《植物生理学报》 2017, 53(2): 185-190

叶片中GUS染色观测油体含量技术体系的建立

胡佳^1,2, 曾文婕², 刘春林^1,2,*

¹湖南农业大学农学院, 长沙410128; ²湖南省作物种质创新和资源利用国家重点实验室培育基地, 长沙410128

通信作者：刘春林；E-mail: liucl100@126.com

摘　要：

植物油体大致可以分为两种: 生殖器官(种子)中的油体和营养器官中的油体。营养器官中的油体数量相对少而小, 且分布弥散, 导致在已有的研究报道中, 有关营养器官中油体的研究很少。为了实现对叶片中油体进行方便有效的观察, 我们将生殖器官与营养器官这两种油体上共有的油体蛋白3 (Oleosin3)对应的启动子与GUS指示基因融合后转化到叶片积累油体的拟南芥突变体中, 经GUS染色成功观察到蓝色斑点。这一研究成功建立了可用于直观检测叶片中油体存在与否的技术体系, 为后续营养器官中油体形成机制的研究提供了有力的技术保障。

关键词：叶片; 油体; 油体蛋白; GUS染色

收稿：2016-09-21   修定：2016-12-08

资助：国家自然科学基金(31571707)。

The establishment of oil body content determination system with GUS staining observation in leaves

HU Jia^1,2, ZENG Wen-Jie², LIU Chun-Lin^1,2,*

¹College of Agriculture, Hunan Agriculture Universiy, Changsha 410128, China; ²Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Changsha 410128, China

Corresponding author: LIU Chun-Lin; E-mail: liucl100@126.com

Abstract:

Oil bodies can be roughly divided into seed oil body and vegetative oil body in plant. In vegetative organs, oil body was poorly reported because of the less, smaller and dispersion of the oil body in vegetative organs. In order to realize the convenient and effective observation of oil bodies in leaf, the promoter of the gene coding for oil body protein Oleosin3 which exists on both reproductive and vegetative oil bodies was fused with GUS gene, and then the cassette was transformed into the Arabidopsis thaliana mutant that can accumulate oil bodies in leaves. By GUS staining, blue spots were observed in mutant leaves. The result indicated that the visual detection technology system for oil body observation in leaf was successfully established, which would provide powerful technical support for the follow-up study of oil body formation mechanism in vegetative organs.

Key words: leaf; oil body; oleosins; GUS staining