U.S. flagAn official website of the United States government
                    Here's how you know **Official websites use .gov**
                            
                            A **.gov** website belongs to an official government organization in the United States.
 **Secure .gov websites use HTTPS**
                            
                            A **lock** (                            
                              A locked padlock                                                      
                            
                            ) or **https://** means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites.


Menu Agricultural Research Service
						
                    U.S. DEPARTMENT OF AGRICULTURE
 Wheat, Sorghum and Forage Research: Lincoln, NE
 close



    Research

                    Research Projects
                                    Publications
                                    Collaborations
                                    News
                People

                    Staff Listing
                                    CareersSearch small
                    Search
            
            ARS Home
                                About ARS
                                Contact Us




ARS Home Plains Area Lincoln, Nebraska Wheat, Sorghum and Forage Research Research Publications at this Location





- ARS National Programs
- Search for a scientific manuscript
- Search for a research project
- ARS Software
- Programs and Projects
- Collaborations
- Subjects of Investigation
- Publications

Title:   Author  item
                                            Palmer, Nathan

                                                         - *Nate*item
                                            Sattler, Scottitem
                                            Saathoff, Aaronitem
                                            Sarath, Gautam
  Submitted to: Biotechnology for Fuels and Chemicals Symposium Proceedings
                                    Publication Type: Abstract Only
                                    Publication Acceptance Date: 4/19/2010
                                    Publication  Date: 4/19/2010
                                    Citation: Palmer, N.A., Sattler, S.E., Saathoff, A.J., Sarath, G. 2010. A Fluorescent Assay for Plant Caffeic Acid O-methyltransferases. Abstract only - Poster presented at the Biotechnology for Fuels and Chemicals Symposium Proceedings in Clearwater, FL, April 19-22, 2010.  Not published elsewhere.Interpretive Summary: We have developed and validated a new enzymatic assay to measure the activity of caffeic acid-O-methyltransferases (COMT) in plants.  Using this method, the native activity in several switchgrass plants differing in lignin content has been documented.  This method will have utility in routinely querying switchgrass plants for COMT activity within a breeding program.
                                Technical Abstract: We have developed a facile, sensitive and continuous assay to measure the activities of plant COMTs using s-adenosyl homocysteine hydrolase as a coupling enzyme and and adeonsine a thiol-specific fluor, Thioglo1, as the detecting reagent. This assay was validated using recombinant sorghum COMT (BMR-12) a key enzyme involved in cell wall lignification, and used to analyze COMT activity in maturing tillers from switchgrass plants.  Data indicated that the calculated Km, Vmax and Kcat values for the recombinant sorghum COMT using different substrates in the fluorescent assay were similar to published values for COMT enzymes isolated and characterized from other plant species using other methods.  For switchgrass, native COMT activity and COMT transcript abundance was greatest in internodes at the top of a tiller and declined in the more basal internodes, although COMT protein as determined by immunoblotting of internode extracts did not appear to change significantly between the internodes along a switchgrass tiller. We have used this assay to determine the level of COMT activity in switchgrass genotypes differing in lignin content. Data on the correlations between activity, protein content and transcript abundance for several switchgrass genotypes will be presented.


Return to top


Connect with ARS Sign up


ARS Home
                            USDA.gov
                            Plain Writing
                            Policies & Links
                            Civil Rights Statements
                            FOIA
                            Accessibility Statement
                            Privacy Policy
                            Non-Discrimination Statement
                            Quality of Information
                            USA.gov
                            WhiteHouse.gov