Orthogonal combination design was adopted in examining the Spirulina platensis (S. platensis) yield and the influence of four factors (Se content, Se-adding method, S content and NaHCO3 content) on algae growth. The results showed that Se content, Se-adding method and NaHCO3 content were key factors in cultivation conditions of Se-enriched S. platensis with the optimal combination being Se at 300 mg/L, Se-adding amount equally divided into three times and NaHCO3 at 16.8 g/L. Algae yield had a remarkable correlation with OD560 and floating rate by linear regression analysis. There was a corresponding relationship between effects of the four factors on algae yield and on OD560, floating rate too. In conclusion, OD560 and floating rate could be served as yield-forming factors.
Bicarbonates ; analysis ; Culture Media ; Cyanobacteria ; growth & development ; Selenium ; analysis ; pharmacology

The effect of CO2 and the manner of CO2 offer on the growth rate and maximual cell density of ultro-high density culture of Chaetoceros mulleri in the photobioreactor were studied in the work. The amount of CO2 offered to the culture was controlled by the parameter of pH value in the culture. Furthermore the growth kinetics of Chaetoceros muller in the photobioreactor was studied. The results showed requirement of CO2 by the cells and the increase of pH in the culture were the key limiting factors to the growth, when a high cell concentration in the culture was reached. The offer of CO2 could improve the statute of CO2, could control the pH in the culture and increase the growth rate and maximum cell density. The results from the experiments of CO2 offer manner showed different efficiency to growth was resulted from differences of CO2 offer manner. The best way is mixing the CO2 and air before the CO2 was offered to the culture.
Bioreactors ; Carbon Dioxide ; pharmacology ; Culture Media ; Culture Techniques ; methods ; Diatoms ; growth & development

OBJECTIVETo research the optimal conditions for the callus induction of anther culture and the plant regeneration of Angelica dahurica var. formosana.

METHODCallus was induced from the anther of A. dahurica from Sichuan province on a MS medium. The effects of callus induction and plant regeneration of different pretreatment hours under low temperature (4 degrees C), different culturing conditions under darkness and illumination, and different culture with different hormone contents and ratios were studied.

RESULTThe results showed that A. dahurica anthers without low temperature pretreatment reached the highest induction rate then under the pretreatment under low temperature (4 degrees C) for two days. The optimal culturing condition was under the darkness. The culturing efficiency reached 38.89% on the medium of MS + 2.0 mg x L(-1) 2,4-D + 1.0 mg x L(-1) 6-BA. The optimum medium for differentiate anther callus was MS + 0.5 mg x L(-1) NAA + 1.5 mg x L(-1) KT + 10 mg x L(-1) AgNO3. 1/2MS medium supplemented with 0.5 mg x L(-1) IBA could well promote seedings to take roots.

CONCLUSIONAn efficient system for callus induction of anther culture and plant regeneration of A. dahurica was preliminarily established.

Angelica ; drug effects ; growth & development ; Culture Media ; chemistry ; pharmacology ; Flowers ; Tissue Culture Techniques ; methods

The effect of sugars, gibberellic acid (GA3) and abscisic acid (ABA) on somatic embryogenesis from internodal explant-derived callus of Tylophora indica (Burm. f.) Merrill has been investigated. Embryogenic calli were produced from internodal explants and the best result was achieved by using MS medium supplemented with 4micromol/L 2, 4-Dichlorophenoxyacetic acid (2, 4-D). Up to 69% of such embryogenic calli differentiated into somatic embryos with an average of 25 embryos per explant (per gram of the calli) on Murashige and Skoog (MS) medium containing 6micromol/L kinetin (Kn). The individual effect of sucrose and glucose together with 6micromol/L Kn was evaluated. There was a significant difference among concentrations of sugar and among kinds of sugar tested in somatic embryogenesis. Sucrose at 200mmol/L with 6micromol/L Kn gave rise to a maximum embryogenesis (71%) with an average of 49 embryos per explant. However, glucose together with 6micromol/L Kn or a combination of glucose, sucrose and 6micromol/L Kn reduced the percentage of embryogenesis culture and the number of embryos per explant. The presence of GA3 and ABA at particular concentrations promoted somatic embryogenesis in T. indica. The addition of 10mol/L GA3 into the 200mmol/L sucrose-containing medium gave a 98% embryogenesis response with an average of 51 embryos per explant. Somatic embryogenesis was significantly enhanced by the addition of 2micromol/L ABA to 200mmol/L sucrose-containing medium. On this medium 95% embryogenesis with an average of 44 embryos per explant was observed. The study reported here indicates that 200mmol/L sucrose with 6micromol/L Kn, 200mmol/L sucrose with 10micromol/L GA3 and 200mmol/L sucrose with 2micromol/L ABA significantly improved somatic embryogenesis in T. indica whereas glucose alone or in combination with sucrose had an inhibitory role. The embryos obtained developed normally and were easily converted into plants.
Abscisic Acid ; pharmacology ; Carbohydrates ; pharmacology ; Culture Media ; Culture Techniques ; Gibberellins ; pharmacology ; Plant Shoots ; embryology ; growth & development ; Tylophora ; embryology ; growth & development

