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1.
新疆杨高效遗传转化系统的建立   总被引:9,自引:0,他引:9  
选择新疆杨(Populus alba L.var.pyramidalis Bge.)为遗传转化受体材料,为建立根癌农杆菌介导新疆杨高效遗传转化系统,从预培养时间、侵染时间、共培养时间、添加乙酰丁香酮(AS)的时机、共培养培养基中添加乙酰丁香酮浓度、侵染菌液的制备方法、外植体继代方式等7个方面优化筛选。结果显示较合适的转化系统为:预培养8h,农杆菌菌液(OD600=0.4)侵染15min,共培养5d,侵染菌液的最优制备方法是液体培养活化农杆菌2次加离心收集菌体重悬,共培养培养基中添加乙酰丁香酮80μmol/L。新疆杨叶盘转化频率可达38.10%。  相似文献   

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不同理化因子对黄芩毛状根诱导的影响   总被引:1,自引:0,他引:1  
张东向  王蕊  张磊 《生物技术》2008,18(1):63-66
目的:利用发根农杆菌1.2556诱导黄芩,得到毛状根.方法:采用共培养法诱导黄芩毛状根,研究不同外植体,不同预培养时间,不同菌液浓度,不同感染时间,乙酰丁香酮,抗生素浓度等条件对转化率的影响.结果:利用预培养2d后的茎段为转化材料,当发根农杆菌浓度在OD600值为0.5时感染10min,转化率最高.在菌液中或培养基中添加100umol/L,乙酰丁香酮可以提高黄芩毛状根的转化效率.培养基中加入250mg/L抗生素Cef能较好地抑制发根农杆菌生长.结论:用共培养法诱导出黄芩毛状根,并确定了最佳诱导条件,以提高黄芩外植体的诱导率.  相似文献   

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紫云英细胞转化条件的研究   总被引:2,自引:0,他引:2  
研究了紫云英(AstragalussinicusL.)细胞遗传转化的条件。根癌农杆菌(Agrobacteriumtum efaciens)经含乙酰丁香酮的低pH/PO3-4 诱导培养后,用来感染紫云英下胚轴原生质体,随后的细胞GUS瞬间表达活性显著提高,间接证明了上述预培养诱导活化了细菌vir基因,促进了T-DNA 向植物细胞转移。在PEG介导的DNA 转移中,较高的pH 和Ca2+ 浓度能够提高细胞GUS活性。质粒DNA 浓度及启动子类型对外源基因在植物细胞内表达也有一定影响。采用外植体-农杆菌共培养法,获得GUS和NPT Ⅱ基因稳定表达的紫云英转化植株  相似文献   

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根癌农杆菌对巴戟天遗传转化的影响因素   总被引:2,自引:0,他引:2  
贺红  林小桦  张桂芳  徐鸿华 《广西植物》2004,24(5):411-413,395,i006
以巴戟天带节茎为材料,研究了根癌农杆菌对巴戟天遗传转化的影响因素。结果表明:外植体感染前先进行2 d预培养,对转化有一定促进作用;外植体与农杆菌共培养时间以3 d为宜;乙酰丁香酮能提高转化效率,抗性芽分化率可达18.0%;外植体与农杆菌共培养后延迟4 d选择,抗性芽分化率有所提高;硝酸银能抑制外植体表面农杆菌的生长,提高GUS阳性芽的比例,硝酸银浓度2 mg/L时,GUS阳性芽比例最高(42.9%)。  相似文献   

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根癌农杆菌介导转化谷子的影响因素   总被引:3,自引:0,他引:3  
利用GUS报告基因,建立了农杆菌介导的谷子(Setaria italica)遗传转化体系,并研究了多种影响因素对转化效率的影响,包括受体基因型、外植体类型、菌液浓度、培养基中乙酰丁香酮的浓度、侵染时间和共培养时间.确定了最优的转化条件为:以谷子幼穗诱导的愈伤组织为外植体,用低浓度的农杆菌菌液侵染30~40min,然后在含有0.1mmol/L乙酰丁香酮的LS培养基上共培养2d.  相似文献   

