无血清培养基干粉的生产工艺优化

为了研究瞬时温度的提高对培养基加工的影响,通过不同的固定温度处理培养基,考察不同温度长时间处理后的培养基对CHO细胞的生长和表达的影响。结果表明,40℃处理后的效果最佳,处理工序与对照工艺相比,大大简化,产能有很大提升,为大规模生产培养基设备的选型和生产参数设置提供理论依据,且提高了设备的生产效率。     

利用铜钱草提升植物光合作用系列实验的效率

在光合作用系列实验中,教材中提到的经典实验材料天竺葵、金鱼藻在冬季难觅,影响实验的开设率。阐述了铜钱草在光合作用系列实验中的应用,并通过改进实验装置、实验步骤等,提高实验效率。     

用家用高压锅消毒微生物培养基

随着新课程标准的实施,生物学实验课的范围更加广泛。初中教材和高中选修课本中,都出现了制备培养基,培养微生物的实验。实验用具、用材的无菌是微生物学实验成败的关键所在。在有条件的学校,一般都采用高压灭菌锅进行培养基灭菌。但在一些农村中学,由于缺乏相关设备,培养基的消毒灭菌就很难进行。于是,我们设想用家用高压锅代替高压灭菌锅来消毒培养基,并对其效果进行初步的探索。     

蘑菇增产效果实验

蘑菇增产效果实验蒋友根（江西省新余市师范学校３３６５２５）１选题目的探索温度、湿度差和不同的覆土厚度对蘑菇产量的影响。２实验过程２．１准备在无菌操作条件下，用小刀切开新鲜蘑菇菌柄，用镊子夹一小块菌柄组织于马铃薯琼脂培养基表面，置２２℃温度下避光培养成...     

“分离以尿素为氮源的微生物”的实验改进

实验过程中,对尿素培养基的配方进行了改进,并思考如何更好地体现实验的重复性原则,如何更好地观察实验结果。改进后实验严谨合理,结果明显。     

不同生长调节剂对狗肝菜愈伤组织诱导和离体快繁的影响

目的 :研究不同生长调节剂对狗肝菜愈伤组织诱导和离体快繁的影响。方法 :狗肝菜不同外植体在附加不同生长调节剂的培养基上诱导愈伤组织 ,比较愈伤组织的诱导率 ;用 3因子 5水平的正交实验 ,比较不同生长调节剂对丛生芽诱导的影响 ;在附加不同生长调节剂的培养基上比较芽增殖倍数 ;附加不同浓度NAA的培养基上比较生根效果。结果 :愈伤组织诱导率相对以叶片最高 ,茎段次之 ,最后为叶柄 ;愈伤组织诱导的最佳培养基为MS 6-BA0 .5 NAA1 .5 ;不同激素对茎段芽诱导的影响次序为 6-BA>KT >NAA ,芽诱导的最佳培养基为MS 6-BA2mg/L KT1mg/L NAA0 .5mg/L ;芽继代增殖的最佳激素组合是MS 6-BA2mg/L NAA2mg/L ,增殖倍数达 3.0 0 ,影响芽继代增殖的因素次序为 6-BA >NAANAA0 .5mg/L的生根效果较好。结论 :附加一定的生长调节剂能提高狗肝菜愈伤组织的诱导率和离体快繁的效率。     

固定化Ralstonia metallidurans CH34降解苯酚的研究

将既能耐抗重金属又能降解苯酚的细菌Ralstonia m etalliduransCH34固定化以提高其降酚效率。首先通过正交实验,得到了固定化该菌种的最优制备条件,然后对固定化细胞的降酚效果进行了研究。结果表明,固定化R.m etalliduransCH34的降酚效果明显优于游离细胞;抗重金属毒性方面也有较大提高;在加入额外碳源(甲苯,柠檬酸)情况下,固定化R.m etalliduransCH34进行苯酚降解时所受影响明显要小于游离态菌。     

一种从土壤样品中选择性分离假诺卡氏菌的方法

为提高假诺卡氏放线菌的分离效率,根据其营养特性和对抗生素的敏感性,设计、检验了5种选择性分离培养基;实验检测了模式菌株在不同培养基上的生长情况,结果表明培养基S1和S2对假诺卡氏菌的生长有显著的选择性。经该方法从韩国、印度尼西亚和中国广西地区不同土样中分离到一些假诺卡氏放线菌。     

