Submitted to: Journal of Microbiological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/30/2015
Publication Date: 12/5/2015
Publication URL: http://handle.nal.usda.gov/10113/61737
Citation: Jones, R.W. 2015. Application of succulent plant leaves for Agrobacterium infiltration-mediated protein production. Journal of Microbiological Methods. 120:65-67.
Interpretive Summary: Plants produce a wide range of important materials, from the food we eat, as well as non food materials such as cotton for clothing and wood for houses. In an effort to improve these materials, new genetic information may be added to plants to test their function and potential use. During a study on using introduced genetic information to produce a product that breaks apart plant materials, we found that the commonly used tobacco plant was quickly damaged. Changing to a thick, succulent leafed plant allowed the product of the new genetic information to accumulate enough to be extracted for further study. This allows for study and development of new plant products. This information will be useful to scientists involved in development of improved, economically enhanced and novel plant products.
Technical Abstract: Infiltration of tobacco leaves with a suspension of Agrobacterium tumefaciens harboring a binary plant expression plasmid provides a convenient method for laboratory scale protein production. When expressing plant cell wall degrading enzymes in the widely used tobacco (Nicotiana benthamiana), difficulties arise due to the thin leaf structure. Two thick leaved succulents, Kalanchoe blossfeldiana and Hylotelephium telephium (stonecrop), were tested as alternatives for protein expression. A glycosyl hydrolase (cel12 ), that causes leaf tissue collapse in tobacco, was successfully produced and isolated from the succulent leaves. A plant encoded cel12 inhibitor protein derived from potato was also successfully produced and shown to be fully functional. While expression was successful in both types of succulent plant, the Kalanchoe was more efficient due to the high extractability of fluids containing the apoplastic proteins under study.