生物反应器在满天星快繁中的应用

利用2.5L柱状气升式生物反应器采用接触法进行了满天星(Gypsophila paniculata)增殖培养。结果表明:外植体接种密度(每个反应器接种外植体个数)为35比密度为25和45更有利于满天星的增殖生长,平均每个生物反应器内可获得252株生长健壮的不定苗;光强为90μmol.m-2.s-1对满天星增殖培养最有利,过高的光强对不定芽和株高有抑制作用;通过小孔隙(15μm)的多孔喷头注入0.1vvm空气有利于满天星增殖培养。     

植物细胞培养的生物反应器

本文介绍了植物细胞培养的特点及其生物反应器的设计原理,概述了植物细胞悬浮培养和固定化细胞系统中各类生物反应器的传氧、混合和流体力学特性与植物细胞生长和次生代谢物生产的关系。     

生物反应器在干细胞培养中的应用研究进展

干细胞是一类具有自我更新能力和多向分化潜能的细胞,在再生医学、药物筛选及毒理学等生物医学领域呈现出诱人的前景。通过目前的干细胞分离培养技术可获得的干细胞数量极少,远远不能满足临床需要,因此体外大规模扩增培养干细胞是亟待解决的问题。该文简述了适用于干细胞培养的各种生物反应器的特点,以及悬浮生物反应器体系在不同类型干细胞群中的研究应用。同时对利用生物反应器培养干细胞过程中几个主要的关键参数进行了阐述,将为干细胞的培养和研究从思路和方法上提供参考。     

连续流生物反应器技术在微生物培养中的应用

自然环境中99%微生物在实验室条件下仍是不能被培养的,称之为"未培养"微生物或微生物"暗物质"。对其进行研究不仅有助于认识环境中微生物代谢多样性,丰富生命之树,同时未培养微生物还蕴含着巨大的新基因和新天然产物资源。但传统培养技术的局限性阻碍了"未培养"微生物资源的开发和利用。虽然随着分子生物学技术的发展,可以直接从环境中获得未培养微生物的遗传信息,分析微生物的广泛代谢多样性,但微生物的生理特征和代谢产物等分析仍然需要建立在研究纯菌株的基础上。目前,已经有很多新颖的培养技术被研发,如原位培养技术、共培养技术和连续流生物反应器培养技术等用于挖掘未培养微生物资源。本文主要介绍了连续流生物反应器培养新技术的发展与改进,探讨了"未培养"微生物培养技术及设备的发展方向,以进一步促进"未培养"微生物资源的开发与利用。     

生物反应器动物细胞培养技术在生物领域的应用及发展趋势

随着我国疫苗市场的迅速发展,疫苗生产技术正经历着从大规模转瓶培养动物细胞疫苗工艺向生物反应器培养动物细胞疫苗工艺的转变。生物反应器培养工艺,具有综合成本低、批量大、批间差小、所获得病毒抗原含量高等优点。不仅提高我国疫苗生产技术、疫苗质量,而且使我国生物产业近一步走向国际化。     

两液相反应器及其在生物反应中的应用

论述了生物反应中两液相反应器的进展及存在的问题，尽管两液相反器有了较多进展，但其仍处于发展阶段，其主要问题是生物催化剂在介面的聚集损失活性等。     

微小型生物反应器及其在生物医药领域的应用展望

微小型生物反应器体积微小但在线分析检测和过程控制功能媲美台式装备。其核心支撑技术包括一次性材料及微加工技术、非接触式光学传感器、自动化以及实验设计(DOE)、数据分析软件与过程控制的整合。由于体积微小、湍流程度和单位能耗较低,微小型反应器内的混合、传质、剪切特性与工业规模设备有一定的区别。现阶段微小型生物反应器主要用于菌株和细胞系筛选和工艺优化,在实现高通量工艺的同时确保了数据的丰度,对缩短研发周期和加速产品上市,尤其是在应对突发性传染性疾病方面有着重要的意义。未来,精准医疗概念的落实也依赖功能柔性化的微小型生物反应器系统。     

封闭式光生物反应器研究进展

国际上80～90年代,封闭式光生物反应器是微藻生物技术的重要研究热点,也是微藻生物技术产业化的关键技术之一。本文较全面地介绍了用于微藻大规模培养的封闭式光生物反应器研究现状。将封闭式光生物反应器分为柱式、管式、板式和光导纤维反应器等类型。工业放大前景的管式和板式光生物反应器采取了典型个案分析的方法,列表比较了典型反应器的主要技术参数,并对它们的技术发展趋势进行了归纳总结。     

