Journal
BIO-PROTOCOL
Volume 10, Issue 24, Pages -Publisher
BIO-PROTOCOL
DOI: 10.21769/BioProtoc.3867
Keywords
Lipid droplet isolation; Triacylglycerides; Caleosin; Leaf senescence; Plant-pathogen interaction
Categories
Funding
- Spanish Ministry of Economy and Competitiveness/FEDER [BIO2015-68130-R]
- Ministry of Science, Innovation and Universities [RTI2018-097102-B-100]
Ask authors/readers for more resources
Lipid droplets (LDs) are neutral lipid aggregates surrounded by a phospholipid monolayer and specific proteins. In plants, they play a key role as energy source after seed germination, but are also formed in vegetative tissues in response to developmental or environmental conditions, where their functions are poorly understood. To elucidate these, it is essential to isolate LDs with good yields, while retaining their protein components. LD isolation protocols are based on their capacity to float after centrifugation in sucrose gradients. Early strategies using stringent conditions and LD-abundant plant tissues produced pure LDs where core proteins were identified. To identify more weakly bound LD proteins, recent protocols have used low stringency buffers, but carryover contaminants and low yields were often a problem. We have developed a sucrose gradient-based protocol to isolate LDs from Arabidopsis leaves, using Tween-20 and fresh tissue to increase yield. In both healthy and bacterially-infected Arabidopsis leaves, this protocol allowed to identify LD proteins that were later confirmed by microscopy analysis.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available