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Studies on developments of cell and tissue culture

细胞和组织培养的进展研究

基本信息

批准号：
13460065
负责人：
KUBO Takafumi
金额：
$ 7.81万
依托单位：
Tokyo University of Agriculture and Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

(1)In subculture of embryogenic cells (ECs) from immature embryo of Pinus densiflora, more effevtve culture conditions were mDCR medimun which added L-glutamine of 1250mg/l and polyvinyl pyrrolidone(PVP) of 1000mg/l, resulting in higher proliferation of ECs with good quality. Furthermore, in formation of somatic embryo from ECs, additions of abscisic acid (ABA) of 30μM, maltose of 10% and polyethylene glycols (PEG) of molecular weight 8000 were effective in the development.(2)In Larix leptolepis, Picea jezoensis and Crypbomeria japonica, characteristics of the culture cells were clarified b**** the morphological observation and a quantitative analysis of endogenous amino acid, using micro-HPLC Consequently, effect of time amino acid was confirmed in the subculture of ECs, and an addition of a certain concentration of amino acid promoted development of ECs with clear and large embryo region and long suspensors. Thus result suggests possibility of somatic embryo formation. In recalcitran … More t species, like Cryptomeria japonica, it is considered that protocols of the analysis and evaluation by monitoring endogenous amino acid contents are effective for selecting the cells with high differential ability in the early stage, which result finally in somatic embryo(3)A suitable condition for isolation of protoplast and an optimum combination of medium and plant hormone for proliferation of protoplast generally vary according to plant species. Micro-plate (24 well) assay method that had been. established for protoplast culture of Populus alba, is adaptable to investigate culture conditions of coniferous tree such as L.leptolepis and C.japonica. By culturing small amount of protoplasts in a well of 96 well micro-plate, an optimal kind and concentration of plant hormone(s), and/or cell density was easily determined.(4)We determined the entire nucleotide sequence of GjNdly with the exception of the 5'-untranslationed promoter region. Although no proline-rich region has previously been detected in the gene product from gymnosperms, we found such a region in the amino-terminal region of the deduced amino acid sequence from Cryptomeria japonica. Less

(1)In对赤松幼胚胚性细胞（EC）进行继代培养，发现在mDCR培养基中添加1250 mg/l的L-谷氨酰胺和1000 mg/l的聚乙烯吡咯烷酮（PVP）对胚性细胞的增殖效果较好。在体细胞胚形成过程中，添加30μM的脱落酸（阿坝）、10%的麦芽糖和分子量为8000的聚乙二醇（PEG）对体细胞胚的形成有促进作用。(2)In通过对日本落叶松、日本云杉和日本柳杉培养细胞的形态学观察和对内源氨基酸的微量高效液相色谱定量分析，阐明了培养细胞的特征。结果表明，氨基酸在EC继代培养过程中具有时间效应，添加一定浓度的氨基酸可促进EC的发育，胚区清晰、大，胚柄长。结果表明，体细胞胚的形成是可能的。在阿西西特兰 ...更多信息对日本柳杉（Cryptomeriajaponica）等植物进行了原生质体培养，认为通过测定内源氨基酸的含量来分析和评价原生质体，是筛选早期分化能力强的细胞的有效方法。（3）原生质体分离的适宜条件和原生质体增殖的培养基与激素的最佳组合因植物种类而异。微孔板（24孔）测定方法。为银白杨原生质体培养建立了一套适合于针叶树如日本落叶松、欧洲落叶松等的原生质体培养条件的研究方法。通过在96孔微板的孔中培养少量原生质体，容易地确定植物激素的最佳种类和浓度和/或细胞密度。(4)We确定了GjNdly的整个核苷酸序列，除了5 ′-未命名的启动子区。虽然没有脯氨酸丰富的区域已被检测到的基因产物从裸子植物，我们发现这样一个区域的氨基端区域推导的氨基酸序列从柳杉。少