Embryo rescue inVicia faba andVicia narbonensis

Embryo rescue in twoVicia faba L. cultivars (‘Polycarpe’ and ‘A-107’) and oneV. narbonensis L. population (A-202) was studied under a 22 ± 2°C/ 16 ± 1° day/night temperature regime. Very young ovules (1.0–1.8 mm long) cultured, in-ovule, on five liquid media remained green for a longer period of time on modified B5, modified Murashige and Skoog and modified Beasley and Ting media than on modified Phillips and Collins and modified Bourgin and Nitsch media. However, no embryo growth or embryo germination was observed. In-ovule culture of older ovules, 6 and 8 days forV. narbonensis and 10 and 14 days forV. faba, on modified B5 liquid medium allowed 6-day-oldV. narbonensis and 14-day-oldV. faba embryos to be rescued. Finally, culture of whole pods of the two species resulted in the rescue of even younger embryos. Thus, plantlets were obtained from as young as 4-day-oldV. narbonensis pods and 11-day-oldV. faba pods.     

二十世纪我国植物学家对植物组织培养的贡献

回顾了上一世纪我国植物组织培养的发展。 1934年以来 ,我国的植物组织培养研究一直与国际发展同步进行。我国学者在离体器官发生、茎尖培养、花药培养、子房培养、胚乳培养、原生质体培养和细胞大量培养等分支领域都取得重要进展。本文在引证我国研究者发表的植物组织培养论文的基础上 ,着重评述了那些被国际同行公认的研究成果。此外 ,还介绍了植物组织培养在我国农业和工业上应用的情况     

二十世纪我国植物学家对植物组织培养的贡献

回顾了上一世纪我国植物组织培养的发展.1934年以来,我国的植物组织培养研究一直与国际发展同步进行.我国学者在离体器官发生、茎尖培养、花药培养、子房培养、胚乳培养、原生质体培养和细胞大量培养等分支领域都取得重要进展.本文在引证我国研究者发表的植物组织培养论文的基础上,着重评述了那些被国际同行公认的研究成果.此外,还介绍了植物组织培养在我国农业和工业上应用的情况.     

猪苓、伴生菌及蜜环菌共培养的形态学研究

本文对猪苓、伴生菌及蜜环菌两两共培养及三者共培养进行了宏观形态观察及细胞学水平上的研究。结果表明,猪苓与伴生菌共培养时,在二者之间形成一致密拮抗线,猪苓菌落表面菌丝分化产生大量菌丝束;猪苓与蜜环菌共培养时,猪苓能阻止蜜环菌菌索对其自身的进一步侵袭,互作区中的双方菌丝及菌索均停止生长;蜜环菌与伴生菌共培养时,蜜环菌能穿透整个伴生菌菌落,在伴生菌菌落下方产生大量分枝;三者共培养后,猪苓对蜜环菌的防御能力有所下降,伴生菌对蜜环菌的耐受力有所提高,蜜环菌产生的新分枝均向伴生菌一侧生长,猪苓与伴生菌之间并不形成致密拮抗线,只可见双方菌丝的白色交融区。 猪苓与伴生菌均能在蜜环菌菌索皮层上形成侵入位点。     

Tapping culture-an improved method for cell suspension culture

Summary A new culture vessel was designed for cell suspension culture. A silicone-convered magnet bar fixed by one end to the side wall of the bottle was held horizontally a short distance from the bottom. A standard type magnetic stirrer was used. In contrast to the conventional horizontal movement of “stirring” in cultures the bar moves vertically with a “tapping” motion. This improvement resulted in less cell injury, higher rate of cell proliferation and formation of fewer bubbles than in the conventional type. Nine cell types were simultaneously cultivated in tapping, stirring and stationary culture. All cell types proliferated more luxuriously in tapping cultures than in stirring cultures. Serial cultivation of cells in tapping cultures was also successful. This work was supported in part by the grants for Cancer Research from the Ministry of Education, Science and Culture, Japan.     

荔枝生物技术研究进展（综述）

从荔枝的组织培养、花药培养和原生质体培养方面综述荔枝生物技术的研究概况,并提出该领域存在问题及发展方向。     

Light requirement and photosynthetic cell cultivation – Development of processes for efficient light utilization in photobioreactors

Although the potential of photosyntheticmicroorganisms for production of various metabolitesand in environmental bioremediation is recognized,their practical application has been limited by thedifficulty in supplying light efficiently tophotobioreactors. Various types of photobioreactorwith high illumination to volume ratios have beenproposed, but most are limited by cost, mass transfer,contamination, scale-up or a combination of these.The problem of light supply to photobioreactorscan be solved by developing photosynthetic cellcultivation systems where light is either substitutedor supplemented. Many strains of photosynthetic cellsare capable of heterotrophic growth under darkconditions and their heterotrophic culture can be usedfor efficient production of biomass and somemetabolites. However, light is absolutely required forefficient production of some metabolites. In suchcases, there is a need to supplement the heterotrophicwith photoautotrophic metabolism. Inphotoheterotrophic (mixotrophic) culture, thephotoautotrophic and heterotrophic metabolisms can beexploited for efficient production of usefulmetabolites but it has many problems such as processoptimization in terms of making a balance between thephotoautotrophic and heterotrophic metabolism. Another promising system is the sequentialheterotrophic/ photoautotrophic cultivation system,where the cells are cultivated heterotrophically tohigh concentrations and then passed through aphotobioreactor for accumulation of the desiredmetabolite(s). Furthermore, cyclicphotoautotrophic/heterotrophic cultivation system canbe used to achieve continuous cell growth underday/night cycles. This involves cultivating thecells photoautotrophically using solar light duringthe day and then adding controlled amount of organiccarbon source during the night for heterotrophicgrowth. In this review, these various systems arediscussed with some specific examples.     

