棉花组织培养中异常苗的发生与转化

研究了棉花 (Gossypium hirsutum L.)组织培养中异常苗发生的类型、规律、影响因素及转化为正常苗的途径及技术。结果表明 ,在棉花组织培养中异常苗发生的频率极高 ,且形式多样 ,大体可分为生长点异常苗、叶片异常苗、胚轴异常苗、联体苗、丛生叶苗、单叶苗、玻璃化苗、白化苗、褐化苗和重新愈伤化苗等 1 0类。其中生长点异常苗和叶片异常苗最为常见。影响异常苗形成的因素很多 ,其中主要有外植体种类、培养基组成、培养方式和培养时间。有的异常苗来源于畸形胚 ,有的异常苗由正常苗转化而来。在合适的条件下异常苗可转化为正常苗 ,但不同培养基对异常苗转化为正常苗的影响不同 ,且不同基因型的异常苗转化为正常苗的能力也有差别。此外 ,从外界调控机制与棉花培养细胞的内在发育机制的相互作用方面讨论了造成棉花组织培养异常苗发生的原因 ;并讨论了降低异常苗产生频率的技术途径和异常苗存在对棉花组织培养的影响     

In vitro induction of genetic variability in cotton (Gossypium spp.)

Summary Through the in vitro culture of excised embryos and ovules, interspecific hybrids have been obtained from cultivated and wild species of Gossypium. The hybrids matured upon transfer to the field. The anthers, ovules and embryos from both the diploid (2n=26) and tetraploid (2n=52) species underwent proliferation, and this response was genotypic. The diploid species invariably showed profuse callusing in comparison with the tetraploid. The callus showed various chromosome numbers, ranging from haploids to hexaploids, and from high polyploidy to aneuploidy. Hybrid callus culture may augment the genetic variability by providing a means for obtaining genetic exchange in interspecific hybrids. The implications of the in vitro induction of genetic variability for cotton improvement are discussed.     

Increasing in vitro microrhizome production of ginger (Zingiber officinale Roscoe)

In this study, using the quadratic saturation 310 D-optimal design method, we examined the effect of kinetin (KT), gibberellic acid (GA), and naphthalene acetic acid (NAA) on microrhizome production in ginger. The effect of GA on rhizome induction was larger than that of KT or NAA. Using simulation and optimality selection for tissue culture, we found that concentrations of GA, KT, and NAA of 1.33–2.35, 0.49–0.66, and 0.62 g/l, respectively, gave a microrhizome weight of over 0.25 g. The optimal conditions for microrhizome production were 80 g/l sucrose, 2 × MS macro-elements, and 1 × MS micro-elements, with a photoperiod of 24L:0D (light/dark). At the same time, 100% survival could be achieved on transfer of the in vitro ginger plantlets with microrhizomes to soil.     

从棉花胚性细胞原生质体培养获得植株再生

原生质体培养植株再生是进行体细胞杂交与基因工程操作的重要基础工作。近年来,国内外已有一些关于棉花原生质体分离与培养的研究报告,1986年 Firoozabady 报道从陆地棉(Ghirsutum)叶肉原生质体培养获得2—3个细胞的微克隆,1987年 Ka~-     

琼脂浓度、pH值及接种苗大小对文心兰试管苗增殖的影响

探讨了培养基中琼脂浓度、pH值和接种苗大小对文心兰试管苗增殖及生长发育的影响。结果表明：培养基中使用的琼脂浓度不仅对文心兰试管苗的增殖有影响,而且对试管苗后期的生长发育也有影响,在琼脂粉使用量达6．0g/L时,增殖率明显降低,后期苗生长发育不良;接种苗的大小对试管苗的增殖率影响较大,小苗易于形成丛生芽苗和原球茎,增殖系数可达7．07,而大苗则不能形成丛生芽苗也无原球茎形成;培养基的pH值在5,2～6．5范围内,试管苗增殖率差异不明显,试管苗的生长发育均正常。     

Sucrose-mediated regulation of fatty acid composition in asexual embryos of Theobroma cacao

Asexual embryos of Theobroma cacao L. cultured in vitro grew to a size and fresh weight equal to mature zygotic embryos in vivo when cultured in liquid medium on a rollodrum apparatus. Changes in the fatty acid composition in the asexual embryos, similar to those which occur in the zygotic embryo during normal maturation, were stimulated as the sucrose concentration in the medium was increased to an optimum at 27% sucrose (w/v). As the sucrose concentration increased, there was a shift in the fatty acid composition from predominantly polyunsaturated to saturated and monounsaturated fatty acids characterized by an increase in the mole percents of stearic (18:0) and oleic (18:1) acids and a decrease in the mole percents of linoleic (18:2) and linolenic acids (18:3).     

