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葡萄基因工程研究进展 总被引:6,自引:0,他引:6
植物基因工程技术为培育优良葡萄品种开辟了一条全新而有效的途径。葡萄基因转化受体系统的建立主要包括器官发生途径和胚状体发生途径,建立良好的受体系统是葡萄基因转化成功的关键,遗传转化途径主要有根癌农杆菌介导的遗传转化和基因枪法。概述了迄今国内外葡萄基因工程的研究进展,着重对葡萄基因转化受体系统的建立、转化的方法、转化植株的筛选和检测、影响葡萄基因转化的主要因素等进行了综述,并展望了葡萄基因工程的发展前景。 相似文献
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苏云金杆菌内毒素蛋白基因转入葡萄胚性愈伤组织细胞及转基因… 总被引:1,自引:0,他引:1
以葡萄的胚性愈伤组织作为农杆菌介导,Ti质粒转化材料,利用共培养法将苏云杆菌内毒素蛋白基因转入葡萄胚性愈伤组织细胞,通过胚状体发生途径再生转基因植株。实验发现:800μmol/L的乙酰丁香酮诱导处理农杆菌和葡萄愈伤组织后可转将化效率提高50倍。 相似文献
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本文就农杆菌介导的玉米遗传转化的技术要点及原理等进行了综述,并对各种影响农杆菌转化玉米效率的关键因子包括农杆菌的菌株与载体、标记基因、受体材料的基因型、来源和发育状态以及组织培养的条件等进行了讨论。 相似文献
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农杆菌介导的玉米遗传转化研究进展 总被引:1,自引:0,他引:1
邓向阳 《基因组学与应用生物学》2010,29(4)
农杆菌介导的转基因法是目前玉米遗传转化的主流方法之一。目前,模式玉米种质幼胚的转化体系已程式化,且开发了新筛选基因和获得不含筛选基因转基因玉米的方法,但是大多数育种骨干自交系转化频率低和转化受体基本上是幼胚。从农杆菌、受体及培养条件多方面各种因素对问题进行分析,多数研究认为针对特定基因型和受体材料建立好的受体再生系统,结合高效率农杆菌转化体系,获得多目的基因聚合(无其它外源片段)的转基因玉米将是农杆菌介导玉米转化体系研究的最终目标。本文主要从农杆菌介导(转基因)法应用于玉米遗传转化的历史、现状、问题等方面进行综述,为同领域的研究者提供一定的参考。 相似文献
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以窄宿主葡萄农杆菌Ag162Ti质粒的T-DNA区tmr、tmsl和ocs基因座位以及T_A-DNA和T_B-DNA片段为探针,对12株我国分离的不同生物型、质粒类型和寄主范围的葡萄根癌农杆菌的引质粒转移DNA(T-DNA)进行Southern杂交分析。在9株生物3型octoplne Ti质粒菌株中,与上述探针均同源。其中窄宿主葡萄根癌农杆菌菌株杂交片段彼此较一致。广宿主葡萄根癌农杆菌菌株的杂交片段彼此差异较大。1株无致瘤能力的生物1型菌株与5个探针均不杂交。1株生物3型nopaline Ti质粒菌株及1株诱导冠瘿瘤中只合成精氨酸的菌株,杂交带的变化也大。由此可见葡萄农杆菌在生物进化过程中其转移DNA呈多态性,成为农杆菌中特殊类群。本分析对葡萄根癌农杆菌致病菌株的鉴定亦有帮助。 相似文献
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高等植物在发根农杆菌介导下的遗传转化 总被引:5,自引:0,他引:5
本文介绍了发根农杆菌的生物学特性、遗传转化的操作技术、RiT-DNA转化体的形态特征以及发根农杆菌转化的利用。与根癌农杆菌不同,农杆碱型发根农杆菌的RiT-DNA含有生长素合成基因、农杆碱、甘露碱合成基因,不含有细胞分裂素合成基因,它的转化体首先是转化根。发根农杆菌介导的植物遗传转化,在次生代谢产物生产,植物抗逆性育种以及细菌与植物进化关系的研究等方面具有广泛的应用前景。 相似文献
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苏云金杆菌内毒素蛋白基因转入葡萄胚性愈伤组织细胞及转基因植株再生的研究 总被引:5,自引:0,他引:5
以葡萄的胚性愈伤组织作为农杆菌介导,Ti质粒转化材料,利用共培养法将苏云金杆菌内毒素蛋白基因转入葡萄胚性愈伤组织细胞,通过胚状体发生途径再生转基因植株。实验发现:80μmol/L的乙酰丁香酮诱导处理农杆菌和葡萄愈伤组织后可将转化效率提高50倍。OD值为0.8的农杆菌菌液稀释8—10信后与在G培养基预培养10天的胚性愈伤组织共培养2—3夭,Ti质粒对葡萄愈伤组织细胞的转化效率可达50%左右。筛选得到的转基因植株在含Km 30 mg/L的选择培养基上继代存活6个月,生长正常;提取叶片染色体DNA做Southern blot,杂交结果为阳性。将转基因植株各部分切段置于含Km 50 mg/L的选择培养基上,能够脱分化产生抗性愈伤组织并能增殖。 相似文献
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农杆菌介导单子叶植物基因转化研究进展 总被引:19,自引:0,他引:19
农杆菌介导基因转化系统是双子叶植物基因转化的普通而有效的手段, 其优点倍受重视,近年来又广泛用于曾被认为不在农杆菌宿主范围之内的单子叶植物的基因转化研究,并在很多重要粮食作物上获得成功,例如水稻、玉米、大麦、小麦等。本文就农杆菌转化的优点,转化机理以及对单子叶植物转化的研究进展作一概述 相似文献
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外源DNA导入在蔬菜分子育种中的研究进展 总被引:1,自引:0,他引:1
随着转基因方法的多样化和技术体系的不断完善,转基因技术在蔬菜育种上的应用取得了显著成果,成为蔬菜种质创新和品种改良的有效途径。介绍了几种不依赖组织培养的外源DNA导入技术,包括花粉管通道法、农杆菌浸渍法、激光微束法、PEG法、电击法和脂质体法,从育种角度比较了这些方法的优缺点,并对其在蔬菜育种中的应用现状、存在问题及应用前景进行了较为全面的综述。 相似文献
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Detection of Horizontal Gene Transfer by Natural Transformation in Native and Introduced Species of Bacteria in Marine and Synthetic Sediments 总被引:15,自引:10,他引:5
