溶液培养及缺素培养实验中番茄幼苗的培养

叙述了溶液培养及缺素培养实验中番茄幼苗的培养过程及注意事项。     

席藻盐适应培养的研究

用０．２ｍｏｌ／Ｌ ＮａＣｌ作选择因子,可直接筛选得到耐０．２ｍｏｌ／Ｌ ＮａＣｌ席藻。在０．２ｍｏｌ／Ｌ ＮａＣｌ培养液中,席藻细胞逐日降解,到第３ｄ达最低点,３ｄ后细胞数上升。细胞降解随ＮａＣｌ浓度提高而加剧。在等渗浓度或等渗浓度以下,席藻能保持慢速增殖。维生素Ｂ１、吲哚乙酸、玉米素、赤霉素都未能提高席藻对盐类的培养适应能力。席藻对不同的盐溶液培养适应性有差异,而对多种盐复合溶液最能适应。     

液体培养蛹虫草虫草素和腺苷的代谢量

为提高虫草素和腺苷的代谢量,用高效液相色谱法作检测手段,以二者含量为检测指标,对液体培养的氮源种类、氮源水平、碳源水平、动态变化、培养体系的虫草素及腺苷含量和总量进行研究,结果表明:不仅蚕蛹粉,豆粕和豆粉也是很好的氮源;培养液以3%氮源、4%碳源为佳;振荡培养8d,培养液中虫草素总量是菌丝体中虫草素总量的6倍多;振荡培养7~9d,可使每瓶培养物的虫草素和腺苷总量达到10mg以上。     

不同培养条件和前体对蛹虫草液体发酵产虫草素的影响

蛹虫草能产生虫草素等多种活性物质。为考察不同液体发酵方式及添加前体物质对虫草素积累的影响,选用蛹虫草08Y1菌株,通过光照振荡、光照静置、黑暗振荡、黑暗静置四种培养条件和添加前体物质（腺嘌呤1g/L+甘氨酸16g/L）,发酵16d后检测虫草素和腺嘌呤含量。结果表明：08Y1菌株在黑暗振荡培养条件下,虫草素积累达1,015.0mg/L,腺嘌呤利用率达98.5%,说明黑暗振荡培养并添加前体物质是提高虫草素产量的有效方法。     

植物生理学实验中溶液培养研究性实验的建立研究

溶液培养实验是植物生理学实验教学中的重要内容,而旧的教学大纲中该部分内容设置单一,不利于学生的理解掌握。笔者在教学实践中,将基础性实验与溶液培养实验有机结合,以溶液培养为技术手段,引导学生开展研究性实验,锻炼学生的动手能力,培养学生的观察能力、综合能力和创新能力,取得了较好的效果。     

果树离体叶片培养再生研究进展

从基本培养基的选择、外植体的选择、培养条件、激素和其它添加物的影响等方面综述了有关的研究成果,并提出了存在问题及今后的研究方向。     

藏红花细胞悬浮培养动力学研究

基于已经建立的藏红花细胞悬浮培养体系,研究了其悬浮培养动力学.结果表明,悬浮培养时细胞的生长周期约为20 d,在20 d时生物量达到最大,为12.3 g DW/L,但是藏红花素的合成周期大约为28 d,在28 d时藏红花素的含量和产量达到最大,分别为95.8 mg/g和0.92 g/L.藏红花素的积累与细胞的生长之间的关系为半生长偶联型,为其生物反应器放大培养提供了参数等理论依据.     

铜、锌对水稻幼苗生长及超氧物歧化酶的影响

用缺Cu或Zn,与缺Cu、Zn的木村B营养溶液培养杂交水稻威优48(V_(48)),威优49(V_(49))与常规稻竹系26(ZX_(26))幼苗至七叶期。缺Cu的稻苗,上部新生叶(六、七叶)先端扭曲,叶尖枯黄。缺Zn与缺Cu、Zn的稻苗,六、七叶脉间全部褪绿,植株矮小,根系衰退。凡是缺素培养的稻苗,根与叶的超氧物歧化酶(SOD)活性均显著减弱,叶绿体基粒与间质片层明显减少,缺Cu、Zn的叶绿体还出现较大的淀粉“液泡”。经凝胶电泳,显示出水稻SOD有4组活性带,缺Cu或Zn与缺Cu、Zn使迁移率较高的3组活性带的活性明显减弱。     

蛹虫草菌丝产虫草素液体培养条件的研究

通过对蛹虫草菌丝产虫草素液体培养条件的研究,明确蛹虫草菌丝产虫草素的适宜碳源及浓度,适宜氮源及浓度,最适pH值,最适培养温度,最适转速以及最适培养时间,以便应用于虫草素的工厂化生产。结果表明,蛹虫草菌丝产虫草素的条件：适宜碳源为D-果糖,最适浓度为10g/L;适宜氮源为蛋白胨,最适浓度为15g/L;最适初始pH为7,最适培养温度为24℃,最适转速为180r/min,最适培养时间为9d,其培养液虫草素含量可达到0.537g/L。     

