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目的:诱变筛选阿魏菇多糖高产菌,探索食用菌的离子束和激光复合诱变育种方法。方法:尝试用阿魏菇菌丝单细胞为靶材,以离子束注入和激光辐照为复合诱变手段,采用营养缺陷型筛选办法定性初筛,摇瓶发酵定量复筛。结果:通过离子束注入诱变,获得了2株菌丝体多糖产量分别达到551.80mg/L和659.46mg/L、较1号出发菌株提高了46.5%和75.2%的多糖高产菌PFPH-1和PFPH-2;在此基础上,以激光辐照为复合诱变手段,获得了1株菌丝体多糖产量达到762.50 mg/L、较出发菌株PFPH-2提高了15.63%的多糖高产菌PFPH-3。结论:离子束和激光复合诱变育种方法在改良阿魏菇多糖高产性状方面诱变功效显著。 相似文献
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离子束注入阿魏菇生物效应及诱变育种方法 总被引:1,自引:0,他引:1
目的:考察离子束注入阿魏菇的生物效应,探索大型真菌阿魏菇的离子束诱变育种简易方法。方法:分别采用能量5keV~30keV、剂量1.5×10^15cm~~1.5×10^16cm。的N+束注入青河野生阿魏菇ACK的分生孢子和菌丝单细胞,镜检,洗脱初筛,转接复筛。结果:离子束注入对阿魏菇分生孢子及菌丝单细胞刻蚀作用明显,细胞存活率能量、剂量效应显著;获得了一种梯次注入、营养缺陷性或目标性状高通量定性初筛和目标性状定量复筛的离子束诱变育种方法。结论:该方法可操作性强,是一种简便易行适用于大型真菌的诱变育种方法。 相似文献
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工业微生物物理诱变育种技术的新进展 总被引:2,自引:0,他引:2
物理诱变技术是当今工业微生物育种中最重要、最有效的技术之一。传统的物理诱变技术主要有紫外线、X射线、γ射线诱变等,它们已在包括青霉素、"-淀粉酶等几乎所有的工业微生物菌种的诱变选育中发挥了巨大的作用。多数菌株在多次重复使用传统诱变源时往往出现抗性饱和的现象。太空环境、离子束、激光等是20世纪70~80年代逐渐兴起的新型诱变技术,因它们具有诱变谱广和在一定程度上能克服菌株对传统诱变源的抗性饱和等优点,而广受工业微生物育种工作者的欢迎。现就空间、离子束、激光等诱变育种技术的作用特点、诱变机理、应用及前景进行阐述。 相似文献
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【目的】建立里氏木霉(Trichoderma reesei)高产突变菌株的快速筛选方法,选育出高产内切葡聚糖酶的突变株。【方法】对里氏木霉T306菌株的初筛培养基进行优化,建立快速筛选方法;通过紫外诱变手段选育内切葡聚糖酶高产突变菌株,并对突变菌株的产酶培养基进行优化。【结果】在初筛培养基中添加浓度为0.1%(W/V)的乳糖、蛋白胨及脱氧胆酸钠有利于菌株的筛选。诱变后筛选出菌落形态发生明显变化的内切葡聚糖酶高产突变株0516,其羧甲基纤维素酶活力(CMC酶)较出发菌株提高了38.9%。其产酶培养基经优化后,得到最适碳、氮源分别为:乳糖1.50%、硫酸铵0.14%、尿素0.05%、蛋白胨0.10%,优化后CMC酶活力达64.2 U/mL,较优化前提高了2.3倍。【结论】建立了里氏木霉高产突变菌株的快速筛选方法,通过紫外诱变育种获得了产内切葡聚糖酶能力高且遗传稳定的突变株0516。 相似文献
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以一株由青藏高原牦牛粪中分离出的链霉菌为出发菌株,对其培养特性、产酶条件和酶学性质进行初步研究.通过重离子诱变,筛选出遗传稳定的高产菌株.结果表明,该菌以玉米芯和麸皮(1∶1)为碳源能高效地诱导木聚糖酶的胞外分泌,其最适培养基和培养条件氮源为酵母膏、初始pH7、培养温度为25℃,在此条件下,第4天酶活力达到峰值3480.25 U/mL.说明该酶能够利用农业废弃物高效生产木聚糖酶.该菌株所产木聚糖酶的最适反应温度为15℃、pH4,属低温酸性木聚糖酶.经重离子诱变后,筛选出一株高产菌株SZ10-7,其酶活力可达5 338.42 U/mL. 相似文献
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梅岭霉素产生菌抗药性突变标志诱变筛选模型的初步研究 总被引:3,自引:0,他引:3
本文通过梅岭霉素 (Meilingmycin)产生菌南昌链霉菌NS 4 1 80菌株孢子对 6种抗生素敏感性测定 ,采用诱变剂EMS四种不同诱变剂量对菌株孢子进行诱变处理 ,诱变处理的孢子涂布在含致死浓度链霉素的高氏平板上。然后从抗药性突变标志菌株中进一步筛选梅岭霉素高产菌株。在 150 0多个抗药性突变株中通过初筛获得了比诱变出发菌株的产素能力提高 50 %以上菌株。通过诱变剂量分别与抗药性突变率和突变株产素产量的变势统计分析表明 ,菌株抗药性突变与产素突变密切相关 ,产素突变的EMS诱变剂量高于抗药性突变诱变剂量 ,在 0 .0 3mol/LEMS剂量作用下 ,菌株致死率为 99.4 3% ,抗药性突变率为 0 .0 4 4 0 % ,建立了梅岭霉素产生菌抗药性突变标志诱变推理性筛选模型。为南昌链霉菌高产菌种选育研究作了有益的尝试 ,并有助于其它链霉菌属的抗生素产生菌育种研究。 相似文献
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Song EJ Jeong HJ Lee SM Kim CM Song ES Park YK Bai GH Lee EY Chang CL 《Journal of microbiological methods》2007,68(2):430-433
The clinical efficacy of a spoligotyping method for discriminating Mycobacterium tuberculosis strains was evaluated. Among the strains other than Beijing or Beijing like family, 30 different spoligotypes out of 39 strains were produced, which included 4 strains not having IS6110 sequence. The oligonucleotide chip-based spoligotyping technique is useful for early screening and detection of clonal proximity of M. tuberculosis isolates. 相似文献