Technology for cell cycle research with unstressed steady-state cultures

A culture system for performing cell cycle analyses on cells in undisturbed steady-state populations was designed and tested. In this system, newborn cells are shed continuously from an immobilized, perfused culture rotating about the horizontal axis. As a result of this arrangement, the number of newborn cells released into the effluent medium each generation is identical to the number of cells residing in the immobilized population, indicating that one of the two new daughter cells is shed at each cell division. Thus, the immobilized cells constitute a continuous, steady-state culture because the concentrations, locations and microenvironments of the cells in the culture vessel do not vary with time. In tests with mouse L1210 lymphocytic leukemia cells, about 10⁸ newborn cells were produced per day. This new culture system enables a multiplicity of cell cycle analyses on large numbers of cells assured to be from populations in steady-state growth.

A simple device for planting replicate mammalian cell cultures

Summary The design and use of a unit for planting uniform inocula for replicate cultures are described. Its design permits continuous gassing of suspensions of mammalian cells with humidified CO₂, thus stabilizing the pH (±<0.05 pH unit) of culture media buffered with sodium bicarbonate. The unit can be readily modified to deliver different volumes; identical samples can be dispensed simply and rapidly, with minimal cell damage and chance of microbial contamination. Quantitative data regarding sample uniformity and growth subsequent to planting with this unit are presented.     

A simple device for planting replicate mammalian cell cultures

Summary The design and use of a unit for planting uniform inocula for replicate cultures are described. Its design permits continuous gassing of suspensions of mammalian cells with humidified CO₂, thus stabilizing the pH (±<0.05 pH unit) of culture media buffered with sodium bicarbonate. The unit can be readily modified to deliver different volumes; identical samples can be dispensed simply and rapidly, with minimal cell damage and chance of microbial contamination. Quantitative data regarding sample uniformity and growth subsequent to planting with this unit are presented.     

A method for growing stationary plant suspension cell cultures

Summary Stationary culture of plant cell suspensions has been achieved. Slurries, produced when small amounts of agar (0.1–0.4%) were added to culture media, were used to suspend plant cells. Growth proceeded more slowly than in standard shake culture, but cells remained viable for months of culture. This method of growing plant cells in stationary culture should be useful for general applications including long-term cell culture, shipment of cultures, and physiological, molecular biological, and pathological studies. Mention of a trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products or vendors that may also be suitable. Editor’s Statement This procedure for growing stationary suspension cultures in an agar slurry should be useful for shipping suspensions and for long-term maintenance of little used or back-up cultures.     

黄瓜矿棉无土育苗效应的研究

本文报道了利用矿棉进行黄瓜无土育苗的效应及生长发育的研究结果。于1986年秋季进行了试验研究,两种育苗法的黄瓜幼苗干重对数值与时间的回归方程分别为: 矿棉培土培黄瓜幼苗LnW=6.0376 0.1068t LnW=5.8500 0.0960t 经F值检验达极显著。两种育苗法在生长量上的差距显著,试验结束时,矿棉培幼苗于重高于土培幼苗近两倍;经幼苗各器官分析,矿棉培幼苗根系效应最大,高出土培幼苗的3-4倍;其次是叶,高出2倍;再次是茎轴1.5倍左右。经矿棉育苗的幼苗质量好,移栽成活率高,有利于植物根系舒伸生长,证明矿棉是一种良好的植物育苗基质。本文详细地讨论了利用矿棉进行无土育苗的优越性和发展前景。     

北京协和医院文化建设的理念和实践

为了继承和发扬协和优良传统,协和医院近年来大力开展文化建设,创新性提出了“待病人如亲人,提高病人满意度;待同事如家人,提高员工幸福感”新办院理念,并通过提炼协和精神、开展理念教育、学习典范人物、加强制度建设等,让协和文化这一宝贵精神财富代代传承,发扬光大。     

我院医院文化建设发展的途径探索

基于西安医学院第一附属医院医院文化建设实践,探索通过人才文化、创新文化、制度文化等主要途径,实现医院文化的落地生根,使医院文化为增强凝聚力、提高医疗质量和技术服务水平、提升医院核心竞争力发挥更加积极的作用。

     

胡萝卜细胞培养产胡萝卜素的研究

从鲜红胡萝卜主根部选取外植体,用MS型培养基诱导出愈伤组织,继代培养,周期22d,接着进行细胞的液体悬浮培养。液体培养20d时,测定β-胡萝卜素含量达41．7mg／L,占细胞干重的0．35％。培养细胞中含胡萝卜素是胡萝卜根部的3．38倍,色素比产率提高15．4倍。     

