香蕉试管苗混倍性变异的研究

首次报道了香蕉试管苗变异中一种重要的类型－混倍性变异体,并且以形态变异、染色体数量变异、同工酶（ＭＤＨ,ＥＳＴ,ＰＯＸ）变异和内源激素（ｉＰＡｓ和ＩＡＡ）变异等项检测为主要手段,研究了这种混倍性变异体在试管繁殖过程中的变异性质。用混倍性变异作外植体培养时,细胞水平染色体数量畸变具有起点高的前１５代增长快的特点,同工酶变异率比正常有显著增加。检测了用混倍性变异体作外植体培养所得的大棚苗的染色体数量畸     

玉米胚性愈伤组织的长期继代及其染色体分析

对５种基因型幼胚诱导的愈伤组织继代培养表明,玉米胚性愈伤组织的长期继代受基因型,培养基成分,激素,培养条件的影响。适时继代,逐代筛选对胚性保持起重要作用。适当降低培养温度（１２±２℃）有利于愈伤组织的保存和胚性保持,可以减少愈伤组织长期继代所需的物质和工作量。长期继代培养的胚性愈伤组织,胚状体发生能力和植株再生率无显著变化,但正常苗的再生频率显著下降。观察愈伤组织细胞染色体发现：（１）基因型对不同倍性细胞的比例有明显影响。（２）随着继代时间的延长,二倍体细胞下降,四倍体和非二倍体细胞增多。（３）愈伤组织中出现多种染色体结构变异,这些结构变异有可能导致非整倍体细胞的形成。     

新疆紫草的组织培养及其染色体分析

新疆紫草(Arnebia euchroma)的胚轴、子叶、根都产生愈伤组织。不同来源的外植体产生苗的能力不同,胚轴来源的愈伤组织分化苗最多,而且苗生长旺盛。根的愈伤组织只产生不定根。新疆紫草的核型是 K(2n)=14=2m 8sm 2sm(SAT) 2st。随着继代培养代数增多,染色体变异的种类和频率以及染色体数目显著增加。如果继代培养时间较长,染色体数目变异的范围更广,而且出现染色体结构变异。     

新疆紫草的组织培养及其染色体分析

新疆紫草(Arnebia euchroma)的胚轴、子叶、根都产生愈伤组织。不同来源的外植体产生苗的能力不同,胚轴来源的愈伤组织分化苗最多,而且苗生长旺盛。根的愈伤组织只产生不定根。新疆紫草的核型是K(2n)=14=2m+8sm+2sm(SAT)+2st。随着继代培养代数增 多,染色体变异的种类和频率以及染色体数目显著增加。如果继代培养时间较长,染色体数目变异的范围更广,而且出现染色体结构变异。     

伊贝母愈伤组织在继代培养过程中的染色体变异和分化频率的研究

本文报道了伊贝母愈伤组织在不同培养基上继代培养的的染色体变异和分化频率。结果表明,随着继代培养的进程,二倍体细胞频率逐渐减少。亚二倍体和超二倍体细胞频率迅速增加。同时还观察到比率不等的亚单倍体、单倍体,超三倍体和超四倍体细胞。于是培养物最终变成为只有少量整倍体而多数为非整倍体染色体数的混倍体细胞群体。不同培养基对染色体变异的效应是2,4-D+KT>IAA+KT>NAA+KT。分化频率随继代次数的增加而下降。同时还观察到了各种类型的染色体结构变异和有丝分裂异常。根据试验结果,对继代培养中染色体变异的原因以及与分化频率的关系进行了讨论。     

香蕉试管苗变异因素的初步研究

本文初步研究了香蕉离体培养中,培养基内不同激素浓度、继代培养数,以及３个品种（品系）对试管苗变异的影响。结果表明：６３１和台湾８号比威廉斯（Ｗｉｌｌｉａｍｓ）稳定,培养条件相同,培养代数多时,变异率明显提高。外源激素的配比试验表明：ＩＢＡ∶ＢＡ为０．５∶４．０（ｍｇ／Ｌ）为最佳配方,当ＩＢＡ∶ＢＡ达到２．０∶１０（ｍｇ／Ｌ）时,异常苗发生率最高。用田间变异株根尖细胞染色体观察结果,发现其染色体数不是增多就是严重缺失。变异株的过氧化物酶同工酶酶谱经电泳分析,表明在迁移率为０．７处的一条带纹消失,而正常对照苗则稳定存在。     

