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1.
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°C.
Supported by Doctoral Initial Fund of Ludong University (No.43304) 相似文献
2.
Mature amphioxus (Branchiostoma belcheri tsingtauense) collected by trawl in June, 1994 from Qingdao Shazikou sea area bottom sands, were cultured under controlled indoor conditions, where they spawned and their eggs were fertilized and hatched into larvae which gradually settled at the bottom and dug into the sands after 40-50 days culture in a water trough outdoors. The total survival rate of the larvae(LSR)was 5.5% before they went into the sands, 1.9% in 4 months, 0.7% in 10 months. The survival rate of the young fish which had dug into the sands (YSR) was 35.6% in 4 months, and 12.6% in 10 months. The amphioxus number tended to be constant from the 11th month on. In the first five months after incubation, the amphioxus body length increased by an average of about 1.5 mm a month, and about 0.4 mm a month from Dec. to May of next year. The maximum length after ten months was 24 mm; the average was about 11 mm. After two years culture in the water trough, the maximum length could reach 34-35 mm, when the amphioxus gonads began to develop. 相似文献
3.
为研究生物酶采油解堵剂中产蛋白酶菌株的初、复筛选及培养条件优化,从大庆原油样品中筛选菌种,通过水解酪素的透明圈实验及福林酚测蛋白酶酶活的方法进行菌株的初、复筛选;以蛋白酶酶活为优化指标,采用单因素实验对筛选的产蛋白酶菌株的培养基及培养条件进行优化,优化最适培养基:可溶性淀粉为15g/L,蛋白胨为20g/L,酵母膏为20g/L,NaCl为1.0g/L,CaCl2为0.02g/L,Na2HPO4为0.2g/L,NaH2PO4为0.1g/L;在初始pH为6.0、接种量为5%(体积分数)、温度为31℃、摇床转速为160r/min的条件下,培养72h后,菌株的蛋白酶酶活为551.0U/mL,为复筛选菌株的蛋白酶酶活的22.92倍,即为菌株生长繁殖及代谢的最佳条件,能够获得更高的蛋白酶酶活,有利于后续实验的进行.结果表明:菌株产蛋白酶对原油作用效果为发酵液表面张力从作用前的56.2mN/m降低到作用后的30.5mN/m,表面张力显著降低,还有降解降黏原油等效果,具有一定的研究价值. 相似文献
4.
ZHU Luying ZHANG Xuecheng REN Xueying ZHU Qinghua 《中国海洋大学学报(英文版)》2008,7(1):83-88
The effects of temperature, initial pH, salinity of culture medium, and carbon and nitrogen sources on growth and docosahexaenoic acid （C22：6 n-3, DHA） production from Schizochytrium limacinum OUC88 were investigated in the present study. The results revealed that the optimal temperature, initial pH and salinity level of the medium for DHA production were 23 ℃, 7.0 and 18, respectively. Glucose was proved the best carbon source for the growth and DHA production from S. limacinum. Among the nitrogen sources tested, soybean cake hydrolysate, a cheap by-product, was found to be effective for the accumulation of DHA in S. limacinum cells. In addition, increasing the concentration of carbon sources in the medium caused a significant increase in cell biomass; however, accumulation of DHA in cells was mainly stimulated by the ratio of C/N in the medium. Under the optimal culture conditions, the maximum DHA yield achieved in flasks was 4.08 g L^-1 after 5 d of cultivation 相似文献
5.
研究分离自对虾养殖池的一株促藻生长细菌的最佳培养条件,通过单因子实验得到该菌株的最佳碳源为葡萄糖,最佳氮源为酵母膏,初始pH值为7.5,最佳氯化钠质量浓度5 g/L;在其他培养条件不变的情况下,通过对酵母膏、葡萄糖、初始pH值做三因子三水平的正交实验,得到培养基最佳组合为:酵母膏10 g/L,葡萄糖1 g/L,初始pH值7.5。 相似文献
6.
Arctic sea ice in the polar region provides a cold habitat for microbial community. Arctic sea ice microorganisms are revealed to be of considerable importance in basic research and potential in biotechnological application. This paper investigated the culture condition and extraceIlular hydrolase of 14 strains of different Arctic sea ice bacteria. The results showed that optimal growth temperature of strains is 15 ℃ or 20 ℃. The optimal pH is about 8.0. They hardly grow at acid condition. 3 % NaCl is necessary for better growth. These strains have different abilities in producing amylase, protease, eellulase and lipase. Pseudoalteronomas sp. Bsi429 and Pseudoalteronomas sp. Bsi539 produced both cellulose, protease and lipase. These results provide a basis for further developing and exploiting the cold adapted marine enzyme resources. 相似文献
7.