The diatom Nitzschia laevis is a good alternative source of eicosapentaenoic acid (EPA). Besides strategies for high cell density culture, EPA productivity may be further improved by herbicides. The effect of the herbicide quizalofop-p-ethyl on the growth and EPA production was studied in this paper. As the solvent of the herbicide, DMSO was proved to inhibit the growth and EPA production of N. laevis. The concentration of DMSO in the medium should not exceed 0.2%. Quizalofop-p-ethyl could cause morphology damage to the N. laevis cells. With the increasing concentration of quizalofop-p-ethyl from 0 mmol/L to 0.4 mmol/L, the dry cell weight production decreased, while at the same time, the lipid content of the dry cell mass increased. When treated with 0.1 mmol/L quizalofop-p-ethyl, the EPA content increased from 3.00% to 3.58% (of dry cell weight, DW), and the proportion of EPA (20:5) in total fatty acids (TFA) increased from 25.15% to 32.88% . These results indicated that the herbicide quizalofop-p-ethyl could stimulate the accumulation of EPA; therefore it might be useful for selecting algae colonies that overproduce EPA.
Culture Media ; Culture Techniques ; Diatoms ; growth & development ; metabolism ; Eicosapentaenoic Acid ; biosynthesis ; Herbicides ; pharmacology ; Propionates ; pharmacology ; Quinoxalines ; pharmacology

OBJECTIVETo study tissue culture and plant regeneration of Withania somnifera.

METHODLeaves of W. somnifera were used for explants, effects of different plant growth substances on callus and shoot induction were studied, different medium and plant growth substances for rooting induction was optimized.

RESULT AND CONCLUSIONThe best plant growth substances combination for callus induction was MS + 1.0 mg x L(-1) 2,4-D + 0.1 mg x L(-1) KT. The optimal medium for germination was MS + 1.0 mg x L(-1) 6-BA + 0.1 mg x L(-1) NAA. The best medium and plant growth substances combination for rooting induction was 1/2MS + 0.5 mg x L(-1) NAA, transplant survival rate of plantlet reached 92% in humus soil-pearlite (1:1).

Culture Media ; pharmacology ; Plant Growth Regulators ; pharmacology ; Regeneration ; drug effects ; Tissue Culture Techniques ; Withania ; drug effects ; growth & development

OBJECTIVETo study the effects of different factors on buds and microtuber. These factors included plant growth substances and sucrose.

METHODstems were selected as explants. The effects of three kinds of factors were studied by orthogonal design method including sucrose, 6-BA, NAA on the buds and microtuber induction. The data were analyzed with range analysis and vadance analysis. RESULT AND CONDUSION: The optimal media to induce many buds from stems were MS + 6-BA 1 mg x L(-1) + NAA 1 mg x L(-1) + sucrose 3%, the effect of the three factors was in sequence of sucrose >6-BA > NAA. The optimal media to induce microtuber from stems were MS+6-BA 1.5 mg x V1 +NAA 1.5 mg x L(-1) + sucrose 5%, the effect of the three factors was in sequence of sucrose >6-BA > NAA.

Culture Media ; Dioscorea ; drug effects ; growth & development ; Plant Growth Regulators ; pharmacology ; Sucrose ; pharmacology ; Tissue Culture Techniques ; methods