异源八倍体小冰麦体细胞无性系的建立及其染色体变异

从５个异源八倍体小冰麦（Ｔｒｉｔｉｃｕｍ －Ａｇｒｏｐｙｒｏｎ）的叶片、幼穗及成熟胚诱导愈伤组织,建立体细胞无性系,获得大量再生植株。附加一个冰草染色体组的异源八倍体小冰麦杂种无性系中３７．５％ 表现变异,其中非整倍体植株变异较多,很多变异的再生植株形态与小麦近似,同时出现一定数量染色体重排、交换、易位、断裂、融合等变异。结果表明,通过杂种无性系变异进行染色体基因转化及遗传修饰是一条可行的途径。实验还观察了小冰麦愈伤组织分化过程中绿点的形成过程,首次提出两种类型绿点,即芽绿点和根绿点,并描述了两者的差异     

金不换群三七液对哺乳动物致突变和致畸安全性评价

研究了金不换鲜三七液特殊毒理学效应的致突变性。以小鼠骨髓细胞染色体畸变试验,小鼠睾丸减数分裂染色体畸变及小鼠致畸试验为指标,研究金不换鲜三七液的安全性。结果：（１）小鼠骨髓细胞染色体畸变试验：低,中,高３个剂量组小鼠肌髓细胞染色体畸变率分别为０．７％,０．２％和０．９％,与对照组相比无显著差异。阳性对照组染色体畸变率大大增高。（２）小鼠睾丸减数分别细胞染色体畸变;在本实验条例上,小鼠睾丸细胞染色体     

半夏茎尖培养及块茎的品质改良

离体培养半夏Ｐｉｎｅｌｌｉａ ｔｅｒｎａｔａ 茎尖获得小植株。用其叶柄作外植体在ＭＳ添加２,４－Ｄ 和ＫＴ的培养基上诱导出愈伤组织,继代培养中挑选出一种表面呈颗粒状、易分散、生长快的愈伤组织。在ＭＳ添加ＫＴ０．５ ｍ ｇ／Ｌ, ＮＡＡ ０．２ ｍ ｇ／Ｌ的培养基上,叶柄可直接分化形成小植株,愈伤组织通过形成小块茎的途径产生完整植株。块茎顶端芽分化过程中,先形成的叶原基或幼叶总是覆盖着后面的叶原基而出现一种依次叠套的特殊结构。液体浅层培养对试管苗的增殖速率比固体培养快１ 倍。叶柄和愈伤组织的小植株分化率均在７０％ 以上,移栽成活率为１００％ 。试管苗的块茎产量（鲜重）比对照（用小块茎繁殖）的净块茎产量（鲜重）高１０３％ 。试管苗块茎的总生物碱含量为０．３４４％ ,野生和人工栽培半夏块茎的总生物碱含量分别为０．２６４％ 和０．２０３％ 。     

籼稻花药培养中胚性细胞的利用和继代研究

诱发籼稻花药培养形成胚性细胞,并建立胚性细胞系,通过胚胎发育途径,在两个月内,由8块胚性细胞团经切割培养增殖获得再生绿苗12690株和白化苗若干。继代培养一年的胚性细胞团再生苗率仍高达90％,但畸变率高,移栽不易成活;检查胚性细胞532个,染色体变化呈多样性,但仍可分为12个同源染色体组。     

基于幼化效果的山杏组培繁殖技术研究

以山杏为试验材料,选取不同部位、不同树龄、不同继代次数的组培苗,研究基于丛生芽诱导率和组培苗生根率、形态指标、生理指标3个层次的幼化效果。结果显示:(1)随着山杏外植体取材部位从树冠上部、中部、下部的依次下降,其丛生芽诱导率和组培苗生根率依次升高且部位间差异显著,其组培苗的株高、叶长、叶宽、根系数量和根系长度也随外植体取材部位下降而增加,但仅根系数量和根系长度在部位间差异显著,但其组培苗和母树叶片SOD活性、POD活性和MDA含量却逐渐降低,且组培苗叶片始终低于母树叶片。(2)随着取材母树的树龄从10a、4a、1a的不断降低,组培苗上述各项指标值均依次升高,1a树龄丛生芽诱导率及组培苗生根率、根系数量、根系长度分别为80%、96.6%、4.3条、2.40cm,极显著高于相应的10a和4a树龄的组培苗。(3)从初代培养到1次、2次继代培养,随着继代次数的增加,各类组培苗的丛生芽诱导率、株高、叶长和叶宽值不断增大,且以2次继代培养的增幅较大;同时相应组培苗叶片SOD活性、MDA含量和POD活性则呈下降趋势,但仍表现为2次继代的组培苗幼化效果较好。研究认为,各层次幼化效果指标在组培苗中表现出相同的变化规律。随着外植体在母树上取材部位的下移、取材母树年龄的降低以及组培苗继代次数的增加,山杏丛生芽诱导率和组培苗生根率、形态指标、生理指标表现出一致的变化趋势,其组培幼化效果愈来愈佳。     

