An Efficient Clearing Protocol for the Study of Seed Development in Tomato (Solanum lycopersicum L.)

doi:10.3791/64445

	An Efficient Clearing Protocol for the Study of Seed Development in Tomato (Solanum lycopersicum L.)
	Feng, Yixuan 1; Wang, Tai1,2 ; Liu, Lingtong
	2022
发表期刊	JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
ISSN	1940-087X
期号	187
摘要	Tomato (Solanum lycopersicum L.) is one of the major cash crops worldwide. The tomato seed is an important model for studying genetics and developmental biology during plant reproduction. Visualization of finer embryonic structure within a tomato seed is often hampered by seed coat mucilage, multi-cell-layered integument, and a thick-walled endosperm, which needs to be resolved by laborious embeddingsectioning. A simpler alternative is to employ tissue clearing techniques that turn the seed almost transparent using chemical agents. Although conventional clearing procedures allow deep insight into smaller seeds with a thinner seed coat, clearing tomato seeds continues to be technically challenging, especially in the late developmental stages. Presented here is a rapid and labor-saving clearing protocol to observe tomato seed development from 3 to 23 days after flowering when embryonic morphology is nearly complete. This method combines chloral hydrate-based clearing solution widely used in Arabidopsis with other modifications, including the omission of Formalin-Aceto-Alcohol (FAA) fixation, the addition of sodium hypochlorite treatment of seeds, removal of the softened seed coat mucilage, and washing and vacuum treatment. This method can be applied for efficient clearing of tomato seeds at different developmental stages and is useful in full monitoring of the developmental process of mutant seeds with good spatial resolution. This clearing protocol may also be applied to deep imaging of other commercially important species in the Solanaceae.
学科领域	Multidisciplinary Sciences
DOI	10.3791/64445
收录类别	SCI
语种	英语
WOS关键词	ENDOSPERM DEVELOPMENT ; FRUIT-GROWTH ; EMBRYO ; AUXIN ; GIBBERELLIN
WOS研究方向	Science Citation Index Expanded (SCI-EXPANDED)
WOS记录号	WOS:000909465300044
出版者	JOURNAL OF VISUALIZED EXPERIMENTS
文献子类	Article
出版地	CAMBRIDGE
资助机构	National Natural Science Foundation of China [31870299] ; Youth Innovation Promotion Association of the Chinese Academy of Sciences
作者邮箱	ltliu@ibcas.ac.cn
引用统计
文献类型	期刊论文
条目标识符	http://ir.ibcas.ac.cn/handle/2S10CLM1/29057
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, Peoples R China 2.Univ Chinese Acad Sci, Coll Life Sci, Beijing, Peoples R China 3.Chinese Acad Sci, Innovat Acad Seed Design, Beijing, Peoples R China
推荐引用方式 GB/T 7714	Feng, Yixuan,Wang, Tai,Liu, Lingtong. An Efficient Clearing Protocol for the Study of Seed Development in Tomato (Solanum lycopersicum L.)[J]. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS,2022(187).
APA	Feng, Yixuan,Wang, Tai,&Liu, Lingtong.(2022).An Efficient Clearing Protocol for the Study of Seed Development in Tomato (Solanum lycopersicum L.).JOVE-JOURNAL OF VISUALIZED EXPERIMENTS(187).
MLA	Feng, Yixuan,et al."An Efficient Clearing Protocol for the Study of Seed Development in Tomato (Solanum lycopersicum L.)".JOVE-JOURNAL OF VISUALIZED EXPERIMENTS .187(2022).

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