Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38437
Yukai Yang, Yangjie Xie, Mingyi Cai, Linjiang Jian, Qingkai Chen, Bao Lou, Zhiyong Wang
In order to establish a procedure to induce artificial gynogenesis for genetic improvement and sex control studies in Nibea albiflora,ultraviolet irradiation was used to genetically inactivate the sperm and cold shock was used to induce the chromosome duplication of the eggs.Relative DNA content in newly-hatched larvae of candidate haploid gynogens,candidate diploid gynogens and the control groups from normal fetilization were mensurated,and paternity test was made using SSR markers for confirmation of gynogenesis.The gynogenetic diploid was successfully induced by activating egg development with UV-irradiated sperm combined with cold shock to prevent extrusion of the second polar body.UV irradiation time range was preliminarily determined by an insemination trial with the sperms irradiated by different doses of UV.The fertilized egg hatchability exhibited typical Hertwig effect when sperm irradiation time was 0-100 seconds at a UV intensity of 3 800 μW/(cm2·s).All newly-hatched larvae exhibited typical haploid syndrome when the UV irradiation time was over 60 seconds.In addition,we designed an orthogonal experiment to further optimize UV irradiation time,cold shock initiation time and cold shock duration time.The orthogonal experiment results showed that the hatching rate was highest(16%)in the following conditions:sperm irradiation time was 60 seconds,cold shock temperature was 3-4 ℃,cold shock initiation time was 2 minutes after fertilization and cold shock duration time was 10 minutes.The larvae of the optimal group have the same morphological characteristics and cellular DNA content with normal diploids.A further paternity test with five compatible SSR primers of Larimichthys crocea proved that the larvae in the optimal group were all gynogenetic diploids without paternal specific alleles.
{"title":"Induction and identification of gynogenesis in Nibea albiflora","authors":"Yukai Yang, Yangjie Xie, Mingyi Cai, Linjiang Jian, Qingkai Chen, Bao Lou, Zhiyong Wang","doi":"10.3724/SP.J.1231.2013.38437","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38437","url":null,"abstract":"In order to establish a procedure to induce artificial gynogenesis for genetic improvement and sex control studies in Nibea albiflora,ultraviolet irradiation was used to genetically inactivate the sperm and cold shock was used to induce the chromosome duplication of the eggs.Relative DNA content in newly-hatched larvae of candidate haploid gynogens,candidate diploid gynogens and the control groups from normal fetilization were mensurated,and paternity test was made using SSR markers for confirmation of gynogenesis.The gynogenetic diploid was successfully induced by activating egg development with UV-irradiated sperm combined with cold shock to prevent extrusion of the second polar body.UV irradiation time range was preliminarily determined by an insemination trial with the sperms irradiated by different doses of UV.The fertilized egg hatchability exhibited typical Hertwig effect when sperm irradiation time was 0-100 seconds at a UV intensity of 3 800 μW/(cm2·s).All newly-hatched larvae exhibited typical haploid syndrome when the UV irradiation time was over 60 seconds.In addition,we designed an orthogonal experiment to further optimize UV irradiation time,cold shock initiation time and cold shock duration time.The orthogonal experiment results showed that the hatching rate was highest(16%)in the following conditions:sperm irradiation time was 60 seconds,cold shock temperature was 3-4 ℃,cold shock initiation time was 2 minutes after fertilization and cold shock duration time was 10 minutes.The larvae of the optimal group have the same morphological characteristics and cellular DNA content with normal diploids.A further paternity test with five compatible SSR primers of Larimichthys crocea proved that the larvae in the optimal group were all gynogenetic diploids without paternal specific alleles.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"1297-1303"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69967144","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38438
Jian Linjiang, Yang Yukai, Liu Xiande, Chen Qing-kai, W. Zhiyong
To understand the feasibility of cross breeding between Larimichthys crocea ♀ and Nibea miichthioides ♂,two hybrid families were built(LN1 and LN2).Ploidy of F1 hybrids was identified.Moreover,ten microsatellite loci were used for genetic relationship analysis between F1 hybrids and their parents.The result showed that L.crocea(♀)and N.miichthioides(♂)can be successfully crossed and hybrid offspring of normal survival were produced.Survival rate of hybrid offspring at the age of 45 days post hatch reached about 30%.However,fertilization rate(29.0%,32.6%)and hatching rate(75.0%,76.7%)of two hybrid families were significantly lower than those in large yellow croaker families(P0.05).Appearance of juvenile hybrid fish is visibly different from those of large yellow croaker,which has characteristics of both parents:slender body,sharp blunt head and their body sides are covered with black-brown spots.The results of DNA relative content investigation and genetic relationship analysis demonstrated that more than 90% of the hybrid offspring were hybrid diploid,in addition,a small number of those were hybrid triploid and gynogenesis diploid.Till the age of 45 days,growth rate of both hybrid diploid and hybrid triploid was slower than that of large yellow croaker.These findings will provide reference for exploitation and management of F1 hybrids of L.crocea(♀)and N.miichthioides(♂).
