Abstract [Objective] The paper was to prepare and detect the extended DNA fibers of Chinese cabbage. [Method] Chinese cabbage nuclei was first successfully isolated by chopping young leaves with a blade, then nuclei w...Abstract [Objective] The paper was to prepare and detect the extended DNA fibers of Chinese cabbage. [Method] Chinese cabbage nuclei was first successfully isolated by chopping young leaves with a blade, then nuclei were lysed by SDS to release DNA, and DNA fibers were dragged and extended with a coverslip. [Result] The results of Fiber-FISH with genomic DNA and 25S rDNA as probes showed that DNA fiber size as long as about 1.93 Mb could be measured and the number of 25S rDNA copies region were estimated to be 258 and 687 in Chinese cabbage genome. DNA fibers prepared by this method showed equally spread parallel thread with clear background, and were suitable for FISH analysis. [Conclusion] The study would accelerate Chinese cabbage genome mapping and organization analysis.展开更多
Background:The use of image analysis to understand the structure of chromosome and chromatin is critical to the study of genetic evolution and diversification.Furthermore,a detailed chromosome map and the structure of...Background:The use of image analysis to understand the structure of chromosome and chromatin is critical to the study of genetic evolution and diversification.Furthermore,a detailed chromosome map and the structure of chromatin in the nucleus may contribute to the plant breeding and the study of fundamental biology and genetics in organisms.Results:In plants with a fully annotated genome project,such as the Leguminosae species,the integration of genetic information,including DNA sequence data,a linkage map,and the cytological quantitative chromosome map could further improve their genetic value.The numerical parameters of chromocenters in 3D can provide useful genetic information for phylogenetic studies of plant diversity and heterochromatic markers whose epigenetic changes may explain the developmental and environmental changes in the plant genome.Extended DNA fibers combined with fluorescence in situ hybridization revealed the highest spatial resolution of the obtained genome structure.Moreover,image analysis at the nano-scale level using a helium ion microscope revealed the surface structure of chromatin,which consists of chromatin fibers compacted into plant chromosomes.Conclusions:The studies described in this review sought to measure and evaluate chromosome and chromatin using the image analysis method,which may reduce measurement time and improve resolution.Further,we discussed the development of an effective image analysis to evaluate the structure of chromosome and chromatin.An effective application study of cell biology and the genetics of plants using image analysis methods is expected to be a major propeller in the development of new applications.展开更多
基金Supported by National Natural Science Foundation of China (30471182)Youth Science Foundation of Hebei Province (C2010000738)~~
文摘Abstract [Objective] The paper was to prepare and detect the extended DNA fibers of Chinese cabbage. [Method] Chinese cabbage nuclei was first successfully isolated by chopping young leaves with a blade, then nuclei were lysed by SDS to release DNA, and DNA fibers were dragged and extended with a coverslip. [Result] The results of Fiber-FISH with genomic DNA and 25S rDNA as probes showed that DNA fiber size as long as about 1.93 Mb could be measured and the number of 25S rDNA copies region were estimated to be 258 and 687 in Chinese cabbage genome. DNA fibers prepared by this method showed equally spread parallel thread with clear background, and were suitable for FISH analysis. [Conclusion] The study would accelerate Chinese cabbage genome mapping and organization analysis.
基金This work was supported by“e-ASIA Joint Research Program”,“Development of nano-visualization for structural analyses of genetic materials and early infection process for further innovation of functional bio-nanotechnology”from SICORP,and the Mobility Plus Project(No.JPJSBP 120203507)(to NO)This work was also supported by“The 51st Research Grants in the Natural Sciences”from The Mitsubishi Foundation and Nanotechnology Platform of MEXT(No.JPMXP09F20OS0004)(to NO)。
文摘Background:The use of image analysis to understand the structure of chromosome and chromatin is critical to the study of genetic evolution and diversification.Furthermore,a detailed chromosome map and the structure of chromatin in the nucleus may contribute to the plant breeding and the study of fundamental biology and genetics in organisms.Results:In plants with a fully annotated genome project,such as the Leguminosae species,the integration of genetic information,including DNA sequence data,a linkage map,and the cytological quantitative chromosome map could further improve their genetic value.The numerical parameters of chromocenters in 3D can provide useful genetic information for phylogenetic studies of plant diversity and heterochromatic markers whose epigenetic changes may explain the developmental and environmental changes in the plant genome.Extended DNA fibers combined with fluorescence in situ hybridization revealed the highest spatial resolution of the obtained genome structure.Moreover,image analysis at the nano-scale level using a helium ion microscope revealed the surface structure of chromatin,which consists of chromatin fibers compacted into plant chromosomes.Conclusions:The studies described in this review sought to measure and evaluate chromosome and chromatin using the image analysis method,which may reduce measurement time and improve resolution.Further,we discussed the development of an effective image analysis to evaluate the structure of chromosome and chromatin.An effective application study of cell biology and the genetics of plants using image analysis methods is expected to be a major propeller in the development of new applications.
