VIGS(Virus-induced gene silencing),a method for posttranscriptional gene silencing,is an effective technique for investigating the activities of genes in plants.Since there is no report for available VIGS system in St...VIGS(Virus-induced gene silencing),a method for posttranscriptional gene silencing,is an effective technique for investigating the activities of genes in plants.Since there is no report for available VIGS system in Styrax japonicus,the application of a VIGS approach that results in a gene knockdown to study gene function is limited.In this study,we compared the characteristics that could affect the viability of VIGS in S.japonicus,including the acetosyringone(AS)concentration,the Agrobacterium’s optical density and the inoculation method.The stable reference genes of S.japonicus were selected to validate the gene’s knockdown by quantitative PCR.As a result,we successfully constructed 2 VIGS systems based on TRV virus:vacuum with AS concentration of 200μmol·L^(-1)and OD600of 0.5,and friction-osmosis with AS concentration of 200μmol·L^(-1)and OD600of 1.0,which silencing efficiency was 83.33%and 74.19%,respectively.The successfully applied VIGS method provides a rapid and effective reverse gene functional analysis approach in S.japonicus to identify unknown gene functions.展开更多
在半精制饲料中分别添加0、0.35%、0.70%、1.05%、1.40%、1.75%苏氨酸,制成苏氨酸实际梯度为1.05%、1.35%、1.65%、2.00%、2.42%、2.65%的6组等能等氮饲料(44.67%粗蛋白质,21.65 k J/g总能),对初始体重为(333.93±6.60)g的鲈鱼(Late...在半精制饲料中分别添加0、0.35%、0.70%、1.05%、1.40%、1.75%苏氨酸,制成苏氨酸实际梯度为1.05%、1.35%、1.65%、2.00%、2.42%、2.65%的6组等能等氮饲料(44.67%粗蛋白质,21.65 k J/g总能),对初始体重为(333.93±6.60)g的鲈鱼(Lateolabrax japonicus)在海水浮式网箱(1.5 m×1.5 m×2.0 m)中进行了70 d的喂养实验,研究其对苏氨酸的最适需求量。结果显示,鲈鱼成活率在89.58%–95.83%之间,各处理组之间无显著差异(P>0.05);随着饲料中苏氨酸水平的升高,鲈鱼的特定生长率(SGR)显著增加(P<0.05),且在2.00%苏氨酸饲料组出现最大值,但随着苏氨酸水平的继续升高,SGR呈减小的趋势;饲料效率(FE)随饲料中苏氨酸水平的升高呈先增加后减小的趋势,2.00%苏氨酸组的FE显著高于1.05%组及2.65%组(P<0.05);随着饲料中苏氨酸水平的升高,蛋白质沉积率(PPV)呈先增加后减小的趋势,且于2.00%苏氨酸组出现最大值;肝脏谷草转氨酶、谷丙转氨酶活性随饲料中苏氨酸水平的升高呈先增加后减小的趋势;饲料中不同水平苏氨酸对鱼体粗蛋白、粗脂肪、粗灰分无显著影响(P>0.05)。以特定生长率、饲料效率及蛋白质沉积率为评价指标,经二次回归分析得出,鲈鱼对饲料中苏氨酸的最适需求量分别为占饲料干重的1.84%、1.87%及1.83%,占饲料蛋白质的4.11%、4.18%及4.09%。展开更多
A homologue of the lower vertebrates translationally controlled tumor protein (TCTP) was cloned from the marine fish Japanese sea perch (Lateolabrax japonicus) by the technology of homology cloning. The full-length cD...A homologue of the lower vertebrates translationally controlled tumor protein (TCTP) was cloned from the marine fish Japanese sea perch (Lateolabrax japonicus) by the technology of homology cloning. The full-length cDNA sequence of the sea perch TCTP gene contained a 5' untranslated region (UTR) of 47 bp, a 3' UTR of 433 bp, and a putative open reading frame (ORF) of 510 bp encoding a polypeptide of 170 amino acids. The deduced amino acid sequence of the sea perch TCTP gene showed a high similarity to that of zebrafish, rohu, rabbit, chicken and human. Sequence analysis revealed there were a signature sequence of TCTP family, an N-glycosylation site, and five Casein kinase phosphorylation sites in the sea perch TCTP. The temporal expression of TCTP genes in healthy and lipopolysaccharide (LPS) challenged fishes was measured by semi-quantitative reverse transcription-PCR (RT-PCR). The results indicated that LPS could up-regulate the expression of sea perch TCTP in the examined tissues, including head-kidney, spleen and liver.展开更多
The metabolic rate of Japanese sea bass, Lateolabrax japonicus (C & V), was estimated in laboratory at temperature 25.2±0.5℃. The fresh weight of the fish was 4.64-52.28 g (average of 17.81±0.33 g). The...The metabolic rate of Japanese sea bass, Lateolabrax japonicus (C & V), was estimated in laboratory at temperature 25.2±0.5℃. The fresh weight of the fish was 4.64-52.28 g (average of 17.81±0.33 g). The routine metabolism was related to body weight by the exponential equation: R r =14.966 W 0.74 ( r =0.934). The rate of feeding metabolism increased linearly with food consumption. Feeding metabolic rate was 1.8-2.4 times the routine metabolic rate.展开更多