生物反应器培养转基因鱼腥藻的研究

在反应器中研究了转人TNF-α基因鱼腥藻7120(Anabaena sp．PCC7120,pDC-TNF)的培养。结果表明气升式反应器适合于转基因鱼腥藻的培养。气升式反应器中通气量和光照是主要的影响因素,观察到1L罐中最适通气量为60～75L／h,最适光照强度为1200lx,此时在25℃混养,光照时间／黑暗时间为12h／12h,15d生物量干重大于3g／L,TNF表达水平约占总可溶蛋白的22％,达到了摇瓶培养水平。实验发现添加维生素B1 300μg／L、B12 200μg／L和生物素4μg／L时,生产周期为12d,缩短20％,表达水平相同。培养过程通入含有5％CO2的空气,能促进生长,缩短生产周期,但收获生物量不受影响。从添加维生素和通入CO2的培养结果证明反应器中培养时,光照是限制性因素,当反应器系统一定时,最终生物量有一个最大值,如需进一步提高产量,必须设法改变光照系统。     

光生物反应器培养微藻研究进展

微藻具有固定CO2和净化有机废水的能力,在环保、食品饲(饵)料、医药和生物能源开发等领域备受关注,但规模化培养及其产业化仍是研究的难点,亟待解决。就常用于大规模培养微藻的光生物反应器的特点和结构进行了综述。其中,封闭式微藻光生物反应器能够较好地调控藻种的培养条件、不易遭受污染,藻种的纯度容易控制,但培养规模小,生产成本较高;而开放式微藻光生物反应器无法精确控制藻种生长环境,但生产规模大、产量高、生产成本低,因此应用广泛。最佳的方法是综合两者优点,即首先利用封闭式微藻光生物反应器进行中试放大,大量繁殖藻种,然后投入开放式微藻光生物反应器内进行大规模商业生产,此方法有望成为微藻光生物反应器的发展方向,以期为微藻大规模培养提供参考借鉴。     

白檀离体快繁技术

以白檀(Symplocos paniculata)幼嫩茎段为实验材料, 通过对启动培养、增殖、生根培养及移栽的影响因子进行研究, 初步建立了白檀的组织培养体系。结果表明: 白檀外植体最适灭菌方案为0.1%升汞3分钟, 无菌苗获得率达81%; 最适初代启动培养基为1/2MS+30 g∙L^-1蔗糖+8 g∙L^-1琼脂, 出芽率达86.83%; 增殖最适培养基为1/2MS+1.0 mg∙L^-1 6-BA+0.02 mg∙L^-1 IBA+30 g∙L^-1蔗糖+8 g∙L^-1琼脂, 增殖系数达3.57; 最适生根培养基为WPM+0.5 mg∙L^-1 IBA+0.5 mg∙L^-1 NAA+20 g∙L^-1蔗糖+2 g∙L^-1 AC+8 g∙L^-1琼脂, 生根率达93%; 炼苗后, 移入园土:草炭土=1:1 (v/v)的基质中, 成活率达83%。     

重瓣满天星植株再生及其复壮和移栽

重瓣满天星 (Gypsophila paniculata)花蕾在附加有不同激素的 MS培养基上 (0～ 3mg/L6 - BA,0～ 1mg/L NAA,0～ 1mg/L 2 ,4 - D)均可长出再生植株 ,其中以 MS+6 - BA1mg/L效果最佳。试管苗复壮以 OM +6 - BA 1mg/L +2 ,4 - D 1mg/L效果最好。经复壮的试管苗较粗壮 ,但直接移植仍不能成活 ,必须在加盖旋松的培养瓶中进行溶液培养。2周后 ,小苗根系进一步发育扩展 ,然后逐步揭盖 ,锻炼幼苗抗干能力 ,移栽成活率可达 80 %。     

Uptake,accumulation and phytoextraction efficiency of cesium in Gypsophila paniculata

Abstract

To evaluate the phytoextraction efficiency of Gypsophila paniculata from Cs-contaminated soils and analyze the mechanism of Cs accumulation in G. paniculata, we analyzed the characteristics of Cs bioaccumulation and subcellular distribution, in addition to its chemical forms in the plant under hydroponic conditions. The results showed that total Cs content in the aboveground parts and the entire plant were as high as 6137.32?mg·kg^?1?dry weight and 7338.49?mg·kg^?1?dry weight, respectively, after 17?days in the 50?mg·L^?1 Cs treatment. The BCF was between 2.35 and 3.38. The TF was between 1.00 and 2.46 in G. paniculata. Subcellular distribution of Cs in the plant was as follows: soluble fraction?>?cell wall?>?organelles. Inorganic Cs (F-ethanol) and water-soluble Cs (F-dH2O) were the main types of Cs in G. paniculata. Further studies show that the phytoextraction efficiency can reach 10.30–11.91% planting a season of G. paniculata under potted conditions. The results suggested that G. paniculata, a perennial, drought-tolerant herb, was a high-accumulator of Cs, which may have potential uses in phytoremediation of Cs-contaminated soil.     

Adventitious shoot regeneration from leaf explants of Gypsophila paniculata L.