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AIMS: The aim was to simplify the cumbersome conventional process of isolating virulent bacilli, which involves isolating all bacilli strains from a source followed by screening for strains that are effective for bio-control of mosquito vectors. METHODS: A new simplified technique involving eight steps was devised for screening soil samples for the presence of mosquito-pathogenic bacilli before isolating individual strains. RESULTS: Using the new technique, we obtained eight bacilli strains (KSD1-8) showing pathogenic activity against mosquito larvae from three out of 10 soil samples screened. These strains were characterized, identified and the main bioassay tests were performed with three most promising strains (KSD-4, KSD-7 and KSD-8), and their pathogenic activity against Anopheles stephensi Liston, Culex quinquefasciatus, Say and Aedes aegypti Linnaeus compared well with commercial reference strains of B. thuringiensis israelensis and B. sphaericus. SIGNIFICANCE AND IMPACT OF THE STUDY: The new technique of screening soil samples for the presence of virulent pathogenic strains of bacilli against mosquito larvae proved quick, efficient and cost effective. 相似文献
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Zhou X Wang Z Jiang K Wei Y Lin J Sun X Tang K 《Prikladnaia biokhimiia i mikrobiologiia》2007,43(4):490-494
A total of 38 endophytic fungus strains were isolated from Taxus chinensis var. mairei by aseptic technique. Genomic DNA was extracted from isolated endophytic fungi and subjected to polymerase chain reaction (PCR) analysis for the presence of Taxus taxadiene synthase (TS) gene, a rate-limiting enzyme gene in the taxol biosynthetic pathway. Twelve out of 38 isolated endophytic fungus strains showed PCR positive for the ts gene. Subsequently, taxol and its related compounds were extracted from culture filtrates and mycelia of the PCR positive strains, separated by column chromatography and analyzed by High Performance Liquid Chromatography and Mass Spectrum. The analysis result showed that 3 strains could produce taxol and its related compounds at the detectible level. This study indicates that molecular detection of the ts gene is an efficient method for primary screening of taxol or its related compounds-producing endophytic fungi which can improve prominently screening efficiency. 相似文献
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Iu V Vertiev I A Shaginian M V Stepanova Iu V Ezepchuk 《Molekuliarnaia genetika, mikrobiologiia i virusologiia》1985,(3):38-40
A solid phase variant of radioimmunoassay has been elaborated for screening toxin-producing strains of E. coli and V. cholerae grown on agar plates. The method is based on the ability of cholera-like toxins to be absorbed on nitrocellulose filters and their further identification with the use of homologous sera and [125I]-A protein from staphylococci. Sensitivity of the method reaches 20 pg. The proposed technique permits identification of intracellular enterotoxin and is aimed at a massive screening of E. coli strains, NAG-vibrios and V. cholerae strains for toxin production. 相似文献
