The tender stems from new Lycium barbarum L. cultivar ＂Ningqi 3＂ released by Ningxia Academy of Agricultural and Forestry Sciences were regarded as explants to investigate the vitrification of Lycium barbarum plantl...The tender stems from new Lycium barbarum L. cultivar ＂Ningqi 3＂ released by Ningxia Academy of Agricultural and Forestry Sciences were regarded as explants to investigate the vitrification of Lycium barbarum plantlets in tissue culture under different concentrations of 6-BA, sucrose, agrose, culture temperature, and illumination duration with MS as basic medium. The results show that the conditions for maximal proliferation coefficient and min- imal vitrification are as following： the basic medium with 0.2 mg/L 6-BA, 3% sucrose and 0.65% agarose; culture at 25℃; 12 h/d（ daylight lamp, 2 000 lx） illumination.展开更多
Amphioxus, a cephalochordate, is an important model fish for studies in evolution and comparative biology. A successful cell culture from amphioxus tissues in vitro would help understanding some basic issues. To deter...Amphioxus, a cephalochordate, is an important model fish for studies in evolution and comparative biology. A successful cell culture from amphioxus tissues in vitro would help understanding some basic issues. To determine the optimal culture conditions for proliferation of amphioxus cells, primary cultures were initiated from buccal cirri, tail, gill, gut and metapleural fold of amphioxus Branchiostoma belcheri tsingtauense. The media tested were L-15, F-12, M 199, MEM, DMEM, PRMI 1640 and LDF, each was supplemented with 20% fetal bovine serum. The optimal conditions include tail tissue cultured in L-15 or F-12 with supplement of 20% FBS and 1.5% NaCl at about 25℃.展开更多
To establish a micropropagation system of three Laurencia complex species （Laurencia okamurai, Laurencia tristicha, and Chondrophvcus undulatus） by tissue culture techniques, we studied the regeneration characterist...To establish a micropropagation system of three Laurencia complex species （Laurencia okamurai, Laurencia tristicha, and Chondrophvcus undulatus） by tissue culture techniques, we studied the regeneration characteristics and optimal culture conditions of axenic algal fragments cultured on solid medium and in liquid medium. Regeneration structures were observed and counted regularly under a reverse microscope to investigate the regeneration process, polarity and optimal illumination, and temperature and salinity levels. The results show that in most cultures of the three species, we obtained bud regeneration on solidified medium with 0.5% agar and in liquid medium. Rhizoid-like regeneration was filamentous and developed from the lower cut surface of fragments in L. okamurai, but was discoid and developed from the apical back side of bud regeneration in L. tristicha and C. undulatus. Regeneration polarity was localized to the apical part of algal fronds in all three species, and on fragments cut from the basal part of algae buds could develop from both the upper and the lower cut surfaces. Buds could develop from both the medullary and the cortical portions in L. okamurai and C. undulatus, while in L. tristicha, buds only emerged from the cortex. The optimal culture conditions for L. okamurai were 4 500 lx, 20℃ and 35 （salinity）; for C. undulatus, 4 500 lx, 20℃ and 30; and for L. tristicha, 4 500 lx, 25℃ and 30.展开更多
Anti-lipopolysaccharide factors （ALFs） are important antimicrobial peptides that are isolated from some aquatic species. In a previous study, we isolated ALF genes from Chinese mitten crab, Eriocheir sinensis. In th...Anti-lipopolysaccharide factors （ALFs） are important antimicrobial peptides that are isolated from some aquatic species. In a previous study, we isolated ALF genes from Chinese mitten crab, Eriocheir sinensis. In this study, we optimized the production of a recombinant ALF by expressing E. sinensis ALF genes in Eseherichia coli maintained in shake-flasks. In particular, we focused on optimization of both the medium composition and the culture condition. Various medium components were analyzed by the Plackett-Burman design, and two significant screened factors, （NH4）2SO4 and KH2PO4, were further optimized via the central composite design （CCD）. Based on the CCD analysis, we investigated the induction start-up time, the isopropylthio-D-galactoside （IPTG） concentration, the post-induction time, and the temperature by response surface methodology. We found that the highest level of ALF fusion protein was achieved in the medium containing 1.89 g/L （NH4）2SO4 and 3.18 g/L KH2PO4, with a cell optical density of 0.8 at 600 nm before induction, an IPTG concentration of 0.5 mmol/L, a post-induction temperature of 32.7~C, and a post-induction time of 4 h. Applying the whole optimization strategy using all optimal factors improved the target protein content from 6.1% （without optimization） to 13.2%. We further applied the optimized medium and conditions in high cell density cultivation, and determined that the soluble target protein constituted 105% of the total protein. Our identification of the economic medium composition, optimal culture conditions, and details of the fermentation process should facilitate the potential application of ALF for further research.展开更多
基金Fund for the Transformation of Scientific and Technological Achievements in China (2006GB2G300311)National Natural Science Foundation of China (30760127)~~
文摘The tender stems from new Lycium barbarum L. cultivar ＂Ningqi 3＂ released by Ningxia Academy of Agricultural and Forestry Sciences were regarded as explants to investigate the vitrification of Lycium barbarum plantlets in tissue culture under different concentrations of 6-BA, sucrose, agrose, culture temperature, and illumination duration with MS as basic medium. The results show that the conditions for maximal proliferation coefficient and min- imal vitrification are as following： the basic medium with 0.2 mg/L 6-BA, 3% sucrose and 0.65% agarose; culture at 25℃; 12 h/d（ daylight lamp, 2 000 lx） illumination.
