濒危植物珙桐愈伤组织的诱导及悬浮细胞培养初探

以珙桐（Davidia involucrate）无菌苗的下胚轴为外植体,以LSD分析和正交设计与分析为主要研究方法。诱导和筛选愈伤组织,进行悬浮培养,初步建立了珙桐悬浮培养体系,并对珙桐悬浮培养中生物量和pH值的变化进行了初步研究。结果表明：KT和2,4-D的组合诱导出的愈伤组织更适合进行细胞悬浮培养,最佳愈伤组织诱导培养基为WPM＋KT 1.0 mg/L＋2,4-D 0.5 mg/L＋Vc 1.0 mg/L;悬浮细胞培养中,接种量和Ca2＋浓度是影响生物量的主要因素,最佳接种量为25 g/L,最佳Ca2＋浓度为标准WPM培养基中Ca2＋浓度的2倍;黑暗条件下,珙桐悬浮培养生物量变化曲线呈＂S＂型,在21 d时可获得最大生物量;pH值的变化与生物量的变化有一定的相关性。     

毛白杨愈伤组织悬浮细胞耐盐性研究

把生长在全盐含量为 0 .361%的土壤中的毛白杨 ( Populus tomentosa Carr.)愈伤组织悬浮细胞培养在含不同浓度 Na Cl的液体培养基中 ,定期测量各组的细胞密度 ,根据生长曲线研究其耐盐性。结果显示 ,与不加 Na Cl的液体培养基相比 ,生长于含较低浓度 Na Cl( 2 .7‰和 5.5‰ )液体培养基中的细胞生长较好 ,而在含较高浓度 Na Cl( 8.2‰和 10 .9‰ )的液体培养基中 ,细胞生长受到较强抑制。     

茅苍术愈伤组织诱导及其细胞悬浮培养研究

采用正交试验法研究了不同的植物生长调节剂对茅苍术叶柄、叶片和根茎愈伤组织诱导的影响,结果表明,不同外植体在各自的最佳培养条件下,叶柄、叶片和根茎愈伤组织的诱导率分别为99.0%、83.5%和71.5%,以叶柄的培养效果最好,其中2,4-D对茅苍术愈伤组织的诱导具有极显著的效果,在各种植物生长调节剂组合中,诱导叶柄愈伤组织形成的最佳组合为0.4mg·L-1NAA、4.0mg·L-12,4-D和0.4mg·L-1KT,培养20d左右,诱导率达到99.0%。此外,将茅苍术叶柄细胞悬浮培养至18d时,细胞量、多糖和苍术素的含量均达到最大值,分别为9.07g·L-1、15.68mg·L-1和19.62ug·L-1。     

翅果油树茎段愈伤组织和芽发生的组织的学研究

本文对翅果油树大宫灯型茎段培养在ＭＳ附加６－ＢＡ较高,ＮＡＡ较低浓度的培养基上培养０－３０ｄ的组织学变化进行了研究。创伤对其愈伤组织的形成有明显的刺激作用,培养３－４ｄ切口处的皮层细胞,形成层细胞,韧皮部薄壁细胞以及髓组织细胞,甚至表皮细胞均脱化分化开始分裂;     

药用植物刺山柑愈伤组织诱导及细胞生长代谢特征研究

本文研究了不同外植体及激素对刺山柑愈伤组织诱导的影响,不同激素配比对愈伤组织增殖培养以及悬浮细胞的生长与代谢特征.结果表明:以刺山柑叶片作为诱导愈伤组织的材料,效果较佳;愈伤组织诱导和继代的适宜培养条件是分别是MS+0.5 mg/L 2,4-D+1.0mg/L6-BA和MS+1.0mg/L2,4-D+1.5mg/L6-BA.刺山柑悬浮培养细胞的生长周期为30天左右,细胞生长曲线呈"S"形,生物量增长2.8倍左右;细胞生长周期内,碳源消耗规律表现为蔗糖和可溶性总糖的浓度持续降低,而还原糖则表现为先升高后降低;过氧化物酶活测定显示酶活水平与蔗糖浓度的高低呈一定程度的正相关.     