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研究了产酯酶微生物的筛选,包括筛选模型、酶活力检测方法及菌株的分布。对30多份土样以及实验室保存的菌种进行了大量的筛选,以添加三醋酸甘油酯、乳酸乙酯酯类物质对土样等样品富集,采用添加显色剂溴甲酚紫的快速简便平板显色法,观察水解变色圈直径和菌落直径的大小进行初筛。获得两者直径之比相对大的菌株174株,采用平板打孔检测法和摇瓶发酵比色法测酶活力相结合进行复筛,最终得到酯酶活力较高的24株菌株。就初筛和复筛方法及结果加以比较分析,复筛菌株做不同底物的酶活力检测,建立了一个有效、简便及快速的微生物酯酶的筛选模型。并对酯酶产生菌的立体选择专一性进行了初步考察。 相似文献
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Yuanming Yin Chenjin Jiang Zhengyu Wang Genfu Wu Peihua Chen 《Biotechnology Techniques》1993,7(1):51-56
Summary With the technique of rotating liquid culture system in 7000 bottomless micro-holes, mutant strains having urease activity higher than that of the original strain were screened out in a medium with urea as a sole nitrogen source. A strain having a lysine yield of 12.5% and urease activity 20% higher than those of the original strain was isolated. The technique is suitable for screening strains where visual methods are available to estimate the amount of metabolite. 相似文献
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Cellular fatty acid analysis as a potential tool for predicting mosquitocidal activity of Bacillus sphaericus strains. 总被引:1,自引:0,他引:1
E Frachon S Hamon L Nicolas H de Barjac 《Applied and environmental microbiology》1991,57(11):3394-3398
Gas-liquid chromatography of fatty acid methyl esters and numerical analysis were carried out with 114 Bacillus sphaericus strains. Since only two clusters harbored mosquitocidal strains, this technique could be developed in screening programs to limit bioassays on mosquito larvae. It also allows differentiation of highly homologous strains. 相似文献
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Cellular fatty acid analysis as a potential tool for predicting mosquitocidal activity of Bacillus sphaericus strains. 总被引:2,自引:1,他引:1
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Gas-liquid chromatography of fatty acid methyl esters and numerical analysis were carried out with 114 Bacillus sphaericus strains. Since only two clusters harbored mosquitocidal strains, this technique could be developed in screening programs to limit bioassays on mosquito larvae. It also allows differentiation of highly homologous strains. 相似文献
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A rapid and simple microdilution technique on 96-well microplate based on turbidimetry was optimized and validated for screening of antimicrobial activity against erythromycin-resistant bacterial strains of Streptococcus pyogenes and Staphylococcus simulans isolated from Finnish patients. Using S. pyogenes ATCC 12351 as reference strain the developed method was evaluated by reproducibility measurements and using parameters typically employed for screening methods, i.e. signal-to-background, signal-to-noise and a screening-window coefficient, the Z' factor. The method was further used for screening a group of natural compounds and their synthetic derivatives against resistant bacterial strains. Of these, octyl and dodecyl gallates, and usnic and ursolic acids were the most active. The described method is a rapid, homogeneous, cost-effective and easy-to-perform system for screening of new potential antimicrobial agents in drug discovery. 相似文献