, which were identified by pathological anatomy, DF- 1 cell culture and RT- PCR. And then they were named HB1002, HB1003 and HB1009, respectively. [ Result] Test sequence analysis showed, the length of gp85 gene was 921 bp, consistent with expect result; the nucleotide homology between the three isolates was in 97.7% -99.7% ,the homology of amino acid was in 95.1% - 99%. The nucleotide homology between HPRS - 103 and the three isolates was in the 94.1% - 94.8% ; and the nucleotide homolo- gy between other ALV -J and the three isolates was in 87.6% -97.3%. The phylogenetic trees analysis showed that the homology of JS09GY6 vi- rus and the three isolates was nearest, in 95.2% -97.3%. [ Conclusion] In the test, the three strains of virus which were isolated were ALV- J.展开更多
Background:Salmonella pullorum is one of the most harmful pathogens to avian species.Magnolol and honokiol,natural compounds extracted from Magnolia officinalis,exerts anti-inflammatory,anti-oxidant and antibacterial ...Background:Salmonella pullorum is one of the most harmful pathogens to avian species.Magnolol and honokiol,natural compounds extracted from Magnolia officinalis,exerts anti-inflammatory,anti-oxidant and antibacterial activities.This study was conducted to evaluate the effects of dietary supplemental magnolol and honokiol in broilers infected with S.pullorum.A total of 360 one-day-old broilers were selected and randomly divided into four groups with six replicates:the negative control group(CTL),S.pullorum-infected group(SP),and the S.pulloruminfected group supplemented with 300 mg/kg honokiol(SPH)or magnolol(SPM).Results:The results showed that challenging with S.pullorum impaired growth performance in broilers,as indicated by the observed decreases in body weight(P<0.05)and average daily gains(P<0.05),along with increased spleen(P<0.01)and bursa of Fabricus weights(P<0.05),serum globulin contents,and the decreased intestine villus height and villus/crypt ratios(P<0.05).Notably,supplemental magnolol and honokiol attenuated these adverse changes,and the effects of magnolol were better than those of honokiol.Therefore,we performed RNA-Seq in ileum tissues and 16S rRNA gene sequencing of ileum bacteria.Our analysis revealed that magnolol increased the α-diversity(observed species,Chao1,ACE,and PD whole tree)and β-diversity of the ileum bacteria(P<0.05).In addition,magnolol supplementation increased the abundance of Lactobacillus(P<0.01)and decreased unidentified Cyanobacteria(P<0.05)both at d 14 and d 21.Further study confirmed that differentially expressed genes induced by magnolol and honokiol supplementation enriched in cytokine-cytokine receptor interactions,in the intestinal immune network for IgA production,and in the cell adhesion molecule pathways.Conclusions:Supplemental magnolol and honokiol alleviated S.pullorum-induced impairments in growth performance,and the effect of magnolol was better than that of honokiol,which could be partially due to magnolol’s ability to improve the intestinal microbial and mucosal barrier.展开更多
The roots of legume plant play a crucial role in nitrogen fixation. However, the transcriptomes of different cell types of legume root and their functions remain largely unknown. Here, we performed single-cell RNA seq...The roots of legume plant play a crucial role in nitrogen fixation. However, the transcriptomes of different cell types of legume root and their functions remain largely unknown. Here, we performed single-cell RNA sequencing and profiled more than 22,000 single cells from root tips of Lotus japonicus, a model species of legume.We identified seven clusters corresponding to seven major cell types, which were validated by in situ hybridization. Further analysis revealed regulatory programs including phytohormone and nodulation associated with specific cell types, and revealed conserved and diverged features for the cell types. Our results represent the first single-cell resolution transcriptome for legume root tips and a valuable resource for studying the developmental and physiological functions of various cell types in legumes.展开更多