Adventitious shoots were successfully regenerated from leaf explants of Gypsophila paniculata L. The efficiency of shoot regeneration for cv. Arbel was tested on 18 media based on Murashige and Skoog basal medium containing different concentrations of thidiazuron or 6-benzylaminopurine in combination with naphthaleneacetic acid. Both explant age and that of the cuttings used as leaf donors affected the regeneration efficiency. The highest efficiency of adventitious shoot regeneration was obtained with the oldest leaves originating from the youngest cutting analyzed; on thidiazuron-containing medium, shoots regenerated on average from 67% of the leaves, with an average of seven shoots per explant. This regeneration procedure was suitable for all six commercial cultivars studied. Regenerated shoots elongated, rooted and successfully acclimatized to the greenhouse where they were grown to flowering. Received: 25 July 1998 / Revision received: 11 November 1996 / Accepted: 30 November 1996     

Effect of Cytokinins and Cytokinin Antagonists on in vitro Cultured Gypsophila paniculata L.

This study deals with the effects of two cytokinins [kinetin (Kin) and N-(2-chloro-4-pyridyl)-N-phenylurea (CPPU)] and cytokinin antagonists [2-chloro-4-cyclobutyl-amino-6-ethylamino-1,3,5-triazine (ACK1) and N-(4-pyridyl)-O-(4-chlorophenyl)carbamate (ACK2)] in concentration of 1 μM on in vitro cultured Gypsophila. The application of Kin and CPPU stimulated bud opening and increased fresh and dry masses. Cytokinin antagonists reduced the number of sprouted buds and bud fresh and dry masses. In plants treated with CPPU the chloroplasts possessed well developed membrane system, which covered almost the entire chloroplasts volume. In ACK2 treated plants, the plastid apparatus in each cell was represented by two types of chloroplast in which the inner membrane system was differently organized. Cell wall adjacent chloroplasts possessed structure similar to the controls. In inner located chloroplasts part of thylakoids were semi-concentrically arranged and partially destructed. This revised version was published online in July 2006 with corrections to the Cover Date.     

UDP-Glucuronosyltransferase activity is correlated to saponin production in Gypsophila paniculata root in vitro cultures

UDP-Glucuronosyltransferase (UGT, EC 2.4.1.17) activity was detected in excised root cultures of Gypsophila paniculata. The UGT activity (up to 0.8 nkat mg^–1 protein) correlated to the total saponin content (2 mg g^–1 dw) during the exponential phase of the batch culture. This gives rise to the hypothesis of a direct relationship in the biosynthetic pathway regulation between the enzyme activity and the saponin biosynthesis.     

Intergeneric somatic hybrid plantlets between Dianthus barbatus and Gypsophila paniculata obtained by electrofusion

Hypocotyl-derived protoplasts of Dianthus barbatus that had been pretreated with iodoacetamide were fused electrically with cell suspension culture-derived protoplasts of Gypsophila paniculata that could divide to form callus but could not regenerate shoots under the culture conditions used in this study. Electrofusion-derived calli which produced shoots were selected as putative somatic hybrids, and plantlets were subsequently regenerated from 2 of these selected calli. These plantlets, which in vitro produced flowers precociously, were identified as intergeneric somatic hybrids by nuclear ribosomal DNA analysis. Normal plants have not been established up to the present.     

Improved micropropagation of Gypsophila paniculata with bioreactor and factors affecting ex vitro rooting in microponic system

Studies on the mass production of high-quality plantlets in Gypsophila paniculata L. using a bioreactor and microponic system (a hydroponic system in which micropropagation shoots are planted) indicated that both aeration treatments, in which bioreactors were aerated from the top of explants by sparger (AS) and by tub (AT), were more effective than unaerated treatment for shoot proliferation and growth, and the maximum shoots (15.7 shoots per explant) with low hyperhydricity rate (2.9%) were found in the AS group. The ex vitro culture was more efficient for rooting when compared to the in vitro culture; the better shoot and root growth was obtained in the ex vitro culture, with rooting rate reaching 100% after 20 d of culture, but only 65% of in vitro shoots rooted; all stomata of ex vitro shoots closed, and their length was more than their width, but the stomata in in vitro shoots were all opened, the length close to the width. Furthermore, the stomata numbers were less in ex vitro (67.8) than in vitro (267.2). The survival rate of ex vitro plants reached 83.3% when plantlets derived in vitro and ex vitro were transferred to pots, while only 23.3% of in vitro plantlets survived. During ex vitro rooting with the microponic system, foam as the supporter material, 90 μmol?m^?2?s^?1 of light, and 80 shoots of planting density were favorable for shoot and root growth. The combination of bioreactor and microponic systems is an efficient way to produce high-quality plantlets of G. paniculata. Their application can reduce costs during large-scale industrial production.     

满天星试管苗与其玻璃化苗的RAPD指纹图谱分析

采用分离群体分组分析法(BSA),用100个随机引物对满天星的正常苗和玻璃化苗进行RAPD分析的结果表明,7个随机引物扩增出多态性差异条带。再用上述7个引物分别对试管苗及其玻璃化苗个体进行DNA的PCR扩增的结果显示,引物J20在2种苗中出现差异条带。