Feasibility studies of large scale production of human anti-tetanus toxoid monoclonal antibodies

The feasibility of large scale production of human anti-tetanus toxoid monoclonal antibody for therapeutic use was evaluated using a human heterohybridoma. The effects of duration of subculture, transition from static to agitated culture conditions and the level of serum concentration were studied. The level of antibody secreted by the clone decreased with increasing length of subculture and decreasing serum concentration. The clone exhibited heterogeneity in expression of surface IgG after 2 or 7 weeks of subculture in static culture conditions irrespective of the serum concentration. However, a prolonged duration of subculture (9 weeks) in 3% serum medium had an effect on the expression of surface IgG both in static and agitated culture conditions. With respect to total (surface and intracellular) IgG, two distinct cell populations were observed. On long term subculture (9 weeks) in low serum medium (3% FCS), there was a decrease in the population which was the high synthesizer. In addition, when these cells were cultivated in agitated spinner flasks, a defect in secretion of antibodies was observed. Thus a general fall in the amount of antibody in the supernatant of agitated cultures was due to decrease in antibody synthesis as well as the defect in secretion of antibodies.     

Effect of fetal calf serum on the cadmium clastogenicity

Inconsistent results among reports on cadmium genotoxicity revealed that certain confounding factors might significantly influence the outcomes of assessment. In Chinese hamster ovary (CHO-W8) cells, chromosome aberration induced by six different cadmium compounds was found positively associated with intracellular cadmium concentration. A parallel association was also observed among different CHO strains treated with same cadmium compound, the cadmium acetate. Both the cadmium-induced chromosome aberration and cadmium uptake were influenced by the presence of fetal calf serum (FCS). The presence of 10% FCS during the 2 h treatment period greatly retarded the cellular cadmium uptake, and concurrently reduced the chromosome aberration induction. Other factors such as specific cadmium anion involved and the duration of cadmium treatment period in the investigation also influenced the assessment results of cadmium-induced chromosome aberration. In the protocol with a 2 h pulse treatment, cadmium acetate, chloride and sulfate induced more chromosome aberration than cadmium nitrate, carbonate and oxide. When cadmium was present in the culture of the entire treatment period for 18 h, the results went the opposite way. Cadmium nitrate, carbonate and oxide induced significant chromosome aberration, while other three cadmium compounds gave negative results. Cadmium compounds did not induce significant SCE at the same dose level that yielded significant chromosome aberration induction, either in the protocol with the short pulse or long treatment period.     

人胸腺上皮细胞培养及其分泌产物

本文通过降低培养基中血清含量,向RPMI 1640培养基中补加三碘甲腺原氨酸而获得一种人胸腺网状上皮细胞占优势生长的培养物。在此培养基中细胞经传代培养长达90天,仍维持正常形态特征。胸腺组织在培养14天后,新生细胞的突起形成网状结构,细胞化学检查和电镜观察表明具有丰富的分泌颗粒,囊泡及张力原纤维束和桥粒等上皮细胞特征。收集合并细胞培养液,经部分纯化后检查其生物活性,表现出具有促进玫瑰花结形成和降低胸腺细胞TdT活性的作用,说明培养细胞的分泌产物具有胸腺激素活性。根据形态学,细胞化学和生物活性检测结果,我们倾向于认为该培养物主要为网状上皮细胞。     

Regeneration of whole plants from protoplasts isolated from tissue-cultured shoot primordia of garlic (Allium sativum L.)

Protoplasts derived from tissue-cultured shoot primordia of garlic (Allium sativum L.) initiated successive cell divisions within 4 days and formed small individual calli (0.2mm in diameter) after 5 weeks of culture on Gamborg's B5 medium supplemented with 0.1% casein hydrolysate, 1mg/1 1-naphthaleneacetic acid and 1mg/1 6-benzylaminopurine. Plating efficiency was roughly 5% at the density of 1x10⁴ protoplasts/ml of medium. Adventitious buds developed from the calli during subsequent subculture on Gamborg's B5 medium supplemented with 40mg/l adenine and 10% coconut milk. When transferred to the same medium without supplements, these buds grew into shoots and rooted. The regenerated garlic plantlets were successfully transferred to the greenhouse and grew into whole plants.     

