芍药组织培养中地下芽污染的克服

针对芍药地下芽组织培养中污染严重的问题,对不同外植体的选择及不同消毒处理方法进行了研究。结果表明:芍药的地下芽培养应该选取生长健壮、饱满紧实的中等大小芽子;对于不同大小的地下芽应采取不同时长的HgCl2消毒处理,以5~8 min为宜。对地下芽进行消毒处理时要剥去外面数层鳞片且至少保留2层鳞片;芍药地下芽以一步灭菌法为宜,太多的处理增加了其感染细菌的概率且无法保证灭菌彻底。     

二乔杜鹃无菌株系建立的初步研究

以二乔杜鹃顶芽为外植体,探讨不同花色类型、不同消毒时间、培养基中不同生长调节剂组合对其无菌株系建立的影响。结果表明,以0.1%升汞作表面消毒剂,顶芽外植体的最适消毒时间为10 min;在相同消毒条件下,白花类型的污染率低于红色类型,红花类型外植体的褐变率低于白花类型;含较高浓度ZT的初代培养基中外植体的褐变率较低。     

“胡萝卜的组织培养”消毒方法的优化

为了尽可能在高中开展胡萝卜的组织培养实验,选取胡萝卜根为外植体,从改进外植体消毒方法、缩短消毒时间2个方面进行优化。实验结果表明:胡萝卜根经多次消毒组合:70%酒精浸泡30 s+20%次氯酸钠溶液浸泡10 min+酒精灼烧12 s处理后,污染率降至5.55%,实验操作总时间约20 min。     

晚松不同外植体腋芽诱导的初步研究北大核心CSCD

以一年生晚松实生苗春梢不同部位的茎段和种子萌发的顶芽两种材料为外植体,用浓度为0.1%的升汞对外植体进行消毒处理,选择消毒效果较好的外植体作为无菌材料,然后优化最适合腋芽诱导的培养基类型和激素水平组合。结果表明,0.1%的升汞消毒处理10 min的春梢中部茎段最佳,污染率最低,为0.8%,其腋芽诱导的最佳培养基和激素浓度为:MS+2.5 mg/L 6-BA+0.2 mg/L NAA。而种子的最佳消毒时间为4 min,污染率为2.4%,在含有0.7 mg/L NAA和1.0 mg/L 6-BA的MS培养基中诱导率最高。本文建立了由春梢茎段和种子诱导腋芽的有效方法,为晚松的快速繁育提供理论依据和技术支持。     

过氧化氢干雾对生物安全柜微环境表面消毒效果的研究

生物安全实验室微环境消毒是控制实验室污染的重要环节。过氧化氢广泛用于病原微生物实验室微环境消毒,但其对不同病原微生物的消毒效果有待研究。本文研究了过氧化氢干雾（粒径％10μm）以不同消毒程序对生物安全柜表面常见微生物的消毒效果。结果显示,在生物安全柜内采用优化的消毒程序（发散循环8次,每次1min,达60ppm后,静置消毒2h）,过氧化氢干雾可完全杀灭1×106CFU枯草芽胞杆菌、嗜热脂肪芽胞杆菌、金黄色葡萄球菌、表皮葡萄球菌、耻垢分枝杆菌,以及1×106CFU大肠埃希菌。然而,当金黄色葡萄球菌、表皮葡萄球菌、耻垢分枝杆菌浓度达1×107CFU时,过氧化氢干雾无法完全杀灭。因此,建议在进行过氧化氢干雾消毒时,应先用消毒剂处理,以期彻底杀灭生物安全柜微环境中污染的病原微生物。     

花梨木组织培养外植体消毒方法初步探讨

针对在花梨木(Dalbergia odofifera T. Chen)组织培养过程中外植体的有效消毒方法进行研究。外植体消毒主要以2年生以上实生苗的茎段为材料,开展了茎段外植体次氯酸钠(5%,10%,15%)和升汞(0.05%,0.1%,0.2%)3个浓度水平的消毒试验。在此基础上,开展了外植体消毒的时间试验,次氯酸钠消毒时间为10 min、20 min、30 min,升汞消毒时间为5 min、10 min、15 min。结果表明：茎段外植体采用10%浓度的次氯酸钠消毒效果较好,最佳消毒方式75%的酒精浸泡2 min,10%次氯酸钠消毒20 min,该方法污染率最低、存活力最高。     