一种从土壤样品选择性分离假诺卡氏菌的方法

为提高假诺卡氏放线菌的分离效率,根据其营养特性和对抗生素的敏感性,设计、检验了５种选择性分离培养基;实验检测了模式菌株在不同培养基上的生长情况,结果表明培养基Ｓ１和Ｓ２对假诺卡氏菌生长有显著的选择性。经该方法从韩国、印度尼西亚和中国广西地区不同土样中分离到一收诺卡氏放线菌。     

酿酒酵母S.cerevisiae高密度培养条件优化研究

考察了培养基组成和培养条件对酿酒酵母Saccharomyces cerevisiae发酵的影响。以TB培养基为初始培养基,通过正交实验设计优化培养基组成,确定了影响酵母细胞产量最主要的因素是葡萄糖,最适培养基组成为:酪蛋白胨15 g/L,酵母粉25 g/L,葡萄糖30 g/L,KH2PO42.4g/L,K2HPO4.3H2O 16.34 g/L。并确定了最佳培养条件:温度30℃,转速150 r/min。采用优化培养基及培养条件下进行发酵,菌液最高OD600值和细胞密度分别达15.82和2.03×108/mL,比优化前分别提高24.2%和22.0%。     

Effect of genotype,induction medium,carbohydrate source,and polyethylene glycol on embryogenesis in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) anther culture

This study conducted two experiments involving in vitro anther culture of Zea mays L. The first experiment tested 46 maize genotypes, including inbred lines, single and three-way cross hybrids, and line A188 as control, in three different induction basal media (IMSS, N6 and YPm) for their androgenic responses. The results showed that the embryos were established 2–3 weeks after the anthers of the few responsive genotypes were cultured. Most responsive genotypes produced embryos in at least one of the three basal media; therefore, genotype is more important than the type of medium for androgenesis in maize. The mean number of anthers that developed to embryo ranged from 19 embryos per Petri dish in YPm medium for the cross (DH5 × DH7) genotype to 0 for some maize genotypes. In the second experiment, this research reports for the first time the effect of carbohydrates and polyethylene glycol (PEG) as a non-metabolized osmoticum on the embryogenesis anther culture of maize. The genotype DH5 × DH7 was used for this experiment, and the media were varied by altering sucrose, maltose, and PEG concentrations. Results showed that the maximum embryogenesis (32 embryos per Petri dish) was obtained by YPm basal medium supplemented with 60 gl^?1 sucrose + 0.0125 M PEG and 30 gl^?1 sucrose + 30 gl^?1 maltose + 0.0125 M PEG. The lowest rate of embryogenesis was observed in YPm basal medium with 60 gl^?1 maltose and 0.0125 or 0.025 M PEG. Sucrose or a high concentration of maltose was found to be necessary for embryogenesis in anther culture of maize. Therefore, the addition of low levels of PEG and/or different sugars in the experimental design appeared to improve the protocol currently available in the world, especially for anther embryo yield and haploid plant regeneration in maize.     

A Broad Spectrum Tissue Culture Experiment with Tobacco (Nicotiana tabacum) Pith Tissue Callus

The composition and rationale of a broad spectrum tissue culture experiment involving 81 different media are described. Tobacco calluses sub-cultured on media in this experiment were induced to form six main types of callus depending on the concentration levels of minerals, auxins, cytokinins, and sucrose, growth factors and amino acids in the medium. Only nine of the 81 media inhibited callus growth, and growth (increase in mass divided by initial mass of fresh matter) varied considerably reaching a maximum of nearly 200 after eight weeks on a medium high in minerals, cytokinins, sucrose, growth factors and amino acids and low in auxins. Five media induced regeneration from the calluses. This experiment is suggested as a potentially fruitful introductory test for many new or unresolved tissue culture situations.     

Comparison of genetic algorithms for experimental multi-objective optimization on the example of medium design for cyanobacteria

In this work, two different genetic algorithms were applied to improve culture media composition for the autotrophic cyanobacteria Synechococcus PCC 7942. Biomass yield and conversion of the asymmetric reduction of 2', 3', 4', 5', 6'-pentafluoroacetophenone were considered as simultaneous objectives, resulting in a multi-objective optimization problem. Even when similar performances of both algorithms were observed, it could be shown that a novel strength pareto approach was able to achieve remarkable results with a reduced number of experiments (160 instead of 320). Handling a high number of media components (13), their concentrations were adjusted, delivering high improvements in comparison to the standard BG 11 culture media. The quality of the Synechococcus biocatalyst could be increased up to fivefold compared to the initial state of the optimization.     

