草麻黄细胞悬浮培养体系的建立

目的:建立草麻黄悬浮培养体系.方法:采用组织培养的方法探讨了水解酪蛋白(CH)、基本培养基、取材时间、和摇床转速对麻黄愈伤组织悬浮培养的影响.结果:MS基本培养基、300mg/l水解酪蛋白、继代25d的愈伤组织、转速110r/min是愈伤组织悬浮培养的适宜条件,愈伤组织细胞增殖量分别为0.76g、0.80g、0.80g、0.80g.结论:初步选择出草麻黄愈伤组织细胞的悬浮培养条件,为麻黄细胞扩大培养及有效成分提取奠定基础.     

紫茉莉悬浮细胞培养体系的建立

为建立紫茉莉(Mirabilis jalapa L.)悬浮细胞培养体系,以紫茉莉无菌苗叶片诱导的愈伤组织为材料,筛选紫茉莉悬浮细胞的适宜培养体系。结果表明,紫茉莉愈伤组织在MS+2,4-D 1 mg L-1+KT 0.5 mg L-1的液体培养基中悬浮继代培养3~4次,能得到稳定的悬浮细胞系。培养基的pH值为5.5~5.9,蔗糖浓度为30 g L-1更适合悬浮细胞的生长。紫茉莉悬浮细胞的生长曲线大致呈S型。最佳继代培养时间是10 d,培养液的体积为40 mL时,接种量为7.5 mL,可以较好地保持悬浮细胞系。1 L培养液中可提取分泌蛋白(0.42±0.15) g。这些有助于对悬浮细胞提取分泌蛋白的研究。     

沙枣细胞悬浮培养体系的建立

分别以沙枣幼嫩叶片,茎段为外植体诱导沙枣愈伤组织,选择生长旺盛的松散愈伤组织进行细胞悬浮培养,探讨了培养基种类和激素组合对其产生的影响.结果表明,最适的沙枣愈伤组织诱导培养基为NT(包括0.1 mg·L-1BA和0.1 mg·L-1TDZ);沙枣悬浮细胞在MS,NT,WPM,B5和IS培养基中均可生长,其中在NT培养基中生长状态最好,呈现生长均一的绿色疏松状态.激素组合对沙枣细胞生长影响很大,其中附加BA 0.1 mg·L-1和TDZ 0.1 mg·L-1组合的NT液体培养基可获得大量繁殖速度快、生长均匀一致的悬浮细胞,细胞增长系数达5.73.HPLC测定结果证明沙枣细胞中含有一定量的浓缩鞣质达5.2022 mg/gDW,这为利用沙枣悬浮细胞生产次生代谢产物提供了一条有效途径.     

水翁悬浮细胞系的建立及其悬浮培养的生长特性

建立了水翁悬浮细胞系,并对其悬浮培养的生长特性作了初步探讨。以水翁新生芽尖作为外植体,接种于添加有不同浓度和配比的生长调节物质及各种附加物的MS固体培养基中,诱导培养产生初代愈伤组织;挑选Ⅰ和Ⅱ型的愈伤组织进行继代培养改良,考察愈伤组织的生长状况和统计生长量来决定最佳继代培养基的配方和得到适合悬浮培养的愈伤组织;将以上得到的愈伤组织转接于最佳继代液体培养基中,于24±1℃,120r/min条件下振荡培养,筛选分散度好、较均匀、生长快、色浅透明的细胞作为种子传代,数次传代后得到性能良好的悬浮细胞系;以细胞生长量(鲜重)为指标,绘制了水翁悬浮细胞的生长曲线。研究表明:2.0mg/L的2,4-D的诱导率最高(92%,初代愈伤组织为Ⅰ型),Ⅱ型愈伤组织的最高诱导率为75%;最佳的继代培养基配方为MS 0.5mg/L 2,4-D 0.5mg/L 6-BA 1.0mg/L IAA 0.5mg/L IBA 0.5mg/L NAA 0.1mg/L KT 700mg/L LH,形成Ⅱ型愈伤组织的生长量可达3.28g/瓶(鲜重);液体继代培养3代后,可得到性能良好的悬浮细胞系;水翁悬浮细胞的生长曲线表明,最佳接种期为培养后的16~18d。     

