蝴蝶兰组织培养快繁技术研究

本试验对蝴蝶兰组织培养不同阶段的适用培养基和激素配比水平进行了筛选,结果表明:在蝴蝶兰花梗腋芽诱导中,选用花宝(N-P-K:6.5-6-19)2.5 g/L(克/升)为基础培养基,添加BA3.0 mg/L(毫克/升)有利于对花梗腋芽诱导;原球茎增殖培养以较低的无机盐浓度为好,采用1.0g/L(克/升)花宝为基础培养基,添加2.0 mg/L(毫克/升)的6-BA和0.5 mg/L(毫克/升)的NAA,原球茎的增殖系数2个月内能达5倍以上,是最佳的原球茎增殖组合;以2.7 g/L(克/升)的花多多1号(N-P-K:20-20-20)为基础培养基,添加0.1 mg/L 6-BA 0.5 mg/L(毫克/升)NAA,植株生根率高,根数多,生根长,是较为理想的生根培养基.     

不同添加物对金针菇几种胞外酶活性的影响

以棉子壳培养基为对照,向棉子壳培养基中分别添加淀粉、尿素、石膏,研究在金针菇整个生长发育过程中培养基中胞外纤维素酶、半纤维素酶、淀粉酶、果胶酶、过氧化物酶、漆酶活性的变化规律、呼吸消耗及绝对生物学效率。结果表明:(1)添加尿素能明显提高金针菇的绝对生物学效率;而添加淀粉、石膏不能提高金针菇的绝对生物学效率;(2)纤维素酶、半纤维素酶和β-葡萄糖苷酶在金针菇菌丝生长阶段活性较低,子实体发育期活性较高。淀粉酶、果胶酶、过氧化物酶在菌丝生长阶段活性较高,子实体发育期活性较低,漆酶活性变化趋势不明显;(3)金针菇具有完整的胞外纤维素酶系;(4)不同添加物只影响酶活性的相对大小,而对胞外酶的活性变化趋势影响较小。     

高灌蓝莓微体繁殖技术研究初报

从美国引进高灌蓝莓无毒试管苗，进行组织培养微繁技术研究，在改良WPM添加玉米素0．5－2.0mg/L的增殖培养基上，新梢增殖50倍以上，在1／2改良WPM添加IBA0.1mg/L的生根培养基上，生根率达30％－705，采用1：1的草炭土与河沙做基质移栽炼苗，成活率50％以上，移栽到砂壤土中，当年生长量可达50cm，翌春有60％的植标开花结果，山东沙石山区的微酸性土壤适合北美高灌蓝莓的生长发育。     

TDZ在4种观赏花卉芽增殖中的应用

为了探讨噻重氮苯基脲(TDZ)在4种观赏花卉(石斛兰,卡特兰、绿巨人、火鹤)芽增殖中的作用,提高芽增殖效率。试验在传统培养基基础上添加TDZ,分析不同浓度TDZ对4种观赏花卉芽增殖的影响。结果表明:石斛兰、卡特兰、绿巨人和火鹤芽增殖效果最好的培养基分别为:MS+6-BA 1.0 mg/L+NAA 0.1 mg/L+TDZ 0.04 mg/L、1/2MS+6-BA 0.2 mg/L+NAA 0.1 mg/L+TDZ 0.01 mg/L、MS+6-BA 3.0 mg/L+2,4-D 0.5 mg/L+TDZ 0.02 mg/L和MS+6-BA 1.5 mg/L+TDZ 0.04 mg/L。说明在4种观赏花卉芽增殖中添加TDZ能够提高分化效率。     

大花蕙兰原球茎诱导、增殖与分化影响因素研究

以大花蕙兰试管组织为试材,MS为基本培养基,研究了不同添加物6-BA、NAA、IBA、活性炭,对大花蕙兰原球茎诱导、增殖和分化的影响。结果表明:适宜原球茎诱导、增殖的最佳培养基为MS+1.00mg·L~(-1) 6-BA+0.1mg·L~(-1) NAA,适宜原球茎分化的最佳培养基为MS+1.0mg·L~(-1) 6-BA+0.2mg·L~(-1) NAA,在培养基中添加0.3%活性炭有利于原球茎的增殖和分化。     