To demonstrate the fructose metabolism pathway in Bifidobacterium Longum NCC2705 and to construct its fermentation model, we explored the comparative proteome cultivating the strain on glucose or fructose, based on a proteomic reference map of B. longum NCC2705 constructed earlier. Then, we used matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry and electro-spray ionization tandem mass spectrometry (ESI-MS/MS) for differently expressed proteins identification. Furthermore, with semi-quantitative RT-PCR we determined the distinctively expressed proteins at the level of transcription. Proteomic comparison of glucose- and fructose-grown cells demonstrated much similarity. On the page of fructose there were all the enzymes and proteins that exist during the process of glucose degradation. We observed a greater variation of more than three-fold for the identified 9 spots representing 5 protein entries by MALDI-TOF MS. The sugar-binding protein specific to fructose (BL0033) and an ABC transporter ATP binding protein (BL0034) showed higher expression level from cells grown on fructose. It was also determined by semi-quantitative RT-PCR subsequently. BL0033 time course and concentration experiments showed that the induction time correlated to higher fructose concentration, and increased expression of BL0033. Fructose was catabolized via the same degradation pathway as glucose at the level of proteomics. BL0033 was induced by fructose. All results suggest that the uptake of fructose into the cell may be conducted by a specific ABC transport system, in which BL0033 and BL0034 as components might have played an important role.
Bifidobacterium ; chemistry ; genetics ; metabolism ; Culture Media ; Fermentation ; Fructose ; pharmacology ; Glucose ; pharmacology ; Proteome ; analysis ; genetics ; Proteomics ; methods

We studied medium alkalinization in Salvia miltiorrhiza suspension cultures treated with salicylic acid and the effect of Ca2+ in this process through application of calcium channel antagonists (Verapamil, LaCl3, LiCl, 2-APB) and ionophore A23187. The results show that salicylic acid could induce significant medium alkalinization in S. miltiorrhiza culture. Verapamil and LaCl3 or LiCl and 2-APB, two different groups of calcium channel antagonist, significantly inhibited the medium alkalinization induced by salicylic acid. However, the suppression effect of verapamil or LaCl3 on medium alkalinization induced by salicylic acid was higher than that of LiCl or 2-APB. When two types of calcium channel inhibitor (LaCl3 and 2-APB) were used together, the medium alkalinization induced by salicylic acid was completely suppressed and even reduced the pH in medium. On the other hand, A23187 could promote the medium alkalinization. Based on the results above, we speculated that salicylic acid could induce significant medium alkalinization in S. miltiorrhiza culture, depending on the calcium from both extracell and intracell. Moreover, calcium from extracell plays a more dominant role in this process. Reveal of relationship in this research between Ca2+ and medium alkalinization can provide theory evidence for mechanism of the plant secondary metabolism.
Calcimycin ; pharmacology ; Calcium ; chemistry ; Calcium Channel Blockers ; pharmacology ; Calcium Ionophores ; pharmacology ; Cell Culture Techniques ; Culture Media ; chemistry ; Salicylic Acid ; pharmacology ; Salvia miltiorrhiza ; metabolism ; Verapamil ; pharmacology

To study the effects of serum-free murine bone marrow endothelial cell conditioned medium (mBMEC-CM) on the growth of bone marrow endothelial cells, mBMEC-CM was collected and ultrafiltrated by Centriprep-10. The retentate of mBMEC-CM [molecular weight (MW)>10 kDa] and the filtrate of mBMEC-CM (MW<10 kDa) were obtained. The effect of bone marrow conditioned media, their components and exogenous cytokines on the formation of endothelial cell colonies were observed. The effect of bone marrow conditioned media, their components and exogenous cytokines on the proliferation of murine bone marrow endothelial cells were determined by [(3)H]-thymidine incorporation. The method of hybridizing to the Atlas cDNA array was used to determine the expression of cytokine mRNAs in bone marrow endothelial cells. The results obtained are as follows: vWF was expressed in bone marrow endothelial cells. The original mBMEC-CM and MW>10 kDa component of mBMEC-CM promoted the proliferation of bone marrow endothelial cell colonies and increased [(3)H]-thymidine incorporation of bone marrow endothelial cells. The MW<10 kDa component did not affect the production of endothelial cell colonies and did not increase [(3)H]-thymidine incorporation of endothelial cells. Six cytokines (IL-6, IL-11, SCF, GM-CSF, VEGF, bFGF) promoted the proliferation of bone marrow endothelial cell colonies. VEGF, bFGF and SCF increased [(3)H]-thymidine incorporation of bone marrow endothelial cells. According to the results of the Atlas cDNA array, GM-CSF,TGF-beta,BMP-2, bFGF, SCF, endothelin-2, thymosin beta10, MSP-1, connective tissue GF, PDGF-A chain, MIP-2 alpha, PlGF, neutrophil activating protein ENA-78, INF-gamma, IL-1, IL-6, IL-13, IL-11, inhibin-alpha mRNAs were expressed in endothelial cells. These results suggest that murine bone marrow endothelial cell conditioned medium promotes the proliferation of bone marrow endothelial cells.
Animals ; Bone Marrow Cells ; cytology ; Cell Line ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media, Conditioned ; pharmacology ; Culture Media, Serum-Free ; pharmacology ; Endothelial Cells ; cytology ; Hematopoiesis ; physiology ; Mice