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。     

红叶加拿大紫荆离体快繁技术研究

研究了基本培养基、生长调节剂配比、培养条件等对加拿大紫荆继代增殖的影响及不定根诱导和驯化移栽技术.结果表明:加拿大紫荆继代增殖宜选用DKW或WPM基本培养基附加40.0 g/L的白砂糖;生长素NAA较IBA、IAA更适于继代培养;间隔1～2代使用I.0 mg/L TDZ取代1.0～2.0 mg/L BA可明显提高繁殖系数和出苗量.27.5℃左右的温度有利于继代增殖,30℃高温、20℃低温及接种后黑暗培养均不适于生长.在1/2MS或1/2DKW附加1.0 mg/L IAA及25.0 g/L白砂糖的生根培养基上,生根率达80%,温室驯化移栽成活率达89%,可满足快繁要求.     

生姜离体叶片愈伤组织悬浮细胞系建立与植株再生

以‘莱芜大姜’为试材,研究了生姜离体叶片愈伤组织的诱导以及细胞悬浮系建立与植株再生。结果表明,以生姜试管苗叶片为外植体,接种到MS+1.0 mg/L 2,4-D+0.5 mg/L 6-BA+30 g/L蔗糖的培养基上,可有效诱导出生长迅速、质地疏松的愈伤组织。将获得的愈伤组织接种到MS+0.15 mg/L 2,4-D+6.0 mg/L 6-BA+30 g/L蔗糖的液体培养基上,25℃黑暗条件下震荡培养25-30 d,可建立分散性好、生长迅速的悬浮细胞系,细胞悬浮系培养的适宜参数为:初始接种量为1.0-1.5 g,继代培养的适宜间隔期为15 d,继代培养液体培养基更新比例为3/4。将悬浮细胞接种到固体培养基MS+0.2 mg/L NAA+10.0 mg/L 6-BA+30 g/L蔗糖上可获得再生植株。     

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。     

‘正午’牡丹微繁殖体系的建立

本研究以‘正午’牡丹腋芽为外植体,建立了高效的牡丹微繁殖体系。腋芽的初始培养基为WPM+0.5mg/L BA+0.2mg/L GA3,培养50d后,一个丛生芽平均可切分为13个繁殖体用于增殖培养;增殖培养基为WPM[1668mg/L Ca(NO3)2·4H2O]+0.5mg/L BA+0.2mg/L GA3,以35d为一个继代培养周期,增殖率为3.0,共继代培养7次;生根培养时,先将无根苗在复壮培养基[1/2MS(296mg/L CaCl2)+0.5g/L活性炭]上培养20d,再转入根诱导培养基[1/2 MS(296 mg/L CaCl2)+1.0 mg/L腐胺+1.0 mg/L IBA]培养30d,最后转入根形成培养基[1/2 MS(296mg/L CaCl2)+4.0g/L活性炭]培养20d,其生根率达77.2%;驯化与移栽基质为珍珠岩∶蛭石∶草炭土=1∶1∶1,组培苗移栽成活率高达92.1%。这表明以‘正午’牡丹腋芽建立的微繁殖体系具备规模化商业生产的价值。     

诺丽茎段愈伤组织诱导优化及细胞悬浮系的建立

为获取诺丽茎段中的次生代谢物并为建立遗传转化体系奠定基础,以诺丽茎段(无腋芽)为外植体诱导愈伤组织,并建立细胞悬浮系,对影响愈伤组织的诱导及细胞悬浮系的因子进行了研究。结果表明:愈伤组织诱导的最优培养基是MS+1.0mg/L6-BA+0.1mg/L2,4-D;悬浮培养的最佳培养基为MS+1.0mg/L6-BA+0.1mg/L2,4-D+3%蔗糖,pH为5.85,当初始接种量为37.5g/L、摇床转速为110r/min且(25±2)℃暗培养时,悬浮细胞生长良好,生长速率最大;诺丽茎段悬浮细胞生长曲线呈"S"型,最适继代周期为12–20 d;培养过程中,培养基的pH呈先下降后缓慢升高的变化趋势,诺丽茎段愈伤组织悬浮细胞培养的最适pH在4.5–5.0之间。文中成功建立了以诺丽茎段为外植体的稳定的细胞悬浮系。     

