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1.
影响根癌农杆菌介导的香蕉基因转化早期的主要因素   总被引:18,自引:1,他引:18  
用含质粒载体pCAMBIA2301的根癌农杆菌(Agrobacterium inoculation)转化“641”香蕉(Musa AAA Cavendish subgroup cv.64-1)的薄片外植体,通过测定GUS基因瞬时表达率,对转化早期的主要影响因素进行了研究。结果表明:农杆菌EHA105较适合于介导转化,将薄片置于固体高渗培养基上培养4h后,通过真空减压法使农杆菌接种于其上,获得了较高的瞬时表达率(41.67%),是对照(8.33%)的5倍,农杆菌悬液稀释5倍用于转化较好,共培养3d为宜,薄片外植体越靠近根部,瞬时表达率越高。而外植体预培养会降低瞬时表达率。  相似文献   

2.
大白菜遗传转化体系的建立   总被引:1,自引:0,他引:1  
系统研究了根癌农杆菌LBA4404介导的大白菜遗传转化的若干因素,成功地建立了大白菜快速有效的遗传转化体系,并获得了大白菜抗性植株。实验结果表明,大白菜的不同基因型、苗龄、外植体类型,菌液浓度、浸染时间,共培养和抗生素浓度对转化频率均有一定影响,其最高转化频率达4.0%。抗性植株经PCR和Southern blot检测表明,目的基因已整合进大白菜基因组中。  相似文献   

3.
农杆菌介导抗虫基因转化芥菜型油菜的研究   总被引:7,自引:0,他引:7  
用农杆菌介导法对芥菜型油菜进行了转化,并对影响根癌农杆菌转化效率的因素进行了研究,建立了以油菜子叶柄为外植体的转化体系.用携带蝎毒素(BmKITs)和几丁质酶(chi)双价基因的农杆菌转化油菜,共获得了抗卡那霉素的再生苗153株,移栽成活21株.对成活植株进行了PCR分子检测,证实有外源基因的整合.转基因植株形态正常,开花、结实.  相似文献   

4.
用LBA4404/pCDH,Agl 1/pUNN2和Ag;P(无质粒)3种菌株分别转化小麦品种济南177、99P、核生3号幼胚诱导的愈伤组织以及济南177的幼胚.其中以胚性愈伤组织为外植体,获得了转基因植株.PCR和PCR-Southem分析证实转化植株中包含了外源基因.通过染色体分析,发现胚性愈伤组织在农杆菌LBA4404侵染后形成的染色体削减比经Agl1侵染和未经感染的对照形成的染色体削减程度更大,甚至发现较多的染色体断片.分析了农杆菌菌株,材料的基因型,外植体类型,培养时间和温度以及筛选的时间和周期对于转化效率的影响.  相似文献   

5.
农杆菌介导东方百合“西伯利亚”遗传转化体系建立   总被引:1,自引:0,他引:1  
以东方百合"西伯利亚"(Lilium seberia)为材料,建立了百合再生和农杆菌介导的遗传转化体系.采用LBA4404和GV3101菌株对鳞片来源的愈伤组织进行侵染,并以鳞片作为对比转化材料,两种菌株携带有相同的双元载体pCAMBIA1301.研究结果表明,农杆菌侵染愈伤组织外植体转化率较高;乙酰丁香酮(AS)的添加能大大提高根癌农杆菌转化百合的效率.另外,共培养时间、不同菌株均对转化效率有一定的影响.筛选后获得的抗性植株经PCR检测和GUS组织化学染色证实外源基因已经整合到基因组DNA中并获得表达,转化率为5%~8%.  相似文献   

6.
以向日葵无菌苗的子叶节为外植体,通过农杆菌介导法,对其遗传转化条件进行了研究,建立了向日葵子叶节农杆菌介导的遗传转化体系.结果表明:在农杆菌浸染前(浸染浓度为OD600=0.6)进行2 d的预培养、浸染8 min的外植体在MS+1.2 mg/L 6-BA+0.03 mg/L IAA的培养基上转化效率较高.PCR检测初步证明,LycB基因已整合到向日葵再生植株中.  相似文献   