吕庆芳 《广东海洋大学学报》2003,23(4):57-61
以粉蕉的吸芽为试材 ,进行了离体快速繁殖研究 ,结果表明 :以改良MS +BA4 .0mg/L(单位下同 ) +AD5.0 +NAA0 .1作诱导培养基 ,外植体褐变率低 ,芽的诱导率高 ,且芽的生长良好 ;以改良MS +BA3- 6 +NAA0 .1为继代培养基 ,可以获得较高的不定芽增殖率 ,在增殖过程中 ,给予一定的低温处理和变化调整BA的用量 ,可以保持较高的增殖率 (2 .6倍以上 ) ,而且芽的长势好 ;生根培养以 1/ 2MS +IBA1.0 +NAA0 .2 +AC1为宜。 相似文献
8.
As seedling culture is a crcial factor for successful cultivation of Gelidium, the authors researched tis-sue culture technology for producing seedlings. The merphogeny and experirmental ecology were observedand studied fully in 2-5 mm isoIated tissue fragments, appearaare of branching creepersand attaching structure and new erect seedlings production and development were studied. Fragments weresown on bamboo slice and vinyon rope. The seedlings were cultured 20-30 days indoor, then cultured inthe sea, where the density of erect seedlings was 3- 19 seedlings/cm~2, growth rate was 3.84%/day. The frondarising from seedlings directly was up to 10 cm per year. The ecological conditions for regenerated seedlingsare similar to the natural ones. The regenerated seedlings are sintable for raft culture in various sea areas. 相似文献
9.
西津水库网箱养殖区浮游植物特征及水质营养状态 总被引：2，自引：0，他引：2
于2006年10月至2008年7月对西津水库米埠坑网箱养殖区浮游植物进行两个年度共8次调查。第1年发现浮游植物6门53属,冬季出现种属最多,达到43属,春、夏季最少,为35属,浮游植物年平均数量为35.18×104 L-1,平均生物量为1.07 mg·L-1。第2年度采集到浮游植物6门47属,以春季出现种属最多,达到40属,夏季最少,为29属,第2年浮游植物年平均数量低于第1年,为19.77×104 L-1,生物量高于第1年,达到1.22 mg·L-1。综合营养指数评价结果显示,米埠坑网箱养殖区处于中营养水平,有向富营养化过渡的趋势。 相似文献
10.
During a survey for marine microalgal resources, we isolated a rare marine euglenoid from the coastal waters of Qingdao, China in 2009, and established a pure culture. Electron microscopic and molecular phylogenetic (18S rDNA and 16S rDNA sequences) analyses revealed a close affinity with Eutreptiella gymnastica, a bloom-forming species. Different culture conditions were monitored to understand optimal E. gymnastica growth characteristics. The optimal growth conditions in a batch culture of this isolate were 20°C, 160 μmol photons/(m2×s) of white light, and a salinity of 10-31. Nutrient experiments demonstrated that growth increased dramatically with a phosphorus concentration greater than 72 μmol/L. Understanding the effect of culture conditions on E. gymnastica may help understanding the blooming mechanism of this alga in its natural environment. 相似文献
11.
Algal biotechnology has advanced greatly in the past three decades. Many microalgae are now cultivated to produce bioactive substances. Odontella aurita is a marine diatom industrially cultured in outdoor open ponds and used for human nutrition. For the first time, we have systematically investigated the effects of culture conditions in cylindrical glass columns and fiat-plate photobioreactors, including nutrients （nitrogen, phosphorus, silicon, and sulfur）, light intensity and light path, on O. aurita cell growth and biochemical composition （protein, carbohydrate, β-1,3-glucan, lipids, and ash）. The optimal medium for photoautotrophic cultivation of O. aurita contained 17.65 mmol/L nitrogen, 1.09 mmol/L phosphorus, 0.42 mmol/L silicon, and 24.51 mmol/L sulfur, yielding a maximum biomass production of 6.1-6.8 g/L and 6.7-7.8 g/L under low and high light, respectively. Scale-up experiments were conducted with fiat-plate photobioreactors using different light-paths, indicating that a short light path was more suitable for biomass production of O. aurita. Analyses of biochemical composition showed that protein content decreased while carbohydrate （mainly composed of 15-1,3-glucan） increased remarkably to about 50% of dry weight during the entire culture period. The highest lipid content （19.7% of dry weight） was obtained under 0.11 mmol/L silicon and high light conditions at harvest time. Fatty acid profiles revealed that 80% were Cx4, C~6, and C20, while arachidonic acid and eicosapentaenoic acid （EPA） accounted for 1.6%-5.6% and 9%-20% of total fatty acids, respectively. High biomass production and characteristic biochemical composition profiles make O. aurita a promising microalga for the production ofbioactive components, such as EPA and D-1,3-glucan. 相似文献
12.