{"title":"The cross breeding and genetic analysis of hybrids of Larimichthys crocea(♀)and Nibea miichthioides(♂)","authors":"Jian Linjiang, Yang Yukai, Liu Xiande, Chen Qing-kai, W. Zhiyong","doi":"10.3724/SP.J.1231.2013.38438","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38438","url":null,"abstract":"To understand the feasibility of cross breeding between Larimichthys crocea ♀ and Nibea miichthioides ♂,two hybrid families were built(LN1 and LN2).Ploidy of F1 hybrids was identified.Moreover,ten microsatellite loci were used for genetic relationship analysis between F1 hybrids and their parents.The result showed that L.crocea(♀)and N.miichthioides(♂)can be successfully crossed and hybrid offspring of normal survival were produced.Survival rate of hybrid offspring at the age of 45 days post hatch reached about 30%.However,fertilization rate(29.0%,32.6%)and hatching rate(75.0%,76.7%)of two hybrid families were significantly lower than those in large yellow croaker families(P0.05).Appearance of juvenile hybrid fish is visibly different from those of large yellow croaker,which has characteristics of both parents:slender body,sharp blunt head and their body sides are covered with black-brown spots.The results of DNA relative content investigation and genetic relationship analysis demonstrated that more than 90% of the hybrid offspring were hybrid diploid,in addition,a small number of those were hybrid triploid and gynogenesis diploid.Till the age of 45 days,growth rate of both hybrid diploid and hybrid triploid was slower than that of large yellow croaker.These findings will provide reference for exploitation and management of F1 hybrids of L.crocea(♀)and N.miichthioides(♂).","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"801-808"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69967251","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38465
Chengsong Zhang, Fuhua Li, J. Xiang
In order to ascertain the optimal and measurable indices used in artificial breeding of Exopalaemon carinicauda,the effects of morphometric attributes on body weight of E.carinicauda were analyzed by path analysis.Besides body weight,ten morphometric attributes,including the body length,total length,carapace length,carapace width,carapace height,first abdominal segment height,first abdominal segment width,abdomen length,number of upper frontal eminence spin and number of lower frontal eminence spin were recorded for 177 four-month-old E.carinicauda.The correlation coefficients among all attributes were calculated,and the path coefficients and determination coefficients were calculated by taking the body weight as a dependent variable,and the other ten morphometric attributes as independent variables in path analysis.The data indicated that the path coefficients of four morphometric attributes(body length,carapace width,carapace height,first abdominal segment width)to body weight reached significant difference(P0.01)level.Among above four significant factors for body weight,body length showed the greatest effect on body weight with the direct effect efficiency of 0.468* *.The direct effects of total length,carapace length,first abdominal segment height,abdomen length,number of upper frontal eminence spin and number of lower frontal eminence spin on body weight were quite low,therefore,they were regarded as insignificant factors influencing the body weight.The stepwise regression analysis established a multiple regression equation on the effects of body length,carapace width,carapace height,first abdominal segment width on body weight with partial regression coefficients of-3.277,0.470,0.980,0.528 and 1.942 separately.Judging from the high multiple-correlation coefficient(R2=0.976),the main attributes(body length,carapace width,carapace height,first abdominal segment width)determining the body weight have been selected.