藏红花细胞悬浮培养体系的建立及优化

基于诱导的藏红花细胞系,通过摇瓶法,优化了其液体培养基、接种量和种龄等培养条件,以建立藏红花细胞悬浮培养体系。结果表明,将生长在固体培养基上的藏红花愈伤组织接种在MS液体培养基(添加了2mg/L2,4-D,1mg/L6-BA和300mg/LCH)中,于(22±0.3)℃,120r/min的摇床上,暗培养30d,便可获得藏红花的悬浮细胞系。经优化其培养基、接种量和种龄,将种龄为20d的细胞系,按照5%接种量接种在液体B5培养基(添加了2mg/LNAA,1mg/L6-BA和300mg/LCH)中,于(22±0.3)℃,120r/min的摇床上,培养36d,细胞生物量(13.4g/L)和藏红花素产量(0.91g/L)均达到最高。本研究建立的藏红花细胞悬浮培养体系为其生物反应器放大培养奠定了基础。     

薰衣草细胞悬浮培养体系的优化

采用正交设计方法对影响薰衣草悬浮细胞生长的因素进行了优化研究,筛选了最有利于薰衣草悬浮细胞生长的培养条件:含有0.025mg/L2,4-D、0.5mg/L6-BA、40g/L蔗糖的B5培养基和120r/min的转速,在此条件下培养15天,悬浮活细胞密度可达到4.4×105个/ml;经过3个月的培养,悬浮细胞的分化率可达到90%以上。通过分析薰衣草细胞对残糖的消耗规律,探讨了蔗糖浓度影响活细胞密度的原因。     

白桦单细胞培养体系的建立

以白桦花药（单核靠边期）、种子（萌发）及再生苗的茎段诱导愈伤组织,在NT1、NT2和B5三种不同的培养基中进行悬浮培养,筛选出优良的白桦悬浮体系,而后进行单细胞分离的结果表明,白桦花药愈伤组织在B5培养基上生长迅速,分散性好,适合于细胞分离。分离得到的单细胞用不同方式培养,建立了白桦单细胞两步培养法。     

中麻黄的组织培养

1植物名称中麻黄（Ephedraintermedia）。2材料类别幼茎。3培养条件愈伤组织诱导及其继代培养基：(1)MS＋KT0．05mg·L－1＋2．4D1．1mg·L－1（单位下同）;（2）芽分化培养基：MS＋BA3＋IAA0．2;（3）生根培养基：MS＋BA0．06＋2,4-D1.1。以上培养基均加30g·L-1蔗糖,9g·L-1琼脂,pH值5.8左右,高压灭菌。培养温度(25±）℃,光照12h·d－1,光照度2000lx.4生长与分化情况4.1愈伤组织诱导及继代培养无菌操作下,切取中麻黄幼苗节间幼茎5mm大小,置培养基（1）上。15d后,淡黄绿色愈伤组织开始生长,愈伤组织疏松易碎。再过1…     

中麻黄的化学成分研究

研究中麻黄(Ephedra intermedia Schrenk et C.A.Mey.)干燥草质茎的化学成分及其抗哮喘活性。应用硅胶柱色谱、Sephadex LH-20柱色谱、Toyopreal HW-40C柱色谱以及半制备型高效液相色谱等多种手段对中麻黄草质茎的50%丙酮提取物进行化学成分研究,从中分离鉴定了22个化合物,根据其理化性质和波谱学数据鉴定其结构,分别为4-甲氧基肉桂酸(1)、肉桂酸(2)、ω-hydroxypropioguaiacone(3)、threo-8S-7-methoxysyringylglycerol(4)、3-(2,4′-dihydroxy-3′,5′-dimethoxyphenyl)propanoic acid(5)、8-hydroxy-9-methyl-7-(4-hydroxy-3,5-dimethoxyphenyl)-7-propanone(6)、3-羟基-1-(4-羟基-3,5-二甲氧基苯基)-1-丙酮(7)、2-羟基-1-(4-羟基-3-甲氧基)苯基-1-丙酮(8)、异咖啡酸甲酯(9)、trans-syringin(10)、cis-syringin(11)、(4R)-4-hydroxy-4-(2-hydroxypropan-2-yl)cyclo-hex-1-ene-1-carboxylic acid(12)、7-羟基-α-松油醇(13)、去氢吐叶醇(14)、脱落酸(15)、黑麦草素内脂(16)、(E)-3-(3-hydroxybut-1-enyl)-2,4,4-trimethylcyclohexa-2,5-dienone(17)、3-(3′-hydroxybutyl)-2,4,4-trimethylcyclohexa-2,5-dienone(18)、对异丙基苯甲酸(19)、p-cymen-7-yl β-D-glucopyranoside(20)、acyclic 7-hydroxygeranic acid(21)、7,7-dimethylbicyclo[3.1.1]hept-2-ene-1-carboxylic acid(22),其中化合物3～6、8、10～14、17～19、21、22为首次从该植物中分离得到,同时经体外抗哮喘活性筛选,结果显示化合物11和14能够显著抑制β-氨基己糖苷酶的释放。     