The laying quail is a worldwide breed which exhibits high economic value. In our current study, the vas- oactive intestinal peptide receptor-1 （VIPR-1） was selected as the candidate gene for identifying traits of eg...The laying quail is a worldwide breed which exhibits high economic value. In our current study, the vas- oactive intestinal peptide receptor-1 （VIPR-1） was selected as the candidate gene for identifying traits of egg produc- tion. A single nucleotide polymorphism （SNP） detection was performed in 443 individual quails, including 196 quails from the H line, 202 quails from the L line, and 45 wild quails. The SNPs were genotyped using a polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP）. Two mutations （G373T, A313G） were detected in all the tested quail populations. The associated analysis showed that the SNP genotypes of the VIPR-1 gene were sig- nificantly linked with the egg weight of G373T and A313G in 398 quails. The quails with the genotype GG always exhibited the largest egg weight for the two mutations in the H and L lines. Linkage disequilibrium （LD） analysis in- dicated that G373T and A313G loci showed the weakest LD. Seven main diplotypes from the four main reconstructed haplotypes were observed, indicating a significant association of diplotypes with egg weight. Quails with the hlh2 （GGGT） diplotype always exhibited the smallest egg weight and largest egg number at 20 weeks of age. The overall results suggest that the alterations in quails may be linked with potential major loci or genes affecting reproductive traits.展开更多
Retinoid X receptor(RXR) α is a member of the nuclear hormone receptor superfamily that mediates the biological effects on several hormones,vitamins,and regulates lipid,glucose and energy metabolism.In this study,the...Retinoid X receptor(RXR) α is a member of the nuclear hormone receptor superfamily that mediates the biological effects on several hormones,vitamins,and regulates lipid,glucose and energy metabolism.In this study,the tissue expression profiles of the bovine RXRαgene and association analysis of single nucleotide polymorphisms(SNPs) with growth traits were carried out in 413 Chinese native cattle.The expression profile was analysed in ten Jiaxian cattle tissues by real-time PCR,and the results showed that RXRαgene was abundantly expressed in adipose tissue and spleen,moderately expressed in heart,liver,lung,kidney,muscle and testis.Meanwhile,three SNPs(T27919A,T28139C and G28142A) and five haplotypes were identified.Haplotype with TTG was dominant with frequency of 69.1%.Chi-square test showed all populations were in Hardy-Weinberg equilibrium(P>0.05) at the three sites except Jiaxian cattle at G28142A site and Qinchuan cattle at T27919A site.Statistical analysis of combined sites showed that the individuals with TTGA genotype had significantly higher heart girth than those with TAGG genotype(P<0.05) and the animals with AAGA genotype had higher body weight than those with TAGG genotype(P<0.05) in T27919A-G28142A site.Heart girth,abdominal circumference and body weight of individuals with TCAG genotype were exceedingly higher than those with TTGG(P<0.01),TTGA and TCGG(P<0.05) in T28139C-G28142A site.For T27919A-T28139C site,the individuals of TCTA and TCTT genotype had significantly higher heart girth and lower height at hip cross than those with TTTA(P<0.05),and the body weight of TCAA and TCTT genotype individuals was higher than those with TTTA(P<0.05).In conclusion,these results provided evidence that the polymorphisms of RXRαgene were associated with growth traits and might apply to Chinese indigenous yellow cattle breeding program as a possible candidate for marker-assisted selection(MAS).展开更多
Beef and mutton production has been aided by breeding to integrate allelic diversity for myostatin(MSTN),but a lack of diversity in the MSTN germplasm has limited similar advances in pig farming.Moreover,insurmountabl...Beef and mutton production has been aided by breeding to integrate allelic diversity for myostatin(MSTN),but a lack of diversity in the MSTN germplasm has limited similar advances in pig farming.Moreover,insurmountable challenges with congenital lameness and a dearth of data about the impacts of feed conversion,reproduction,and meat quality in MSTN-edited pigs have also currently blocked progress.Here,in a largest-to-date evaluation of multiple