魔芋无土栽培营养液配方研究初报

分别采用全MS、1／2MS、1／5MS、1／10MS(含大量元素、微量元素和铁盐)、1／10MS大量元素营养液(不含微量元素和铁盐)定期浇灌用蛭石栽培的不同级别的魔芋,前50d生长状况没有差异,50～80d以1／2MS～1／10MS营养液为佳,80～110d时1／5MS～1／10MS生长较好;魔芋在缺乏微量元素的营养液中后期生长不良,提前倒苗;产量分析结果表明：35g以上的种芋以1／2MS最高,35g以下的种芋以1／10MS最高,魔芋的耐盐浓度在2．25％。以下,高盐浓度对块茎的形成有毒害作用,造成产量大幅度下降,添加微量元素和铁有较好的增产作用。     

魔芋试管芋营养液栽培生产原原种技术研究

在温室对魔芋试管芋立体多架层营养液栽培生产原原种技术进行了研究。以不同质量分数和比例的N、P、K、Ca、Mg等大量元素和MS培养基的微量元素配制成营养液,对魔芋试管芋进行立体多架层无土栽培试验,结果表明:生长状况和原原种的产量均以N∶P∶K∶Ca:Mg的比例为4∶1∶2.4∶0.2∶0.7的Ⅴ号营养液最高,光照强度以2000~6000 Lux为宜。该试验为魔芋营养液的配方研制及栽培管理技术提供了理论依据。     

魔芋微型试管芋(拟球茎)繁殖技术研究

在魔芋愈伤组织的基础上,研究了培养基组成、培养条件等因素对试管芋形成的影响。魔芋愈伤组织在添加6-BA3.0 mg.L-1、NAA0.5 mg.L-1、IBA0.5 mg.L-1、KT5.0 mg.L-1的MS培养基上,20 d后形成大量不定芽。待不定芽生长至约0.5~1 cm时,将温度控制在15~20℃之间,不定芽基部膨大,2个月后形成多达10个以上的微型种芋,种芋之间以愈伤组织相连。将微型种芋连同愈伤组织取出,栽培到适当的基质中,浇灌营养液使基质保持湿润,自然条件下萌发,出苗率在90%以上,叶片多达18片,当年每块微型种芋可形成5 g以上的多个子芋。     

哈蜜瓜幼胚体培养

植物名称:哈蜜瓜(Cucumis melo var Saccharinus Naudin) 材料类别:授粉后10天的幼胚——球形～心形胚期,带三分之一的种体培养。培养条件:诱导幼胚生长及产生愈伤组织的基本培养基为MS附加(单位为mg/1)①KT0.5,BA0.5,IAA1;②KT1,IAA?诱导愈伤组织再分化培养基为改良MS附加不同浓度的激素,其配方为:①改良MS(变有机物为B_5的成分及用量)附加 KT2,硫酸腺嘌呤3,赤霉素 5,②1/2 MS大量元素、微量元素、铁盐,附加BA10。诱导生根培养基为N_6大量、MS微量、铁盐(以上元素都减半),B_5的碘和钼、有机物,MS的肌醇,硝酸铵297,附     

麝香石竹(康乃馨)试管苗的开花

植物名称:麝香石竹(Dianthus caryophyllus)材料类别:茎尖芽及幼苗培养条件:选用4种培养基,即①MS-1:0.5MS大量+微量元素;②MS-2:完全的MS基本成分;③MS-3:0.25MS大量+微量元素;④怀特(White)     

无机盐用量调整对太行菊不定芽增殖与生长的影响

以太行菊不定芽为材料,通过调整MS培养基中大量元素、微量元素、铁盐、Ca2+、K+的用量与配比,研究MS培养基不同的无机盐用量水平对太行菊不定芽增殖和生长的影响。结果表明,太行菊不定芽的增殖与生长适应较高含量的MS大量元素,降低培养基中的MS大量元素用量,不定芽分化数量减少,增殖系数降低,并且不定芽长势弱,叶片出现黄枯。单独加倍MS培养基中的K+(KNO3与KH2PO4)含量,可促进太行菊不定芽的增殖与生长,不定芽增殖系数明显高于对照及其他处理。增加或降低MS培养基中的Ca2+含量,对太行菊不定芽的增殖与生长影响不大;去掉培养基中的微量元素,不定芽的增殖与生长明显受到抑制,不定芽分化数量减少,且不定芽的叶片枯死严重。减半培养基中的铁盐含量,对不定芽的生长影响较小;但加倍培养基中的铁盐含量,对不定芽的增殖和生长均产生强烈的抑制作用。     