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影响根癌农杆菌介导的木霉菌遗传转化因素分析   总被引:10,自引:0,他引:10       下载免费PDF全文
采用根癌农杆菌介导的转化方法,以木霉菌分生孢子为受体材料,对影响转化效率的主要因素进行了分析。结果表明,农杆菌菌株类型、初始菌液量、分生孢子浓度、共培养时间以及乙酰丁香酮的诱导等因素对转化效率都具有重要的影响。通过对这些因素的分析,基本得出了根癌农杆菌转化系统对木霉菌遗传转化的特点和规律,为将该转化系统用于其它丝状真菌的遗传转化提供了重要参考。  相似文献   

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采用根癌农杆菌介导的转化方法,以木霉菌分生孢子为受体材料,对影响转化效率的主要因素进行了分析。结果表明,农杆菌菌株类型、初始菌液量、分生孢子浓度、共培养时间以及乙酰丁香酮的诱导等因素对转化效率都具有重要的影响。通过对这些因素的分析,基本得出了根癌农杆菌转化系统对木霉菌遗传转化的特点和规律,为将该转化系统用于其它丝状真菌的遗传转化提供了重要参考。  相似文献   

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影响农杆菌介导的大豆子叶节遗传转化的因素   总被引:1,自引:0,他引:1  
利用携带pCAMBIA1301质粒(含hpt和gus基因)的超毒根癌农杆菌菌株EHA105对大豆子叶节外植体进行遗传转化,研究了影响农杆菌介导的大豆子叶节遗传转化的因素。研究结果表明.农杆菌侵染液和共培养培养基中添加200μmok/L乙酰丁香酮和50mg/L抗坏血酸可以有效促进农杆菌对大豆子叶节的转化。农杆菌与子叶节共培养后羧苄青霉素(250mr/L)和头孢霉素(100mg/L)结合使用能有效抑制农杆菌过度繁殖并提高转化芽诱导频率;在转化细胞的分化和转化芽伸长过程中,改进的筛选策略可以明显改善对转化芽的筛选效果,从而提高转化频率。应用优化后的转化体系.获得了3个国内大豆主栽品种的转基因植株,PCR阳性植株频率为3.8%~7.6%。转化植株叶片总DNA的PCR和Southern blot实验表明,T-DNA上的外源基因已经整合到大豆基因组中。  相似文献   

9.
除虫菊发状根的诱导及培养条件优化   总被引:1,自引:0,他引:1       下载免费PDF全文
以除虫菊(Pyrethrum cinerariifolium Trey.)无菌苗为外植体,研究除虫菊发状根的诱导、培养条件优化,并对发状根中的除虫菊素进行检测和生物活性测定。结果显示,乙酰丁香酮能促进除虫菊下胚轴和子叶发状根的诱导,当乙酰丁香酮浓度为150 μmol/L时除虫菊下胚轴和子叶的诱导率为对照的2.29倍和2.66倍,预培养6 d时,下胚轴发状根诱导率为对照的2.25倍,发根农杆菌A4的诱导率均高于ATCC15834,愈伤组织较适合发状根的诱导,愈伤组织侵染后适合在无激素的MS培养基上进行发状根诱导,250 mL三角瓶中添加50 mL MS培养基较适合发状根的生长。对除虫菊发状根进行PCR检测发现,发根农杆菌含有的Ri T-DNA的rolB基因已整合进入发状根基因组中。通过GCMS检测发现,愈伤组织中除虫菊素的6种成分均未检测到,而发状根中检测到瓜菊素Ⅰ、茉酮菊素Ⅰ和茉酮菊素Ⅱ 3种成分,发状根对粘虫的拒食作用明显优于愈伤组织。本研究为通过组织培养方式生产除虫菊素奠定了基础。  相似文献   

10.
利用根癌农杆菌介导转化技术成功将潮霉素抗性基因转入发白红曲菌中,优化了抗生素浓度,发白红曲菌孢子浓度,根癌农杆菌浓度,共培养温度及时间,以及乙酰丁香酮浓度等转化条件,最终转化效率可达52个转化子/105个红曲孢子.将转化子在含有潮霉素B的培养基继代培养5代,得到了多株稳定的转化子,对部分转化子进行PCR鉴定,结果进一步...  相似文献   

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Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.  相似文献   

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Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.  相似文献   

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Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.  相似文献   

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正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases  相似文献   

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