Genetic diversity of two wild Penaeus monodon populations sampled from the coastal waters of Qinglan ( Hainan Province of China, HN) and Malaysia (KD), and the F1 generation of a Thailand broodstock population (C...Genetic diversity of two wild Penaeus monodon populations sampled from the coastal waters of Qinglan ( Hainan Province of China, HN) and Malaysia (KD), and the F1 generation of a Thailand broodstock population (CP) were examined by vertical polyacrylamide gel electrophoresis. Of 21 loci encoded by ten enzymes, 11 were polymorphic. The mean proportions of polymorphic loci of HN, KD and CP were 36. 36%, 45.45% and 50.00% , with the average heterozygosities of 0.135, 0.181 and 0.191, and the effective numbers of alleles per loci were 1. 300, 1. 330 and 1. 329, respectively. The divergent indexes of HN, KD and CP were0.023, 0.124, and 0.117, respectively. The genetic distance between the two wild populations was 0.005, and the gene differentiation coefficient was also very low (0.014). The results indicated that the F1 generation population had a higher genetic diversity than the two wild populations, and that the HN population had the lowest one. There was no significant differentiation between HN and KD populations and all populations existed with a slight heterozygote excess.展开更多
Study on shrimp miRNAs was limited and just 7 mature miRNA sequences of Marsupenaeus japonicus are deposited in mir Base database. In this study, miRNAs and their target gene candidates were computationally identified...Study on shrimp miRNAs was limited and just 7 mature miRNA sequences of Marsupenaeus japonicus are deposited in mir Base database. In this study, miRNAs and their target gene candidates were computationally identified from shrimp Penaeu s monodon and then experimentally validated. Using 39 908 expressed sequence tags(ESTs) and 21 124 genome survey sequences(GSSs) of P. monodon(pmo) as reference dataset, a comprehensive approach based on inter-species homolog search was employed to investigate the candidate miRNAs(i.e. pmo-miRNA). A total of eight miRNAs belonging to 7 families were computationally identified and five out of them were subsequently validated by PCR and sequencing. Of these, pmo-miR-4961a, pmo-miR-4961b, pmo-miR-4979 and pmo-miR-3819 were first identified from shrimps. Both the mature pmo-miRNAs and the corresponding precursors were conserved among different species. Based on perfect or near-perfect match to the target region, the target gene candidates of pmomiRNAs were predicted from 10 331 mRNA sequences of P. monodon. A total of 20 genes were predicted as the targets of pmo-miR-4961a, pmo-miR-4961b, pmo-miR-4979 and pmo-miR-6492. Experimental validation by dual luciferase reporter assay confi rmed the targeting between 3 pmo-miRNAs and one or two of their target genes, especially the pmo-miR-4979 which could significantly down-regulate the expression of target gene(JR226772). This study updates the miRNAs and their targets in P. monodon and lays a solid foundation for future RNAi study.展开更多
The hepatopancreatic necrosis disease(HPND)is a common shrimp disease.Aquaculture of the black tiger shrimp Penaeus monodon has suffered with the frequent outbreak of HPND.Understanding the molecular mechanisms of hos...The hepatopancreatic necrosis disease(HPND)is a common shrimp disease.Aquaculture of the black tiger shrimp Penaeus monodon has suffered with the frequent outbreak of HPND.Understanding the molecular mechanisms of host response to HPND is of pivotal importance for improving its farming performance.In this study,the RNA-Seq platform was utilized to investigate transcriptomic changes of the hepatopancreas in pond-cultured P.monodon with HPND symptoms.A total of 62071 genes with an average length of 953 bp were obtained,and a lot of simple sequence repeat(SSR)loci related to these genes were identified.Totally,5204 differentially expressed genes(DEGs)were detected between healthy shrimp and HPND shrimp,with 3399 genes upregulated and 1,805 genes down-regulated.These genes had a wide range of biological functions,and several well-known immunerelated genes including integrin alpha 5,integrin beta 1,C type lectin and catalase were among the DEGs.Cell signaling pathways including the extracellular matrix(ECM)receptor interaction,phagosome and lysosome pathways were significantly upregulated in HPND-infected shrimps,indicating their involvement in the immune responses of shrimp against HPND.The data obtained in this study offers new insights into the molecular mechanisms of shrimp host response to HPND disease,and provides a resource for molecular marker development and genetic breeding studies of P.monodon.展开更多