{"title":"Path analysis of effects of morphometric attributes on body weight of Exopalaemon carinicauda","authors":"Chengsong Zhang, Fuhua Li, J. Xiang","doi":"10.3724/SP.J.1231.2013.38465","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38465","url":null,"abstract":"In order to ascertain the optimal and measurable indices used in artificial breeding of Exopalaemon carinicauda,the effects of morphometric attributes on body weight of E.carinicauda were analyzed by path analysis.Besides body weight,ten morphometric attributes,including the body length,total length,carapace length,carapace width,carapace height,first abdominal segment height,first abdominal segment width,abdomen length,number of upper frontal eminence spin and number of lower frontal eminence spin were recorded for 177 four-month-old E.carinicauda.The correlation coefficients among all attributes were calculated,and the path coefficients and determination coefficients were calculated by taking the body weight as a dependent variable,and the other ten morphometric attributes as independent variables in path analysis.The data indicated that the path coefficients of four morphometric attributes(body length,carapace width,carapace height,first abdominal segment width)to body weight reached significant difference(P0.01)level.Among above four significant factors for body weight,body length showed the greatest effect on body weight with the direct effect efficiency of 0.468* *.The direct effects of total length,carapace length,first abdominal segment height,abdomen length,number of upper frontal eminence spin and number of lower frontal eminence spin on body weight were quite low,therefore,they were regarded as insignificant factors influencing the body weight.The stepwise regression analysis established a multiple regression equation on the effects of body length,carapace width,carapace height,first abdominal segment width on body weight with partial regression coefficients of-3.277,0.470,0.980,0.528 and 1.942 separately.Judging from the high multiple-correlation coefficient(R2=0.976),the main attributes(body length,carapace width,carapace height,first abdominal segment width)determining the body weight have been selected.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"809"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69967289","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38463
Yitao Huang, Xiuhong Cai, Ziping Zhang, Guodong Wang, Z. Zou, Shuhong Wang, Yilei Wang
Allograft inflammatory factor-1(AIF-1)is a kind of calcium-binding protein with EF-hand domain.AIF-1 mRNA is robustly induced by IFN-γ in murine macrophages.It is involved in transplant rejection,immune inflammatory reaction,non-inflammatory injury and so on.In this study,a molluscan AIF-1 gene,HdAIF-1,was cloned for the first time from Haliotis diversicolor.Its full-length cDNA sequence is 942 bp,with a 456 bp open reading frame encoding a protein of 151 aa.Quantitative real-time PCR results indicated that HdAIF-1 could be detected in all examined tissues,with the highest level in hemolymph and gill.Under thermal stress,HdAIF-1 was up-regulated in gill significantly at temperature-rise period and achieved the highest level when up to 31 ℃.However,the expression level of HdAIF-1 in hemolymph and hepatopancreas did not show significant difference between thermal group and control group from the first phase to the fourth,and its expression level was up-regulated significantly at 96 h in these two tissues.Under hypoxia stress,the expression level of HdAIF-1 in hemolymph showed no significant difference between control and exposed groups.However,it was down-regulated at 24 h and up-regulated at 192 h remarkably in gill.After Vibrio parahaemolyticus challenge,the HdAIF-1 expression level achieved the highest level at 3 h in hemolymph,reduced to the control level at 12 h,and then up-regulated significantly at both 24 h and 48 h.These results suggest that HdAIF-1 may play important roles as an immune factor under different stresses.
{"title":"Molecular cloning and expression of allograft inflammatory factor 1 in Haliotis diversicolor under stresses","authors":"Yitao Huang, Xiuhong Cai, Ziping Zhang, Guodong Wang, Z. Zou, Shuhong Wang, Yilei Wang","doi":"10.3724/SP.J.1231.2013.38463","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38463","url":null,"abstract":"Allograft inflammatory factor-1(AIF-1)is a kind of calcium-binding protein with EF-hand domain.AIF-1 mRNA is robustly induced by IFN-γ in murine macrophages.It is involved in transplant rejection,immune inflammatory reaction,non-inflammatory injury and so on.In this study,a molluscan AIF-1 gene,HdAIF-1,was cloned for the first time from Haliotis diversicolor.Its full-length cDNA sequence is 942 bp,with a 456 bp open reading frame encoding a protein of 151 aa.Quantitative real-time PCR results indicated that HdAIF-1 could be detected in all examined tissues,with the highest level in hemolymph and gill.Under thermal stress,HdAIF-1 was