晚松不同外植体腋芽诱导的初步研究

以一年生晚松实生苗春梢不同部位的茎段和种子萌发的顶芽两种材料为外植体,用浓度为0.1%的升汞对外植体进行消毒处理,选择消毒效果较好的外植体作为无菌材料,然后优化最适合腋芽诱导的培养基类型和激素水平组合。结果表明,0.1%的升汞消毒处理10 min的春梢中部茎段最佳,污染率最低,为0.8%,其腋芽诱导的最佳培养基和激素浓度为:MS+2.5 mg/L 6-BA+0.2 mg/L NAA。而种子的最佳消毒时间为4 min,污染率为2.4%,在含有0.7 mg/L NAA和1.0 mg/L 6-BA的MS培养基中诱导率最高。本文建立了由春梢茎段和种子诱导腋芽的有效方法,为晚松的快速繁育提供理论依据和技术支持。     

大蒜愈伤组织诱导条件初报

以白皮大蒜和大头蒜为材料,报道了大蒜品系、外植体的低温前处理、外植体大小及取材种类不同对愈伤组织诱导的影响。结果表明：（１）无论是４℃低温保藏或室温保存前处理,大头蒜的消毒效果均优于白皮大蒜,４℃低温前处理可降低外植体出愈污染;（２）只有带表皮的外植体才可诱导愈伤组织,外植体较大（０．５ｃｍ×０．３ｃｍ×０．２ｃｍ）,继代期愈伤组织产生量较多;外植体偏小（０．２ｃｍ×０．１ｃｍ×０．１ｃｍ）,极少产生愈伤组织;（３）４℃冷藏１８ｄ对大头蒜愈伤组织诱导有一定影响,但对白皮大蒜影响不大。冷藏可推迟出愈１～２ｄ。     

降低卷丹百合组培褐变技术研究

以卷丹百合Lilium lancifolium鳞片为外植体,采用1/2MS+0.8 mg·L~(-1) 6-BA+0.2 mg·L~(-1) NAA为基本培养基,研究不同消毒时间的外植体消毒效果,探讨聚乙烯吡咯烷酮(PVP)、抗坏血酸(V?)、活性炭(AC)以及光、暗两种培养条件对卷丹百合组织培养过程中褐变的影响。结果表明,用75%酒精消毒30 s、0.1%HgCl_2消毒10 min、2%NaClO消毒6 min的消毒效果最好,污染率为33.33%,成活率和诱导率均为66.67%。在培养基中加入PVP、V?、AC和暗培养均能减轻褐变的发生,其中以4.5 mg·L~(-1) PVP处理对卷丹百合褐变的抑制效果最好,外植体褐变率最低为30%,愈伤组织诱导率为70%。     

猴面包树的离体快繁条件筛选

该文以猴面包树(Adansonia digitata)种子为外植体,首先筛选合适的种子预处理及消毒方法,然后经过启动培养获得无菌外植体后在增殖培养基中进行丛生芽诱导,将丛生芽切成单株进行生根壮苗培养,最终建立猴面包树离体快繁技术体系.结果表明:75％酒精浸泡3 min+0.1％升汞消毒15 min消毒效果较佳,污染率为...     

香蕉组织培养过程中内生菌污染的控制

研究了体外培养一种孟加拉传统香蕉(Musa spp.Cv. Kanthali)的茎尖组织。茎尖的原始细胞表面经无菌处理(0.1%HgCl2处理12min) ,接种6~15d后外植体地下茎部分仍有微生物污染(大部分是细菌) ,杀死了85%的外植体。为确定无污染培养基,将等量外植体分别浸泡在含400mg/L氨苄青霉素和200mg/L庆大霉素(两种光谱抗生素)的培养基中1h。结果表明,经抗生素处理的外植体完全没有污染,但培养3周后不能再生。进行二次继代培养后,其中一部分外植体吸收了培养基并胀大,颜色由苍白转变成浅绿或深绿。三次继代培养后数天,不再观察到外植体的生长,所有经抗生素处理过的外植体都开始死亡。在未经抗生素处理的活外植体中,单个茎发育的最佳培养基是:MS 4.0mg/L BA 0.5mg/L KT 15% CW,平均生长时间为18~21d,但再生率很低,只有30%。茎细胞增殖的最佳培养基是:MS 4.0mg/L BA 2.0mg/LIAA 15% CW,每个茎平均只萌发3~4个芽。最后,在添加0.5mg/LIBA的一半浓度的MS培养基中,体外培养茎最大生根率达到90%。     

Comparative surface examinations of morse taper junctions of the MRP-Titan stem shot peened with glass beads]

