Cellulose Synthase-Like Genes:  Physcomitrella as a heterologous expression system for investigating the functions of CESA-like gene products

(Collaborators: Allison Roberts, 
Aaron Liepman & William Willats)
       Cellulose synthase-like (CSL) genes are proposed toencode glycan synthases that polymerize the backbones of non-cellulosic cell wall polysaccharides. Consistent with this hypothesis, CSLA genes are known to encode mannan synthases and a CSLF gene is implicated in mixed-linkage-glucan synthesis. Members of the CSLB, CSLE, CSLG, and CSLH families have not been functionally characterized. The complete genome sequence of the moss Physcomitella patens lacks members of these CSL families. We are using Physcomitrella as a heterologous expression system to investigate the functions of the proteins encoded by CSLB, CSLE, CSLG, and CSLH genes from seed plant species, including rice, poplar, and Arabidopsis. Transgenic Physcomitrella expressing CSLB, CSLE, and CSLG genes from Arabidopsis have been produced. The overall objectives of this collaborative project are to 1) develop additional transgenic lines that express CSLB, CSLE, CSLG, and CSLH genes from rice and poplar, 2) screen transgenic lines for changes in cell wall polysaccharide composition using a novel Comprehensive Microarray Polysaccharide Profiling method, 3) analyze the cell wall polysaccharides of transgenic Physcomitrella for the presence of novel linkages, 4) develop in vitro assays to identify the catalytic activity of the heterologously-expressed CSL proteins, and 5) examine the development and stress response of transgenic Physcomitrella containing novel cell wall polysaccharides. Non-cellulosic cell wall polysaccharides impact the pulping properties of wood, the mechanical properties of textiles fibers, the health effects of dietary fiber, and the efficiency of energy extraction from biofuels.

Funding source: USDA NRI-GCP, Plant Biology
Colaborators: Alison Roberts at University
of Rhode Island, Aaron Liepman, Wayne State University and
William Willats, KÝbenhavns Universitet, Denmark

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