西洋参细胞大量培养的研究

当西洋参细胞培养在Ms培养液中,KN0₃含量提高一倍而去掉NH₄NO₃。时细胞生长速率和皂甙产量分别比在正常培养液中提高65．1％和166．2％。黑节草寡糖素和人参寡糖素均有利于西洋参细胞的生长和皂甙含量的提高,尤其能增加Rg组皂甙的含量。西洋参细胞悬浮培养以生产皂甙收获的最佳时期为培养25天以上,其合成皂甙的高峰在细胞生长的对数期稍后出现。细胞悬浮培养和发酵培养过程中均未见pH值回升的现象。pH值稳定的发酵培养和pH值任其变化的培养相比,其皂甙含量,生长速率和生物量均要高。最后对细胞的培养方式进行了比较。     

Nodulation ofMedicago sativa in solution culture

Summary Calcium and hydrogen ions interacted on nodulation. Increasing acidity from pH 5.6 to pH 4.8 increased the calcium concentration required to nodulate 50% of the plants, from 0.1 mM to 6 mM. Calcium concentration below 0.2 mM or pH below 4.8 inhibited nodulation at all tested levels of the other variable. Root extension and root-hair production were insufficiently affected by calcium or pH to explain reductions in nodule numbers. Initiation of infection, the most acid-sensitive stage of the nodulation process, was also the most calcium-demanding stage at pH 5.2. Once infections were initiated, infection threads still developed and nodules still grew despite transfer of the plants to solutions too low in calcium to have permitted infection to begin. Pretreatments at 0.5 mM and 8 mM calcium at pH 5.2 before inoculation had no significantly different effects on nodulation. Observations on root-hair distribution suggest that developing nodules can suppress further infection by suppressing the emergence of root hairs on newly developing roots.     

Nodulation ofMedicago sativa in solution culture

Summary When nitrate was maintained in continuous supply in solution cultures at concentration 0.02 mM to 2 mM it reduced the numbers of curled root hairs and of nodules. But the addition of indole-3-acetate with the inoculum overcame the nitrate-inhibition of curling without affecting nodulation; and there was no quantitative connection between numbers of curled hairs and of nodules when nitrate treatments of limited duration were applied at different times in relation to inoculationNitrate reduced nodule number to some extent even if present only before inoculation, or only on the first, second or third day after inoculation, so that reduction in nodule number was difficult to attribute solely or chiefly to interference with any one phase of the nodulation process. However, nitrate inhibited formation of infection threads and augmented the proportion of arrested infection threads, to such an extent that this could be important in limiting nodule number.Nitrate also delayed nodulation. The delay occurred early after inoculation and corresponded with delay in the appearance of infection threads.     

Nodulation ofMedicago sativa in solution culture

Summary The time course of increase in nodule number was observed in solution cultures of controlled composition, over periods of 5 to 7 weeks following the inoculation of 8-day-old seedlings. The plants produced successive crops of nodules two to three weeks apart. Maintaining nitrate at concentrations of 0.2 or 0.5mM greatly reduced the number of nodules in the first crop but only slightly or not at all in the larger following crops, relative to the numbers in nil-nitrate control cultures. This was not due to any decrease in the sensitivity of nodulation to inhibition by nitrate. Nodulation in the nil-nitrate controlplants was restrained following their abundant first-crop nodulation. The experiments ceased to be simple tests of effects of nitrate on rate of nodule production as soon as the first crop of nodules appeared. Subsequent nodule production was then inhibited approximately equally, by prior nodulation in the abundantly nodulated control-plants, and by nitrate in the sparsely nodulated plants in the nitrate treatment.When the effect of nitrate was tested on plants comparable in size and in number of previously established nodules, it inhibited nodule production by 70–80%, whether the plants were seedlings or month-old plants, previously nodulated or not.     

A multichannel pH controller for solution culture systems

A multichannelled, computer-controlled system has been developed for maintaining near-constant pH in nutrient solutions. This system will monitor, record and individually maintain pH in up to seven solutions continuously. All components except the software and pump controller were available commercially.In an experiment for which the system was developed, in which six individual solutions were controlled for 20 days, the pH was generally maintained within 0.1 pH unit of the set value. Drift outside this range was generally due to addition of nutrients required to maintain adequate levels. Such drifts were quickly and automatically corrected.This equipment has been subsequently used in several experiments in which it was imperative to maintain pH at a constant level in several solutions simultaneously with actively growing plants over a long period of time.     