培养条件对平贝母愈伤组织分化的影响

以平贝母无菌苗幼茎切段诱导产生的愈伤组织为试材,研究了基本培养基、蔗糖浓度、继代时间及继代周期时间对愈伤组织分化的影响。结果表明,ＭＳ基本培养基有利于不定芽的诱导,Ｎ６培养基有利于体细胞胚的诱导。在ＭＮ培养基上,随着蔗糖浓度的升高,不定芽发生能力下降,体细胞胚发生率升高。继代时间越长,不定芽及体细胞胚的发生能力均下降。体细胞胚胎发生的最佳继代周期时间是２１天,不定芽发生的最佳继代周期时间是２８天。     

Regeneration of plants from the culture of leaves and axillary buds in mulberry (Morus indica L.)

Stem segments, axillary buds and leaves excised from established shoot cultures of Morus indica were soaked in MS liquid medium containing benzyladenine (0.5, 1, 2 mg/1) and were cultured subsequently on semi solid medium of the same composition. Numerous shoot buds differentiated from leaf and axillary buds but stem segments were unresponsive. The shoot buds on isolation and culture developed into plantlets. Callus tissues which developed at the base of the leaf explant upon subculture also differentiated numerous shoot buds.Abbreviations BA benzyl adenine - CM coconut milk - 2, 4-D 2, 4 dichlorophenoxy acetic acid - Kn kinetin - MS Murashige and Skoog - Z zeatin     

继代培养次数对杜梨生根能力和叶形态及其激素水平的影响

为了探索杜梨组培复幼变化规律,对10年生杜梨进行连续继代培养,统计不同继代次数杜梨丛生芽繁殖系数和生根率,观察记录叶片形态变化并测定内源激素含量。结果表明:(1)通过连续继代培养,杜梨丛生芽生根率由0提升到66.70%,繁殖系数由第1代的2.13提升到第10代的4.20。(2)叶片在继代第3次时出现裂刻且随后裂刻程度逐代加深;在继代过程中,丛生芽叶面积和叶脉数显著降低,叶周长和叶形指数呈先下降后上升的变化趋势。(3)丛生芽叶片内源IAA含量在继代第6次时达到46.39 ng·g-1,且显著高于初代丛生芽;随着继代次数的增加,叶片内源ZR呈先上升后下降的变化趋势,内源GA3含量没有发生显著性变化,而内源ABA含量逐渐降低;叶片IAA/ABA和IAA/ZR的值随着继代次数的增加而增加。(4)丛生芽叶片ABA含量和IAA/ZR与其生根率分别呈显著负相关和显著正相关关系,叶片裂刻数和IAA/ABA与生根率均呈现极显著正相关关系,而叶脉数与生根率则呈现极显著负相关关系。研究认为,连续继代培养可显著提高杜梨丛生芽的生根能力,并且与丛生芽叶形和激素含量及其比值有密切的关系,该研究结果为难生根植物无性繁殖以及树木复幼提供了重要技术借鉴。     

Control of pattern of regenerant differentiation and plantlet production from leaflet segments of <Emphasis Type="Italic">Azadirachta indica</Emphasis> A. Juss. (neem)

A process with controlled pattern of regenerant differentiation from leaflet segments leading to production of cloned plants of a 40-year-old tree of Azadirachta indica was developed. A two-step procedure was adopted for containing intervening callusing during regenerant differentiation using modified Murashige and Skoog (MS) medium, where in the first step the explants were subjected to pulse treatments having higher concentration of 6-benzylaminopurine (BAP), while in the second step they were cultured in one-tenth of the initial concentrations of BAP. In the present case, simultaneous differentiation of two types of morphogenetic structures, that is, shoot buds and the meristematic nodules was observed. However, differentiation of higher number of desirable regenerants—the shoot buds and a few meristematic nodules, rather than vice-versa could be controlled by increasing both, the concentration of BAP in pulse treatment and the duration of pulse treatment. In the optimum treatment, where the explants were exposed to 8.88 μM BAP and 81.43 μM adenine hemisulphate for 5 days followed by their transfer to 0.88 μM BAP and 81.43 μM adenine hemisulphate, on an average, 17.4 shoot buds and only 1.6 meristematic nodules were formed from a leaflet. On subculturing, the shoot buds developed into shoots, whereas the meristematic nodules produced three kinds of organized structures that too in varied proportions. Multiplication of shoots was sustained in proliferation medium supplemented with 1.11 μM BAP, 1.43 μM indole-3-acetic acid (IAA) and 135.72 μM adenine hemisulphate. The isolated shoots were rooted and complete plantlets were transferred to potted soil with 100% survival.