up-regulated in gill significantly at temperature-rise period and achieved the highest level when up to 31 ℃.However,the expression level of HdAIF-1 in hemolymph and hepatopancreas did not show significant difference between thermal group and control group from the first phase to the fourth,and its expression level was up-regulated significantly at 96 h in these two tissues.Under hypoxia stress,the expression level of HdAIF-1 in hemolymph showed no significant difference between control and exposed groups.However,it was down-regulated at 24 h and up-regulated at 192 h remarkably in gill.After Vibrio parahaemolyticus challenge,the HdAIF-1 expression level achieved the highest level at 3 h in hemolymph,reduced to the control level at 12 h,and then up-regulated significantly at both 24 h and 48 h.These results suggest that HdAIF-1 may play important roles as an immune factor under different stresses.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"830"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69967603","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38490
L. Liu, Qianghua Xu, Xin-jun Chen, Jie Yan, Wei Yu, Cong-rong Wang
The worldwide climate and marine environmental changes have significant impacts on the distributions and fishery resources abundance fluctuations of Dosidicus gigas in the East Pacific Ocean. It is important to investigate genetic variance and population genetic structure of D. gigas in order to exploit and manage this squid scientifically. In this study,the genetic diversity and the genetic differentiation betw een tw o geographical populations of D. gigas w ere analyzed based on tw o mitochondrial DNA molecular markers. The results show ed that the number of haplotype,haplotype diversities,nucleotide diversities and average nucleotide differences w ere 7,( 0. 397 ± 0. 079),( 0. 001 09 ± 0. 000 96) and 0. 600 respectively in all Cytb sequences of tw o geographical populations,and haplotype diversity in population off the coast of Peru w as only( 0. 282 ± 0. 101) w hich w as significantly low er than that of equatorial w aters. On the other hand,the number of haplotype,haplotype diversities,nucleotide diversities and average nucleotide differences in all COⅠ sequences w ere 17,( 0. 787 ± 0. 051),( 0. 002 90 ± 0. 001 38) and 1. 802 respectively. The pairw ise fixation index F st and median netw ork revealed lack of genetic structure betw een populations of equatorial w aters and off the coast of Peru( Cytb: F st= 0. 055 23,P 0. 05; CO Ⅰ: F st= 0. 005 91,P 0. 05),and this might be caused by equational oceanographic current. The minimum spanning netw ork tree,mismatch distribution analysis and neutrality tests suggested that D. gigas experienced a recent population expansion,possibly 138 900 to 167 900 years ago.
全球气候和海洋环境变化对东太平洋大田螺的分布和渔业资源丰度波动有显著影响。研究巨乌贼的遗传变异和种群遗传结构，对科学开发和管理巨乌贼具有重要意义。本研究利用线粒体DNA分子标记分析了两个地理居群间的遗传多样性和遗传分化。结果表明，单倍型数量、单倍型多样性、核苷酸多样性和平均核苷酸差异分别为7、0。397±0。079),(0。001 09±0。000 96)和0。在2个地理种群的所有Cytb序列中，单倍型多样性分别为600个，秘鲁沿海种群的单倍型多样性仅为(0。282±0。101) w显著低于赤道w水域。另一方面，所有COⅠ序列的单倍型数量、单倍型多样性、核苷酸多样性和平均核苷酸差异w分别为17，(0。787±0。051),(0。002 90±0。001 38)和1。分别为802。配对固定指数F - 1和中位网络显示赤道w水域和秘鲁海岸种群之间缺乏遗传结构(Cytb: F - 1 = 0)。055 23, p 0。05;COⅠ:F st= 0。[05] 91, p . 0。05)，这可能是由相等的海洋洋流引起的。最小跨越网络树、错配分布分析和中性测试表明，巨齿龙经历了最近的种群扩张，可能在13.89万年至16.79万年之间。
{"title":"Comparative analysis of genetic diversity of Dosidicus gigas from eastern Pacific Ocean based on molecular markers of mitochondrial DNA","authors":"L. Liu, Qianghua Xu, Xin-jun Chen, Jie Yan, Wei Yu, Cong-rong Wang","doi":"10.3724/SP.J.1231.2013.38490","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38490","url":null,"abstract":"The worldwide climate and marine environmental changes have significant impacts on the distributions and fishery resources abundance fluctuations of Dosidicus gigas in the East Pacific Ocean. It is important to investigate genetic variance and population genetic structure of D. gigas in order to exploit and manage this squid scientifically. In this study,the genetic diversity and the genetic differentiation betw een tw o geographical populations of D. gigas w ere analyzed based on tw o mitochondrial DNA molecular markers. The results show ed that the number of haplotype,haplotype diversities,nucleotide diversities and average nucleotide differences w ere 7,( 0. 397 ± 0. 079),( 0. 001 09 ± 0. 000 96) and 0. 600 respectively in all Cytb sequences of tw o geographical populations,and haplotype diversity in population off the coast of Peru w as only( 0. 282 ± 0. 101) w hich w as significantly low er than that of equatorial w aters. On the other hand,the number of haplotype,haplotype diversities,nucleotide diversities and average nucleotide differences in all COⅠ sequences w ere 17,( 0. 787 ± 0. 051),( 0. 002 90 ± 0. 001 38) and 1. 802 respectively. The pairw ise fixation index F st and median netw ork revealed lack of genetic structure betw een populations of equatorial w aters and off the coast of Peru( Cytb: F st= 0. 055 23,P 0. 05; CO Ⅰ: F st= 0. 005 91,P 0. 05),and this might be caused by equational oceanographic current. The minimum spanning netw ork tree,mismatch distribution analysis and neutrality tests suggested that D. gigas experienced a recent population expansion,possibly 138 900 to 167 900 years ago.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"91 1","pages":"1618"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69967882","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38528