九谷幼穗培养及植株再生

植物名称:谷子(Setaria italica)九谷品种。材料类别:大田栽培的九谷取用穗长约2～15cm的幼穗。培养条件:(1)愈伤组织的诱导采用组合的培养基,即PC-l2的大量元素和微量元素+MS生长物质+1/2MS铁盐,附加500mg/l(单位下同)水解乳蛋白(LH)、5％蔗糖、0.5％(v/v)酵母浸出     

几种培养基及光照对蝴蝶兰叶片外植体褐变的影响

蝴蝶兰叶片外植体在MS纸桥培养基上培养10 d发生褐变率较低,1/2 MS次之,MS、B₅和N₆培养基外植体褐变率最高。MS培养基中铁盐加倍或微量元素缺乏造成外植体严重褐变,培养基积累很多褐色物质;减少琼脂含量,外植体褐变加重;添加柠檬酸可减轻外植体褐变;而添加活性炭、抗坏血酸或PVP对减轻外植体褐变无显著作用。光照强度增加外植体褐变严重。     

云南火焰兰种子无菌萌发和试管育苗

对云南火焰兰进行无菌播种技术试验.结果表明,适宜萌动的培养基为1/4MS 椰子水20% 蛋白胨1 g L-1 蔗糖10 g L-1,播种30 d时萌动率可达93.1%;适宜萌发的培养基为VW 椰子水10%,培养75 d时萌发率达36.6%.盐浓度较高的MS培养基不适合种子萌发和原球茎生长,低盐的KC培养基上种子能较好地萌发和成苗;添加椰子水可明显提高种子萌动率,并有利于原球茎的生长和分化.光培养和暗培养对种子萌动的影响差异不明显,但暗培养3周会对种子后期发育不利.添加150 g L-1的苹果匀浆有利于幼苗生长.试管苗移栽成活率在95%以上.     

红边朱蕉茎尖的离体培养和快速繁殖

植物名称:红边朱蕉(Cordyline terminalis cv."Reel edge”)。材料类别:茎尖。培养条件:(1)生长培养基为MS附加6BA0.3和NAA0.1;(2)增殖培养基为1/2MS大量元素、MS微量元素、有机物和铁盐,附加6BA0.5～1.0;(3)生根培养基为l╱2MS,附加NAA0.03。以上培养基均加3％蔗糖,pH为5.8。培养温度26±2℃,光照度3000 lx,每天光照12～14小时。生长与分化情况:4～6月间,取红边朱蕉茎尖,     

绞股蓝悬浮细胞的原生质体再生植株

绞股蓝(Gynostemma pentaphyllum (Thumb)Mak．)是葫芦科多年生草本药用植物,现已得到广泛的开发利用,本文首次报道了绞股蓝悬浮细胞的原生质体再生植株。     

枯草芽孢杆菌中性β—甘露聚糖酶的产生及性质

由土壤中分离出一株产中性β甘露聚糖酶的枯草芽孢杆菌（Ｂａｃｉｌｕｓｓｕｂｔｉｌｉｓ）,编号ＢＭ９６０２。该菌在液体培养条件下,产生中性β甘露聚糖酶。多糖能作为碳源,而单糖不能作为碳源;有机氮源优于无机氮源。产酶最适培养基组成：魔芋粉４％,牛肉蛋白胨和酵母膏各１％。产酶最适培养条件：培养基起始ｐＨ８５,３５℃,振荡培养３６ｈ。以槐豆胶为底物,培养滤液中性β甘露聚糖酶活力为９６ＩＵ／ｍＬ。酶在ｐＨ５０～１００和５０℃下稳定;作用最适条件为ｐＨ６０和５０℃;水解魔芋粉和槐豆胶均产生寡聚糖。     

High-yield production of valepotriates by hairy root cultures of Valeriana officnalis L. var. sambucifolia Mikan