基金Supported by Doctoral Initial Fund of Ludong University (No.43304)
文摘Amphioxus, a cephalochordate, is an important model fish for studies in evolution and comparative biology. A successful cell culture from amphioxus tissues in vitro would help understanding some basic issues. To determine the optimal culture conditions for proliferation of amphioxus cells, primary cultures were initiated from buccal cirri, tail, gill, gut and metapleural fold of amphioxus Branchiostoma belcheri tsingtauense. The media tested were L-15, F-12, M 199, MEM, DMEM, PRMI 1640 and LDF, each was supplemented with 20% fetal bovine serum. The optimal conditions include tail tissue cultured in L-15 or F-12 with supplement of 20% FBS and 1.5% NaCl at about 25℃.
基金Supported by the fund from Jiangsu Key Laboratory of Marine Biotechnology (No. 2006HS006)Huaihai Institute of Technology, Lianyungang, China211-Project Funding of Soochow University (No. 14134908)
文摘To establish a micropropagation system of three Laurencia complex species （Laurencia okamurai, Laurencia tristicha, and Chondrophvcus undulatus） by tissue culture techniques, we studied the regeneration characteristics and optimal culture conditions of axenic algal fragments cultured on solid medium and in liquid medium. Regeneration structures were observed and counted regularly under a reverse microscope to investigate the regeneration process, polarity and optimal illumination, and temperature and salinity levels. The results show that in most cultures of the three species, we obtained bud regeneration on solidified medium with 0.5% agar and in liquid medium. Rhizoid-like regeneration was filamentous and developed from the lower cut surface of fragments in L. okamurai, but was discoid and developed from the apical back side of bud regeneration in L. tristicha and C. undulatus. Regeneration polarity was localized to the apical part of algal fronds in all three species, and on fragments cut from the basal part of algae buds could develop from both the upper and the lower cut surfaces. Buds could develop from both the medullary and the cortical portions in L. okamurai and C. undulatus, while in L. tristicha, buds only emerged from the cortex. The optimal culture conditions for L. okamurai were 4 500 lx, 20℃ and 35 （salinity）; for C. undulatus, 4 500 lx, 20℃ and 30; and for L. tristicha, 4 500 lx, 25℃ and 30.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2006AA100311)
文摘Anti-lipopolysaccharide factors （ALFs） are important antimicrobial peptides that are isolated from some aquatic species. In a previous study, we isolated ALF genes from Chinese mitten crab, Eriocheir sinensis. In this study, we optimized the production of a recombinant ALF by expressing E. sinensis ALF genes in Eseherichia coli maintained in shake-flasks. In particular, we focused on optimization of both the medium composition and the culture condition. Various medium components were analyzed by the Plackett-Burman design, and two significant screened factors, （NH4）2SO4 and KH2PO4, were further optimized via the central composite design （CCD）. Based on the CCD analysis, we investigated the induction start-up time, the isopropylthio-D-galactoside （IPTG） concentration, the post-induction time, and the temperature by response surface methodology. We found that the highest level of ALF fusion protein was achieved in the medium containing 1.89 g/L （NH4）2SO4 and 3.18 g/L KH2PO4, with a cell optical density of 0.8 at 600 nm before induction, an IPTG concentration of 0.5 mmol/L, a post-induction temperature of 32.7~C, and a post-induction time of 4 h. Applying the whole optimization strategy using all optimal factors improved the target protein content from 6.1% （without optimization） to 13.2%. We further applied the optimized medium and conditions in high cell density cultivation, and determined that the soluble target protein constituted 105% of the total protein. Our identification of the economic medium composition, optimal culture conditions, and details of the fermentation process should facilitate the potential application of ALF for further research.