药用植物刺山柑愈伤组织诱导及细胞生长代谢特征研究

本文研究了不同外植体及激素对刺山柑愈伤组织诱导的影响,不同激素配比对愈伤组织增殖培养以及悬浮细胞的生长与代谢特征。结果表明:以刺山柑叶片作为诱导愈伤组织的材料,效果较佳;愈伤组织诱导和继代的适宜培养条件是分别是MS 0.5mg/L 2,4-D 1.0mg/L6-BA和MS 1.0mg/L2,4-D 1.5mg/L6-BA。刺山柑悬浮培养细胞的生长周期为30天左右,细胞生长曲线呈"S"形,生物量增长2.8倍左右;细胞生长周期内,碳源消耗规律表现为蔗糖和可溶性总糖的浓度持续降低,而还原糖则表现为先升高后降低;过氧化物酶活测定显示酶活水平与蔗糖浓度的高低呈一定程度的正相关。     

银杏幼嫩茎段培养诱导愈伤组织及其细胞学研究

将银杏幼嫩茎段培养于含NAA和BA的MS和改良MS培养基中获得球型胚。培养基类型对愈伤组织的诱导和分化有明显影响,MS培养基优于DCR培养基。所有的MS和改良MS培养基上有胚性细胞的分化。在含有4mg／L的NAA的MS和改良MS培养基中,有三细胞和四细胞的原胚出现,并在MS 2mg／L NAA 0．1mg／L BA的培养基中观察到球形胚的分化。     

油松胚珠愈伤组织诱导和悬浮细胞系的建立

雌配子体处于游离核时期的油松胚珠以含不同浓度和配比生长调节剂的培养基诱导产生愈伤组织,并建立了悬浮细胞系.这一悬浮细胞系生长快,分散性好,稳定均一,适用于研究其生理、生化和细胞周期调控.     

药用植物人参愈伤组织的诱导培养与悬浮细胞体系建立

人参(Panax ginseng C.A Meyer)是五加科人参属植物,其主要药效活性成分为人参皂苷,具有广泛的药理作用。为建立人参愈伤组织诱导培养体系,以探索实现工程化细胞大规模生产人参皂苷的有效途径。本研究以人参根切片作为外植体,成功诱导出质地松疏、生长迅速、易于分散、启动较早的淡黄色透明状的愈伤组织,建立了适合于长期继代培养的人参细胞系,为深入开展人参干细胞分化及人参皂苷生物合成分子调控研究提供了理想的模型与基础。     

西洋参愈伤组织悬浮培养基质中蔗糖和无机元素的消耗状况

植物离体培养所采用的基本基质大多数是广谱性的,各种营养成分是否被充分利用因植物种类不同而异,这方面的研究报道较少[1～3],尤其在西洋参(PanaxquinquefoliumLinn.)芽胞愈伤组织悬浮培养中尚未见报道。本实验通过对西洋参愈伤组织培养基中蔗糖和无机元素消耗的研究,确定培养周期中碳源和无机元素的用量,为西洋参大规模细胞培养中碳源和无机元素及时补充和起始供给浓度的确定提供参数。1　材料与方法1.1　供试材料与培养条件采用本实验室在1999年筛选的西洋参芽胞愈伤组织无性系作为实验材料。先将固体培养基上的愈伤组织转入含有20m…     

菊芋试管苗的初代培养及愈伤组织不定芽诱导

菊芋（Helianthus tuberosus Linn.）为菊科（Asteraceae）向日葵属（Helianthus Linn.）多年生草本植物,耐寒、耐旱、耐贫瘠、耐盐碱[1];其地下块茎富含菊糖,还可通过发酵生产乙醇,在功能性食用多糖及生物能源方面的开发潜力巨大。菊芋主要通过块茎进行无性繁殖,其种子成活率和发芽率均很低[2],严重阻碍了菊芋的杂交育种。近年来以植物组织培养为基础的一系列现代育种技术为菊芋的种质改良提供了新途径,但由于菊芋的愈伤组织难以诱导不定芽或体胚发生,导致以农杆菌转化为主的转基因育种技术的应用受到限制。     