茶条槭悬浮培养体系的建立与没食子酸合成的优化条件

初步建立茶条槭（Acer ginnala）细胞悬浮培养体系：以茶条槭子叶为外植体,接种于WPM培养基中,对茶条槭愈伤组织进行诱导和继代培养。悬浮培养中,每代增长指数达到11．6,没食子酸含量达到1．518％。通过对比NT、IS、WPM、B5和MS培养基所含成分对茶条槭愈伤组织悬浮培养的影响,综合考虑悬浮细胞的生长速率和有效成分的含量,确定WPM为基本培养基。WPM培养基大量元素的浓度对细胞的生长和没食子酸的积累有显著影响,其浓度越高,促进作用越明显。3倍浓度的大量元素最有利于没食子酸的积累。蔗糖浓度为10g·L^-1最适于没食子酸的积累,浓度为30g·L^-1最适于茶条槭细胞生长和没食子酸合成。     

盐碱地种植麻黄试验研究

麻黄在干旱区硫酸盐,氯化物盐碱地中种植,其土壤0-30cm土层含盐量1.2%以下生长良好,初步化验分析,麻黄在盐碱地中种植其麻黄碱含量有所提高,同时可获得较大的社会、生态、经济效益。     

2种麻黄光合及其耐逆性分析

在河西临泽小泉子麻黄大田种植试验区和民勤沙生植物园中草药种植区,对多年生中麻黄(Ephedra intermedia Schrenk ex Mey.)和草麻黄(E.sinica Stapf)的光合速率、蒸腾速率及其影响因子进行测试分析,结果表明:(1)中麻黄和草麻黄的光合速率日变化均为单峰曲线,峰值均出现在11:00时,分别为12.098和11.560μmol.m-2.s-1。(2)中麻黄蒸腾速率日变化为单峰曲线,峰值在11:00时,为25.992 5 mol.m-2.s-1;草麻黄蒸腾速率日变化呈双峰曲线,峰值分别出现在11:00时与15:00时,峰值为26.280 0和24.3600 mol.m-2.s-1。(3)2种麻黄光合速率与光量子通量密度、大气温度、水汽压亏缺及蒸腾速率之间均呈显著或极显著正相关关系,与大气CO2浓度和胞间CO2浓度之间均呈显著或极显著负相关关系,与其余因子相关不显著。(4)2种麻黄水分利用效率对比分析显示,中麻黄水分利用效率值(平均0.9022)高于草麻黄(平均0.4532),表明中麻黄与草麻黄相比在生长过程中是以较低的蒸腾强度和相对较高的光合速率值来适应干旱荒漠环境的,比草麻黄更具抗干旱性。     

Does proliferation capacity of lymphocytes depend on human blood types?