Nodulation and growth ofPhaseolus vulgaris in solution culture

Summary Conditions and techniques for achieving good nodulation ofPhaseolus vulgaris L. in continuously aerated solution were developed from greenhouse experiments.If nodules had been established, their growth and activity and the growth of the plant were at least as good in solution culture as in gravel culture. Nodule formation was observed within 10 days of inoculation in small volumes of solution culture (1 liter). In large volumes (19 liters), similarly prompt nodulation occurred only if the plants were inoculated before or immediately after the seedlings were transferred to the solution from gravel or vermiculite; and the nodules were restricted to the roots that had been present at the time of transfer. Delayed inoculation, 2 days after transfer to large volume solutions, led to sparse nodulation observed only after 3 weeks. Delay or inhibition of nodulation in large volumes of solution could not be explained by failute of bacteria to colonize roots or by sparsity of root hairs.Nodule initiation in solution culture was severely inhibited at pH below 5.4. An additional problem in growing N₂-dependent bean in solution culture was the buildup of Cl^– to toxic levels in the plant in nitrate-free media, even at solution concentrations as low as 0.4 mM Cl^–. Daily addition of 0.5 to 1.0 mg N per plant delayed nodule growth and activity slightly, but increased plant growth and alleviated the severe N-deficiency that otherwise developed before the onset of N₂-fixation.     

Cell culture media impact on drug product solution stability

To enable subcutaneous administration of monoclonal antibodies, drug product solutions are often needed at high concentrations. A significant risk associated with high drug product concentrations is an increase in aggregate level over the shelf‐life dating period. While much work has been done to understand the impact of drug product formulation on aggregation, there is limited understanding of the link between cell culture process conditions and soluble aggregate growth in drug product. During cell culture process development, soluble aggregates are often measured at harvest using cell‐free material purified by Protein A chromatography. In the work reported here, cell culture media components were evaluated with respect to their impact on aggregate levels in high concentration solution drug product during accelerated stability studies. Two components, cysteine and ferric ammonium citrate, were found to impact aggregate growth rates in our current media (version 1) leading to the development of new chemically defined media and concentrated feed formulations. The new version of media and associated concentrated feeds (version 2) were evaluated across four cell lines producing recombinant IgG₄ monoclonal antibodies and a bispecific antibody. In all four cell lines, the version 2 media reduced aggregate growth over the course of a 12 week accelerated stability study compared with the version 1 media, although the degree to which aggregate growth decreased was cell line dependent. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:998–1008, 2016     

Maintaining exponential growth, solution conductivity, and solution pH in low-ionic-strength solution culture using a computer-controlled nutrient delivery system

Studies of plant nutrient requirements in solution culture have often used nutrient concentrations many-fold higher than levels found in fertile soils, creating an artificial rooting environment that can alter patterns of nutrient acquisition. The relative addition rate (RAR) technique addresses this problem by providing nutrients in exponentially increasing quantities to plant roots in solution culture. A computer-controlled RAR nutrient delivery system has been developed to reduce workload and to facilitate more frequent nutrient additions (4x daily) than is possible with manual additions. In initial experiments, a minimum background solution containing 500 microM nitrogen and all other essential nutrients in optimal proportions was required for the healthy growth of Triticum aestivum. This requirement was reduced to 50 microM nitrogen when calcium in the background solutions was increased to 400 microM. Varying the abundance of ammonium and nitrate in both background and delivery solutions provided a means of controlling plant-induced pH changes in growth solutions. In optimized solutions, plant relative growth rates (RGR) in the order of 0.2 g g(-1) plant d(-1) were maintained over a 22 d experimental period. Variation in RARs provided a means of growing plants with varying RGRs under relatively constant conditions of solution electrical conductivity and pH.     

Screening sorghum for tolerance to excess manganese in solution culture

This study was conducted to define traits to screen sorghum (Sorghum bicolor L. Moench) genotypes for tolerance to excess Mn. Visual Mn toxicity symptoms, net and total root lengths, shoot and root dry matter yields, and shoot and root Mn concentrations were determined for plants grown in nutrient solutions (pH 4.5) at different levels of Mn (0, 3, 6, 9 and 12 mM above the initial 18 M) to assess plant responses to excess Mn. Dry matter yields showed greatest variability among genotypes, and was an effective trait to evaluate sorghum for tolerance to excess Mn. Reductions in dry matter yields did not occur until Mn levels were above 3 mM. Levels of Mn between 3 and 6 mM could effectively be used to screen sorghum for genotypic differences to excess Mn. Manganese levels above 6 mM were too severe to allow good genotypic differentiation. Of genotypes tested, NB9040 and Wheatland showed good tolerance and SC283 and ICA-Nataima were sensitive to excess Mn.