Summary Hairy root cultures of Valeriana officinalis var. sambucifolia were established by infection of sterile plantlets with Agrobacterium rhizogenes strain R1601 The transformed roots were grown in 10 different, hormone-free liquid media and the isovaltrate, valtrate, didrovaltrate, isovaleroxyhydroxydidrovaltrate content was quantified by HPLC. Valepotriates were entirely retained inside the root tissues. The highest overall valepotriate content (10.3 % dry wt), 4 times the amount found in the roots of 9-month-old nontransformed plants, was observed in half strength Gamborg B5 medium supplemented with 2 % sucrose. The hairy roots cultured in Murashige and Skoog liquid medium supplemented with 2 % sucrose for 50 days produced over 44 mg/g dry wt valepotriates.Abbreviations MS Murashige and Skoog medium (Murashige and Skoog 1962) - B5 Gamborg B5 medium (Gamborg 1970) - WP McCown's woody plant medium (Lloyd and McCown 1980) - 1/2 MS-2 half strength MS+2 % sucrose - 1/4 B5-2 quarter strength B5+2% sucrose - MS-7 full strength MS+7% sucrose - YMB Yeast mannitol broth (Hooykaas et al. 1977) - IVAL isovaltrate - VAL valtrate - IVHD isovaleroxyhydroxydidrovaltrate - DI didrovaltrate (Fig. 1)     

甘蓝下胚轴原生质体再生植株

经纯化后,甘蓝下胚轴原生质体的产量为1．5×10⁶g^-1(Fw),采用液体浅层培养的方法进行培养。2～3d后,发生第一次分裂,第10天,统计分裂频率为6l％,5周内形成大量的细胞团和小愈伤组织,统计植板率为1．1％,把小愈伤组织转到与原生质体培养基相同激素的MS固体培养基上增殖。当愈伤组织长到3～5mm大小时,接到分化培养基上,芽分化率为46.7％．分化出来的芽长到3～4cm长时,从基部切下,插入生根培养基,两星期左右即可长成完整植株。     

Determination of salirasib (S-trans,trans-farnesylthiosalicylic acid) in human plasma using liquid chromatography-tandem mass spectrometry

A liquid chromatography/tandem mass spectrometric (LC/MS/MS) assay was developed for the quantitative determination of salirasib (S-trans,trans-farnesylthiosalicylic acid, FTS) in human plasma. Sample pretreatment involved liquid-liquid extraction with methyl t-butyl ether of 0.5-mL aliquots of lithium heparin plasma spiked with the internal standard, S-trans,trans-5-fluoro-farnesylthiosalicylic acid (5-F-FTS). Separation was achieved on Waters X-Terra C(18) (50 mm x 2.1 mm i.d., 3.5 microm) at room temperature using isocratic elution with acetonitrile/10 mM ammonium acetate buffer mobile phase (80:20, v/v) containing 0.1% formic acid at a flow rate of 0.20 mL/min. Detection was performed using electrospray MS/MS by monitoring the ion transitions from m/z 357.2-->153.0 (salirasib) and m/z 375.1-->138.8 (5-F-FTS). Calibration curves were linear in the concentration range of 1-1000 ng/mL. A 5000 ng/mL sample that was diluted 1:10 (v/v) with plasma was accurately quantitated. The values for both within day and between day precision and accuracy were well within the generally accepted criteria for analytical method (<8.0%). This assay was subsequently used for the determination of salirasib concentrations in plasma of cancer patients after oral administration of salirasib at a dose of 400 mg.     

Optimization of protoplast isolation from NaCl stressed primary, secondary and tertiary calli derived from mature seeds of Bangladeshi indica rice cultivar Binnatoa

A standardized protocol was developed for the isolation of protoplasts from salt stressed primary, secondary and tertiary calli of the moderately salt tolerant indica rice land race Binnatoa. Calli were induced from mature seeds using MS2 callus induction media supplemented with 0, 50 and 100 mM NaCl. Subsequently cultures were maintained in the same medium for 1–2 passages with or without salt stress at the same concentrations. Large numbers of protoplasts (about 1.57–2.10×10⁵/ml) with high viability were isolated from both control and salt stressed (50 and 100 mM NaCl) secondary and tertiary calli compared with the primary calli. The mannitol concentraion in the isolation and washing media was gradually increased, based on salt concentrations in which 13–15% was the most compatible for the control and 50 mM NaCl stressed calli and 17% for the 100 mM NaCl stressed calli. Isolated protoplasts at a density of 1–1.5×10⁵/ml were cultured in MS1₁₅ medium by liquid culture or agarose droplets. The first division of protoplasts was observed 4–5 days after culture using either method. The agarose droplet method led to sustained division of protoplasts and microcolonies formed within 2 weeks. Although no microcalli or protocalli were observed the procedure described provides a method for the isolation of salt-stressed protoplasts in indica rice which avoids the need for laborious and time-consuming suspension cultures. Subsequent regeneration from calli, derived from these protoplasts is reported in a further publication.     