Ethylene production by carob (Ceratonia siliqua) callus cultures on varying media

Carob callus from hypocotyl segments produced ethylene in different amounts which were related to the composition of the medium and age of the callus. Both light and darkness stimulated high levels of ethylene production. No correlation was found between growth rate and ethylene production under dark conditions. In the light, a significant correlation was found, indicating that ethylene production and growth rate follow one another. Culture medium was the most important factor in controlling the growth rate and ethylene production. The highest values of ethylene production were obtained on media showing highest callus growth rate. These studies seem to indicate that most of the ethylene produced is a by-product of metabolic changes during carob callus development, though, under certain conditions, the initial evolution could regulate growth.     

文心兰试管苗分化及育苗技术研究

文心兰原球茎分化苗过渡驯化培养试验表明,改良1号+椰子汁100g/L的培养基有利于原球茎分化成苗,能促进分化苗生长健壮、整齐度好。壮苗与生根培养试验表明,以改良1号为基本培养基,加入香蕉泥100g/L、活性炭1g/L,在较强光照(2 000~3 000 lx)下培养,植株各项生长指标表现较好,有利于后期炼苗成活;以三角瓶为定瓶容器培养,植株接种效率最高、污染率最低;选取3cm高的植株定瓶,可显著提高文心兰工厂化组培育苗的工作效率。     

4种红景天植物的组织培养研究

以大花红景天、云南红景天、长鞭红景天和库页红景天的茎和叶为外植体进行组织培养,结果表明：大花红景天以茎为外植体诱导芽效果最好,其它3种红景天以叶为外植体诱导芽效果最好。云南红景天和长鞭红景天适合的芽诱导激素组合是0．1mg／L NAA和2．5mg／L 6-BA的组合,在该激素水平下两种红景天的出芽频率分别达到71％和84％;大花红景天和库叶红景天适合的芽诱导激素组合是0．5mg／L NAA和2．5mg／L 6-BA的组合,在该激素水平下两种红景天的出芽频率均达到80％。长鞭红景天和库叶红景天在添加IBA的培养基上容易生根形成完整植株,生根率分别达到87％和73％;经过炼苗后,长鞭红景天再生苗能够成功移栽,成活率达66％。     

盾叶薯蓣胚乳再生体系的建立及其染色体倍性鉴定

以盾叶薯蓣（Dioscorea zingiberensis）的胚乳为外植体,研究了不同植物生长调节剂对胚乳愈伤组织诱导及植株再生的影响,并鉴定了再生植株。结果表明：愈伤组织诱导形成的适宜培养基为MS＋2.0mg·L^–12,4-D＋0.5mg·L^–16-BA,不定芽分化的适宜培养基为MS＋2.0mg·L^–16-BA＋0.1mg·L^–1NAA,生根的适宜培养基为1/2MS＋0.3mg·L^–1NAA;再生植株炼苗移栽后,成活率可达80%;对获得的再生植株腋芽生长点进行染色体制片观察,发现染色体数目为20的细胞占观察细胞总数的10%,染色体数目为21–29的细胞占16%,染色体数为30的细胞占74%;获得了三倍体植株。     

不同红掌品种的叶片、叶柄和茎段愈伤组织的诱导及植株再生

红掌幼嫩叶片表面灭菌的最佳方法是：0．01％KMnO4 10min,0．05％链霉素、0．05％制菌霉素和0．05％头孢唑林钠各灭菌20min,然后用75％乙醇30s和HgCl2 2min,污染率为零。Anthurrium adraeanum “Rosetta“和A.adraeanum “Oilcloth-flower”愈伤组织诱导率和芽分化率显著高于A.adraeanum “Atlanta”.A.adraeanum “Fantasis”,A.adraeanum“Afrikanerin”。不同品种茎段、叶柄、叶片的愈伤组织诱导率和芽分化率有明显差异,大多数品种是幼茎段＞叶柄＞叶片。幼苗移栽在草煤或血竭渣基质中,成活率可达97％。     