MSTN-edited pig populations,we demonstrated a practical alternative edit-site-based solution that overcomes the major production obstacle of hindlimb weakness.We also provide long-term and multidomain datasets for multiple breeds that illustrate how MSTN-editing can sustainably increase the yields of breed-specific lean meat and the levels of desirable lipids without deleteriously affecting feed-conversion rates or litter size.Apart from establishing a new benchmark for the data scale and quality of genome-edited animal production,our study specifically illustrates how gene-editing site selection profoundly impacts the phenotypic outcomes in diverse genetic back-grounds.展开更多
基金supported in part by the National Basic Research Program of China(973 Program,2012CB124701)National Natural Science Foundation of China No.81170047,81370151(to DG)+6 种基金Shenzhen overseas high-level talentsinnovation program No.YFZZ20111009(to DG)Shenzhen Nanshan Core Technology Program No.KC2013JSJS0020AShenzhen Municipal Basic Research Program No.JCYJ20130329120507746(to KK)Postdoctoral Science Foundation of China No.2013 M542203(to KK)Hubei Province Research and Development Project No.2011BBB080(to KY)Project supported by the Key Natural Science Foundation of Hubei Province,China No.2012FFA067(to YT)the Opening Subject of Hubei Key Laboratory of Animal Embryo and Molecular Breeding No.2012ZD156(to KY)
文摘Background： Porcine reproductive and respiratory syndrome virus （PRRSV）, and particularly its highly pathogenic genotype （HP-PRRSV）, have caused massive economic losses to the global swine industry. Results： To rapidly identify HP-PRRSV, we developed a direct reaL-time reverse transcription polymerase chain reaction method （dRT-PCR） that could detect the virus from serum specimen without the need of RNA purification Our dRT-PCR assay can be completed in 1.5 h from when a sample is received to obtaining a result. Additionally, the sensitivity of dRT-PCR matched that of conventional reverse transcription PCR （cRT-PCR） that used purified RNA The lowest detection limit of HP-PRRSV was 6.3 TCIDs0 using dRT-PCR. We applied dRT-PCR assay to 144 field samples and the results showed strong consistency with those obtained by cRT-PCR. Moreover, the dRT-PCR method was able to tolerate 5-20% （v/v） serum. Conclusions： Our dRT-PCR assay allows for easier, faster, more cost-effective and higher throughput detection of HP-PRRSV compared with cRT-PCR methods. To the best of our knowledge, this is the first report to describe a real-time RT-PCR assay capable of detecting PRRSV in crude serum samples without the requirement for purifying RNA. We believe our approach has a great potential for application to other RNA viruses.
基金Plan of Hubei Province Science and Technology Research and Development Project:the Research and Demonstration of Comprehensive Prevention and Control Technology on Poultry Avian Influenza and other Major Diseases YJN0065the Construction Fund of Modern Agriculture Industry Technology System CARS-42-G11
文摘[Objective] To isolate three strains of avian leukosis virus subgroup J（ALV -J） , and then amplify and sequence the gp85 gene. [ Method] Three strains of ALV- J were isolated from Hubei Province, which were identified by pathological anatomy, DF- 1 cell culture and RT- PCR. And then they were named HB1002, HB1003 and HB1009, respectively. [ Result] Test sequence analysis showed, the length of gp85 gene was 921 bp, consistent with expect result; the nucleotide homology between the three isolates was in 97.7% -99.7% ,the homology of amino acid was in 95.1% - 99%. The nucleotide homology between HPRS - 103 and the three isolates was in the 94.1% - 94.8% ; and the nucleotide homolo- gy between other ALV -J and the three isolates was in 87.6% -97.3%. The phylogenetic trees analysis showed that the homology of JS09GY6 vi- rus and the three isolates was nearest, in 95.2% -97.3%. [ Conclusion] In the test, the three strains of virus which were isolated were ALV- J.
基金supported by the project of Hubei innovation center of agricultural science and technology(grant number 2016-620-000-001-028)National Natural Science Foundation of China(31702309)the Youth Fund of Hubei Academy of Agricultural Sciences(2019NKYJJ03).