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Both naturally occurring marine sediments and artificial sediments were used as supports for natural transformation of marine bacteria. While transformation of Pseudomonas stutzeri ZoBell suspended in artificial seawater was not detected when recipient cells and rifampin resistance DNA were loaded onto sterile sediment columns, transformation could be detected at frequencies 4 to 20 times that of spontaneous resistance when recipient cells and rifampin resistance DNA were loaded onto sterile sediment columns. Treatment of these columns with DNase I reduced transformation frequencies to levels comparable to those of spontaneous-resistance frequencies. Sediments with higher organic contents supported higher frequencies of transformation than did those with lower amounts of organic matter. Transformation was also detected when recipient cells and DNA were loaded on columns prepared from nonsterile sediments, although the frequencies of transformation were lower than when sterile sediments were used. Finally, nonsterilized sediments that were not supplemented with laboratory strains did not support detectable levels of transformation in sediment columns, but when these same sediments were transferred to filters and placed on complex media, transformation was detected at a frequency three times that for spontaneous resistance. This transformation frequency was partially reduced to levels near that for spontaneous resistance by the addition of DNase I to sediment filters. These results indicate that marine sediments facilitate the uptake and expression of exogenous DNA by transformable marine bacteria and that sediments are a more likely niche for natural transformation than the water column in the marine environment. 相似文献
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Effects of toxicity, aeration, and reductant supply on trichloroethylene transformation by a mixed methanotrophic culture 总被引:9,自引:0,他引:9
The trichloroethylene (TCE) transformation rate and capacity of a mixed methanotrophic culture at room temperature were measured to determine the effects of time without methane (resting), use of an alternative energy source (formate), aeration, and toxicity of TCE and its transformation products. The initial specific TCE transformation rate of resting cells was 0.6 mg of TCE per mg of cells per day, and they had a finite TCE transformation capacity of 0.036 mg of TCE per mg of cells. Formate addition resulted in increased initial specific TCE transformation rates (2.1 mg/mg of cells per day) and elevated transformation capacity (0.073 mg of TCE per mg of cells). Significant declines in methane conversion rates following exposure to TCE were observed for both resting and formate-fed cells, suggesting toxic effects caused by TCE or its transformation products. TCE transformation and methane consumption rates of resting cells decreased with time much more rapidly when cells were shaken and aerated than when they remained dormant, suggesting that the transformation ability of methanotrophs is best preserved by storage under anoxic conditions. 相似文献
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J. C. Sanford K. A. Skubik B. I. Reisch 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1985,69(5-6):571-574
Summary Experiments were conducted to test the validity of previous reports of pollen-mediated plant transformation utilizing genomic donor DNA. Multiple Mendelian markers were employed in Zea mays L. and Lycopersicon esculentum Mill, to detect transformation events. Pollen from multiple recessive (recipient) lines was incubated with genomic DNA from multiple dominant (donor) lines, under various conditions. Treated pollen was subsequently used for pollinations on multiple recessive females, and resulting seeds were screened for transformation events. Over 200 crosses were made in tomato, and over 80 crosses were made in corn. Over 600 resulting seedlings were tested in tomato and over 800 seeds were screened in corn. Because multiple markers were used, 4,937 potential transformation events were screened. No clear-cut transformation events were observed. Therefore, using well-defined multiple markers, we have been unable to confirm the earlier claims of high efficiency pollen-mediated transformation employing genomic donor DNA. 相似文献