The t-SNARE protein SNAP-25 (synaptosome-associated protein of 25 kDa) plays an essential role in regulating fusion between the vesicle and plasma membranes during exocytosis. To clone and characterize SNAP-25 gene, t...The t-SNARE protein SNAP-25 (synaptosome-associated protein of 25 kDa) plays an essential role in regulating fusion between the vesicle and plasma membranes during exocytosis. To clone and characterize SNAP-25 gene, the first step in the functional study of SNARE proteins in marine teleostean, was to obtain the cDNA of sea perch SNAP-25 (SPsn25) by RT-PCR and RACE-PCR amplification of a Japanese sea perch. The full-length cDNA of 831bp contains a CDS of 615 bp, coding 204 amino acid residues, and a 5′UTR of 219bp. Bioinformatic analysis revealed that SPsn25 corresponds with SNAP-25a isoform and shares 91.1% identity with SNAP-25a of a goldfish and a zebrafish. The SPsn25 expression in both mRNA and protein levels in the Japanese sea perch had been identified through semi-quantitative RT-PCR and Western Blot assay. Together, these data again confirmed the nerve tissue specificity of the fish SNAP-25 gene expression.展开更多
Information on survival and growth during the early life stage is essential to understand the mechanism of interannual variations in fish recruitment.Chub mackerel Scomber japonicus is a commercially important pelagic...Information on survival and growth during the early life stage is essential to understand the mechanism of interannual variations in fish recruitment.Chub mackerel Scomber japonicus is a commercially important pelagic fish widely distributed in the northwestern Pacific.Its catch showed large fluctuations with changes in distribution and migration under climate change and strong fishing.We determined the hatch dates and growth rates of young-of-the-year of chub mackerel through otolith microstructure using samples collected in the Oyashio water in autumn 2018.Results show that the ages of young chub mackerel ranged between 120 and 180 d,and the estimated hatch date lasted from midJanuary to late May with a peak from mid-March to mid-April.Average otolith daily increment width during the early life stages(from hatching to 25 d)showed an increasing trend.Chub mackerel grows slowly in the first 10 d,and then grows faster during the 10thto 25thd.Three groups with dissimilar growth histories and migration routes were identified using unsupervised random forest clustering analysis,but all eventually converge on the same nursery ground.The faster growth of young-of-the-year chub mackerel leads to better recruitment due to the hypothesis of growth-dependent mortality.Most chub mackerels hatched in March and April,the spawning period is longer and earlier,which could lead to strong year classes.These findings on population composition and life history traits of young-of-the-year of chub mackerel provide valuable information on its recruitment processes during the period of stock recovery.展开更多
Lymphocystis disease virus(LCDV) infects target cells by attaching to a 27.8 k Da receptor(27.8R) protein in flounder Paralichthys olivaceus, and anti-27.8R monoclonal antibodies(MAbs) have been developed. However, th...Lymphocystis disease virus(LCDV) infects target cells by attaching to a 27.8 k Da receptor(27.8R) protein in flounder Paralichthys olivaceus, and anti-27.8R monoclonal antibodies(MAbs) have been developed. However, the 27.8R existence in tissues of sea bass(Lateolabrax japonicus) and its role in LCDV infection have remained unclear. In this study, the results of western blotting demonstrated that the same 27.8R was shared by flounder and sea bass. LCDV-free sea bass individuals were intramuscularly injected with LCDV, and viral copies were detected in tissues from 3 h post infection and showed a time-dependent increase during 9 days infection. Distribution and synthesis