杜鹃兰原球茎增殖培养条件

以杜鹃兰种子萌发的原球茎为试材,研究不同培养基、不同植物生长调节物质(6-BA、NAA和IBA)、活性炭、温度及光照强度对杜鹃兰原球茎增殖的影响。结果表明:1/2MS培养基为适于原球茎增殖的基本培养基;添加一定浓度的6-BA、NAA和IBA均有利于原球茎的增殖,IBA的浓度为1.0mg·L~(-1)时,原球茎增殖率较高,40d可达170.07%;适量添加活性炭对原球茎增殖有促进作用;温度及光照强度的控制对原球茎增殖是必需的。杜鹃兰原球茎增殖的适宜培养条件为1/2MS+1.0mg·L~(-1) 6-BA+1.0mg·L~(-1) IBA+0.5g·L~(-1)活性炭,温度为15℃,光照强度为2 000lx。     

梨矮化中间砧S5离体增殖的研究

采用L16(44)正交试验对梨矮化中间砧S5进行了离体增殖研究,结果表明,不同种类的培养基及不同配比的植物生长调节剂对试管苗增殖的效果不同.MS培养基较QL、AS、WPM培养基更有利于S5茎的增殖,中等浓度的细胞分裂素(BA)与较低浓度的生长素(IBA)、赤霉素(GA3)配合使用效果更理想.综合茎质量、茎数量以及茎长度等指标认为,MS 2.0mg/0L BA 0.1 mg/L IBA 1 mg/L(毫克/升)GA3是S5试管苗茎增殖的最佳培养基.     

不同激素配比对白及愈伤组织诱导、增殖和分化的影响

以白及为试材,以MS和1/2MS为基本培养基,研究了愈伤组织诱导、增殖和分化培养基中的最佳激素种类和浓度配比。结果表明:在MS中添加6-BA 1 mg·L~(-1)和2,4-D3mg·L~(-1)时白及愈伤的诱导率最高,达89.89%。2,4-D是诱导白及愈伤生成的关键激素,而在基本培养基中单添加6-BA时并不能诱导愈伤组织的生成,同时添加2,4-D和6-BA能有效促进愈伤的诱导,且愈伤诱导率随着2,4-D浓度的增加而逐渐增加,但优质愈伤率也随之降低。添加6-BA 0.1mg·L~(-1)和2,4-D 2mg·L~(-1)的增殖培养基中愈伤细胞的增殖率最高,30d的增殖倍数达7.457。在分化培养基中添加NAA和TDZ有利于白及愈伤组织分化,分化率随着二者浓度的增加而增加,且NAA的促分化效果优于TDZ。MS+2,4-D 2mg·L~(-1)+NAA 1mg·L~(-1)为最佳分化培养基,分化30d时其分化率为86.67%。     

梨矮化中间砧S_5离体增殖培养基的研究

采用4×4正交试验对梨矮化中间砧S5进行了离体增殖研究。结果表明,不同种类的培养基及不同配比的植物生长调节剂对试管苗增殖的效果不同。MS培养基较QL、AS、WPM培养基更有利于S5茎的增殖,中等浓度的细胞分裂素(BA)与较低浓度的生长素(IBA)、赤霉素(GA3)配合使用效果更理想。综合茎质量、茎数量以及茎长度等指标认为,MS+2.0mg/LBA+0.1mg/LIBA+1.0mg/LGA3是S5试管苗茎增殖的最佳培养基。     