Comparative pharmacokinetics of liquid and lyophilized formulations of IV RhIG immune globulin.

To compare the pharmacokinetics, safety, and tolerability of the liquid and lyophilized formulations of Rh(0)(D) immune globulin intravenous (human) (IV RhIG) administered intramuscularly (IM) and intravenously (IV). In 2 randomized, parallel arm, blinded, phase I studies, 142 healthy adult volunteers received a single dose of either the liquid or lyophilized formulation administered IM (300 microg in Study 1; 15 microg/kg in Study 2) or IV (50 microg/kg in Study 1). Pharmacokinetics (area under the curve [AUC}, C(max), t(1/2), T(max)) and safety data were assessed over 84 days. The 2 formulations had comparable pharmacokinetics following both IM and IV administration. The ratios (90% confidence intervals) for AUC and C(max) treatment means were, for most assessments, within the predefined FDA acceptance range of 80%-125%, demonstrating the bioequivalence of the liquid and lyophilized formulations. Both formulations were equally well tolerated. Study results demonstrate comparable safety and pharmacokinetic profiles of the liquid and lyophilized formulations of IV RhIG. These findings suggest that the liquid formulation will be therapeutically equivalent to the lyophilized formulation but in a more convenient ready-to-use dosage form that may also reduce preparation errors.     

Quantification of carvedilol in human plasma by high-performance liquid chromatography coupled to electrospray tandem mass spectrometry: application to bioequivalence study

A rapid, sensitive and specific method to quantify carvedilol in human plasma using metoprolol as the internal standard (IS) is described. The analyte and the IS were extracted from plasma by liquid-liquid extraction using a diethyl-ether solvent. After removed and dried the organic phase, the extracts were reconstituted with a fixed volume of acetonitrile-water (50/50; v/v). The extracts were analyzed by a high performance liquid chromatography coupled to electrospray tandem mass spectrometry (HPLC-MS/MS). Chromatography was performed isocratically on Alltech Prevail C18 5 microm analytical column, (150 mm x 4.6 mm i.d.). The method had a chromatographic run time of 3.5 min and a linear calibration curve over the range 0.1-200 ng ml(-1) (r2>0.997992). The limit of quantification was 0.1 ng ml(-1). This HPLC-MS/MS procedure was used to assess the bioequivalence of two carvedilol 25 mg tablet formulations (carvedilol test formulation from Laboratórios Biosintética Ltda and Coreg from Roche Químicos e Farmacêuticos S.A standard reference formulation). A single 25 mg dose of each formulation was administered to healthy volunteers. The study was conducted using an open, randomized, two-period crossover design with a 2-week wash-out interval. Since the 90% CI for C(max) and AUCs ratios were all inside the 80-125% interval proposed by the US Food and Drug Administration Agency, it was concluded that carvedilol formulation elaborated by Laboratórios Biosintética Ltda is bioequivalent to Coreg formulation for both the rate and the extent of absorption.     

Plant Regeneration from Protoplast of Peucedanum praeruptorum Dunn

Calll were initiated from the seedling segment of Peucedanum praeruptorum Dunn and subcultured on the MS agar medium with 0.5 mg/L 2,4-D. Cell suspension culture with a lot of embryogenic cell clumps was obtained in liquid medium. Protoplasts were isolated from the cell clumps in enzyme mixture solution containing 1.5% Onozuka R-10, 0.3% Macerozyme R-10, 0.5% helicase, 5 mmol/L CaCl2 and 0.6 mol/L mannital, at pH 5.6 and shaking for 5- hours at 25℃. Helicase is necessary for isolation. After purified by washing, the protoplasts were cultured in liquid medium containing 1 mg/L 2,4-D +0.5 mg/L zeatin. First cell division was observed after four days. Large cell clumps were formed after thirty days. Microcalli of 1 mm in size was formed after about fifty days, and continued to grow on the MS solid medium containing 0.5 mg/L 2,4-D and 200 mg/L casein hydrolysate, and later differentiated into embryoids when transferred to MS agar medium with 0.1 mg/L zeatin. Eventually, embryoids developed into whole plantlets on the MS solid medium without phytohormones.