烟草NC89叶肉细胞原生质体培养再生植株及影响因素的研究

生长１２０天的ＮＣ８９无菌苗叶片在Ｏｎｏｚｕｋａ Ｒ－１０和Ｍａｃｅｒｏｚｙｍｅ Ｒ－１０的混合酶液中,酶解、收集、纯化,在修改的ＮＴ培养基上,应用液体－固体双层培养,原生质体经生长、分裂,形成愈伤组织,在多种分化培养基上,分化成再生植株。试验中对传统的酶解、收集方法进行了改进并对原生质体的分裂和愈伤组织的分化条件作了研究,对影响分化的多种因素作了深入的比较和探索,同时,本试验表明,叶片的生长状态     

Rapid regeneration of <Emphasis Type="Italic">Phaseolus angustissimus</Emphasis> and <Emphasis Type="Italic">P. vulgaris</Emphasis> from very young zygotic embryos

A rapid regeneration protocol for proembryos of Phaseolus angustissimus as young as 1 day after pollination (DAP) involving pod culture for 1 week followed by embryo culture for 2 weeks and embryo germination for 1 or 2 weeks is provided. Optimization of the media was conducted with pods collected 3 DAP. The best pod culture medium was composed of basal medium [(Phillips and Collins 1979) salts with (Geerts et al. 2001) vitamins], 1000 mg l⁻¹ glutamine, 1000 mg l⁻¹ casein hydrolysate, 3% sucrose and 0.5% agar. Embryo culture medium consisted of basal medium with 500 mg l⁻¹ glutamine, 250 mg l⁻¹ casein hydrolysate, 1.9 μM ABA, 3% sucrose and 0.5% bacto-agar. Embryos developed into plantlets on germination medium containing basal medium with 0.25 μM BA, 3% sucrose and 0.7% bacto-agar. Fertile, normal plants were recovered from direct embryogenesis and from micrografted embryo-derived shoots. Embryos obtained from pods collected 3 DAP regenerated plantlets at a rate of 29.3%, while embryos from pods collected 2 DAP and 1 DAP regenerated at rates of 20.2 and 4%, respectively. A second accession of P. angustissimusregenerated at a rate of 26.2%. Using this 5-week protocol for P. vulgaris resulted in a plantlet regeneration rate of 12.5%.     

海水组培法培育芦荟耐盐品系研究

以库拉索芦荟(Aloe vera)组培苗为材料,在培养基中添加不同浓度海水对芦荟进行盐胁迫驯化培养.结果显示:(1)在50%海水处理的组培植株性状分离最明显,且有部分芦荟苗生长旺盛;盐胁迫驯化前芦荟(库拉索)与盐胁迫驯化后芦荟('南盐1号')的组培苗在分化过程中存在明显的差异,库拉索芦荟的芽分化率、平均分芽数、心叶生长速率及生根系数等指标均明显低于同比例海水处理下的'南盐1号';'南盐1号'芦荟苗在0～20%海水条件下组培时,芦荟分化率随着海水浓度的加大而降低,而30%海水组培处理时芦荟芽增长数上升至淡水组培水平,且平均心叶数增加速率最大,根生长变化特征与其芽分化生长基本一致.(2)移栽入圃后用50%天然海水灌溉时,'南盐1号'芦荟植株根系发达,生长正常,而库拉索芦荟则盐胁迫症状明显;经一年大田海水灌溉比较试验发现,'南盐1号'的地上部与根部的鲜重在淡水灌溉下与库拉索芦荟没有差异,而在30%和60%比例的海水灌溉下均显著高于库拉索芦荟;用30%的海水灌溉培养下,'南盐1号'地上部产量与淡水灌溉的产量无显著差异,而库拉索芦荟的地上部产量却比淡水处理降低15%～20%.研究表明,通过海水组织培养能筛选出芦荟耐盐株系,且其耐盐分化株系筛选的最适浓度为50%海水.