共查询到19条相似文献,搜索用时 93 毫秒
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深黄被孢霉3.3410原生质体的制备和再生研究 总被引:2,自引:0,他引:2
目的:为了获得数目较多且稳定性较好的深黄被孢霉原生质体,对影响深黄被孢霉原生质体制备和再生的因素进行了研究.方法:以培养20h的幼嫩菌丝体为材料,在1.5%纤维素酶+0.5%的蜗牛酶+1%溶菌酶混合酶作用下,以0.8mol/L MgSO4·7H2O作渗透压稳定剂于30℃酶解3h,然后用血球计数板计数,计算原生质体产量.结果:在上述条件下,原生质体浓度可达到5.5 × 107个/ml.并对该原生质体在高渗培养基上进行再生实验,其再生率达到25.40%.为深黄被孢霉原生质体诱变及融合育种奠定了基础. 相似文献
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本研究以卵孢小奥德蘑液体培养菌丝作为实验材料,利用单因子变量法探索研究了菌丝培养时间、酶浓度、酶解时间、酶解温度、稳渗剂类型对卵孢小奥德蘑原生质体制备的影响,并对原生质体再生培养基进行选择和优化。通过荧光染色,利用激光共聚焦显微镜和流式细胞仪对原生质体的制备过程、得率和活力进行研究。结果表明,将卵孢小奥德蘑菌丝在液体培养基中培养5d收集菌丝体,以甘露醇作为渗透压稳定剂,在溶壁酶浓度2%、30℃条件下酶解5h,获得的原生质体得率最高,达2.0×10 7个/mL;通过流式细胞仪分析,约57.69%的原生质体细胞为活细胞;在RM培养基中再生效果最好,再生率为(0.103±0.025)%。研究结果可以为卵孢小奥德蘑育种与食用菌原生质体制备再生提供研究基础。 相似文献
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γ——亚麻酸生产菌深黄被孢霉原生质体的形成和再生 总被引:8,自引:2,他引:6
采用2%的溶壁酶加4%的蜗牛酶,从深黄被孢霉的菌丝中获得了大量的原生质体,同时对菌丝的培养时间,渗透压稳定性,酶解系统和酶解温度等因素了进行了系统的观察,从而获得了制备深黄被孢霉原生质体的最适条件,并对该原生质体在高渗培养基上进行了再生实验,其再生率为32%。 相似文献
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桃褐腐病菌(Monilia fructigena)原生质体制备及再生条件 总被引:3,自引:0,他引:3
以桃褐腐病菌(Monilia fructigena)为供试菌株,研究了酶系组成、液体培养基、菌龄、酶解温度、酶解时间对原生质体制备的影响,以及等渗液、固体再生培养基、酶解时间对原生质体再生的影响。结果表明:Fries(1/2)液体培养基培养24h,在10mg/mL崩溃酶+5mg/mL纤维素酶+20mg/mL蜗牛酶+10mg/mL溶菌酶的混合酶液中28°C酶解4h为桃褐腐病菌原生质体制备的最佳条件。采用液体再生涂布平板法,以含Ca2+的STC为等渗液的液体培养基和含蔗糖及Ca2+的Fries(1/2)固体培养基为桃褐腐病菌原生质体再生的最佳条件。经过观察与测定,再生菌株保持了原有的培养性状和致病性,接种桃果实后发病率为100%。 相似文献
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紫杉醇产生菌Nodulisporium sylviforme原生质体诱变研究 总被引:5,自引:0,他引:5
对酶系组成、pH、酶解温度和酶解时间等影响树状多节孢原生质体制备和再生的因素和原生质体诱变进行了研究。结果表明 ,原生质体制备和再生的最佳条件为 :用pH5.5~ 6.0的0.7mol/LNaCl配制由 3%溶壁酶 + 3%蜗牛酶 + 1 %的溶菌酶 + 3%纤维素酶组成的复合酶系 ( 1ml酶液/2 5 0mg湿菌体 ) ,在 30℃恒温水浴条件下酶解 6h ;然后 ,将获得的原生质体过滤洗涤后 ,在含0.7mol/ LNaCl的PDA再生培养基上 ,采用双层平板培养法再生制备到的原生质体。树状多节孢紫杉醇产生菌原生质体诱变的最佳条件为 :30w紫外灯、距离 30cm、照射 5 0s;UV + 0.6%LiCl复合诱变、照射时间 40s,诱变菌株经初筛和复筛 ,选出了两株高产紫杉醇的原生质体诱变菌株———UV40-19和UL50-6,其产量从出发菌株紫杉醇的产量 ( 314.07μg /L)分别提高至 376.38μg/L和392.63μg/L。 相似文献
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采用2%的溶壁酶加4%的蜗牛酶,从深黄被孢霉的菌丝中获得了大量的原生质体。同时对菌丝的培养时间、渗透压稳定剂、酶解系统和酶解温度等因素进行了系统的观察,从而获得了制备深黄被孢霉原生质体的最适条件。并对该原生质体在高渗培养基上进行了再生实验,其再生率为32%。 相似文献
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采用2%的溶壁酶加4%的蜗牛酶,从深黄被孢霉的菌丝中获得了大量的原生质体。同时对菌丝的培养时间、渗透压稳定剂、酶解系统和酶解温度等因素进行了系统的观察,从而获得了制备深黄被孢霉原生质体的最适条件。并对该原生质体在高渗培养基上进行了再生实验,其再生率为32%。 相似文献
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大球盖菇原生质体再生及单核化特性的研究* 总被引:1,自引:0,他引:1
大球盖菇原生质体再生条件及单核化特性结果。原生质体再生速度极快,涂布平板3d后肉眼即可见明显的再生菌落形成,在PGPM再生培养基上再生率为0.97~2.0%,渗稳剂种类对再生率无明显影响,但可影响再生菌落形态,液体预培养1~2d,再生率明显下降;大球盖菇原生质体单核化率高达77.6%,且再生双核体和再生单核体在形成再生菌落时无时间差,其生长速度亦无快慢之分,液体预培养可显著减少单核化率,再生单核体中存在亲本两种交配型,但二者的比率不为1。 相似文献
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E. S. Dias E. F. Araújo W. V. Guimarães R. M. C. Muchovej 《World journal of microbiology & biotechnology》1996,12(6):625-628
Experiments were performed with the mycorrhizal fungus Suillus granulatus to define the parameters for production and regeneration of protoplasts. Protoplasts were released at frequencies between 1 and 3×107/ml from mycelium 3 to 7 days old. The best osmotic stabilizer for protoplast release was MgSO4 (0.7 m). To optimize protoplast release and regeneration an enzyme (Novozym 234) concentration 1.7 mg/ml was chosen, with a digestion time of 1 to 2 h. Regenerated colonies formed mycorrhizae within 60 days after inoculation in Pinus caribaea var. hondurensis seedlings. 相似文献
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Zheng Ma Jinxiu Liu Xiaozhen Lin Xuping Shentu Yalin Bian Xiaoping Yu 《Folia microbiologica》2014,59(2):93-97