7.
超声波辅助的发根农杆菌对黄瓜的遗传转化   总被引:14,自引:0,他引:14  
由于超声波处理可在外植体上产生许多微小伤口,从而提高农杆菌与外植体之间的接触面积,提高农杆菌对外植体的转化频率。用野生型发根农杆菌(Agrobacterium rhizogenes)A4菌株感染黄瓜(Cucumis sativus L.) 无菌苗的子叶和下胚轴,建立了黄瓜的发根遗传转化体系。同时,对超声波处理时间、外植体放置方向、预培养时间、外植体类型、乙酰丁香酮的添加与否等影响因素进行了研究。结果表明,采用外植体在添加0.5mg/L NAA和2mg/L 6-BA的MS培养基上预培养4d,超声波处理2s,共培养3d,在预培养共培养期间子叶上表皮向下可以取得最佳的转化效果,发根诱导率为94.7%。乙酰丁香酮对发根诱导频率影响不明显。冠瘿碱检测表明,发根确定能合成农杆碱。  相似文献   

8.
本文回顾了根瘤农杆菌的玉米遗传转化的发展历史 .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 ,受体材料的基因型和发育程度 ,培养环境 ,培养基成分等因素进行了综述 .对发展现状和未来趋势做了讨论 .  相似文献   

9.
通过非洲菊(Gerbera hybrida)组织培养建立起适合于农杆菌介导的基因转化受体系统,非洲菊叶柄外植体在MS 3mg/L BA 0.01mg/L NAA培养基中直接诱导芽生长,在MS 0.1mg/L NAA培养基中生根培养,获得再生植株,诱导率达57%。带有云南矮牵牛(Petunia hybrida)二氢黄酮醇3’5’羟化酶cDNA的pEH表达载体与农杆菌(Agrobacterium tumefaciens)LBA4404融合后,与非洲菊叶柄外植体共培养3d,通过含有卡那霉素25mg/L和羧苄菁霉素500mg/L的选择培养,诱导出转基因非洲菊。经PCR和Southern分子杂交检测,证明目的基因已整合入非洲菊的基因组中,转基因非洲菊花色、花型及叶型都有显变化。  相似文献   

10.
利用根癌农杆菌LBA4404介导,建立了丝状真菌简青霉(Penicillium simpli-cissimum)H5的遗传转化系统.潮霉素抗性筛选、PCR和Southern blot分子鉴定等结果表明:筛选获得的转化子能够稳定遗传,外源的T-DNA以单拷贝随机整合到简青霉的基因组中.实验初步研究了农杆菌浓度、乙酰丁香酮浓度和共培养时间等因素对转化效率的影响,经过优化后转化体系的效率可达50个转化子/105个孢子.农杆菌介导的遗传转化方法在简青霉上的运用,将为研究该菌的基因工程改造提供强有力的工具.  相似文献   

11.
Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.  相似文献   

12.
理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。  相似文献   

13.
As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.  相似文献   

14.
正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-  相似文献   

15.
We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-  相似文献   

16.
正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that  相似文献   

17.
For molecular systems,the quantum-mechanical treatment of their responses to static electromagnetic fields usually employs a scalar-potential treatment of the electric field and a vector-potential treatment of the magnetic field.Although the potential for each field separately is associated with the choice of an(unphysical)origin,the precise choice of the origin for the electrostatic field has little consequences for the results.This is different for the  相似文献   

18.
Franck-Condon factors bridge the gap between theoretical modeling and experimental observations for molecular electronic spectroscopy and electron transfer.Under the displaced harmonic oscillator approximation,multidimensional Franck-Condon factors are decomposed into a product of many one-dimensional(1D)Franck-Condon(FC)factors,and each 1D-FC factor is associated with one Huang-Rhys factor that determines the leading contribution of  相似文献   

19.
<正>"The Journal of Shanghai Normal University:Mathematics"is published by Shanghai Normal University as regular issues of The Journal of Shanghai Normal University each year from 2014 in English.The editors-in-chief of the issues are professors Yuhao Cong and Maoan Han.The Journal of Shanghai Normal University was started in 1958 with  相似文献   

20.
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