在静态和动态培养条件下杂交瘤细胞的生长和代谢

通过对WuT3杂交瘤细胞在静态和动态培养条件下的比较,发现细胞生长和代谢有很大不同。在静态培养条件下,细胞培养周期较长,细胞密度较高;但在动态培养条件下,细胞代谢更加旺盛,葡萄糖、氨基酸等营养物质的消耗加快,乳酸、氨、丙氨酸等代谢产物的比生成速率较大。     

Scale-up of suspension and anchorage-dependent animal cells

Alternative culture processes for laboratory scale-up (to 20 L) are described for both suspension and anchorage-dependent cells. Systems range from simple multiple culture units such as the roller bottle, through stirred suspension and microcarrier unit bioreactors, to highly sophisticated perfusion culture capable of maintaining cells at densities of about 10⁸/mL. Critical parameters in scale-up are discussed, and the advantages and disadvantages of each culture system are critically evaluated.     

Microalgal mass culture systems and methods: Their limitation and potential

Cultivation of microalgae using natural and man-made open-ponds istechnologically simple, but not necessary cheap due to the high downstream processing cost. Products of microalgae cultured in open-pondscould only be marketed as value-added health food supplements, specialityfeed and reagents for research. The need to achieve higher productivityand to maintain monoculture of algae led to the development of enclosedtubular and flat plate photobioreactors. Despite higher biomassconcentration and better control of culture parameters, data accumulatedin the past 25 years have shown that the illuminated areal, volumetricproductivity and cost of production in these enclosed photobioreactors arenot better than those achievable in open-pond cultures. The technicaldifficulty in sterilizing these photobioreactors has hindered their applicationfor the production of high value pharmaceutical products. The alternativeof cultivating microalgae in heterotrophic mode in sterilizable fermentorshas achieved some commercial success. The maximum specific growth ratesof heterotrophic algal cultures are in general slower than those measured inphotosynthetic cultures. The biomass productivity of heterotrophic algalcultures has yet to achieve a level that is comparable to industrialproduction of yeast and other heterotrophic microrganisms. Mixotrophiccultivation of microalage takes advantage of their ability to utilise organicenergy and carbon substrates and perform photosynthesis concurrently. Moreover, production of some algal metabolites is light regulated. Futuredesign of sterilizable bioreactors for mixotrophic cultivation of microalgaemay have to consider the organic substrate the main source of energy andlight the supplemental source of energy, a change in mindset.     

High density culture of mouse-human hybridoma cells using a perfusion culture apparatus with multi-settling zones to separate cells from the culture medium

Mouse-human hybridoma X87X cells were cultivated using a novel perfusion culture apparatus provided with three-settling zones to separate the cells from the culture medium by gravitational settling. The maximum viable cell density in a serum-free culture medium attained 3.0×10⁷ cells/ml, when the specific perfusion rate was set to 2.3 vol day^-1, and monoclonal antibody was continuously produced. These results were almost the same as those in the perfusion culture vessel with one settling zone and revealed that the process with a plurality of settling zones is a promising one for scale-up of a gravitation type of perfusion culture vessel.     

<i>In Vitro</i>-Propagation of <i>Agave tequilana</i> Weber cv. azul in a Temporary Immersion System

In Mexico, there is a need to produce large quantities of plantlets for the establishment and replanting of blue (cv. azul) agave production areas. Most of these plots are within the origin denomination area (DOT, Spanish acronym) of the distilled product of this plant, known as tequila. The objective of this study was to develop an in vitro-propagation protocol for Agave tequilana Weber cv. azul using segmented stems in both: solid and liquid media. A disinfection and in vitro technique were developed to obtain shoots, through plantlets collected in commercial plots, which attained 100% surface-disinfection and budding rate. At the multiplication stage, the effects of 6-Benzylaminopurine (BA) (0.0, 4.4 and 13.2 μM) and kinetin (0.0, 9.4, 18.8 and 37.6 μM) were evaluated on lateral-shoot production of segmented sagittal stems. These were cultivated on Murashige & Skoog (MS) medium, with the addition of 3.0% sucrose and 8 g L⁻¹ agar. It was observed that BA and kinetin increased the number of shoots per explant, obtaining up to 18 and 26, respectively. Furthermore, it was found that just the sagittal segmentation of explants increased axillary budding. On the other hand, segmented-stem bases were grown in MS liquid medium with 3.0% sucrose, inside a RITA® system, programmed by a 5 min immersion step with a frequency of every 4 h. The effect of Indole−3-Acetic acid (IAA) (0.57, 2.9, 5.7 μM) was evaluated, while maintaining a concentration of BA (13.2 μM). It was observed that the greatest concentration of IAA led to the formation of more than 20 buds per explant. These results offer a new methodology to increase the efficiency of A. tequilana Weber cv. azul-in vitro multiplication by sagittal segmentation of stems and the addition of BA and/or IAA.