Growth and Maintenance of an Embryogenic Cell Culture of Daylily {Hemerocallis) on Hormone-free Medium

The division of chloroplasts in Trebouxia potteri was studiedby electron microscopy. At the beginning of chloroplast division,vesicles with fine fibrils (FVs) and ER attach to the isthmusof the chloroplast. Then, filaments appear around the isthmusparallel to the direction of constriction and seem to contractin order to decrease the diameter of the isthmus. It is suggestedthat the FVs are involved in the formation of the filamentsand that the ER is involved in the contraction of the filaments.At the final stages of the division of the chloroplast, thefilaments decompose. FVs are partially surrounded and decomposedby lysosomal sheets. For the next cycle of division of the chloroplast,the recovery of substances from decomposed filaments by functionalFVs seems a realistic possibility. chloroplast division, division apparatus, division cycle, transmission electron microscopy, Trebouxia potteri     

葡萄子房胚性细胞悬浮系的建立及其生长特性研究

由葡萄栽培品种“蜜汁”子房诱导的愈伤组织,经不同浓度２,４－Ｄ试验表明,０．５ｍｇ／Ｌ的２,４－Ｄ最有利于形成胚性细胞;该浓度下形成的质地紧密适度的愈伤组织经２个月的悬浮振荡培养,成功地建立了分散性好、生长旺盛的胚性细胞悬浮系;并通过培养过程中生长曲线的测定,确定了细胞悬浮系的生长特性。     

New Culture Medium for Maintenance of Tsetse Tissues and Growth of Trypanosomatids*

SYNOPSIS. A new culture medium (SM), based on the amino-acid composition of tsetse hemolymph and containing fetal bovine serum, was designed for the maintenance of tsetse organs and the cultivation of various trypanosomatids. For optimum growth 20% (v/v) serum was required. The medium supported prolonged peristalsis of the alimentary tract and salivary glands of pre-emerged Glossina morsitans morsitans. In established cultures, derived from bloodstream forms of pleomorphic Trypanosoma brucei brucei and Trypanosoma brucei rhodesiense strains, inocula of ～ 10⁶ procyclics/ml yielded 4–5 × 10⁷ organisms/ml after 4 or 5 days of incubation at 28 C. Bloodstream forms of a cloned monomorphic T. b. brucei strain were also able to transform into procyclics, which, however, multiplied at a lower rate, with maximum yields of ～ 2 × 10⁷ after 5 days. Cultures of Trypanosoma congolense and of a nearly monomorphic Trypanosoma brucei gambiense strains could be established in SM medium only in the presence of tsetse alimentary tract. The procyclic trypomastigotes of these species, adapted to SM medium and able to grow in it without Glossina organs, gave maximum populations of ～ 4.5 × 10⁷ cells/ml. Promastigotes of Leishmania donovani, cultivated routinely in a diphasic Table's medium, multiplied actively upon being transferred into SM medium, producing yields of ～ 4 × 10⁷ cells/ml.     

Solasodine Yield in Callus Cultures of Solanum Iaciniatum Ait. Selected for Growth on Hormone-free Medium

Callus autotrophic for both auxin and cytokinin was selectedfrom a hormone-requiring culture of Solanum laciniatum Ait.The control of solasodine yield, callus growth and adventitiousshoot initiation by some exogenous growth regulators and/orlight in the habituated culture was determined. Solasodine concentrationvaried from 0.3 to 1.4 mg g^–1 d.wt. Solanum laciniatum, callus, habituation, solasodine     

细胞实验中细胞生长及换液处理对实验结果的影响

利用所设计的核酶抑制转染了HBV的HEPG2细胞内病毒的复制,对所产生的病毒抗原的量进行统计学处理,从而观察到细胞生长和细胞培养液换液对结果的影响。结果表明,换液处理组与未换液处理组的抑制率不同;在96h,未经换液的孔抑制率出现了负值。细胞在80h后数目减少,镜检显示细胞状态不佳。应用细胞生长曲线和细胞换液处理对于实验结果有一定的修正作用。     