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Effects of toxicity, aeration, and reductant supply on trichloroethylene transformation by a mixed methanotrophic culture. 总被引:14,自引:12,他引:2
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The trichloroethylene (TCE) transformation rate and capacity of a mixed methanotrophic culture at room temperature were measured to determine the effects of time without methane (resting), use of an alternative energy source (formate), aeration, and toxicity of TCE and its transformation products. The initial specific TCE transformation rate of resting cells was 0.6 mg of TCE per mg of cells per day, and they had a finite TCE transformation capacity of 0.036 mg of TCE per mg of cells. Formate addition resulted in increased initial specific TCE transformation rates (2.1 mg/mg of cells per day) and elevated transformation capacity (0.073 mg of TCE per mg of cells). Significant declines in methane conversion rates following exposure to TCE were observed for both resting and formate-fed cells, suggesting toxic effects caused by TCE or its transformation products. TCE transformation and methane consumption rates of resting cells decreased with time much more rapidly when cells were shaken and aerated than when they remained dormant, suggesting that the transformation ability of methanotrophs is best preserved by storage under anoxic conditions. 相似文献
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Zang X Liu B Liu S Arunakumara KK Zhang X 《Journal of microbiology (Seoul, Korea)》2007,45(3):241-245
This study was conducted to determine the optimal conditions for introduction of exogenous DNA into Synechocystis sp. PCC 6803. Of the three transformation techniques studied, electroporation, ultrasonic transformation and natural transformation, natural transformation showed the highest efficiency. Additionally, this study demonstrated that the higher plasmid concentration and longer homologous recombining fragments resulted in a greater number of transformants. For successful transformation, the lowest concentration of plasmid was 0.02 microg/ml, and the shortest homologous recombining fragment was 0.2 kb. Use of Synechocystis sp. PCC 6803 in the logarithmic growth phase resulted in two-fold higher transformation rate than that of the same organism when cells in the latent phase or the plateau phase were used for transformation. Pretreatment of the host strain, Synechocystis sp. PCC 6803, with EDTA (2 mM) for two days prior to transformation increased the transformation efficiency by 23%. Additionally, incubation of the cells and DNA for 5 h under light conditions increased the transformation efficiency by two orders of magnitude. Moreover, recovery treatment of the cells before they were plated onto antibiotic medium also increased the transformation efficiency. 相似文献
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