Ying-jiao Liang, R. Guan, Wen-shu Huang
The acetic acid extract of the spleen from Japanese eel,Anguilla japonica,was separated by tangential flow ultrafiltration with 10 and 3 ku fibre columns,and the two protein samples of the molecular weight(Mw)10 ku and the Mw3 ku were purified by using the cationic exchange liquid chromatography together with reverse-phase high-performance liquid chromatography(RP-HPLC).The results showed that three compositions(the protein samples of Mw10 ku,Mw10 ku and Mw3 ku)were separated from the acid extract with two fibre columns.The samples of Mw10 ku and Mw3 ku exhibited stronger antibacterial activity than the protein sample of the Mw10 ku.The protein sample with the Mw10 ku was purified by the cationic exchange liquid chromatography and using pH 9 buffer solution in RP-HPLC,and we obtained an antibacterial peptide with the molecular weight of 1 391.82 u when determined by MALDI-TOF MS.The pH 9 and the pH 2 buffer solutions were applied in RP-HPLC of the factions obtained from cationic exchange liquid chromatography of the protein sample with the Mw3 ku,and we also obtained an antibacterial peptide with the molecular weight of 839.19 u when determined by MALDI-TOF MS.This study suggests that the main antibacterial components in the spleen of A.japonica are the proteins of low molecular weight.
{"title":"Isolation of antibacterial peptides with low molecular weight from the spleen of Anguilla japonica","authors":"Ying-jiao Liang, R. Guan, Wen-shu Huang","doi":"10.3724/SP.J.1231.2013.38528","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38528","url":null,"abstract":"The acetic acid extract of the spleen from Japanese eel,Anguilla japonica,was separated by tangential flow ultrafiltration with 10 and 3 ku fibre columns,and the two protein samples of the molecular weight(Mw)10 ku and the Mw3 ku were purified by using the cationic exchange liquid chromatography together with reverse-phase high-performance liquid chromatography(RP-HPLC).The results showed that three compositions(the protein samples of Mw10 ku,Mw10 ku and Mw3 ku)were separated from the acid extract with two fibre columns.The samples of Mw10 ku and Mw3 ku exhibited stronger antibacterial activity than the protein sample of the Mw10 ku.The protein sample with the Mw10 ku was purified by the cationic exchange liquid chromatography and using pH 9 buffer solution in RP-HPLC,and we obtained an antibacterial peptide with the molecular weight of 1 391.82 u when determined by MALDI-TOF MS.The pH 9 and the pH 2 buffer solutions were applied in RP-HPLC of the factions obtained from cationic exchange liquid chromatography of the protein sample with the Mw3 ku,and we also obtained an antibacterial peptide with the molecular weight of 839.19 u when determined by MALDI-TOF MS.This study suggests that the main antibacterial components in the spleen of A.japonica are the proteins of low molecular weight.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"1236"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69968008","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38496
Xi Xie, Dongfa Zhu, Xiaoyu Cui, Jie Tang, Xier Qiu
To study the regulatory role of farnesoic acid O-methyl transferase(FAMeT)in molting process of crustaceans,the full-length FAMeT cDNA(GenBank accession number:KC192659)of Portunus trituberculatus is cloned by using reverse transcript PCR(RT-PCR)and rapid-amplification of cDNA ends(RACE).FAMeT cDNA contains a 201 bp 5′-untranslated region(5′-UTR),a 318 bp 3′-untranslated region(3′-UTR)and a 825 bp opening reading frame,which encodes 274 amino acid residues.Alignment of deduced amino acid sequence with FAMeT amino acid sequences of other crustaceans revealed that it shares the highest identity with P.pelagicus among the identities ranging from 75% to 97%.The amino acid residues consist of two copies of CF(CPAMD8/FAMeT)domain,which are the hallmark domain of FAMeT and are present in all crustacean FAMeTs.Quantitative real-time PCR(qRT-PCR)was used to quantify the relative expression level of FAMeT in different tissues and at molting stages in P.trituberculatus.The results showed that FAMeT was expressed in various tissues.The mRNA level of FAMeT was the highest in taoracic ganglia,followed by gill and mandibular organ(MO).During the molting process,the expression of FAMeT in MO increased to the maximum at D1 stage,then gradually decreased to the minimum at D4 stage.The results suggest that FAMeT plays an important role in molting regulation in P.trituberculatus.