多孔微载体无血清培养rCHO细胞生产u-PA

在30L搅拌式反应器中无血清培养分泌尿激酶型纤溶酶原激活剂(u-PA)的DNA重组CHO细胞,定期部分更换Cytopore多孔微载体,使生长在多孔微载体中的细胞不断更新繁殖,解决大规模细胞培养中的细胞凋亡问题。在91d连接换液培养过程中,细胞密度可维持在(1.3～2.6)×107/mL,活细胞比率维持在90%以上。在7.5L搅拌罐中培养细胞,利用外部周期性压力振荡刺激并结合载体更新技术,可减轻密度效应对细胞生长和表达的影响,在一定程度上提高细胞在高密度培养条件下的表达水平。在67d连续换液培养中,细胞最高密度为2.64×10⁷/mL,活细胞比率维持在95%以上。与稳压操作相比,利用周期变压刺激技术可提高产量10%～20%,且可降低葡萄糖厌氧代谢生成乳酸的转化率,利用4步纯化工艺,从含u-PA约135g的2100 L上清中获得约80guPA(单链比例约为90%)。     

多孔微载体无血清培养rCHO细胞生产u-PA

在30L搅拌式反应器中无血清培养分泌尿激酶型纤溶酶原激活剂(u-PA)的DNA重组CHO细胞,定期部分更换Cytopore多孔微载体,使生长在多孔微载体中的细胞不断更新繁殖,解决大规模细胞培养中的细胞凋亡问题。在91d连接换液培养过程中,细胞密度可维持在(1.3～2.6)×107／mL,活细胞比率维持在90％以上。在7.5L搅拌罐中培养细胞,利用外部周期性压力振荡刺激并结合载体更新技术,可减轻密度效应对细胞生长和表达的影响,在一定程度上提高细胞在高密度培养条件下的表达水平。在67d连续换液培养中,细胞最高密度为2.64×107／mL,活细胞比率维持在95％以上。与稳压操作相比,利用周期变压刺激技术可提高产量10％～20％,且可降低葡萄糖厌氧代谢生成乳酸的转化率,利用4步纯化工艺,从含u-PA约135g的2100L上清中获得约80gu-PA(单链比例约为90％)。     

Detection of an Embryogenic Cell Antigen in Carrot

Monoclonal antibodies were raised against proteins in a microsomalfraction from carrot embryogenic cells. The presence of a 31-kDaembryogenic cell antigen detected by one antibody (ID 11) wasdemonstrated in embryogenic cells but not in other plant materials,such as non-embryogenic cells, somatic embryos, crown gallsand hairy roots. The antigen was also present in organ segmentsthat carried somatic embryos having been induced by exposureto various stresses. In non-embryogenic cells, the antibodyrecognized a small amount of a 32-kDa antigen. Both the 31-and the 32-kDa antigens accumulated in carrot seeds during theirdevelopment and then disappeared after germination. (Received April 16, 1990; Accepted July 16, 1990)     

大花萱草‘金娃娃’雌雄蕊发育进程及其与花蕾长度的相关性研究

采用石蜡切片法对大花萱草'金娃娃'(Hemerocallis hybridus cv.'Stella de oro')不同花蕾长度时雌雄蕊的形态及其发育时期进行观察,以探讨花营长度与雌雄蕊发育进程的相关性.结果显示:(1)'金娃娃'雄蕊发育正常,小孢子发生及雄配子体的发育过程与常见单子叶植物类似,成熟花粉属二胞型;雌蕊子房具多胚珠,但各胚珠中雌配子体发育进程不同步,出现大量无胚囊或胚囊内核发育紊乱、解体的现象而导致雌配子体发育不正常,大田结实率不到5%且种子干瘪不能发芽.(2)'金娃娃'花蕾长度与花粉发育时期具有相关性,为花蕾长度作为组织培养外植体取材外形标准提供丁植物胚胎学依据.     