Distribution of injected Vibrio anguillarum in body of Penaeus monodon was studied with immunohistochemical method. Bacteria could be detected throughout the experiment in some individuals; however in lymphoid tissue,...Distribution of injected Vibrio anguillarum in body of Penaeus monodon was studied with immunohistochemical method. Bacteria could be detected throughout the experiment in some individuals; however in lymphoid tissue, gill, heart and haemolymph of all vibrio injected shrimp, the bacteria could be observed only 5 min after injection. The bacteria density in haemolymph, haemolymph of the hepatopancreas and gills decreased with time. In the lymphoid organ and heart, the bacteria density was the highest 48 h after injection, then decreased. Nodules could be formed in the heart, lymphoid organ and injection site.展开更多
The patterns of isozyme variation throughout the life cycle of the giant tiger prawn, Penaeusmonodon were described for ten enzyme systems (G3PD, MDH, LGG, GPI, MPI, FBP, PGM, LDH,PGD, and PROT). The zymograms showh l...The patterns of isozyme variation throughout the life cycle of the giant tiger prawn, Penaeusmonodon were described for ten enzyme systems (G3PD, MDH, LGG, GPI, MPI, FBP, PGM, LDH,PGD, and PROT). The zymograms showh little variation in the number of isozymic loci expressed duringlarval development compared with results reported for other penaeid prawns. There were specific ontogenicpatterns at some loci. As larvae developed beyond the embryonic stage, there was tendency for intensityof staining and complexity of banding patterns to increase.展开更多
mRNA was isolated from the hepatopancrease of shrimp Penaeus monodon with a PolyAT-tract System 1000 Kit. By using mRNA as template, double - strand cDNA with EcoR I/Xho I ends was synthesized by using a ZAP Express c...mRNA was isolated from the hepatopancrease of shrimp Penaeus monodon with a PolyAT-tract System 1000 Kit. By using mRNA as template, double - strand cDNA with EcoR I/Xho I ends was synthesized by using a ZAP Express cDNA Synthesis Kit. The cDNA was inserted into the lambda ZAP Express vector predigested with EcoR l/Xho Ⅰ, and the recombinant DNA was in vitro packaged into lambda phage with GigapackⅢ Gold packaging extracts. These recombinant phages were then used to transfect E. coli XL1 - Blue MRF', and finally a cDNA expression library was constructed. The library is 7.2 × 10~5pfu in capacity and its recombination ratio is higher than 99 % . The size of the inserted cDNAs was determined by EcoR l/Xho I digestion of 9 phagemids prepared by in vivo excision of plaques selected randomly from amplified cDNA library . The longest inserted cDNA is about 1.6 kb in length. The complete sequence (about 1.2 kb) of actin cDNA was amplified from the library by PCR reveals that this library contains full-length cDNAs of Penaeus monodon hepatopancreas and is available for screening and expression of shrimp genes.展开更多
A 3 x 4 two-factor factorial experimental design with three replications per treatment was conducted to determine the effects three temperatures viz., room temperature (29°C ± 1°C) and elevated temperat...A 3 x 4 two-factor factorial experimental design with three replications per treatment was conducted to determine the effects three temperatures viz., room temperature (29°C ± 1°C) and elevated temperatures (33°C and 35°C ± 0.5°C) and four salinities (25, 30, 33 and 35 ppt) on survival of postlarvae tiger prawn Penaeus monodon under laboratory conditions. Results showed that temperature had a greater influence on survival of postlarvae P. monodon than salinity and low water temperature (29°C) gave better larval survival than higher water temperatures (33°C and 35°C). In addition, the PL1 had better tolerance to high temperatures than PL15. At the termination of the experiment, the best survival of PL1 was found at 29°C for all salinities tested with an average of 95.8%, while the best survival of PL15 was also found at 29°C for all salinities tested with an average of 70.6%. Based on survival at temperatures and salinities tested, the best salinity and temperature combination for the culture of P. monodon PL1 and PL15 were 29°C at 25, 30, 33 and 35 ppt, and 29°C at 25 and 30 ppt, respectively.展开更多
This study monitored the present status ofPenaeus monodon post larvae (PL.) of Kholpatua river in Satkhira. The survey was conducted in Kholpatua river from January 2014 to December 2014 and was compared with the te...This study monitored the present status ofPenaeus monodon post larvae (PL.) of Kholpatua river in Satkhira. The survey was conducted in Kholpatua river from January 2014 to December 2014 and was compared with the ten years old secondary data. The results showed that there was a 41.57% decrease in number of catches ofP. monodon PL. over the course of time from ten years ago to the year of 2014. Although the number of active fishermen's boats have increased from 250 to 360 in ten years. The study also showed that the severe shrimp seed collection activities reduced the availability ofP. monodon PL. at Satkhira district, which has a severe negative impact on the availability of total catches. To control such decrease of P. monodon PL. in Bangladesh, more intensive study should be done.展开更多
基金The National Natural Science Foundation of China under contract No.30471322.