Establishment of embryogenic callus and high protoplast yielding suspension cultures of sugarcane (Saccharum spp. hybrids)

For 18 sugarcane cultivars, four distinct callus types developed on leaf explant tissue cultured on modified MS medium, but only Type 3 (embryogenic) and Type 4 (organogenic) were capable of plant regeneration. Cell suspension cultures were initiated from embryogenic callus incubated in a liquid medium. In stage one the callus adapted to the liquid medium. In stage two a heterogeneous cell suspension culture formed in 14 cultivars after five to eight weeks of culture. In stage three a homogeneous cell suspension culture was developed in six cultivars after 10 to 14 weeks by selective subculturing to increase the proportion of actively dividing cells from the heterogeneous cell suspension culture. Plants were regenerated from cell aggregates in heterogeneous cell suspension cultures for up to 148 days of culture but plants could not be regenerated from homogeneous cell suspension cultures. High yields of protoplasts were obtained from homogeneous cell suspension cultures (3.4 to 5.2 × 10⁶ protoplasts per gram fresh weight of cells [gfwt^-1]) compared to heterogeneous cell suspension cultures (0.1 × 10⁶ protoplasts gfwt^-1). Higher yields of protoplasts were obtained from homogeneous cell suspension cultures for cultivars Q63 and Q96 after regenerating callus from the cell suspension cultures, then recycling this callus to liquid medium (S-cell suspension cultures). This process increased protoplast yield to 9.4 × 10⁶ protoplasts gfwt^-1. Protoplasts isolated from S-cell suspension cultures were regenerated to callus and recycled to produce SP-cell suspension cultures yielding 6.4 to 13.2 × 10⁶ protoplasts gfwt^-1. This recycling of callus to produce S-cell suspension cultures allowed protoplasts to be isolated for the first time from cell lines of cultivars Q110 and Q138.     

Cryopreservation of wheat suspension culture and regenerable callus

Wheat (Triticum aestivum L. cv. Norstar) suspension cultures and regenerable calli initiated from immature embryos can be cryopreserved in liquid nitrogen temperature (–196°C) by slow freezing (0.5°C/min) in the presence of a mixture of DMSO and sucrose or sorbitol. Cold hardening or ABA treatment before cryopreservation increased the freezing resistance and improved the survival of wheat suspension culture in liquid nitrogen. Callus culture, established from immature embryos, prefrozen in 5% DMSO and 0.5M sorbitol survived liquid nitrogen storage and resumed plant regeneration after thawing. The results confirm the feasibility of long term preservation of wheat embryo callus by cryopreservation and retention of plant regeneration ability.Abbreviations ABA Abscisic acid - 2,4-D 2,4-Dichlorophenoxyacetic acid - DMSO Dimethylsulfoxide - LN Liquid nitrogen - TTC 2,3,5-triphenyltetrazolium chloride NRCC No. 23850.     

Plant regeneration from highly embryogenic callus,cell suspension and protoplast cultures of Trifolium fragiferum

Using various media, tissue and protoplast cultures plant regeneration systems were developed for Trifolium fragiferum (2n=16). (L.). The best media for induction of embryogenic cultures were based on Kao (1977) or Kao and Michayluk (1975). Somatic embryogenesis was observed in cultures derived from green leaf mesophyll protoplasts of branching plants, somatic embryo protoplasts and cell suspension protoplasts, leaflets and various explants of immature zygotic embryos. The process of somatic embryogenesis was maintained for over two years on Murashige and Skoog's (1962) medium supplemented with 0.5 mg l^-1 benzyladenine and 0.05 mg l^-1 naphthaleneacetic acid. These long term cultures were capable of regenerating plants that were fertile and produced seeds. These results were compared with those from protoplast, tissue and organ culture of other species of the Trifolium genus. This revised version was published online in June 2006 with corrections to the Cover Date.     