INTRODUCTION: Shot peening is widely used for surface treatment of hip implants. Shot peening with steel balls followed by a cleaning process with glass beads is used for introduction of negative stress in the production of morse taper junctions of the MRP-Titan stem. An increasing number of publications in maxillofacial surgery and orthopaedic surgery show that there is a significant contamination of Alumina or glass blasted surfaces. Latest research suggested an association between contaminant particles with early loosening of endoprostheses (third body wear). The aim of this study is to evaluate the amount and the effects of surface contamination with glass particles on morse taper junctions of implants and explants of the MRP-Titan stem. MATERIAL AND METHOD: The surface of morse taper junctions of the MRP-Titan stem (5 original-package implants and explants each) are analysed for glass particle contamination. A field emission scanning electron microscopy (LEO 1525) is used for the detection of the glass-particles on the implant surface with a backscattered electron detector. The relative surface area covered by particles was calculated by means of an image analyzing software (analySIS, Soft Imaging System GmbH). RESULTS: The surface of the implants showed a considerable contamination with glass particles with a mean of 6.67 +/- 0.82% compared to 2.06 +/- 0.74% on the surface of the explants. The difference was statistically significant (p<0.0001). DISCUSSION: The results of this study show that there is a relative high percentage of contamination with glass particles on shot peened morse taper junctions of the MRP-Titan stem. This contamination is significantly lower on the surface of the explants. With respect to third body wear and osteolysis in total hip arthroplasty further studies are necessary to minimize contamination while maintaining adequate surface quality.     

Explant preparation affects culture initiation and regeneration of Panax ginseng and Panax quinquefolius

The influence of explant preparation on culture initiation and regeneration was investigated. Explant preparation was defined as the application of surface disinfection and homogenization to ginseng roots and embryos. Surface disinfection significantly affected culture initiation and subsequent embryogenesis. A high success rate (80–97%) in culture initiation was associated with explants from non-surface disinfected root tissue. The cumulative contamination rate after 8 months was below 4%; in contrast, a contamination rate of 85% was observed in disinfected root explants. For non-disinfected explants, visible callus was induced in 1 week, adventitious root and somatic embryos were formed in 2 and 6 months, respectively. Explants from disinfected roots required 1 month to induce visible callus, 5–7 months for adventitious roots and 10 months for somatic embryogenesis. For germinating embryos, disinfected embryos required double the time for embryogenesis than non-disinfected ones. It was considered that surface disinfection imposed a stress and subsequently a carry-over effect on explants.     

Control of <Emphasis Type="Italic">In vitro</Emphasis> contamination of explants from greenhouse- and field-grown trees

Summary Controlling fungal and bacterial contamination of woody plant material can be extremely difficult. Isothiazolone biocides and sodium dichloroisocyanurate have been used singly and in combination to reduce microbial contamination in bud explants derived from greenhouse- and field-grown citrus trees. Explants from greenhouse-grown trees were effectively disinfested (<5% vs. 85% contamination) using a ‘standard’ disinfestation (SD) procedure followed by culture on medium containing 5 ml l⁻¹ Plant Preservative Mixture, or by the SD procedure but substituting 300 ppm sodium dichloroisocyanurate for 48 h for the treatment with 1.05% NaOCl in the SD procedure. Disinfestation of explants from field-grown trees was less effective than explants from the greenhouse, but was improved (10% vs. 47% contamination) by using a combination of the SD procedure with 1.05% NaOCl, or 100 or 300 ppm sodium dichloroisocyanurate followed by culture onto medium containing 5 ml l⁻¹ Plant Preservative Mixture. Mention of a trademark, warranty, proprietary product, or vendor does not constitute a guarantee by the US Department of Agriculture and does not imply its approval to the exclusion of other products or vendors that may also be suitable.     

Transformation of blueberry without antibiotic selection

Transformation of the blueberry cultivar North Country (Vaccinium corymbosum × V. angustifolium) was achieved using the disarmed Agrobacterium tumefaciens isolate LBA4404, containing a binary vector with an intron-containing β-glucuronidase (GUS) marker gene. Plant regeneration was carried out in the absence of antibiotics due to their toxicity to blueberry explants. Agrobacterium contamination was controlled by dipping the explants in antibiotics and rinsing in sterile distilled water. Once regeneration had been achieved, the plantlets were placed on to medium containing the antibiotic ticaricillin at 250 mg litre^-1 to control and try to eliminate any remaining Agrobacterium. Selection of regenerating explants expressing GUS was achieved by growing the plant material for 2 days on a medium containing 4-methyl umbelliferyl glucuronide (MUG), and examining the medium under UV light to detect fluorescent activity. From the 19 explants showing signs of regeneration, seven produced fluorescent patches on MUG medium. From the selected explants, five plantlets were found to express the GUS gene as detected by fluorometric and histochemical analysis. PCR was used to confirm the presence of the GUS and/or NPTII marker genes after 2 years in culture. Bacterial contamination isolated from plant material (which appeared free of contamination) was examined for GUS activity and analysed using PCR with GUS and NPTII specific primers, but no positive results were obtained.     