of 27.8R in sea bass tissues were investigated by using anti-27.8R MAbs as probes. It was found that 27.8R was distributed in all the tested tissues. The levels of 27.8R protein were highest in gill and skin, then a bit lowly in stomach, head kidney and heart, followed by spleen, intestine, blood cells, gonad and liver, and least in kidney and brain in healthy sea bass. Upon LCDV infection, 27.8R synthesis was up-regulated in each tissue, and higher in the tissues with higher LCDV copies. The 27.8R and LCDV were detected in some peripheral blood leukocytes but not in red blood cells. These results suggested that 27.8R was widely distributed in sea bass tissues, and it served as a receptor and correlated with tissue tropism of LCDV infection. Furthermore, leukocytes had the potential of being a LCDV carrier and were responsible for a systemic infection of LCDV in sea bass.展开更多
Sea cucumber(Apostichopus japonicus)is an excellent model for investigating effects of bottom-dwellers on carbon mig-ration and transformation.However,the molecular mechanism of respiratory metabolism process variatio...Sea cucumber(Apostichopus japonicus)is an excellent model for investigating effects of bottom-dwellers on carbon mig-ration and transformation.However,the molecular mechanism of respiratory metabolism process variation caused by feeding rations is poorly understood.In this study,treatment groups set as 1%(about 0.63g),3%,and 7%of total body weight(named F1,F3 and F7 groups respectively).The potential molecular mechanisms behind the functions of respiratory tree and body wall were investigated by RNA-Seq.A total of 52411 expressed genes were identified from 89342 expressed transcripts.The results showed 759,254 and 334 genes were up-regulated,and 334,445 and 992 genes were down-regulated in respiratory tree of F1 vs.F3,F1 vs.F7 and F3 vs.F7,respectively.Meanwhile,2070,1601 and 896 genes were up-regulated,and 1303,1337 and 1144 genes were down-regulated in body wall between F1 vs.F3,F1 vs.F7 and F3 vs.F7,respectively.Differentially expressed genes were enriched in salivary secretion and ECM-receptor interaction pathways in respiratory tree,and in various types of N-glycan biosynthesis,ribosome and sphingolipid metabolism pathways in body wall.These results suggested respiratory tree and body wall were involved in activation of respiratory metabolisms in response to different feeding rations.Our research provided valuable knowledge for physiological differences in res-piratory metabolism.展开更多
基金supported by the Subject of Key R&D Plan of Shandong Province(Major Scientific and Technological Innovation Project)“Mining and Accurate Identification of Forest Tree Germplasm Resources”(Grant Nos.2021LZGC02303)Science&Technology Specific Projects in Agricultural High-tech Industrial Demonstration Area of the Yellow River Delta(Grant No.022SZX16)。
文摘VIGS(Virus-induced gene silencing),a method for posttranscriptional gene silencing,is an effective technique for investigating the activities of genes in plants.Since there is no report for available VIGS system in Styrax japonicus,the application of a VIGS approach that results in a gene knockdown to study gene function is limited.In this study,we compared the characteristics that could affect the viability of VIGS in S.japonicus,including the acetosyringone(AS)concentration,the Agrobacterium’s optical density and the inoculation method.The stable reference genes of S.japonicus were selected to validate the gene’s knockdown by quantitative PCR.As a result,we successfully constructed 2 VIGS systems based on TRV virus:vacuum with AS concentration of 200μmol·L^(-1)and OD600of 0.5,and friction-osmosis with AS concentration of 200μmol·L^(-1)and OD600of 1.0,which silencing efficiency was 83.33%and 74.19%,respectively.The successfully applied VIGS method provides a rapid and effective reverse gene functional analysis approach in S.japonicus to identify unknown gene functions.