As seedling culture is a crucial factor for successful cultivation ofGelidium, the authors researched tissue culture technology for producing seedlings. The morphogeny and experimental ecology were observed
and studied fully in 2–5 mm isolated tissue fragments. Regeneration, appearance of branching creepers and attaching structure
and new erect seedlings production and development were studied. Fragments were sown on bamboo slice and vinylon rope. The
seedlings were cultured 20–30 days indoor, then cultured in the sea, where the density of erect seedlings was 3–19 seedlings/cm2, growth rate was 3.84% day. The frond arising from seedlings directly was up to 10 cm per year. The ecological conditions
for regenerated seedlings are similar to the natural ones. The regenerated seedlings are suitable for raft culture in various
sea areas. 相似文献
13.
CHEN Xin WANG Chunlan XU Jiakun WANG Fang JIANG Yihui CHEN Yixuan and ZHAO Xianyong 《中国海洋大学学报(英文版)》2020,19(1):209-215
Lipase from Antarctic krill,with a molecular weight of 71.27kDa,was purified with ammonium sulfate precipitation and a series of chromatographic separations over ion exchange(DEAE)and gel filtration columns(Sephacryl S-100),resulting in 5.2%recovery with a 22.4-fold purification ratio.The optimal pH and temperature for enzyme activity were 8.0 and 45℃,respectively.Purified lipase had Km and Vmax values of 3.27mmolL−1 and 2.4Umg−1,respectively,using p-nitrophenyl laurate as the substrate.Lipase activity was enhanced by adding Ca2+and Mg2+ions in the concentration ranges of 0–0.5mmolL−1 and 0–0.3mmolL−1,respectively,while the activity was inhibited by a further increase in these ion concentrations.Fe3+and Cu2+ions showed obvious inhibitory effects on enzyme activity,and the inhibition rates were 71.8%and 53.3%when the ion concentrations were 0.5mmolL−1. 相似文献
14.
研究无乳链球菌(Streptococcus agalactiae)ZQGY08的培养与生长特性,探讨脑心浸液(Brian Heart Infusion,BHI)培养基的营养因子、初始pH值、溶氧、温度等因素对无乳链球菌生长的影响,并采用正交实验法对无乳链球菌培养基营养因子添加量进行优化。结果表明,无乳链球菌生长的最适初始pH为7～8,培养温度为28～37℃,振荡转速为180～200 r/min。在BHI培养基中添加20 mg/mL的酵母粉和2 mg/mL的葡萄糖即可在对数期获得最大细菌量。 相似文献
15.
波吉卵囊藻的固定化培养及利用 总被引：3，自引：0，他引：3
以褐藻酸钙为固定化载体,探讨了胶球直径、不同接种量、CaCl2溶液浓度、褐藻酸钠浓度等条件对波吉卵囊藻(Oocystisborgei)固定化培养及对养殖水体水质的影响。结果表明:波吉卵囊藻固定化培养的最好条件是微藻接种量为1×106 mL-1,CaCl2溶液的质量浓度为30g/L,褐藻酸钠溶液的质量浓度为20g/L。在此条件下制备的固定化藻珠,波吉卵囊藻的生物量从1 83×10-2 mg/粒增加到17. 46×10-2 mg/粒,增加了近10倍,生长率相对较高,证明了它的生理活性不会因固定化而受干扰。引入固定化的波吉卵囊藻不但可以降低养殖水体中氨氮、亚硝酸氮等有害因子的浓度,还能提高水中溶解氧含量,使水体环境长时间处于良好的动态平衡状态。 相似文献
16.
To establish a micropropagation system of three Laurencia complex species (Laurencia okamurai, Laurencia tristicha, and Chondrophycus 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°C and 35 (salinity); for C. undulatus, 4 500 lx, 20°C and 30; and for L. tristicha, 4 500 lx, 25°C and 30. 相似文献
17.
