中国科学院机构知识库网格
Chinese Academy of Sciences Institutional Repositories Grid
Methods to isolate a large amount of generative cells, sperm cells and vegetative nuclei from tomato pollen for "omics" analysis

文献类型:期刊论文

作者Lu, Yunlong1,2; Wei, Liqin1; Wang, Tai1
刊名Frontiers in plant science
出版日期2015-06-02
卷号6页码:9
关键词Solanum lycopersicum Generative cell Sperm cell Vegetative nuclei Isolation Percoll density gradient centrifugation
ISSN号1664-462X
DOI10.3389/fpls.2015.00391
通讯作者Wang, tai(twang@ibcas.ac.cn)
英文摘要The development of sperm cells (scs) from microspores involves a set of finely regulated molecular and cellular events and the coordination of these events. the mechanisms underlying these events and their interconnections remain a major challenge. systems analysis of genome-wide molecular networks and functional modules with high-throughput "omics" approaches is crucial for understanding the mechanisms; however, this study is hindered because of the difficulty in isolating a large amount of cells of different types, especially generative cells (gcs), from the pollen. here, we optimized the conditions of tomato pollen germination and pollen tube growth to allow for long-term growth of pollen tubes in vitro with scs generated in the tube. using this culture system, we developed methods for isolating gcs, scs and vegetative cell nuclei (vn) from just-germinated tomato pollen grains and growing pollen tubes and their purification by percoll density gradient centrifugation. the purity and viability of isolated gcs and scs were confirmed by microscopy examination and fluorescein diacetate staining, respectively, and the integrity of vn was confirmed by propidium iodide staining. we could obtain about 1.5 million gcs and 2.0 million scs each from 180 mg initiated pollen grains, and 10 million vn from 270 mg initiated pollen grains germinated in vitro in each experiment. these methods provide the necessary preconditions for systematic biology studies of sc development and differentiation in higher plants.
WOS关键词MALE GAMETOPHYTE DEVELOPMENT ; TUBE GROWTH ; SMALL RNA ; IN-VITRO ; ARABIDOPSIS-THALIANA ; NICOTIANA-TABACUM ; FLOWERING PLANTS ; GENE-EXPRESSION ; MATURE POLLEN ; ZEA-MAYS
WOS研究方向Plant Sciences
WOS类目Plant Sciences
语种英语
WOS记录号WOS:000357012900002
出版者FRONTIERS RESEARCH FOUNDATION
URI标识http://www.irgrid.ac.cn/handle/1471x/2376402
专题中国科学院大学
通讯作者Wang, Tai
作者单位1.Chinese Acad Sci, Inst Bot, Key Lab Plant Mol Physiol, Beijing 100093, Peoples R China
2.Univ Chinese Acad Sci, Beijing, Peoples R China
推荐引用方式
GB/T 7714
Lu, Yunlong,Wei, Liqin,Wang, Tai. Methods to isolate a large amount of generative cells, sperm cells and vegetative nuclei from tomato pollen for "omics" analysis[J]. Frontiers in plant science,2015,6:9.
APA Lu, Yunlong,Wei, Liqin,&Wang, Tai.(2015).Methods to isolate a large amount of generative cells, sperm cells and vegetative nuclei from tomato pollen for "omics" analysis.Frontiers in plant science,6,9.
MLA Lu, Yunlong,et al."Methods to isolate a large amount of generative cells, sperm cells and vegetative nuclei from tomato pollen for "omics" analysis".Frontiers in plant science 6(2015):9.

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来源:中国科学院大学

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