In vitro human lymphocyte culture methodology is well established yet certain confounding factors such as age, medical history as well as individual’s blood type may potentially modulate in vitro proliferation response. These factors have to be carefully evaluated to release reliable test report in routine cytogenetic evaluation for various genetic conditions, radiation biodosimetry, etc. With this objective, the current study was focused on analyzing the proliferation response of lymphocytes drawn from 90 individuals (21-29 years) with different blood types. The proliferation response was assessed in the cultured lymphocytes by cell cycle, mitotic index (MI), and nuclear division index (NDI) after stimulation with phytohaemagglutinin (PHA). To investigate the toxic effect on proliferation, MI was calculated in representative samples of each blood type were X-irradiated. The results showed that there was no significant difference among the cell cycle phases of lymphocytes in different blood types (P > 0.05). Similarly, both MI and NDI of lymphocytes derived from different blood types also did not show significant difference ( P > 0.05). The extensive interindividual variation within and among the blood types is likely responsible for the lack of significant difference in lymphocyte proliferation. Although spontaneous proliferation efficiency of lymphocytes of different blood types after PHA stimulation was grossly similar, the MI observed after radiation exposure showed a significant difference ( P < 0.05) indicating a differential proliferation response among the blood types. Our results suggest that the blood types did not have any impact on PHA-induced proliferation; however, a specific differential lymphocyte proliferation observed after radiation exposure needs to be considered.     

洗涤剂对蚕豆根和叶片的作用

研究了家用洗衣粉对蚕豆根生长、细胞分裂和叶片叶绿素含量的影响。结果表明:不同浓度的洗衣粉均能抑制蚕豆根的生长,降低根尖细胞有丝分裂指数,诱发细胞产生高频率的微核且是明显的时间与剂量效应关系和叶绿素含量明显下降。     

Syrian hamster lymphocyte cytogenetic technique and effects of blood storage on mitotic index

Summary The Syrian hamster has a diploid chromosome complement similar to humans in both number (2N=44) and morphology. For comparative mutagenic studies with humans, a repeatable lymphocyte chromosome technique involving laboratory mammals is desirable. The reported hamster lymphocyte cytogenetic technique appears technically uncomplicated and repeatable while providing a sufficient number of metaphase cells for quantiative analysis. In order to determine the effects of storing whole-blood (in culture media) for varying time periods at 8°C prior to adding a mitogen, the mitotic index was calculated following a 48-h culture period. Results indicated the storage up to six days can still result in a mitotic index adequate for cytogenetic analyses.     

Annual mitotic activity in the intestinal epithelium of the musselCrenomytilus grayanus

The seasonal dynamics of cell reproduction in the intestinal epithelium of the musselCrenomytilus grayanus are described in detail. Mitotic indices in the intestinal epithelium varied throughout the year from 0.005 to 0.26% (averaged data) and from 0.003 to 0.37% (individual data). Cyclic seasonal changes were found in the mussel’s intestinal epithelium. In general, the average values of mitotic activity in the intestinal epithelium were low (the mitotic index was 0.13%); there was a rise in activity in late April–June and September and a decline in July–August and especially in January–March. The winter-early spring period was characterized by a profound inhibition of cell reproduction and the transition of cells to the resting state. An outburst of proliferation occurred in the spring, due to a manifold increase in the number of cells in the mitotic cycle. The musselC. grayanus may be a good model for the study of the two extreme states of proliferation and their alternation in marine animals in nature. The diel dynamics of mitotic activity in the intestinal epithelium were followed during the most active growth period (May). The mitotic index (MI) varied during the day within a narrow range, deviating from the daily average value by no more than one third; no pronounced diel rhythm was found. Optimum water temperatures for cell reproduction ranged from 5 to 18°C.     

Effects of the Application of Different Concentrations of NaN3 for Different Times on the Morphological and Cytogenetic Characteristics of Barley （Hordeum vulgate L.） Seedlings

Sodium azide （NAN3） has been used in many biological studies as a mutagen. In the present study, the morphological and cytogenetic effects of NaN3 on barley （Hordeum vulgare L.） seedlings were investigated. Seeds of barley were treated with different concentrations of NaN3 （0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 mmol/L） and applied for different periods of time （3 and 4 h）. Parameters investigated, except for the mitotic index, were determined on Days 7 and 14. Increasing concentrations of NaN3 affected germination rates on Days 7 and 14 following application for 3 and 4 h. Although the length of the roots and leaves was affected by treatment with NaN3 on Day 14 of the germination period, coleoptile length was affected by NaN3 treatment on Day 7. Increasing concentrations of NaN3 and increased treatment period decreased the mitotic index compared with the untreated control. The inhibitory effects of NaN3 on the mitotic index indicate that NaN3 can have genotoxic and mutagenic effects on barley seedlings.