文摘Genetic diversity of two wild Penaeus monodon populations sampled from the coastal waters of Qinglan ( Hainan Province of China, HN) and Malaysia (KD), and the F1 generation of a Thailand broodstock population (CP) were examined by vertical polyacrylamide gel electrophoresis. Of 21 loci encoded by ten enzymes, 11 were polymorphic. The mean proportions of polymorphic loci of HN, KD and CP were 36. 36%, 45.45% and 50.00% , with the average heterozygosities of 0.135, 0.181 and 0.191, and the effective numbers of alleles per loci were 1. 300, 1. 330 and 1. 329, respectively. The divergent indexes of HN, KD and CP were0.023, 0.124, and 0.117, respectively. The genetic distance between the two wild populations was 0.005, and the gene differentiation coefficient was also very low (0.014). The results indicated that the F1 generation population had a higher genetic diversity than the two wild populations, and that the HN population had the lowest one. There was no significant differentiation between HN and KD populations and all populations existed with a slight heterozygote excess.
基金Supported by the National Natural Science Foundation of China(Nos.31172391,31472274)the Fundamental Research Funds for Central Universities(No.201762003)+2 种基金the Scholarship Foundation for Excellent Scientists of Shandong Province(No.BS2011SW054)the National HighTech R&D Program of China(863 Program)(No.2012AA10A402)the Open Funds of Institute of Biodiversity and Evolution,Ocean University of China(No.201362017)
文摘Study on shrimp miRNAs was limited and just 7 mature miRNA sequences of Marsupenaeus japonicus are deposited in mir Base database. In this study, miRNAs and their target gene candidates were computationally identified from shrimp Penaeu s monodon and then experimentally validated. Using 39 908 expressed sequence tags(ESTs) and 21 124 genome survey sequences(GSSs) of P. monodon(pmo) as reference dataset, a comprehensive approach based on inter-species homolog search was employed to investigate the candidate miRNAs(i.e. pmo-miRNA). A total of eight miRNAs belonging to 7 families were computationally identified and five out of them were subsequently validated by PCR and sequencing. Of these, pmo-miR-4961a, pmo-miR-4961b, pmo-miR-4979 and pmo-miR-3819 were first identified from shrimps. Both the mature pmo-miRNAs and the corresponding precursors were conserved among different species. Based on perfect or near-perfect match to the target region, the target gene candidates of pmomiRNAs were predicted from 10 331 mRNA sequences of P. monodon. A total of 20 genes were predicted as the targets of pmo-miR-4961a, pmo-miR-4961b, pmo-miR-4979 and pmo-miR-6492. Experimental validation by dual luciferase reporter assay confi rmed the targeting between 3 pmo-miRNAs and one or two of their target genes, especially the pmo-miR-4979 which could significantly down-regulate the expression of target gene(JR226772). This study updates the miRNAs and their targets in P. monodon and lays a solid foundation for future RNAi study.
基金This study was funded in part by the Natural Science Foundation of Guangdong Province(No.2018A030310049)2019 Annual Guangdong Provincial Special fund of Nan-hai Economic Shrimp Breeding and Culture Laboratory(No.2319412525)+2 种基金the Nanhai Scholar Project of Guangdong Ocean University,Fangchenggang Science and Technol-ogy Plan Project(No.AD19008017)the R&D Program of Key Areas in Guangdong Province(No.2020B0202010009)2019 Provincial Financial Special Fund Con-struction Project of Guangdong Ocean University(No.231619003).
文摘The hepatopancreatic necrosis disease(HPND)is a common shrimp disease.Aquaculture of the black tiger shrimp Penaeus monodon has suffered with the frequent outbreak of HPND.Understanding the molecular mechanisms of host response to HPND is of pivotal importance for improving its farming performance.In this study,the RNA-Seq platform was utilized to investigate transcriptomic changes of the hepatopancreas in pond-cultured P.monodon with HPND symptoms.A total of 62071 genes with an average length of 953 bp were obtained,and a lot of simple sequence repeat(SSR)loci related to these genes were identified.Totally,5204 differentially expressed genes(DEGs)were detected between healthy shrimp and HPND shrimp,with 3399 genes upregulated and 1,805 genes down-regulated.These genes had a wide range of biological functions,and several well-known immunerelated genes including integrin alpha 5,integrin beta 1,C type lectin and catalase were among the DEGs.Cell signaling pathways including the extracellular matrix(ECM)receptor interaction,phagosome and lysosome pathways were significantly upregulated in HPND-infected shrimps,indicating their involvement in the immune responses of shrimp against HPND.The data obtained in this study offers new insights into the molecular mechanisms of shrimp host response to HPND disease,and provides a resource for molecular marker development and genetic breeding studies of P.monodon.
