Esox reieherti Dybowsk genomic microsateUites were developed by using enrichment protocols combined with radioactive hybridization protocol. Four hundred to nine hundred base pair fragments were selected for the whole...Esox reieherti Dybowsk genomic microsateUites were developed by using enrichment protocols combined with radioactive hybridization protocol. Four hundred to nine hundred base pair fragments were selected for the whole genome. DNA PCR amplification after digestion with restriction endonuclease Sau 3A Ⅰ, and (CA)12, (GA)12 probes marked with biotin were used for microsateUite DNA enrichment. The product fragments were connected with carder pGEM-T and transferred into DH5α Escherichia coli competent cells, and radioactive isotope probes marked with γ^-32 p were used for the second hybridization. As a result, a total of 1600 bacteria were obtained in the microsatellite genomic libraries, positive clones accounted for 90.91% before hybridization and 81.25% after hybridization, amounting to 1300. One hundred and ninety-six positive clones were selected for sequencing, and 192 clones included microsateUite sequences. The microsateUite sequences obtained, mono-nucleotide, quad-nucleotide and quint-nucleotide repeat motifs were observed beside double-base-pairs CA/GT, GA/CT. Seventy primers were designed according to the flanking sequences by using software Primer Premier 5.0, and 32 primers were selected to be synthesized. After optimizing PCR reaction conditions, 28 primers were amplified and produced clear purpose bands. The aim of our research was to promote the development and utilization of E. reieherti genomic resource, and lay the foundation for optimizing E. reieherti breeding strain in order to detect the genetic diversity and construct a genetic map.展开更多
In the Pearl River Delta with more than 1000 years of intensive land reclamation history, the developmentof acid sulphate soils has been generally limited in terms of their acid potential (pyrite content) and spatiale...In the Pearl River Delta with more than 1000 years of intensive land reclamation history, the developmentof acid sulphate soils has been generally limited in terms of their acid potential (pyrite content) and spatialextent. This is attributed to the rapid delta progradation, partially resulted from increasing sediment yieldcaused by deforestation within the catchment and the empolderment in the estuarine embayment. Theempolderment practice accompanied by the clearance of mangroves stopped the upward growth of the pyriticlayer on the one hand and limited the vertical accretion of non-pyritic freshwater sediments over the pyriticestuarine sedimellts on the other. In such a case, the Pyritic layer in the area is frequently thin and ofshallow occurrence. Under forced leaching-recharge conditions for the paddy rice cultivation, the leaching ofacid sulphate materials prevails over its production and this leads to a net loss in pyrite oxidation products.Land excaVation for fishpond farming accelerates Pyrite oxidation due to the direct exposure of the pyriticsediments to air on the pond bunds. Severe acidification can intensify the environmental degradation ofestuarine ecosystems.展开更多
文摘Esox reieherti Dybowsk genomic microsateUites were developed by using enrichment protocols combined with radioactive hybridization protocol. Four hundred to nine hundred base pair fragments were selected for the whole genome. DNA PCR amplification after digestion with restriction endonuclease Sau 3A Ⅰ, and (CA)12, (GA)12 probes marked with biotin were used for microsateUite DNA enrichment. The product fragments were connected with carder pGEM-T and transferred into DH5α Escherichia coli competent cells, and radioactive isotope probes marked with γ^-32 p were used for the second hybridization. As a result, a total of 1600 bacteria were obtained in the microsatellite genomic libraries, positive clones accounted for 90.91% before hybridization and 81.25% after hybridization, amounting to 1300. One hundred and ninety-six positive clones were selected for sequencing, and 192 clones included microsateUite sequences. The microsateUite sequences obtained, mono-nucleotide, quad-nucleotide and quint-nucleotide repeat motifs were observed beside double-base-pairs CA/GT, GA/CT. Seventy primers were designed according to the flanking sequences by using software Primer Premier 5.0, and 32 primers were selected to be synthesized. After optimizing PCR reaction conditions, 28 primers were amplified and produced clear purpose bands. The aim of our research was to promote the development and utilization of E. reieherti genomic resource, and lay the foundation for optimizing E. reieherti breeding strain in order to detect the genetic diversity and construct a genetic map.
文摘In the Pearl River Delta with more than 1000 years of intensive land reclamation history, the developmentof acid sulphate soils has been generally limited in terms of their acid potential (pyrite content) and spatialextent. This is attributed to the rapid delta progradation, partially resulted from increasing sediment yieldcaused by deforestation within the catchment and the empolderment in the estuarine embayment. Theempolderment practice accompanied by the clearance of mangroves stopped the upward growth of the pyriticlayer on the one hand and limited the vertical accretion of non-pyritic freshwater sediments over the pyriticestuarine sedimellts on the other. In such a case, the Pyritic layer in the area is frequently thin and ofshallow occurrence. Under forced leaching-recharge conditions for the paddy rice cultivation, the leaching ofacid sulphate materials prevails over its production and this leads to a net loss in pyrite oxidation products.Land excaVation for fishpond farming accelerates Pyrite oxidation due to the direct exposure of the pyriticsediments to air on the pond bunds. Severe acidification can intensify the environmental degradation ofestuarine ecosystems.