Dr. Alison Roberts
Physcomitrella patens, CESA, cellulose synthesis, entry clone, cellulose
Cellulose is very essential to plants because it determines the shape of cells, protects them from pathogens, and helps retain water that is needed for plant functions. It is also the major component of wood, cotton, and paper, which are items we use on a daily basis. Also, it can be used for the synthesis of biofuels. However, cellulose exists as strong fibers, which make it hard to breakdown for biofuel synthesis. If we can understand how cellulose is synthesized we can manipulate its fibers to make them stronger, more flexible, more absorbent, or easier to break down for use as biofuels. Cellulose synthase complexes are observed by electron microscopy in the plasma membrane and Golgi vesicles of algae and plants. However, the different CESA proteins cannot be identified using this technique. Therefore, the number of CESAs in a complex is still unknown as well as their stoichiometry. Regulation of the activity and assembly of the complex are also unknown. A general idea is that there are certain CESAs that are responsible for the formation of the primary cell wall, and others that are responsible for the secondary cell wall in vascular plants. But, the moss Physcomitrella patens has only primary cell walls and still has seven different CESAs. The purpose of this project is to study the properties and characteristics of cellulose synthase 5 (CESA5). Understanding the role of CESA5 in P. patens can lead to a better understanding of the evolution of the cellulose synthase complex and the formation of cellulose. This can ultimately enable us to break down cellulose for biofuels and use it as a possible solution to global warming. During the course of this project, a miniSOG-tagged CESA 5 expression clone was successfully created using Gateway cloning. Polymerase chain reaction and a BP reaction were used to make a tandem-miniSOG entry clone and an LR reaction was used to insert the entry clones into the destination vector. The destination vector was transformed into the moss. Phenotypic analysis will be performed on the transformed lines to gain insight onto the function of CESA 5 in the synthesis of cellulose.