Toyocamycin exhibits effective biological activities for use against plant pathogenic fungi thanks to its structural similarity to nucleoside. It has been recognized as a promising agricultural antibiotic utilized in controlling the occurrence of plant diseases. In our previous study, a strain that was isolated was identified and designated as Streptomyces diastatochromogenes whose major secondary metabolite was toyocamycin, but the production was largely limited. Protoplast transformation is a useful technique in the improvement of streptomycete. In this study, we optimized some key factors necessary for protoplast formation, regeneration, and transformation of S. diastatochromogenes. When mycelium was cultivated in CP medium with 1 % glycine, harvested at 48 h old, and then treated with 3 mg lysozyme/mL in P buffer for 1 h, the greatest regeneration frequency (42.5 %) of protoplasts was obtained. By using 1?×?109/mL protoplasts with polyethylene glycol 1000 at a concentration of 30 % (w/v), the best performance of protoplast transformation efficiency was 4.8?×?103/μg DNA transformants. 相似文献
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A rapid and convenient method for producing protoplasts from 3 d old mycelium of the ascomycete Hypomyces ochraceus is described. The procedure involves a Helix pomatia enzyme preparation and sucrose (20%) for stabilization. Pretreatment with disulfide bond reducing agents reduced the amount of viable protoplasts. Formation of protoplasts and different stages of regeneration were observed by phase contrast microscopy. There was only one type of true regeneration from protoplasts to hyphae in 15-30% gelatine medium by direct forming a germ tube from the original protoplast. Cytological events and physiological conditions are discussed. 相似文献
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Cytologic control of protoplast formation and regeneration in Streptomyces erythraeus, strain BTCC-2
D Liutskaiuva O V Kamzolkina L I Starodubtseva T Todorov V N Danilenko 《Antibiotiki i khimioterapii͡a》1991,36(3):8-11
Experiments on protoplast formation and regeneration in S. erythraeus, strain BTCC-2 (Saccharopolyspora erythrae) were performed under microscopic control at all the stages. It was shown that the highest protoplast titer was provided by the mycelium grown in one step in the absence of glycine. For characterizing the protoplasts formed by the mycelium grown under different conditions, their regeneration capacity was estimated by microscopic examination of the protoplasts after 15-20-hour growth in microchambers and evaluation of the regeneration efficiency 7-10 hours later. Of interest was the fact of spontaneous development of colonies consisting of the protoplast-like cells (L-cells) in 15-20 hours. Such colonies were formed only by the protoplasts grown from the mycelium incubated in one step in the absence of glycine or in the presence of 0.1 per cent of glycine. Such conditions provided also the maximum efficiency of the protoplast regeneration. The long-term storage of protoplasts led to a decrease in their viability. 相似文献
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Protoplast isolation and regeneration in Streptomyces clavuligerus 总被引:10,自引:0,他引:10
The regeneration of streptomycete protoplasts is a major step following genetic manipulations such as fusion and DNA-mediated transformation. Reports of studies on the regeneration of protoplasts from Streptomyces clavuligerus are limited and for this reason the experiments described in this paper were carried out. An investigation of protoplast formation and cytology was made to gain further insight into the loss of protoplast viability in osmotically stabilized support media. Protoplasts with the highest regeneration frequency were isolated from mycelium, grown in a two-stage culture system (without glycine), using lysozyme dissolved in a sucrose osmoticum containing 1% bovine serum albumin. The latter promoted improved protoplast viability. A systematic survey was made of the components of regeneration medium R5, previously used for S. clavuligerus, and other potentially advantageous components and conditions, in an attempt to raise the regeneration frequency of the protoplasts. An improved regeneration medium (R6) and protocol which supported higher and more consistent levels of regeneration of S. clavuligerus protoplasts resulted from these experiments. These improved procedures for protoplast isolation and regeneration proved to be suitable for other streptomycete species. 相似文献