Evaluating methiozolin programs for golf putting greens and investigating potential modes of action

Annual bluegrass is a winter annual grass that is problematic on golf putting greens due to its light green color, prolific seedhead production and intolerance to stress. On creeping bentgrass putting greens, herbicides for annual bluegrass control are limited. A new herbicide, methiozolin, developed by Moghu Research Center, LLC, in Daejeon, South Korea, safely and selectively controls annual bluegrass in creeping bentgrass and several other turfgrass species. Methiozolin typically controls annual bluegrass over several weeks, allowing desirable turfgrass time to grow into areas previously infested by annual bluegrass with little surface disruption. The mode of action of methiozolin is unknown, but has been proposed to act as either a cell wall biosynthesis inhibitor (CBI) or an inhibitor of tyrosine aminotransferase (TAT). Field studies were conducted at Virginia Tech to investigate strategies promoting surface recovery on putting greens following atypically rapid annual bluegrass loss resulting from methiozolin application, intensive core-cultivation as well as potential interactions with plant growth regulators (PGR's), like ethephon. In the rapid annual bluegrass removal study, all treatments receiving additional fertility via synthetic fertilizer with or without trinexapac-ethyl or biostimulant recovered 1 to 3 weeks more quickly than treatments that did not include additional fertility. Addition of the PGR trinexapac-ethyl inconsistently regulated speed of canopy recovery, both increasing and decreasing recovery speed. Under normal maintenance conditions, methiozolin does not negatively influence putting green recovery, however, if the putting green is exposed to droughty conditions, methiozolin can reduce recovery time by several weeks. Core-cultivation should be avoided in conjunction with methiozolin and ethephon applications because when this procedure was conducted on the same day as herbicide application it significantly damaged creeping bentgrass, reducing cover to 19% at 2000 g ai ha⁻¹, compared to the non-treated at 62%. Regarding the question of methiozoling mode of action, laboratory studies supported the claim that addition of exogenous 4-hydroxyphenylpyruvate (4-HPP) alleviates symptoms of methiozolin exposure in lesser duckweed, a model monocot species, but feeding various turfgrass species and annual bluegrass exogenous 4-HPP did not alleviate symptoms. Creeping bentgrass secondary root length and density was not affected by methiozolin, although annual bluegrass, Kentucky bluegrass and perennial ryegrass secondary root lengths were reduced. Based on these data, it does not appear that TAT inhibition is a primary mode of action of methiozolin in turfgrass. Studies were conducted to determine if methiozolin inhibited cell wall biosynthesis in desirable turfgrass species and annual bluegrass. All species exhibited decreased enrichment of ¹³C in cell-wall sugars form ¹³C-glucose in response to methiozolin and a known cell wall biosynthesis inhibitor, indaziflam. Indaziflam and methiozolin at 0.01 µM inhibited ¹³C enrichment of all sugars less than methiozolin at 1.0 µM, for xylose, arabinose and glucose, but not galactose. Addition of 4-HPP increased incorporation of ¹³C into xylose, but had no other influence on ¹³C incorporation into other cell wall sugars. Lack of species specific response indicates that cell wall biosynthesis inhibition is probably not the source of interspecific species responses observed in the field.