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1.
利用扫描电镜对黄槐叶片培养中形态分化过程进行了观察。结果表明:叶片脱分化形成愈伤组织时,细胞最初进行无丝分裂其中有类似酵母的“出芽”方式的无丝分裂;愈伤组织再分化形成不定芽的过程中,最初愈伤组织表面分化出球状体,它为一顶端凹陷,具有分化潜力的细胞团,其顶端可以分化形成不定芽原基,最后发育成不定芽。球状体可以看成是具有形成不定芽能力的繁殖单位,将具有球状体与芽的愈伤组织进行继代培养,可以大量快速繁殖黄槐幼苗。  相似文献   

2.
革新与发明     
这是一项于1995年12月27日新公布的日本发明人申请的中国专利,提供一种流液固相酵母循环发酵制酒法.该方法实质上是使用一座设有液体循环管路和排气口的固相酵母流化床式反应器(如图),在此反应器中装填入固相酵母细胞和酿造原料后,可从反应器上部抽取部分培养液再从反应器下部泵回到反应器内,使之进行液体循环性培养.由于它便于随时排出在发酵过程中产生的CO_2气体,因而可避免CO_2对酵母代谢活性的影响,避免因CO_2气体积聚而减少因相酵母与培养液接触面积和因CO_2气体的压力而导致酵母凝胶挤压粘结对培养液通路的阻塞.既克服了搅拌式反应器对固相酵母的机械破坏,也克服了填充式反应器氨基酸同化率低,酒液味道差的缺点,大大加快了发酵过程和原料中氨基酸的同化,在短期内即可酿制出口味香醇的成品酒.  相似文献   

3.
用光学显微镜和电子显微镜研究热带假丝酵母(Candida tropicalis)的二塑性,菌丝体(M)和单细胞酵母(Y)形态差异很大,M是长管状多,细胞 相连,Y则是球形的单个细胞,内部结构上也体现了与其形态的相适应性,M菌丝顶端大的液泡为菌丝的延伸提供压力,但当顶端生长芽管时,大液泡会分裂成小液泡,部分小液泡进入芽管;菌丝顶端和侧面的泡囊,与其顶端生长和侧枝发生有关,单细胞酵母中,靠近母细胞和子细胞相连的部位,有些囊泡和颗粒物质,可能与细胞的新壁形成和分开有关。  相似文献   

4.
磁场对酿酒酵母微观结构及膜脂流动性的影响   总被引:1,自引:0,他引:1  
以磁场为外加物理场,采用恒定磁场及循环磁处理方式对酿酒酵母进行处理,研究酿酒酵母微观结构及细胞膜流动性的变化.通过透射电子显微镜观察不同磁感应强度恒定磁场处理后酿酒酵母的微观结构.研究发现恒定磁场可以促进酵母细胞的生长,群体中以具有中央大液泡为特征的成熟细胞所占比率增大,同时线粒体呈现适应性膨大、增生.利用原子力显微镜观察循环磁处理下酵母细胞的三维结构,发现细胞壁表面出现皱缩、穿孔,胞质外流.用荧光偏振法对两种处理方式下细胞的膜脂流动性进行测定,发现不同强度恒定磁场处理后,膜脂各向异性下降,流动性增强,0.05T磁感应强度下各向异性值下降3.57%,而循环磁处理后各向异性值比对照提高了8.46%,膜脂流动减慢,证实细胞膜为受到磁场作用的亚细胞结构.  相似文献   

5.
分别用仙客来种苗的子叶和叶柄作为外植体,培养在附加2,4 D、细胞分裂素和细胞激动素的1/2MS培养基上诱导不定芽的发生,研究了愈伤组织和不定芽形成过程中的组织细胞学变化.对两种不同外植体不定芽发生过程中的切片观察表明,两种外植体的组织脱分化始于维管束周围的维管束鞘细胞,随后开始分裂的是与其临近的薄壁细胞并能很快形成胚性分生细胞团.经30d左右的培养,进一步分化形成芽原基.子叶愈伤的芽原基通常出现在愈伤的边缘.叶柄脱分化比子叶快,不定芽可以起源于表层的分生细胞团,也可以由愈伤深处的分生组织分化而形成.  相似文献   

6.
为阐明人参多越冬芽发育形成多茎的生理机制,采用HPLC-MS/MS法对两种类型人参植株在越冬芽发育过程中根茎部位主要内源激素含量进行了检测,分析了各种激素含量变化规律以及激素间相互作用对越冬芽发育数量的影响.结果表明:人参多越冬芽发育是生长素、细胞分裂素相互作用的结果,根茎中低浓度生长素、高浓度细胞分裂素有利于多越冬芽发育;越冬芽发育时活性细胞分裂素是以核苷分子形式存在的tZR、cZR、iPR;赤霉素间接参与越冬芽发育过程,脱落酸拮抗细胞分裂素,对越冬芽发育起抑制作用.根茎中高CKs/IAA值和低浓度赤霉素有利于多越冬芽发育形成多茎.  相似文献   