文摘Background:Salmonella pullorum is one of the most harmful pathogens to avian species.Magnolol and honokiol,natural compounds extracted from Magnolia officinalis,exerts anti-inflammatory,anti-oxidant and antibacterial activities.This study was conducted to evaluate the effects of dietary supplemental magnolol and honokiol in broilers infected with S.pullorum.A total of 360 one-day-old broilers were selected and randomly divided into four groups with six replicates:the negative control group(CTL),S.pullorum-infected group(SP),and the S.pulloruminfected group supplemented with 300 mg/kg honokiol(SPH)or magnolol(SPM).Results:The results showed that challenging with S.pullorum impaired growth performance in broilers,as indicated by the observed decreases in body weight(P<0.05)and average daily gains(P<0.05),along with increased spleen(P<0.01)and bursa of Fabricus weights(P<0.05),serum globulin contents,and the decreased intestine villus height and villus/crypt ratios(P<0.05).Notably,supplemental magnolol and honokiol attenuated these adverse changes,and the effects of magnolol were better than those of honokiol.Therefore,we performed RNA-Seq in ileum tissues and 16S rRNA gene sequencing of ileum bacteria.Our analysis revealed that magnolol increased the α-diversity(observed species,Chao1,ACE,and PD whole tree)and β-diversity of the ileum bacteria(P<0.05).In addition,magnolol supplementation increased the abundance of Lactobacillus(P<0.01)and decreased unidentified Cyanobacteria(P<0.05)both at d 14 and d 21.Further study confirmed that differentially expressed genes induced by magnolol and honokiol supplementation enriched in cytokine-cytokine receptor interactions,in the intestinal immune network for IgA production,and in the cell adhesion molecule pathways.Conclusions:Supplemental magnolol and honokiol alleviated S.pullorum-induced impairments in growth performance,and the effect of magnolol was better than that of honokiol,which could be partially due to magnolol’s ability to improve the intestinal microbial and mucosal barrier.
基金supported by a survey on Central China grassland forage germplasm resources (2017FY100604)National Natural Science Foundation of China (32001395)+1 种基金Chongqing Natural Science Foundation (stc2020jcyjmsxmX0626)The Agricultural Science and Technology Innovation Program (ASTIP) (to Z.L.)。
文摘The roots of legume plant play a crucial role in nitrogen fixation. However, the transcriptomes of different cell types of legume root and their functions remain largely unknown. Here, we performed single-cell RNA sequencing and profiled more than 22,000 single cells from root tips of Lotus japonicus, a model species of legume.We identified seven clusters corresponding to seven major cell types, which were validated by in situ hybridization. Further analysis revealed regulatory programs including phytohormone and nodulation associated with specific cell types, and revealed conserved and diverged features for the cell types. Our results represent the first single-cell resolution transcriptome for legume root tips and a valuable resource for studying the developmental and physiological functions of various cell types in legumes.
基金supported by the Open Project of Hubei Key Laboratory of Animal Embryo and Molecular Breeding(No.2015ZD146),China
文摘The laying quail is a worldwide breed which exhibits high economic value. In our current study, the vas- oactive intestinal peptide receptor-1 （VIPR-1） was selected as the candidate gene for identifying traits of egg produc- tion. A single nucleotide polymorphism （SNP） detection was performed in 443 individual quails, including 196 quails from the H line, 202 quails from the L line, and 45 wild quails. The SNPs were genotyped using a polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP）. Two mutations （G373T, A313G） were detected in all the tested quail populations. The associated analysis showed that the SNP genotypes of the VIPR-1 gene were sig- nificantly linked with the egg weight of G373T and A313G in 398 quails. The quails with the genotype GG always exhibited the largest egg weight for the two mutations in the H and L lines. Linkage disequilibrium （LD） analysis in- dicated that G373T and A313G loci showed the weakest LD. Seven main diplotypes from the four main reconstructed haplotypes were observed, indicating a significant association of diplotypes with egg weight. Quails with the hlh2 （GGGT） diplotype always exhibited the smallest egg weight and largest egg number at 20 weeks of age. The overall results suggest that the alterations in quails may be linked with potential major loci or genes affecting reproductive traits.