植物离体培养中微生物污染的鉴定与控制(综述)

本文综述植物离体培养过程中微生物污染的鉴定与控制的研究进展,包括通过指示培养和菌种鉴别以鉴定污染菌;从保护条件下生长的植株上取材以及材料的预处理,以便有效地控制附生菌和应用抗生素控制内生菌.     

二氧化氯对小水榕外植体消毒效果研究

选取小水榕的绿芽和粗壮嫩芽组织为外植体,研究不同浓度ClO₂对外植体的消毒效果。结果表明:等于或高于500 mg/L的ClO₂溶液能迅速破坏小水榕成熟组织的绿色素;ClO₂溶液浸泡后未经无菌水冲洗的外植体污染率低,但生长缓慢,难以分化丛芽;以粗壮嫩芽组织为外植体的材料经ClO₂或HgCl₂溶液浸泡灭菌都可获得无菌外植体,建立有效的快繁体系。     

观赏凤梨高效离体快繁影响因素的研究

以侧芽为外植体材料,对其组织培养的各个阶段进行了研究。结果表明:品种对外植体的污染率、褐变率、芽诱导率及丛生芽增殖倍数均有显著影响;外源激素的种类和浓度在芽诱导分化、增殖、生根等各个阶段都是主要的影响因素;外植体在1/2MS+NAA0.2mg·L~(-1)+6-BA2mg·L~(-1)上,48d后芽开始分化,诱导率为40%,平均芽分化数为6个,诱导分化效果最好;芽增殖培养阶段,若培养基中仅含6-BA,且浓度在0~4mg·L~(-1)之间,芽增殖倍数随其浓度增大而增大;若培养基中仅含NAA,且其浓度为0.2mg·L-1时,增殖倍数最大;培养基中同时添加6-BA和NAA,比单独添加6-BA或NAA时,芽的增殖效果好,且在6-BA3mg·L~(-1)+NAA0.5mg·L~(-1)的MS培养基上,G.dissitisflora和G.‘Claret’增殖倍数最大,分别为5.24和3.84;在任何相同的培养基上,G.dissitisflora的增殖倍数显著高于G.‘Claret’;小苗在含有NAA0.5mg·L~(-1)+IBA0·5mg·L~(-1)的生根培养基上生长,生根率可达91.03%,平均根数为5.3条/株,生根效果较好。     

In vitro explant culture of mantle epithelium of freshwater pearl mussel

The in vitro culture of nacre secreting pallial mantle explants of freshwater pearl producing mussel, Lamellidens marginalis (Lamarck) included depuration of pearl mussels with different physical and chemical agents to eradicate various commensals, removal of pallial mantle ribbon, aseptic preparation of explants from the ribbon and transfer of those explants into tissue culture petri dishes. Special synthetic tissue culture media enriched with additives viz., inactivated calf fetal serum and antibiotics were poured into plates with explants. The culture plates were incubated at 30 degrees C in a CO2 incubator at 5%, CO2. The cultures could be maintained for 42-45 days without any contamination. After 12 hr epithelial like cells began to migrate out and formed a complete cell sheet surrounding the explant within 12-15 days. The epithelial cells in the culture indicated functional viability as subsequently after 38-40 days of culture, typical aragonitic 'nacre' crystals of CaCO3 could be observed throughout the culture plates.     

Development in vitro of epithelial-cell monolayers derived from fetal rat pancreas.

Purified epithelial-cell monolayers were generated in vitro from explants of fetal rat pancreas. The extent of the development of the epithelial monolayer, as determined by planimetric analysis, was enhanced by the application of two methodological procedures: (a) preincubation of fetal pancreas in situ at 27 degrees C for 5 hr prior to dissection and explantation; and (b) incubation of the explants in medium containing a high concentration (50% to 70%) of fetal bovine serum. By utilizing such culture conditions, sheets of contiguous epithelial cells, with little or no peripheral fibroblastic contamination, were maintained for 9 days. Whereas the majority of cells within the monolayer had morphological characteristics of pancreatic ductal cells, endocrine cells were identified by the specific immunocytochemical localization of insulin and glucagon. In addition, insulin could be detected in the incubation medium throughout the course of experiment. The simplicity of this preparation offers some advantages over other techniques including reduced chance of contamination and reduced cellular damage or death. It provides a model for future studies directed toward developing individual cell strains derived from pancreatic epithelial cells.