氨基酸对荔枝愈伤组织增殖及体胚发生体系优化的研究北大核心CSCD

【目的】探究氨基酸对荔枝愈伤组织增殖及体胚诱导的影响。【方法】采用L9(34)正交设计,分别在妃子笑荔枝愈伤组织继代及体胚诱导阶段添加不同氨基酸,比较其对愈伤组织增殖及体胚诱导的影响。【结果】在愈伤组织增殖阶段培养基中添加氨基酸均显著提高愈伤组织增殖率,最优处理为基础继代培养基添加0.1 g·L^(-1)γ-氨基丁酸+0.2 g·L^(-1)谷氨酰胺,愈伤组织增殖率为初始接种量的14.04倍。与对照相比,氨基酸处理组的胚性愈伤组织淡黄,颗粒较小,细胞呈椭圆形,细胞质浓厚,多为正在分裂状态;非胚性愈伤组织浅白,水渍严重,着生于顶部或者边缘,易于剔除。在愈伤组织增殖阶段培养基中添加γ-氨基丁酸、谷氨酰胺和丙氨酸对后续的体胚发生及萌发影响极显著(p<0.01),最优配方为基础继代培养基添加0.3 g·L^(-1)γ-氨基丁酸+0.4 g·L^(-1)谷氨酰胺+0.05 g·L^(-1)丙氨酸,每克愈伤组织可获得461个体胚,77株组培苗。在体胚发生阶段培养基中添加氨基酸,获得的体胚发生及萌发最优配方为基础诱导培养基添加0.3 g·L^(-1)γ-氨基丁酸+0.4 g·L^(-1)谷氨酰胺+0.05 g·L^(-1)丙氨酸,每克愈伤组织可获得270个体胚,72株组培苗。【结论】氨基酸可调节愈伤组织胚性,提高愈伤组织增殖率,促进体胚发生及萌发。     

Three dimensional tissue culture in culture medium with rhbFGF

AIM:To investigate the behaviour of 3T3 fibroblast and macrophage co-culture on blood fibrin clot or adipose tissue with recombinant basic fibroblast growth factor (rhbFGF).METHODS:MTT method, inverted contrast microscopy, Giemsa staining as well as scanning electron microscope were used in the present study.RESULTS:The effect of rhbFGF on co-culture of 3T3 fibroblast and mouse macrophage on blood fibrin clot in low-serum DMEM with rhbFGF were monitored, and 3T3 fibroblast and macrophage growed well on the blood fibrin blot in low-serum DMEM with rhbFGF. CONCLUSION:The blood fibrin clot, with its low immunogenicity, could be used as a bionic support for three-dimensional tissue culture, and also a physiological carrier to distribute the growth factor rhbFGF for the cells.     

核桃花粉离体萌发的培养基研究

以6a生云新早实核桃的新鲜花粉为试材,采用花粉离体培养法,研究了不同的培养基组分以及温度对核桃花粉萌发的影响。结果表明:1)在单因子试验中,蔗糖、H3BO3和CaCl2在一定质量浓度范围内,对核桃花粉萌发及花粉管生长起促进作用,但超过一定质量浓度时则起抑制作用;而氮、镁和钾对核桃花粉萌发不显著。2)在正交试验中,蔗糖、H3BO3和CaCl2对核桃花粉萌发有极显著的影响,核桃花粉最适的培养基为100g/L蔗糖+10mg/LH3BO3+40mg/LCaCl2,花粉的萌发率可以达到45.24%。3)核桃花粉离体培养的最适温度为25℃,培养温度过高或过低都会抑制核桃花粉的萌发和花粉管生长。     

水溶性生物基磺酸盐营养液在食用菌培养上的应用研究

首次利用水溶性生物基磺酸盐作为主要成分制作一种低成本和营养增强型的食用菌新型营养液,用于糙皮侧耳(平菇)液体培养和栽培。试验结果为:该营养液既适用于糙皮侧耳液体培养,也可作为添加剂用于糙皮侧耳栽培,不仅能促进增产,还可改善食用品质。     