基金supported by the“863"Prijetof China under contract Nos 2001AA628180 and 2002AA626020.
文摘A homologue of the lower vertebrates translationally controlled tumor protein (TCTP) was cloned from the marine fish Japanese sea perch (Lateolabrax japonicus) by the technology of homology cloning. The full-length cDNA sequence of the sea perch TCTP gene contained a 5' untranslated region (UTR) of 47 bp, a 3' UTR of 433 bp, and a putative open reading frame (ORF) of 510 bp encoding a polypeptide of 170 amino acids. The deduced amino acid sequence of the sea perch TCTP gene showed a high similarity to that of zebrafish, rohu, rabbit, chicken and human. Sequence analysis revealed there were a signature sequence of TCTP family, an N-glycosylation site, and five Casein kinase phosphorylation sites in the sea perch TCTP. The temporal expression of TCTP genes in healthy and lipopolysaccharide (LPS) challenged fishes was measured by semi-quantitative reverse transcription-PCR (RT-PCR). The results indicated that LPS could up-regulate the expression of sea perch TCTP in the examined tissues, including head-kidney, spleen and liver.
文摘The metabolic rate of Japanese sea bass, Lateolabrax japonicus (C & V), was estimated in laboratory at temperature 25.2±0.5℃. The fresh weight of the fish was 4.64-52.28 g (average of 17.81±0.33 g). The routine metabolism was related to body weight by the exponential equation: R r =14.966 W 0.74 ( r =0.934). The rate of feeding metabolism increased linearly with food consumption. Feeding metabolic rate was 1.8-2.4 times the routine metabolic rate.
基金the NSFC (No.40476060)Hi-Tech Research and Development Program of China (No. 2002AA629120)
文摘The t-SNARE protein SNAP-25 (synaptosome-associated protein of 25 kDa) plays an essential role in regulating fusion between the vesicle and plasma membranes during exocytosis. To clone and characterize SNAP-25 gene, the first step in the functional study of SNARE proteins in marine teleostean, was to obtain the cDNA of sea perch SNAP-25 (SPsn25) by RT-PCR and RACE-PCR amplification of a Japanese sea perch. The full-length cDNA of 831bp contains a CDS of 615 bp, coding 204 amino acid residues, and a 5′UTR of 219bp. Bioinformatic analysis revealed that SPsn25 corresponds with SNAP-25a isoform and shares 91.1% identity with SNAP-25a of a goldfish and a zebrafish. The SPsn25 expression in both mRNA and protein levels in the Japanese sea perch had been identified through semi-quantitative RT-PCR and Western Blot assay. Together, these data again confirmed the nerve tissue specificity of the fish SNAP-25 gene expression.