大麦花药离体诱导的基因型稳定性和培养基的环境指数分析

本研究以湖北啤酒大麦优良新品种（系）为材料,分析了基因型的稳定性、培养基的环境指数以及基因型与培养基的互作效应.各基因型花药愈伤组织诱导率的稳定性分析表明,E2具有较强的普遍适应性,但稳定性好的材料愈伤组织诱导率并不一定最高,各种配方培养基的环境指数值大小不同,方向也有正有负,基因型效应以及基因型和培养基的互作效应均达极显著水平。     

土壤干旱对黄土高原乡土树种水分代谢与渗透调节物质的影响

以黄土高原4个乡土树种的幼苗为试验材料,采用盆栽方式模拟土壤干旱环境,研究土壤干旱对不同树种水分代谢与渗透调节物质的影响。结果表明,大叶细裂槭、虎榛子叶水势、叶片含水量下降迅速,叶片离体保水能力降幅明显;白刺花、辽东栎则表现为叶水势、叶片含水量缓慢下降,组织相对含水量在中度胁迫下略有上升。白刺花在不同水分处理条件下离体叶片保水力明显高于其它树种。1个树种可溶性糖含量随土壤干旱程度加剧明显增加,可溶性蛋白质含量在树种之间变化较为复杂,无明显规律性。K^ 离子含量和游离脯氨酸含量在中度水分胁迫下均有不同程度升高。白刺花在土壤干旱进程中,可溶性蛋白质含量、K^ 离子含量和游离脯氨酸含量均明显高于其它树种。综合水分代谢和渗透调节物质来看,水分胁迫条件下,白刺花以保持高水势、减少组织水分散失和增加渗透调节物质来提高细胞原生质浓度,增强其抗旱性。     

Effect of epidermal growth factor on preimplantation development and its receptor expression in porcine embryos.

The present study aimed to determine the influence of exogenous epidermal growth factor (EGF) on in vitro preimplantation porcine embryo development and its mRNA expression for EGF receptor (EGFR). Oocytes were aspirated from abattoir ovaries, selected and cultured in defined, protein-free media for 44 hr before in vitro fertilization (IVF). Thirty-six hours after IVF, two-cell stage embryos were selected and treated or cultured until embryo treatment. In experiment 1, compact morulae were selected on day 4 after IVF and randomly allocated into 5 groups: NCSU 23 with PVA as group 1; NCSU 23 with PVA and 0.1 ng/ml, 1.0 ng/ml, 10.0 ng/ml EGF as group 2, 3, 4, respectively; NSCU 23 with 0.4% BSA as group 5. In experiment 2, treatment groups were the same as in experiment 1 except that 0.1% crystallized BSA was added to both washing media and all treatment groups instead of PVA. In experiments 3 and 4, two-cell stage embryos were treated and cultured in the same experimental design as experiments 1 and 2, respectively. RT-PCR was used to detect the mRNA expression of EGF receptor in compact morulae and blastocysts. The PCR products were subjected to direct DNA sequencing. There was no significant improvement in the development rate of embryos from compact morulae to blastocysts in the presence of various EGF concentrations (0.1, 1.0, 10.0 ng/ml) versus without EGF addition. They were all significantly lower than those embryos cultured in the continuous presence of 0.4% BSA. However, when a reduced concentration (0.1%) of crystallized BSA was added to all the treatment groups, a significantly lower rate of embryo development was observed in control media (NCSU23 with 0.1% crystallized BSA) compared with those developed in culture media with 0.4% BSA. With the addition of EGF at 10 ng/ml (with 0.1% BSA), embryo development rates were significantly improved over the control group (P < 0.05) and were as good as those rates in 0.4% BSA culture group. When embryos were selected and treated from the 2-cell stage, they did not develop to blastocyst stages after five more days' culture without any protein (BSA) or growth factor addition. When 0.1% BSA was included in the media, blastocyst formation rates were significantly improved by EGF addition at the concentration of both 1.0 or 10 ng/ml (P < 0.05) as compared to 0.0 or 0.1 ng/ml. EGFR mRNA was detected in both compact morulae and blastocyst stages of porcine embryos and confirmed by direct DNA sequencing. Our results indicate that IVM-IVF porcine embryo developmental rates could be improved by the addition of EGF in the culture media with the presence of a reduced amount of defined BSA (>97% albumin). However, EGF alone was not able to elicit any stimulatory effects on embryo development in the absence of protein supplementation. Further studies are needed to investigate the potential synergistic factors in embryo culture media to eventually define the porcine embryo culture media.     

