Methods to isolate a large amount of generative cells, sperm cells and vegetative nuclei from tomato pollen for omics analysis

doi:10.3389/fpls.2015.00391

	Methods to isolate a large amount of generative cells, sperm cells and vegetative nuclei from tomato pollen for omics analysis
	Lu, Yunlong 1; Wei, Liqin ; Wang, Tai
	2015
发表期刊	FRONTIERS IN PLANT SCIENCE
ISSN	1664-462X
卷号	6
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.
关键词	Solanum lycopersicum generative cell sperm cell vegetative nuclei isolation Percoll density gradient centrifugation
学科领域	Plant Sciences
DOI	10.3389/fpls.2015.00391
收录类别	SCI
语种	英语
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	Science Citation Index Expanded (SCI-EXPANDED)
WOS记录号	WOS:000357012900002
出版者	FRONTIERS RESEARCH FOUNDATION
文献子类	Article
出版地	LAUSANNE
资助机构	Chinese Ministry of Science and Technology [2012CB910504]
作者邮箱	twang@ibcas.ac.cn
作品OA属性	gold, Green Published
引用统计	被引频次：15[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.ibcas.ac.cn/handle/2S10CLM1/25933
专题	中科院植物分子生理学重点实验室
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.
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.
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).

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