Abstract
BackgroundLignocellulosic biomass is an important renewable resource for biofuels and materials. How plants synthesise cellulose is not completely understood. It is known that cellulose synthase complex (CSCs) moving in the plasma membrane synthesise the cellulose. CESA proteins are the core components of CSC. In Arabidopsis, in vitro mutagenesis of proteins followed by complementation analysis of mutants lacking the gene represents an important tool for studying any biological process, including cellulose biosynthesis. Analysis of a large number of plants is crucial for these types of studies.ResultsBy using aspiration rather than centrifugation to remove liquids during various stages of protocol, we were able to increase the throughput of the method as well as minimise the sample loss. As a test case, we determined cellulose content of wild type and secondary wall cesa mutants across the length of primary shoot which was fond to be rather uniform in 7-week-old plants. Additionally, we found that the cellulose content of single mutants was comparable to the higher order mutants.ConclusionsHere we describe a medium-throughput adaptation of Updegraff’s method that allowed us to determine cellulose content of 200 samples each week.
Highlights
Lignocellulosic biomass is an important renewable resource for biofuels and materials
Arabidopsis provides an excellent model system to study cellulose biosynthesis and a vast amount of progress has been made in the identification of genetic components of Arabidopsis cellulose biosynthesis machinery
Cellulose is synthesised by the cellulose synthase complex (CSC) particles moving in the plasma membrane [1,2,3,4]
Summary
By using aspiration rather than centrifugation to remove liquids during various stages of protocol, we were able to increase the throughput of the method as well as minimise the sample loss. We found that the cellulose content of single mutants was comparable to the higher order mutants
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