贡蕉胚性细胞悬浮系的建立和植株再生

鲜食蕉品种的高度不育性和多倍性制约了用传统育种方法培育生产实践中所需的新品种 ,建立稳定的胚性细胞悬浮系是香蕉生物技术育种的前提。以目前国内尚未建立该体系的鲜食蕉品种贡蕉 (AA)未成熟雄花序的第 1～ 15位花梳为外植体 ,对胚性细胞悬浮系的建立和植株再生体系进行了优化。结果表明 ,5～ 6个月的培养后可获得分生小球体和浅黄色、松散易碎的胚性愈伤组织。 9μmol/L 2,4 D对外植体愈伤组织的诱导效果最好 ,诱导率为 40.96 % ,胚性愈伤组织诱导率可达7.45 % ,其中5.79%的胚性愈伤组织来源于第 6～12号位置的花梳。胚性愈伤组织悬浮培养后 ,通过 3个月的筛选和继代培养 ,可得到均质的胚性细胞悬浮系。该培养体系合适继代周期为 15d ,继代时合适的起始接种量为每 30mL培养基加 2mLPCVECS。培养 6个月的胚性细胞在体细胞胚诱导培养基中培养15d后可见到白色半透明体细胞胚的发生 ,体细胞胚诱导率为 2 80× 10³个 mLPCV。成熟体细胞胚的萌发率为 17 2 8% ,其中发育成正常的再生植株的百分率为 14 16 %。     

野葛细胞培养及其生长特性的研究

目的：以野葛不同器官来源的外植体为材料诱导愈伤组织,建立起悬浮细胞培养体系,研究愈伤组织及悬浮细胞的生长特性。方法：采用细胞生长测定、细胞培养物中有效成分含量测定及细胞的观察。结果：野葛细胞培养过程中异黄酮及葛根素含量随细胞生长逐渐积累,在细胞生长静止期含量最高,次生代谢物与细胞生长呈负相关。不同来源的愈伤组织以根愈伤组织中异黄酮及葛根素含量最高,其中总异黄酮含量为：47．60mg/g Dw、葛根素含量为：3．30mg／g Dw,其次叶、茎、子叶,悬浮细胞中含量最低。不同愈伤组织的细胞及悬浮细胞在生长过程中细胞形态有差异。     

应用电导率监测红豆杉细胞悬浮培养中细胞生长与紫杉醇的累积

对中国红豆杉细胞悬浮培养过程中细胞生长和紫杉醇累积及培养液的电导率变化规律进行了研究,实验表明,细胞悬浮培养过程中随着细胞生物量与紫杉醇含量的增加,培养液的电导率逐渐下降,细胞生长和紫杉醇累积曲线与培养液的电导率曲线恰成镜像相关,当生物量达到最高时电导率达到并稳定于最低;紫杉醇累积的高峰滞后于细胞生物量的高峰2～3 d;电导率可作为红豆杉细胞培养中确定生物量和紫杉醇累积高峰期的监测指标.     

大麦胚性细胞悬浮系的建立及其生长特性

由春大麦品种“如车”种胚诱导的松脆型胚性愈伤组织经2个月的悬浮培养,成功建立分散性好、生长速度快的胚性细胞悬浮系。该系细胞直径为1-3mm,由富含淀粉粒的胚性薄壁细胞构成。经不同浓度2,4-D实验,发现2mg/L最适合该细胞系的生长。文中对成功建立大麦胚性细胞悬浮系的关键问题进行了讨论。     

高羊茅胚性悬浮细胞系的快速建立及其单细胞培养

本研究采用高羊茅(Millennium和Hundog Ⅴ)成熟种子为材料,以MS为基本培养基,通过添加2.5 mg/L CuSO4·5H2O,或提高NH4NO3浓度至2.5 g/L等措施均能明显提高愈伤组织的质量,经过3～5个月的筛选获得疏松干燥、颗粒状、生长旺盛、适合悬浮培养的Ⅱ型胚性愈伤组织.悬浮培养初期需用MSⅠ液体培养基进行一个月的启动培养,之后转用MSⅡ继代保持,约2个月左右即建立起来自高羊茅2个品种的3个悬浮细胞系.生长特性测定结果表明,悬浮培养的初始接种量以1.0～3.0 ml/40 ml为宜,生长周期内其pH值不断波动变化,最适范围在5.0～5.7之间.5～8 d为悬浮系的对数生长期,此时细胞分裂旺盛,增殖较快,是分离单细胞的最佳时期.单细胞培养方式以悬浮培养效果最好.     