7.
酒精发酵是一个伴有物质传递的生化反应过程。本文对强制循环发酵液的酒精发酵过程进行了研究。结果表明:采用强制循环发酵液的方法可以促进传质过程,利于糖化酶对作用底物的水解和改善酵母细胞生长状况,为高浓度糖化液发酵生产酒精探索了一条可行的路径,实现了提高发酵效率的目的。  相似文献   

8.
流式细胞仪检测重组毕赤酵母发酵过程中的细胞活性   总被引:1,自引:0,他引:1  
建立了流式细胞仪检测重组巴斯德毕赤酵母(P.p astoris)细胞活性的方法,并在高密度发酵过程中得到应用。使用碘化丙锭染色法测定细胞活性,发现甲醇诱导时酵母细胞活性开始下降,随着细胞密度的增加,由于细胞胁迫和甲醇的影响,细胞活性明显下降,活细胞率最大下降了14%。与此同时,细胞生长减缓,目的蛋白发生降解,表明细胞活性与发酵过程中细胞的生长和目的蛋白生成等参数密切相关。实验证明:流式细胞仪可作为检测毕赤酵母发酵过程中细胞活性参数的一个便捷精确的有力工具。  相似文献   

9.
在芽殖酵母中构建CDC10-mCherry和SPC42-mCherry荧光蛋白, 作为酵母细胞周期不同时期的报告系统。根据酵母细胞中芽环(CDC10-mCherry)的产生和消失以及纺锤体极体(SPC42-mCherry)的复制和分离,可以测量细胞周期中 G1期、S期、Early M期和Late M期的时间长度。在此报告系统的基础上, 分别构建S期细胞周期蛋白CLB5-GFP和M 期细胞周期蛋白CLB2-GFP菌株, 实现单细胞观测, 检验报告系统工作的稳定性和准确性。该报告系统可作为研究酵母细胞周期不同时期时间长度等定量生物学问题的背景菌株。  相似文献   

10.
毕赤酵母作为最成熟的蛋白质过量表达平台微生物之一,广泛应用于酶制剂、功能性大分子等大宗产品的发酵制造.建立毕赤酵母常规温度下(50~60,℃)的热致死动力学模型,有助于指导毕赤酵母发酵制造体系的菌体后处理工艺,增加菌体利用价值.为了建立毕赤酵母在50~60,℃条件下热致死动力学模型,将不同菌体量的毕赤酵母菌液在50、55、60,℃条件下进行不同时间热灭活处理,通过平板培养活细胞计数的方法评价不同处理方式下毕赤酵母的热致死效果,得到的致死曲线用Weibull模型和多元回归方程进行拟合构建热致死动力学模型,并对建立的模型进行数学检验和实用性检验.结果表明:建立的毕赤酵母热致死动力学模型可靠,具有实际使用价值.本研究所建立的模型能较好地模拟不同温度、不同处理时间对不同菌体量的毕赤酵母热致死效果的影响.并可以为毕赤酵母热灭活处理条件的确定提供计算依据.  相似文献   

11.
利用α因子可以使MATa酵母细胞的生长停滞在细胞周期的G1期,获得同步化的YKN-10酵母菌细胞,主要研究在25~200μg·mL-1浓度下的α因子对酿酒酵母YKN-10对数早期细胞作用0~5h的同步化影响,每隔0.5h在荧光倒置显微镜下观察酵母细胞的出芽形态,记录细胞的出芽率,然后用SPSS 17.0对酵母细胞的出芽率进行统计分析,α因子的浓度、α因子作用时间及二者的交互效应均对酵母细胞的平均出芽率有显著影响;α因子浓度在175~200μg·mL-1下培养酵母细胞约3.5~4h时,酵母细胞基本上达到了同步化状态,运用α因子停滞细胞生长法获得了同步化的YKN-10酵母菌群体,为非Δbar1突变菌株的同步化研究提供了参考.  相似文献   

12.
L N Castor 《Nature》1980,287(5785):857-859
Smith and his colleagues have proposed that the duration of the cell cycle is determined by a random transition, analogous to the random decay of a radioactive nucleus, by which a cell passes from an 'A state' within the G1 phase to a 'B phase' that includes the rest of the cycle. The experimental support for this transition probability hypothesis is the tendency of a cumulative plot of differences of cycle times of sibling cells (the beta curve) to be exponential ad parallel to the exponential tail of a cumulative plot of the cycle times themselves (the alpha curve). However, a close examination of four of the most extensive sets of experimental data now suggests that the two beta curves with the steepest slopes may not, in fact, be exponential. These and all the other characteristics of the experimental curves are best matched by computer simulations using a cell-cycle model that will be termed here a G1 rate model. This model is consistent with differences in cell metabolism, rather than a transition at an inherently unpredictable time, being the physiological basis for differences in cycle times within a cell population.  相似文献   