基金Supported by the National Natural Science Foundation of China(No.41930534)the Third Institute of Oceanography through the National Program on Global Change and Air-Sea Interaction(No.GASI-02-PACYDaut)。
文摘Information on survival and growth during the early life stage is essential to understand the mechanism of interannual variations in fish recruitment.Chub mackerel Scomber japonicus is a commercially important pelagic fish widely distributed in the northwestern Pacific.Its catch showed large fluctuations with changes in distribution and migration under climate change and strong fishing.We determined the hatch dates and growth rates of young-of-the-year of chub mackerel through otolith microstructure using samples collected in the Oyashio water in autumn 2018.Results show that the ages of young chub mackerel ranged between 120 and 180 d,and the estimated hatch date lasted from midJanuary to late May with a peak from mid-March to mid-April.Average otolith daily increment width during the early life stages(from hatching to 25 d)showed an increasing trend.Chub mackerel grows slowly in the first 10 d,and then grows faster during the 10thto 25thd.Three groups with dissimilar growth histories and migration routes were identified using unsupervised random forest clustering analysis,but all eventually converge on the same nursery ground.The faster growth of young-of-the-year chub mackerel leads to better recruitment due to the hypothesis of growth-dependent mortality.Most chub mackerels hatched in March and April,the spawning period is longer and earlier,which could lead to strong year classes.These findings on population composition and life history traits of young-of-the-year of chub mackerel provide valuable information on its recruitment processes during the period of stock recovery.
基金the National Natural Science Foundation of China (Nos. 31472295 and 31672685)Science and Technology Development Project of Shandong Province (No. 2014GNC111015)Taishan Scholar Program of Shandong Province
文摘Lymphocystis disease virus(LCDV) infects target cells by attaching to a 27.8 k Da receptor(27.8R) protein in flounder Paralichthys olivaceus, and anti-27.8R monoclonal antibodies(MAbs) have been developed. However, the 27.8R existence in tissues of sea bass(Lateolabrax japonicus) and its role in LCDV infection have remained unclear. In this study, the results of western blotting demonstrated that the same 27.8R was shared by flounder and sea bass. LCDV-free sea bass individuals were intramuscularly injected with LCDV, and viral copies were detected in tissues from 3 h post infection and showed a time-dependent increase during 9 days infection. Distribution and synthesis of 27.8R in sea bass tissues were investigated by using anti-27.8R MAbs as probes. It was found that 27.8R was distributed in all the tested tissues. The levels of 27.8R protein were highest in gill and skin, then a bit lowly in stomach, head kidney and heart, followed by spleen, intestine, blood cells, gonad and liver, and least in kidney and brain in healthy sea bass. Upon LCDV infection, 27.8R synthesis was up-regulated in each tissue, and higher in the tissues with higher LCDV copies. The 27.8R and LCDV were detected in some peripheral blood leukocytes but not in red blood cells. These results suggested that 27.8R was widely distributed in sea bass tissues, and it served as a receptor and correlated with tissue tropism of LCDV infection. Furthermore, leukocytes had the potential of being a LCDV carrier and were responsible for a systemic infection of LCDV in sea bass.
基金supported by the National Natural Science Foundation of China(No.31672657).
文摘Sea cucumber(Apostichopus japonicus)is an excellent model for investigating effects of bottom-dwellers on carbon mig-ration and transformation.However,the molecular mechanism of respiratory metabolism process variation caused by feeding rations is poorly understood.In this study,treatment groups set as 1%(about 0.63g),3%,and 7%of total body weight(named F1,F3 and F7 groups respectively).The potential molecular mechanisms behind the functions of respiratory tree and body wall were investigated by RNA-Seq.A total of 52411 expressed genes were identified from 89342 expressed transcripts.The results showed 759,254 and 334 genes were up-regulated,and 334,445 and 992 genes were down-regulated in respiratory tree of F1 vs.F3,F1 vs.F7 and F3 vs.F7,respectively.Meanwhile,2070,1601 and 896 genes were up-regulated,and 1303,1337 and 1144 genes were down-regulated in body wall between F1 vs.F3,F1 vs.F7 and F3 vs.F7,respectively.Differentially expressed genes were enriched in salivary secretion and ECM-receptor interaction pathways in respiratory tree,and in various types of N-glycan biosynthesis,ribosome and sphingolipid metabolism pathways in body wall.These results suggested respiratory tree and body wall were involved in activation of respiratory metabolisms in response to different feeding rations.Our research provided valuable knowledge for physiological differences in res-piratory metabolism.