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。     

荷叶铁线蕨孢子的离体繁殖（简报）

以荷叶铁线蕨当年生未成熟的孢子为材料.在MS 2,4-D1.0mg/L NAA0.5mg/L培养基上进行愈伤诱导;在MS 6-BA0.5mg/L NAA0.05g/L培养基上进行抽芽诱导与增殖培养,60d为一继代周期,繁殖系数为20～30;在1/2MS 6-BA0.5mg/L NAA0.05mg/L培养基上进行壮苗培养;在1/2MS IBA1.0mg/L培养基上进行生根培养;最后移栽到泥碳中,成活率可达90%.     

不同浓度卡那霉素、潮霉素对楸树试管苗生长的影响

目的:将不同质量浓度的卡那霉素、潮霉素加入楸树培养基中,研究卡那霉素、潮霉素对楸树组培苗生长的影响,以确定抗生素对楸树茎段分化与生根的敏感质量浓度。方法:待楸树继代、生根培养基灭菌后温度降至30~50℃,将不同质量浓度的卡那霉素、潮霉素经抽滤式灭菌加入培养基中,在培养基中接入楸树组培无菌茎段培养,观测茎段继代(增殖芽数、芽长、叶数等)、生根(发根数、根长、芽长等)生长指标并统计分析。结果:楸树组培继代培养基添加卡那霉素质量浓度为100 mg/L时组培瓶苗生长缓慢,浓度为150 mg/L时叶片大部分发白并干枯,茎段基部无愈伤组织形成,瓶苗基本停止生长,楸树继代瓶苗对卡那霉素耐受性范围为100~150 mg/L;添加潮霉素质量浓度为5 mg/L时瓶苗生长较为缓慢,浓度为10 mg/L时叶片开始干枯,茎段基部愈伤组织较小,瓶苗基本停止生长,楸树继代瓶苗对潮霉素耐受性范围为10 mg/L左右。楸树组培生根培养基添加卡那霉素质量浓度为100 mg/L时大部分茎段干枯,少部分为绿但未分化芽与根,浓度为150 mg/L时大部分茎段干枯,极少上部为绿,基部干枯,但未分化芽与根,楸树组培瓶苗生根培养苗对卡那霉素耐受性范围为100~150 mg/L;添加潮霉素质量浓度为5 mg/L时少部分茎段干枯,浓度为10 mg/L时大部分茎段干枯,少部分为绿,茎段未出现芽的分化与根的萌发现象,楸树组培瓶苗生根培养苗对潮霉素耐受性范围为5~10 mg/L。结论:卡那霉素、潮霉素对楸树组培苗生长有明显的抑制作用且与抗生素浓度呈负相关,但低质量浓度(1 mg/L)的潮霉素对楸树继代分化芽数有促进作用;同一抗生素对楸树不同无性系间组培苗生长的影响无显著差异。     

铁皮石斛的组织培养(简报)

以铁皮石斛当年生茎段为材料,在MS＋6-BA 0.1 mg/L＋NAA 0.02 mg/L培养基上进行不定芽诱导培养;在MS＋6-BA 2.5 mg/L＋NAA 0.05 mg/L＋AC（活性炭）1g/L培养基上进行丛生芽诱导与增殖培养,50 d为一个继代周期,繁殖系数为5～8;在1/2 MS＋6-BA 0.5 mg/L＋NAA 0.05 mg/L＋椰子100 g/L＋AC 1g/L培养基上进行壮苗培养;在1/2 MS＋IBA 1.0 mg/L＋AC 1g/L培养基上进行生根培养;最后移栽到小颗粒化树皮中,成活率可达95%。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.