培养基成分对东北红豆杉细胞生长和紫杉醇产量的影响

在东北红豆杉(taxuscuspidate)细胞培养物的基本培养基中加入30mg@L-12.4-D,可以提高细胞培养物的生长量,其相对最高生长量可以达到85.999mgFW/gFW@d;培养基中加入活性炭粉末lg@L-1,可以克服植物细胞生长过程中的褐化问题;适当种类及含量的生长调节剂的加入:6-BA0.5mg@L-1+KT2mg@L-1,可以使植物细胞合成和积累相对较多的紫杉醇(Taxol)。 Abstract:The growth quantity of Taxus Cuspidate's cell culture materials can be increased and the highest quantity will amount to 85.999 mgFW/g·FW·d·if We put 2,4-D (30rog·L-1 )into the basic cuhure medium of it; The difficulty of foxiness during the plants growing process can be overcome if we add acive carbon powder(lgL-1)into the culture medium; The plant cell can synthetize and gather more Taxol comparatively with the addition kf proper and amount of growth regulators:6-BA(0.5mg·L-1) + KT2.     

培养基成分对东北红豆杉细胞生长和紫杉醇产量的影响

在东北红豆杉（taxus cuspidate)细胞培养物的基本培养基中加入30mg.L^-12.4-D,可以提高细胞培养物的生长量,其相对最高生长量可以达到85．999mgFW/gFW.d;培养基中加入活性炭粉末1g.L^-1,可以克服植物细胞生长过程中的褐化问题;适当种类及含量的生长调节剂的加入：6－BA0．5mg.L^-1＋KTmg.L^-1,可以使植物细胞合成和积累相对较多的紫杉醇（Taxol).     

大花萱草组培快繁体系的研究

以大花萱草的6个品种为试材,主要研究了其组织培养中的激素配比、外植体类型、基因型以及不定芽的生根条件。结果表明：适合于大花萱草紫蝶的最佳愈伤组织及不定芽诱导培养基为MS+1 mg·L^-1 6-BA+0.1 mg·L^-1 NAA,出愈率、分化率和平均出芽数分别为63.33%、91.11%和5.86;但是,上述指标在基因型间表现出差异;花茎是诱导不定芽的最佳外植体;1/2MS+0.2 mg·L^-1 NAA是紫蝶较适宜的生根培养基,生根率81.11%,平均生根数6.08;本实验建立了3个品种的再生体系,平均出芽数均在4个以上,其中“金娃娃”最高,是6.88。     

Calcitonin Gene-Related Peptide (CGRP) Stimulates Purkinje Cell Dendrite Growth in Culture

Previous reports described the transient expression during development of Calcitonin Gene-Related Peptide (CGRP) in rodent cerebellar climbing fibers and CGRP receptor in astrocytes. Here, mixed cerebellar cultures were used to analyze the effects of CGRP on Purkinje cells growth. Our results show that CGRP stimulated Purkinje cell dendrite growth under cell culture conditions mimicking Purkinje cell development in vivo. The stimulation was not blocked by CGRP8-37, a specific antagonist, suggesting the activation of other related receptors. CGRP did not affect survival of Purkinje cells, granule cells or astrocytes. The selective expression of Receptor Component Protein (RCP) (a component of CGRP receptor family) in astrocytes points to a role of these cells as mediators of CGRP effect. Finally, in pure cerebellar astrocyte cultures CGRP induced a transient morphological differentiation from flat, polygonal to stellate form. It is concluded that CGRP influences Purkinje cell dendrite growth in vitro, most likely through the involvement of astrocytes.