香菇液体菌种培养基筛选及适宜接种期试验初报

食用菌菌种从形态可以分为液体菌种和固体菌种。试验从4个常用液体菌种发酵培养基配方中筛选出原料易得且发酵效果良好的配方,并得出转接固体(木屑)培养基的液体菌种适宜培养时间为144 h。试验对香菇液体菌种生产及应用具有一定的指导意义。     

Development of a medium for in vitro culture of Galanthus species based on the mineral composition of bulbs

Summary

To optimise conditions for micropropagating Galanthus species, a basal medium (G) was developed based on mineral analyses of G. nivalis, G. nivalis ‘Flore Pleno’ and G. elwesii bulbs. Compared with Murashige and Skoog (MS) medium, the main features of G medium were increased concentrations of Cu ( 30.4), P ( 3.6), Ca ( 1.9), Mg ( 1.3) and S ( 1.2) and reduced levels of Mn ( 0.07), Zn ( 0.59) and K ( 0.65). The efficacy of G medium in supporting bulblet initiation on bulb chip explants, bulblet multiplication (on media supplemented with 30 g l^–1 sucrose, 1.0 mg l^–1 6-benzylaminopurine and 0.1 mg l^–1 naphthalene acetic acid), and bulblet growth (on plant growth regulator-free media with 60 g l^–1 sucrose and 5 g l^–1 activated charcoal) was compared with MS medium over a range of dilutions (full-, ¹?2-, ¹?4-, and ¹?8-strength). Bulblet initiation was superior on G medium for G. nivalis and G. nivalis ‘Flore Pleno’, but inferior for G. elwesii. The choice of basal medium did not influence bulblet multiplication, although multiplication was reduced on both media diluted to ¹?8-strength. G medium supported bulblet growth and rooting better than MS medium, while dilution of either medium reduced bulblet growth and rooting. Using G medium in place of MS medium during bulblet multiplication greatly reduced hyperhydration with G. elwesii, as did dilution of either of the basal media.     

Effect of genotype,explant size,position, and culture medium on shoot generation of Gerbera jamesonii by receptacle transverse thin cell layer culture

An unique procedure for the mass shoot propagation of Gerbera using receptacle transverse thin cell layer (tTCL) culture procedure was developed. Genotype, flower bud age, explant size, position of receptacle tTCLs and culture media were found to affect the success of culture. Ten interspecific crosses of Gerbera showed different shoot regeneration rates and callus induction via receptacle tTCL culture, all of which had shoot regeneration rates higher than 57%. Flower buds collected on the 10th day resulted in 91% shoot regeneration after 6 weeks of culture on basal MS medium [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant. 15, 475–497] supplemented with 0.02 mg l⁻¹ thidiazuron (TDZ), 0.8 mg l⁻¹ adenine and 10% (v/v) coconut water (CW). This was significantly higher than those from flower buds on the 7th and 14th days (22% and 54%), respectively. Shoot regeneration rate was the highest (94–100%) in the middle layers of the receptacle. For mass shoot propagation, shoot clusters were subcultured on half-strength MS medium supplemented with 0.5 mg l⁻¹ indole-3-butyric acid (IBA), 0.5 mg l⁻¹ 6-benzyladenine (BA) and 2.0 mg l⁻¹ kinetin after every 4 weeks. Plantlets formed when single shoots were cultured on half-strength MS medium containing 1 mg l⁻¹ IBA. All plantlets acclimatized well in the greenhouse.     

Shoot tip culture in mango: Influence of medium,genotype, explant factors,season and decontamination treatments on phenolic exudation,explant survival and axenic culture establishment

Summary

Attempts to establish shoot tip culture in mango (Mangifera indica L.) genotypes ‘Alphonso’, ‘Totapuri’, ‘Banganapalli’ and ‘Arka Anmol’ indicated that the problems of phenolic exudation, medium discoloration and explant browning were interrelated and influenced by a number of factors including medium, genotype, explant, season and decontamination treatments. Less phenolic exudation and better explant survival were observed in ½MS medium than full MS and in semi-solid than in liquid medium. Among explant factors, age and shading of shoots, length, thickness and scaling injuries on explant and lower cut petioles coming in contact with medium were important, but not the age of tree and the number of leaves on the shoot. Use of charcoal was beneficial but other adsorbents, antioxidants, dark incubation and submerged culture were not advantageous. The effect of season was prominent with least phenolics, minimum microbial contamination, best explant survival and genotype dependent growth response in current season’s semimature shoots collected during June-August. Deep seated microbial contaminants could not be checked completely. A scheme for establishing in vitro culture is suggested.     

