|Mcgrath, J Mitchell - Mitch|
Submitted to: Journal of Agricultural and Food Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/5/2004
Publication Date: 10/16/2004
Citation: Trebbi, D., McGrath, J.M. 2004. Fluorometric sucrose evaluation for sugar beet. Journal of Agricultural and Food Chemistry. 52(23):6862-6867.
Interpretive Summary: Sucrose is the primary economic product from sugar beet. Measurement of sucrose in beets is currently a laborious and time consuming process, which hinders rapid evaluation of sugar beet roots for their sucrose content. A technique called EFA was developed that will allow more efficient sucrose content determinations. EFA uses well known enzymatic chemistry in a miniaturized format of microtiter plates with a highly sensitive fluorescent detection chemistry to determine sucrose content in dry sugar beet root tissue. The accuracy of EFA was shown to be comparable to the gold standard of High Performance Liquid Chromatography. Breeders and agriculturalists now have a new highly sensitive method to examine sucrose content in sugar beet roots that can be accomplished more rapidly and with greater precision than current methods. The method is applicable to dried root tissues from sugar beet roots of any age, and scientists will now be able to use the method to follow sucrose accumulation during plant development as well as examine the inheritance of sucrose content as a proportion of root dry matter.
Technical Abstract: Sucrose is the economic product from sugar beet. Disease resistance is often available in low sucrose genotypes and, prior to deploying such novel genes as available into the cultivated spectrum, selection for increased sucrose content is required during introgression. The objective of this work was to evaluate a relatively rapid and inexpensive enzymatic-fluorometric microtiter plate assay for sucrose quantification in sugar beet root dry matter, both for progeny testing in the greenhouse and evaluation of field-grown mother roots. As determined using HPLC, sucrose content in diverse populations of sugar and table beet assayed over various developmental stages ranged from 0.213 to 2.416 mmol g-1 dry matter, and these values were used as references for both refractometry and enzymatic-fluorometric assay. As expected, refractometric analysis generally overestimated sucrose content. Enzymatic-fluorometric analyses were reasonably well correlated with HPLC results for young greenhouse-grown root tissues (R2 = 0.976), and less so with older field-grown roots (R2 = 0.605), for unknown reasons. Enzymatic-fluorometric assays may be best deployed for progeny testing young seedlings.