文摘Distribution of injected Vibrio anguillarum in body of Penaeus monodon was studied with immunohistochemical method. Bacteria could be detected throughout the experiment in some individuals; however in lymphoid tissue, gill, heart and haemolymph of all vibrio injected shrimp, the bacteria could be observed only 5 min after injection. The bacteria density in haemolymph, haemolymph of the hepatopancreas and gills decreased with time. In the lymphoid organ and heart, the bacteria density was the highest 48 h after injection, then decreased. Nodules could be formed in the heart, lymphoid organ and injection site.
文摘The patterns of isozyme variation throughout the life cycle of the giant tiger prawn, Penaeusmonodon were described for ten enzyme systems (G3PD, MDH, LGG, GPI, MPI, FBP, PGM, LDH,PGD, and PROT). The zymograms showh little variation in the number of isozymic loci expressed duringlarval development compared with results reported for other penaeid prawns. There were specific ontogenicpatterns at some loci. As larvae developed beyond the embryonic stage, there was tendency for intensityof staining and complexity of banding patterns to increase.
基金This study was supported by the Ocean Science Technology Fundation of State Oceanic Administration Ocean "863" Project under contract No. 2001AA621130.
文摘mRNA was isolated from the hepatopancrease of shrimp Penaeus monodon with a PolyAT-tract System 1000 Kit. By using mRNA as template, double - strand cDNA with EcoR I/Xho I ends was synthesized by using a ZAP Express cDNA Synthesis Kit. The cDNA was inserted into the lambda ZAP Express vector predigested with EcoR l/Xho Ⅰ, and the recombinant DNA was in vitro packaged into lambda phage with GigapackⅢ Gold packaging extracts. These recombinant phages were then used to transfect E. coli XL1 - Blue MRF', and finally a cDNA expression library was constructed. The library is 7.2 × 10~5pfu in capacity and its recombination ratio is higher than 99 % . The size of the inserted cDNAs was determined by EcoR l/Xho I digestion of 9 phagemids prepared by in vivo excision of plaques selected randomly from amplified cDNA library . The longest inserted cDNA is about 1.6 kb in length. The complete sequence (about 1.2 kb) of actin cDNA was amplified from the library by PCR reveals that this library contains full-length cDNAs of Penaeus monodon hepatopancreas and is available for screening and expression of shrimp genes.
文摘A 3 x 4 two-factor factorial experimental design with three replications per treatment was conducted to determine the effects three temperatures viz., room temperature (29°C ± 1°C) and elevated temperatures (33°C and 35°C ± 0.5°C) and four salinities (25, 30, 33 and 35 ppt) on survival of postlarvae tiger prawn Penaeus monodon under laboratory conditions. Results showed that temperature had a greater influence on survival of postlarvae P. monodon than salinity and low water temperature (29°C) gave better larval survival than higher water temperatures (33°C and 35°C). In addition, the PL1 had better tolerance to high temperatures than PL15. At the termination of the experiment, the best survival of PL1 was found at 29°C for all salinities tested with an average of 95.8%, while the best survival of PL15 was also found at 29°C for all salinities tested with an average of 70.6%. Based on survival at temperatures and salinities tested, the best salinity and temperature combination for the culture of P. monodon PL1 and PL15 were 29°C at 25, 30, 33 and 35 ppt, and 29°C at 25 and 30 ppt, respectively.
文摘This study monitored the present status ofPenaeus monodon post larvae (PL.) of Kholpatua river in Satkhira. The survey was conducted in Kholpatua river from January 2014 to December 2014 and was compared with the ten years old secondary data. The results showed that there was a 41.57% decrease in number of catches ofP. monodon PL. over the course of time from ten years ago to the year of 2014. Although the number of active fishermen's boats have increased from 250 to 360 in ten years. The study also showed that the severe shrimp seed collection activities reduced the availability ofP. monodon PL. at Satkhira district, which has a severe negative impact on the availability of total catches. To control such decrease of P. monodon PL. in Bangladesh, more intensive study should be done.