基金supported by The National Natural Science Foundation of China (31172193)the Program for Science & Technology Innovation Talents in Universities of Henan Province (2012HASTIT027)+2 种基金the Natural Science Research Project of Department of Education of Henan (2011A180027)the Hubei Key Laboratory of Animal Embryo Engineering and Molecular Breeding,Institute of Animal Husbandry and Veterinary,Hubei Academy of Agriculture Science (2012ZD119)the twelfth "Five-Year" National Science and Technology Support Project (2011BAD28B04)
文摘Retinoid X receptor(RXR) α is a member of the nuclear hormone receptor superfamily that mediates the biological effects on several hormones,vitamins,and regulates lipid,glucose and energy metabolism.In this study,the tissue expression profiles of the bovine RXRαgene and association analysis of single nucleotide polymorphisms(SNPs) with growth traits were carried out in 413 Chinese native cattle.The expression profile was analysed in ten Jiaxian cattle tissues by real-time PCR,and the results showed that RXRαgene was abundantly expressed in adipose tissue and spleen,moderately expressed in heart,liver,lung,kidney,muscle and testis.Meanwhile,three SNPs(T27919A,T28139C and G28142A) and five haplotypes were identified.Haplotype with TTG was dominant with frequency of 69.1%.Chi-square test showed all populations were in Hardy-Weinberg equilibrium(P>0.05) at the three sites except Jiaxian cattle at G28142A site and Qinchuan cattle at T27919A site.Statistical analysis of combined sites showed that the individuals with TTGA genotype had significantly higher heart girth than those with TAGG genotype(P<0.05) and the animals with AAGA genotype had higher body weight than those with TAGG genotype(P<0.05) in T27919A-G28142A site.Heart girth,abdominal circumference and body weight of individuals with TCAG genotype were exceedingly higher than those with TTGG(P<0.01),TTGA and TCGG(P<0.05) in T28139C-G28142A site.For T27919A-T28139C site,the individuals of TCTA and TCTT genotype had significantly higher heart girth and lower height at hip cross than those with TTTA(P<0.05),and the body weight of TCAA and TCTT genotype individuals was higher than those with TTTA(P<0.05).In conclusion,these results provided evidence that the polymorphisms of RXRαgene were associated with growth traits and might apply to Chinese indigenous yellow cattle breeding program as a possible candidate for marker-assisted selection(MAS).
基金the National Major Transgenic Breeding Project(2008ZX08006-003,2011ZX08006-003,2013ZX08006-003,2014ZX08006-003,and 2016ZX08006-001)the Na-tional Key Basic Research Development Plan(2015CB943100)+1 种基金the Key Projects of the National Natural Science Foundation of China(30830080 and 31330074)China Postdoctoral Foundation Project(2018M631648)。
文摘Beef and mutton production has been aided by breeding to integrate allelic diversity for myostatin(MSTN),but a lack of diversity in the MSTN germplasm has limited similar advances in pig farming.Moreover,insurmountable challenges with congenital lameness and a dearth of data about the impacts of feed conversion,reproduction,and meat quality in MSTN-edited pigs have also currently blocked progress.Here,in a largest-to-date evaluation of multiple MSTN-edited pig populations,we demonstrated a practical alternative edit-site-based solution that overcomes the major production obstacle of hindlimb weakness.We also provide long-term and multidomain datasets for multiple breeds that illustrate how MSTN-editing can sustainably increase the yields of breed-specific lean meat and the levels of desirable lipids without deleteriously affecting feed-conversion rates or litter size.Apart from establishing a new benchmark for the data scale and quality of genome-edited animal production,our study specifically illustrates how gene-editing site selection profoundly impacts the phenotypic outcomes in diverse genetic back-grounds.