Possibility of 3T3 fibroblast growth on blood fibrin clot in culture medium with recombinant human basic fibroblast growth factor

AIM: To investigate the possibility of 3T3 fibroblast growth on blood fibrin clot in culture medium with recombinant human basic fibroblast growth factor (rhbFGF). METHOD: Growth of the cells on blood fibrin clot was studied by phase-contrast, scanning and transmission electron microscopy and by Giemsa stain and MTT assay. RESULTS: The optimal concentration of rhbFGF for proliferation and survival of the cells was 100 ng/mL. The cells also grew on blood fibrin clot scaffold in the low-serum medium containing 100 ng/mL rhbFGF, and a greater number of the cells survived after 48 hours incubation compared to that after 24 hours. The elongated filopodia appeared to bridge the gaps among the fibroblasts after 24 hours incubation. Further incubation to 72 hours, a greater number of platycytes were found to be joined together by lamellopodia. CONCLUSION: 3T3 fibroblasts could grow and survive on blood fibrin clot in the low-serum medium containing rhbFGF, and a combination of blood fibrin clot and rhbFGF may have over proportional effects on wound healing.     

In vitro proliferation and minimum growth storage of fraser photinia: Influences of different medium,sugar combinations and culture vessels

Protocols for the in vitro proliferation and storage of fraser photinia were developed by comparing 6-benzyladenine (BA) concentrations (0.5–4 mg/L) together with different media formulations [Murashige and Skoog (MS) media and Quoirin and Lepoivre (QL) media], sugar combinations (sucrose and mannitol), culture vessels (baby food jars and vitrovents) and methods (synthetic seed technology and slow growth storage). The best responses in terms of both proliferation percentage and multiple shoot formation were obtained in QL medium containing 1 mg/L BA. Synthetic seed production was optimized by encapsulating shoot apices in 3% sodium alginate. Encapsulated shoot apices could be maintained up to 6 months at 4 °C in dark with 91.6% sprouting in MS medium. Microshoots were stored at 4 °C up to 15 months on sucrose and mannitol containing QL medium in both baby food jars and vitrovents without subculture. The stored material could be recovered and multiplied normally in 1 mg/L BA supplemented QL medium. Both in vitro propagated and conserved microshoots were rooted (∼75%) on QL medium with 1 mg/L indole butyric acid (IBA). Optimized synthetic seed and slow growth storage system can be used for short and medium-term storage of fraser photinia germplasm.     

Micropropagation of dahlia in static liquid medium using slow-release tools of medium ingredients

Growth of dahlia shoots in vitro was ca. 4 times faster in liquid medium than on solidified medium. In liquid standard medium (3% sucrose, macroelements according to Driver–Kuniyuki Walnut medium, microelements according to Murashige–Skoog medium, 0.44 μM benzylaminopurine), the major medium ingredients were consumed for 75–80% during the first 6 weeks. Addition of extra ingredients increased growth, demonstrating that the amount of ingredients added at the start of culture was suboptimal. When the extra ingredients were given at the start of the culture, concentrations became too high and therefore inhibitory. When the ingredients were added during the subculture cycle by means of small aliquots of a concentrated solution or by means of slow-release tools, growth was strongly increased. Osmocote gave satisfactory results as a slow-release tool for inorganics. For organic ingredients (sucrose and benzylaminopurine), a novel slow-release tool was developed.