{"title":"Cloning and expression analysis of farnesoic acid O-methyl transferase(FAMeT)gene during molting in Portunus trituberculatus","authors":"Xi Xie, Dongfa Zhu, Xiaoyu Cui, Jie Tang, Xier Qiu","doi":"10.3724/SP.J.1231.2013.38496","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38496","url":null,"abstract":"To study the regulatory role of farnesoic acid O-methyl transferase(FAMeT)in molting process of crustaceans,the full-length FAMeT cDNA(GenBank accession number:KC192659)of Portunus trituberculatus is cloned by using reverse transcript PCR(RT-PCR)and rapid-amplification of cDNA ends(RACE).FAMeT cDNA contains a 201 bp 5′-untranslated region(5′-UTR),a 318 bp 3′-untranslated region(3′-UTR)and a 825 bp opening reading frame,which encodes 274 amino acid residues.Alignment of deduced amino acid sequence with FAMeT amino acid sequences of other crustaceans revealed that it shares the highest identity with P.pelagicus among the identities ranging from 75% to 97%.The amino acid residues consist of two copies of CF(CPAMD8/FAMeT)domain,which are the hallmark domain of FAMeT and are present in all crustacean FAMeTs.Quantitative real-time PCR(qRT-PCR)was used to quantify the relative expression level of FAMeT in different tissues and at molting stages in P.trituberculatus.The results showed that FAMeT was expressed in various tissues.The mRNA level of FAMeT was the highest in taoracic ganglia,followed by gill and mandibular organ(MO).During the molting process,the expression of FAMeT in MO increased to the maximum at D1 stage,then gradually decreased to the minimum at D4 stage.The results suggest that FAMeT plays an important role in molting regulation in P.trituberculatus.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"994"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69968017","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38501
Zhongxiu Liang, Li Jian, Zhijun Tan, Jitao Li, Junping Liang, Hongxing Ge
This study probes into effects of the toxic dinoflagellate Alexandrium tamarense(ATHK),a producer of paralytic shellfish poison,on antioxidant system of Chinese shrimp Fenneropenaeus chinensis,an important mariculture species in China.The shrimp were intramuscularly injected the crude toxin extracted from A.tamarense cells.The dose injection was carried out only one time during the experiment,using extracted solution from 1.4×103 algae cells.Superoxide dismutase(SOD)activity,glutathione-S-transferase(GST)activity and malondialdehyde(MDA)content were analyzed in hepatopancreas and gill at 1,3,6,12,24 and 48 h after stress.SOD activity and GST activity in hepatopancreas and gill increased within 6 h.However,they were inhibited in gill at 12 h and 48 h after stress.MDA content in hepatopancreas had no significant change except at 1 h after stress,but increased in gill with time prolonging.The results indicated that the crude toxin extracted from A.tamarense cells could cause lipid peroxidation in gill of F.chinensis by induction of MDA increase and SOD and GST inactivation.