Fluid shear effects on suspension cultures of Morinda citrifolia

The shear susceptibility of cell suspension cultures of the plant cell Morinda citrifolia was investigated by subjecting the cells to the well-defined shear field generated in turbulent flow through a capillary. Suspensions were circulated using a peristaltic pump and average shear stresses between 25 and 350 N m(-2) were generated in the capillary test section. Control experiments were performed to assess the possible contribution of the peristaltic pump to the observed cell damage. There was clear evidence of pump-induced damage at the more severe test conditions and all viability measurements were corrected accordingly. Both shake flask suspension cultures (aged between 9 and 15 days) and repeated batch fermentation cultures, grown in a stirred tank reactor (STR) under a variety of controlled agitation conditions, were tested in the capillary shear loop. The cell damage incurred was evaluated in terms of suspension viability, as determined by a dye exclusion technique. Viability loss was found to conform closely to a first-order model in which the rate constant was observed to increase with the imposed shear stress. Furthermore, a linear relationship was identified between the specific death constant and the cumulative energy dissipated. Post-shear morphological measurements showed that the chain length distribution is shifted toward markedly lower values. In comparison with shake flask cultures, repeated batch fermentation cultures exhibited a marked increase in sensitivity to capillary shear. Based upon the determined morphological characteristics, this result is primarily attributable to the increased chain lengths characteristic of the repeated batch cultures. (c) 1995 John Wiley & Sons, Inc.     

Berberine synthesis by callus and cell suspension cultures of Coscinium fenestratum

Callus and cell suspension cultures of Coscinium fenestratum were established from sterile petiole segments on Murashige & Skoog (MS) medium, supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D) and benzyl amino purine (BAP). The cells in the culture produced berberine as the major compound. NAA stimulated the product synthesis over 2,4-D. Presence of light inhibited the growth and enhanced the berberine synthesis.Abbreviations BAP 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxy acetic acid - HPLC high pressure liquid chromatography - IAA indole-3-acetic acid - NAA -naphthaleneacetic acid - TLC thin layer chromatography     

Attempts to produce solasodine in callus and suspension cultures of Solanum mauritianum Scop.

Callus cultures of Solanum mauritianum Scop. were initiated from green berry explants on a hormone-free Murashige and Skoog (1962) medium excluding glycine, and containing 0.1 g L^–1 myo-inositol and 3% sucrose. Such cultures contained 10.08±0.59 g g^–1 DW of solasodine, which is equivalent to that in the leaves of mature S. mauritianum plants, but far less than that extracted from the green berries (185 g g^–1 DW). In vitro solasodine productivity could be increased by reducing the strength of the medium by half, substituting 3% glucose for 3% sucrose as carbon source, or by the addition of certain combinations of BA and NAA. Phosphate limitation and alterations in the carbon: nitrogen ratio were not able to increase solasodine productivity. Suspension cultures of S. mauritianum were initiated and maintained in a Murashige and Skoog (1962) medium with the RT vitamins of Khanna and Staba (1968), 0.1 g L^–1 myo-inositol, 3% sucrose and 1 mg L^–1 2,4-D. No solasodine was detectable in these cultures, or slight modifications thereof.Abbreviations BA benzyladenine - NAA naphthaleneacetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - MS Murashige and Skoog's (1962) medium     

Rollinia mucosa cell suspension cultures: Establishment and growth conditions

Different types and concentrations of plant growth regulators were tested in order to obtain the best callus and cell suspension culture growth conditions of Rollinia mucosa (Jacq.) Baill. (Annonaceae). Picloram was shown to be the most efficient for induction and production of friable calluses, independent of the concentration used. Cellular morphology and viability, fresh and dry weights, pH and medium sugar concentration were determined for cell suspension cultures. Dissimilation curves were used for the characterization of the growth of cell suspension cultures. Picloram provided the most rapid growth and produced the highest biomass, with little variation in morphology (differentiated cells). It also provided the highest dissimilation, when compared with cell suspension cultures maintained in media with 2,4-D or NAA + BA + GA₃. Stable cell suspension cultures can be established in MS medium supplemented with 20.8 μM picloram. This revised version was published online in August 2006 with corrections to the Cover Date.