13.
14.
A single double-strand break (DSB) induced by HO endonuclease triggers both repair by homologous recombination and activation of the Mec1-dependent DNA damage checkpoint in budding yeast. Here we report that DNA damage checkpoint activation by a DSB requires the cyclin-dependent kinase CDK1 (Cdc28) in budding yeast. CDK1 is also required for DSB-induced homologous recombination at any cell cycle stage. Inhibition of homologous recombination by using an analogue-sensitive CDK1 protein results in a compensatory increase in non-homologous end joining. CDK1 is required for efficient 5' to 3' resection of DSB ends and for the recruitment of both the single-stranded DNA-binding complex, RPA, and the Rad51 recombination protein. In contrast, Mre11 protein, part of the MRX complex, accumulates at unresected DSB ends. CDK1 is not required when the DNA damage checkpoint is initiated by lesions that are processed by nucleotide excision repair. Maintenance of the DSB-induced checkpoint requires continuing CDK1 activity that ensures continuing end resection. CDK1 is also important for a later step in homologous recombination, after strand invasion and before the initiation of new DNA synthesis.  相似文献   

15.
Mimura S  Seki T  Tanaka S  Diffley JF 《Nature》2004,431(7012):1118-1123
Cyclin-dependent kinases (CDKs) limit the activation of DNA replication origins to once per cell cycle by preventing the assembly of pre-replicative complexes (pre-RCs) during S, G2 and M phases of the cell cycle in the budding yeast Saccharomyces cerevisiae. CDKs inhibit each pre-RC component (ORC, Cdc6, Cdt1/Mcm2-7) by different mechanisms. We show here that the mitotic CDK, Clb2/Cdc28, binds tightly to an amino-terminal domain (NTD) of Cdc6, and that Cdc6 in this complex is unable to assemble pre-RCs. We present evidence indicating that this Clb2-dependent mechanism contributes to preventing re-replication in vivo. CDK interaction with the NTD of Cdc6 is mediated by the cyclin subunit Clb2, and could be reconstituted with recombinant Clb2 protein and synthetic NTD peptides. Tight Clb2 binding occurred only when the NTD was phosphorylated on CDK consensus sites. Human CDKs containing cyclins A, B and E also bound specifically to phospho-NTD peptides. We propose that direct binding of cyclins to phosphopeptide motifs may be a widespread phenomenon contributing to the targeting of CDKs to substrates.  相似文献   

16.
17.
Altschuler SJ  Angenent SB  Wang Y  Wu LF 《Nature》2008,454(7206):886-889
Diverse cell polarity networks require positive feedback for locally amplifying distributions of signalling molecules at the plasma membrane. Additional mechanisms, such as directed transport or coupled inhibitors, have been proposed to be required for reinforcing a unique axis of polarity. Here we analyse a simple model of positive feedback, with strong analogy to the 'stepping stone' model of population genetics, in which a single species of diffusible, membrane-bound signalling molecules can self-recruit from a cytoplasmic pool. We identify an intrinsic stochastic mechanism through which positive feedback alone is sufficient to account for the spontaneous establishment of a single site of polarity. We find that the polarization frequency has an inverse dependence on the number of signalling molecules: the frequency of polarization decreases as the number of molecules becomes large. Experimental observation of polarizing Cdc42 in budding yeast is consistent with this prediction. Our work suggests that positive feedback can work alone or with additional mechanisms to create robust cell polarity.  相似文献   

18.
基于已公布的人体心室肌细胞模型数据建立了一维心室肌细胞模型,仿真了伪心电图及心内膜细胞、心中间膜细胞和心外膜细胞这三种细胞的动作电位.基于构建的模型进行周期实验,针对实验中出现的问题修订模型中的参数.将修订后模型的仿真结果与已经公布的实验数据进行对比,从而验证模型修订的合理性和可靠性.最后基于修订的心室肌细胞模型对与KCNJ2有关的短QT综合征的病理情况进行仿真,分析仿真结果,验证了修订模型的实用性.  相似文献   

19.
The events of cell reproduction are governed by oscillations in the activities of cyclin-dependent kinases (Cdks). Cdks control the cell cycle by catalysing the transfer of phosphate from ATP to specific protein substrates. Despite their importance in cell-cycle control, few Cdk substrates have been identified. Here, we screened a budding yeast proteomic library for proteins that are directly phosphorylated by Cdk1 in whole-cell extracts. We identified about 200 Cdk1 substrates, several of which are phosphorylated in vivo in a Cdk1-dependent manner. The identities of these substrates reveal that Cdk1 employs a global regulatory strategy involving phosphorylation of other regulatory molecules as well as phosphorylation of the molecular machines that drive cell-cycle events. Detailed analysis of these substrates is likely to yield important insights into cell-cycle regulation.  相似文献   

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