{"title":"Effects of the toxic dinoflagellate Alexandrium tamarense on MDA,SOD and GST in hepatopancreas and gill of Fenneropenaeus chinensis","authors":"Zhongxiu Liang, Li Jian, Zhijun Tan, Jitao Li, Junping Liang, Hongxing Ge","doi":"10.3724/SP.J.1231.2013.38501","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38501","url":null,"abstract":"This study probes into effects of the toxic dinoflagellate Alexandrium tamarense(ATHK),a producer of paralytic shellfish poison,on antioxidant system of Chinese shrimp Fenneropenaeus chinensis,an important mariculture species in China.The shrimp were intramuscularly injected the crude toxin extracted from A.tamarense cells.The dose injection was carried out only one time during the experiment,using extracted solution from 1.4×103 algae cells.Superoxide dismutase(SOD)activity,glutathione-S-transferase(GST)activity and malondialdehyde(MDA)content were analyzed in hepatopancreas and gill at 1,3,6,12,24 and 48 h after stress.SOD activity and GST activity in hepatopancreas and gill increased within 6 h.However,they were inhibited in gill at 12 h and 48 h after stress.MDA content in hepatopancreas had no significant change except at 1 h after stress,but increased in gill with time prolonging.The results indicated that the crude toxin extracted from A.tamarense cells could cause lipid peroxidation in gill of F.chinensis by induction of MDA increase and SOD and GST inactivation.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"8 1","pages":"1192"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69968123","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38563
Jiang Qichen, Gui Shuyu, Zhang Wenyi, Z. Chengxiang, Feng Xiaoqing, Tan HongYue, Huang Wenting, Yang Jiaxin, Li Feng
The acute effects of ammonia exposure on selected immunological and metabolic parameters in juvenile red claw crayfish,Cherax quadricarinatus as well as the post-exposure recovery were studied by using Real-time PCR technique and biochemical assays.The crayfish were first exposed to ammonia for 3 d,then half of them were sampled and the other half were moved into aerated tap water for a 7 d post-exposure recovery.The results showed that a 3 d ammonia exposure had significant impact on the activities of ACP,AKP and SOD in the muscle of juvenile C.quadricarinatus.As ammonia concentration increased,the activities of these enzymes significantly decreased.At the highest concentration(16 mg/L),they decreased to 76%,68% and 62% of the control,respectively.The mRNA expression of mitochondrial manganese superoxide dismutase(mMnSOD),cytosolic MnSOD(cMnSOD),extracellular copper/zinc SOD(exCu/ZnSOD)also showed a decreasing trend following increased ammonia concentration.In contrast,the mRNA expression of catalase(CAT),glutathione peroxidase(GPX)and glutathione transferase(GST)in muscle remained unchanged compared to the control.Dissolved protein and triglycerides in hepatopancreas significantly decreased as ammonia concentration increased.At the highest concentration,they decreased to 72% and 59% of the control.AST showed a noble elevation which was 134% of the control in 12 mg/L group.After a 7 d post-exposure recovery,ACP and AKP activities returned to the normal level in ammonia treatments.On the contrary,the activities of SOD and GPX in all ammonia treatments were significantly higher than the control.The expression of antioxidant genes was higher than the control in all ammonia treatments.Metabolic parameters showed no differences between all groups.The results indicate that high levels of ammonia might suppress activities and gene expression patterns of immune-related enzymes,leading to a loss of defence mechanism.The crayfish which had been exposed to ammonia for 3 d could not fully recover after a 7 d post-exposure recovery and oxidative stress still existed in the muscle in C.quadricarinatus.
