Lung Shiu-Cheung, Smith Matthew D, Chuong Simon D X
School of Biological Sciences, University of Hong Kong, Pokfulam, Hong Kong SAR, China;
Department of Biology, Wilfrid Laurier University, Waterloo, Ontario N2L 3C5, Canada;
Cold Spring Harb Protoc. 2015 Oct 1;2015(10):895-9. doi: 10.1101/pdb.prot074559.
Chloroplasts can be isolated from higher plants directly following homogenization; however, the resulting yield, purity, and intactness are often low, necessitating a large amount of starting material. This protocol is optimized to produce a high yield of pure chloroplasts from isolated Arabidopsis protoplasts. The two-part method is a simple, scaled-down, and low-cost procedure that readily provides healthy mesophyll protoplasts, which are then ruptured to release intact chloroplasts. Chloroplasts isolated using this method are competent for use in biochemical, cellular, and molecular analyses.
匀浆后可直接从高等植物中分离出叶绿体；然而，所得的产量、纯度和完整性往往较低，需要大量的起始材料。本方案经过优化，可从分离的拟南芥原生质体中高产率地制备出纯净的叶绿体。这个分为两步的方法是一个简单、规模较小且成本较低的程序，能轻松提供健康的叶肉原生质体，然后将其破碎以释放完整的叶绿体。用这种方法分离出的叶绿体可用于生化、细胞和分子分析。