Shiqi Ji Rui Zhao Qi Yin Yuan Zhao Chenguang Liu Tian Xiao Xiaohua Zhang 《中国海洋大学学报(英文版)》2012,11(1):45-51
A gel microbead (GMD) cultivation method was employed to cultivate microorganisms from an amphioxus breeding zone in Qingdao, P. R. China. The culture results were compared with those by standard plating method. In the GMD-based method, the microcolony-forming GMDs were sorted by fluorescence-activated cell sorting (FACS). To further get pure cultures, a subsequent enrichment culture and a streaking purification procedure were conducted on marine R2A medium. Eighty bacterial strains isolated by the GMD-based method were randomly selected for sequencing. These isolates belonged to Alphaproteobacteria (33%), Gammaproteobacteria (44%), Bacteroidetes (11%), Actinobacteria (5%), Firmicutes (5%), Epsilonproteobacteria (1%), and Verrucomicrobia (1%), the last two groups being usually difficult to culture. The 16S rRNA gene sequences revealed a diverse community with 91.1%-100% of the bacterial rRNAs similarities. Thirteen strains were sharing 16S rRNA gene sequence which was less than 97% similar to any other rRNA genes currently deposited in TYP16S database. Seventy isolates derived from the standard plating method fell into 4 different taxonomic groups: Alphaproteobacteria (9%), Gammaproteobacteria (81%), Bacteroidetes (7%) and Firmicutes (3%) with a 16S rRNA gene sequence similarities between 95.8%-100%, in which only 3 strains were sharing 16S rRNA gene sequence of less than 97%. The results indicated that the GMD-based method with subenrichment culture yielded more taxonomic groups and more novel microbial strains, including members of previously rarely cultured groups, when compared with the standard plating method, and that this method markedly improved the bacterial cultivability. 相似文献
18.
Information regarding antioxidant enzymes in amphioxus remains lacking, and this study was carried out to examine the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) in different genders and tissues of amphioxus Branchiostoma belcheri tsingtauense. Results show that (1) CuZn-SOD, CAT and GPX activities in the whole amphioxus B. belcheri tsing- tauense were basically at the same levels in male and female amphioxus, whereas both T-SOD and Mn-SOD activities in male amphioxus were significantly higher than that in the female (P<0.05); (2) The testis had significantly higher T-SOD and CuZn-SOD activities than the ovary (P<0.05); (3) CuZn-SOD activity was undetectable in the guts of male and female amphioxus; (4) For both male and female am- phioxus, the activities of CAT and GPX in the gonads including testis and ovary were the lowest (P<0.05) among the tissues examined; (5) The gut and gill had the same level GPX activities while the gut had a higher CAT activity; (6) There was no clear difference in CAT and GPX activities in the corresponding tissues between male and female amphioxus. The study on SOD, CAT and GPX activities in different genders and tissues of the protochordate provides data for future comparison of amphioxus antioxidant enzymes with those of invertebrates and vertebrates. 相似文献
19.
比较在0、5、10、15、20、25、30(自然海水)、35、40、45、50等11个盐度梯度下,菊黄东方鲀(Takiugu flavidus)的受精卵发育和仔鱼生长情况,测定仔鱼的不投饵存活系数(SAI).结果表明:1)菊黄东方鲀受精卵在盐度0~50范围内均可孵化,最适盐度20~25.盐度(x)与孵化时间(y)呈正相关关系,表达式为:y=0.345 x+141.1,R2=0.956.2)实验盐度范围内,菊黄东方鲀仔鱼SAI值为1.55~30.10.3)在盐度5~45条件下,仔鱼皆可存活,存活率为3%~27%.其中,在10~30盐度下存活率较高,为20%~27%.经15 d的培育,仔鱼平均全长为6.28~7.34 mm.低盐度对菊黄东方鲀仔鱼生长更有利,在15~20盐度条件下,仔鱼生长较好,最终平均全长为7.30~7.34 mm.4)根据SAI值和生长状况,仔鱼生长和存活的适宜生长盐度为10~30,最适盐度范围为15~20. 相似文献
20.
Wu Xian-han Zhang Shi-cui Wang Yong-yuan Zhang Bao-lu Qu Yan-mei Jiang Xin-ji 《中国海洋湖沼学报》1994,12(4):289-294
Although amphioxus is widespread in temperate and tropical seas, its population is diminishing because of environmental pollution.
To keep the population of this evolutionarily important animal from diminishing, study on its reproduction and development
is necessary. The main findings in this study on the spawning and fecundity of the amphioxus reared in laboratory and its
larval development are as follows. 1. Water temperature markedly affected the spawning. It spawned only when water temperature
reached 21°C. 2. Spawning of the amphioxus in laboratory was markedly extended. Initially, the amphioxus spawned at about
7:00 PM, but spawning time was postponed as spawning days went on. 3. The number of eggs produced by a female ranged from
1400 to 12800, average of 5800. This also represents the fecundity of the amphioxus because it shedded all eggs within the
ovary at a time. 4. During the first few months of life of the amphioxus, its growth rate changed seasonally. The growth rate
in summer and fall was greater than that in winter. 5. The pelagic larva became a benthic adult after 50 days. 6. The amphioxus
reared in laboratory from fertilized eggs could produce fertile eggs and sperms. These findings can be a foundation for measures
to address the problem of diminishing amphioxus population.
Contribution No. 2274 from the Institute of Oceanology, Chinese Academy of Sciences
This work was supported by CNNSF, and Chinese Academy of Sciences. 相似文献