{"title":"Acute effects of ammonia exposure on selected immunological and metabolic parameters in juvenile red claw crayfish(Cherax quadricarinatus)and the post-exposure recovery","authors":"Jiang Qichen, Gui Shuyu, Zhang Wenyi, Z. Chengxiang, Feng Xiaoqing, Tan HongYue, Huang Wenting, Yang Jiaxin, Li Feng","doi":"10.3724/SP.J.1231.2013.38563","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38563","url":null,"abstract":"The acute effects of ammonia exposure on selected immunological and metabolic parameters in juvenile red claw crayfish,Cherax quadricarinatus as well as the post-exposure recovery were studied by using Real-time PCR technique and biochemical assays.The crayfish were first exposed to ammonia for 3 d,then half of them were sampled and the other half were moved into aerated tap water for a 7 d post-exposure recovery.The results showed that a 3 d ammonia exposure had significant impact on the activities of ACP,AKP and SOD in the muscle of juvenile C.quadricarinatus.As ammonia concentration increased,the activities of these enzymes significantly decreased.At the highest concentration(16 mg/L),they decreased to 76%,68% and 62% of the control,respectively.The mRNA expression of mitochondrial manganese superoxide dismutase(mMnSOD),cytosolic MnSOD(cMnSOD),extracellular copper/zinc SOD(exCu/ZnSOD)also showed a decreasing trend following increased ammonia concentration.In contrast,the mRNA expression of catalase(CAT),glutathione peroxidase(GPX)and glutathione transferase(GST)in muscle remained unchanged compared to the control.Dissolved protein and triglycerides in hepatopancreas significantly decreased as ammonia concentration increased.At the highest concentration,they decreased to 72% and 59% of the control.AST showed a noble elevation which was 134% of the control in 12 mg/L group.After a 7 d post-exposure recovery,ACP and AKP activities returned to the normal level in ammonia treatments.On the contrary,the activities of SOD and GPX in all ammonia treatments were significantly higher than the control.The expression of antioxidant genes was higher than the control in all ammonia treatments.Metabolic parameters showed no differences between all groups.The results indicate that high levels of ammonia might suppress activities and gene expression patterns of immune-related enzymes,leading to a loss of defence mechanism.The crayfish which had been exposed to ammonia for 3 d could not fully recover after a 7 d post-exposure recovery and oxidative stress still existed in the muscle in C.quadricarinatus.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"46 1","pages":"1066-1072"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69968289","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-01-01DOI: 10.3724/SP.J.1231.2013.38572
Jinhong Yu, L. Pan, Hui Zhang
The studies on innate immune responses of Litopenaeus vannamei C-type lectin,were conducted by recombinant C-type lectin protein through expression in Escherichia coli.We used cell suspension culture to study the effect of C-type lectin on the cellular immune response of Litopenaeus vannamei.The experiment was divided into two groups:control group and recombinant protein(1.0 mg/mL).After 0,3,6,9,12,24 hours,we harvested the solutions and then assessed parameters respectively.The results showed that SDS-PAGE revealed the molecular weight of LvLec protein was 39 ku,which was consistent with the predicted size.Western-blotting revealed it could bind anti-His antibody,which proved that C-type lectin recombinant protein in Escherichia coli succeeded in expression.For the hemagglutination assay,massive agglutination could be observed after Escherichia coli was incubated with LvLec for 1 h.At the same time,LvLec protein had a significant effect on haemocyte count,cell viability and phagocytic activity of white shrimp Litopenaeus vannamei(P0.05).The extracelluar phenoloxidase activity of the LvLec protein group showed peak changes during 3-12 h,and at 9 h the extracelluar phenoloxidase activity was highest,then returned to the control level(P0.05).Therefore,crustacean C-type lectin not only had the function of hemagglutination,but also activated prophenoloxidase activating system,and probably played an important role in immune response.
{"title":"Prokaryotic expression of C-type lectin gene of Litopenaeus vannamei and immunologic activity evaluation of recombinant protein","authors":"Jinhong Yu, L. Pan, Hui Zhang","doi":"10.3724/SP.J.1231.2013.38572","DOIUrl":"https://doi.org/10.3724/SP.J.1231.2013.38572","url":null,"abstract":"The studies on innate immune responses of Litopenaeus vannamei C-type lectin,were conducted by recombinant C-type lectin protein through expression in Escherichia coli.We used cell suspension culture to study the effect of C-type lectin on the cellular immune response of Litopenaeus vannamei.The experiment was divided into two groups:control group and recombinant protein(1.0 mg/mL).After 0,3,6,9,12,24 hours,we harvested the solutions and then assessed parameters respectively.The results showed that SDS-PAGE revealed the molecular weight of LvLec protein was 39 ku,which was consistent with the predicted size.Western-blotting revealed it could bind anti-His antibody,which proved that C-type lectin recombinant protein in Escherichia coli succeeded in expression.For the hemagglutination assay,massive agglutination could be observed after Escherichia coli was incubated with LvLec for 1 h.At the same time,LvLec protein had a significant effect on haemocyte count,cell viability and phagocytic activity of white shrimp Litopenaeus vannamei(P0.05).The extracelluar phenoloxidase activity of the LvLec protein group showed peak changes during 3-12 h,and at 9 h the extracelluar phenoloxidase activity was highest,then returned to the control level(P0.05).Therefore,crustacean C-type lectin not only had the function of hemagglutination,but also activated prophenoloxidase activating system,and probably played an important role in immune response.","PeriodicalId":15710,"journal":{"name":"Journal of Fisheries of China","volume":"37 1","pages":"1183"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69968452","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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