Genomic stability of Saccharomyces cerevisiae baker's yeasts.

The objective of this study has been to gather data on genomic stability of baker's yeast strains during long-term mitotic growth under restrictive conditions so that comparisons could be made to other studies indicating genomic instability during meiosis. The work describes the analysis of mitotic stability of the nuclear and mitochondrial genomes in the baker's yeast strain V1 during incubation in continuous culture for 190 generations (300 days). The cells were cultured in complete medium containing 2% glucose and 8 to 12% ethanol, as a mutagenic agent specific for mtDNA. The high concentration of ethanol severely limited the growth rate of the cells. DNA samples were monitored for chromosomal pattern, polymorphisms in selected nuclear genes (SUC2, MALIT, ADH1) and mobile genetic elements (Ty1 and Y'), and for RFLPs in mtDNA. The results show that both the nuclear and mitochondrial genomes of grande cells were very stable. However, the frequency of petite mutants in the population varied dramatically during the course of the experiment, reaching as high as 87% petite during the first 27 days of the experiment and declining to 5.8% petite at the end. This decline can be attributed to selection against petite mutants in media containing high concentrations of ethanol. Moreover, when samples and the parental strain were compared at the end of the experiment, no change could be observed in parameters such as their growth rate in different media, capacity to leave doughs, viability in ethanol or frequency of petite mutants. Results therefore indicated that the majority of the cells in the population were very similar to the parental throughout the experiments, with no apparent molecular or phenotypical changes.     

An experimental strategy validated to design cost-effective culture media based on response surface methodology

For any fermentation process, the production cost depends on several factors, such as the genetics of the microorganism, the process condition, and the culture medium composition. In this work, a guideline for the design of cost-efficient culture media using a sequential approach based on response surface methodology is described. The procedure was applied to analyze and optimize a culture medium of registered trademark and a base culture medium obtained as a result of the screening analysis from different culture media used to grow the same strain according to the literature. During the experiments, the procedure quantitatively identified an appropriate array of micronutrients to obtain a significant yield and find a minimum number of culture medium ingredients without limiting the process efficiency. The resultant culture medium showed an efficiency that compares favorably with the registered trademark medium at a 95% lower cost as well as reduced the number of ingredients in the base culture medium by 60% without limiting the process efficiency. These results demonstrated that, aside from satisfying the qualitative requirements, an optimum quantity of each constituent is needed to obtain a cost-effective culture medium. Study process variables for optimized culture medium and scaling-up production for the optimal values are desirable.     

刺葡萄幼胚愈伤组织诱导及其高产原花青素细胞系筛选

以刺葡萄幼胚为材料,研究不同培养方式、培养基配方和培养条件对其愈伤组织诱导的影响,采用正交试验设计法筛选刺葡萄愈伤组织继代增殖的培养基配方,并对继代保持的培养条件和方式进行优化,同时进行了高产原花青素刺葡萄愈伤组织细胞系的筛选研究。结果表明,刺葡萄幼胚以平放的方式接种到MS＋1.0 mg·L^-1 2,4-D或MS＋1.0 mg·L^-1 2,4-D＋0.5mg·L^-1 KT的固体培养基上,在黑暗的条件下,能有效的诱导出愈伤组织,诱导效率为80%;刺葡萄愈伤组织继代增殖以MS＋1.5 mg·L^-1 2,4-D或MS＋1.5 mg·L^-1 2,4-D＋0.5 mg·L^-1 KT的固体培养基为佳,并且采用此两种培养基交替继代培养,在光照条件下能长期保持旺盛且生长一致的刺葡萄愈伤组织;筛选出了紫红色松脆状的高产原花青素的刺葡萄愈伤组织细胞系,培养35 d后每克鲜样的原花青素含量可达1 671.16μg。     

Variation in the abillties of different strains of Drosophila melanogaster to utilize diverse carbon sources

In several experiments, flies of different geographic origins have been tested for their abilities to produce adult progeny on culture media containing primarily one or the other of several sugars. Flies of different origins differ in their abilities to produce progeny on a given sugar; flies of a single origin differ on different sugars. The data reveal a strong strain-sugar interaction as well. Strains of flies maintained for two or more years by mass transfer on culture media containing one or the other of several sugars, including xylose and lactose, adapt to that culture medium. Hybrid flies-either between lines within localities or between localities-are best able to produce offspring on xylose-containing medium.