The herbicide clomazone caused ultrastructural damage to etioplasts of cotton cotyledons. Etioplast envelopes were irregular, prothylakoids were absent or irregular, stroma density was low, and there were abnormal stromal vesicles. Further damage occurred upon exposure to light. Clomazone greatly slowed the conversion of chlorophyllide to chlorophyll in cotton, suggesting that phytol synthesis was affected. Neither synthesis of protochlorophyllide nor phototransformation of protochlorophyllide to chlorophyllide was affected by clomazone. Clomazone completely inhibited carotenoid synthesis without an accumulation of phytoene. However, the sesquiterpenoids hemigossypol and the dimeric sesquiterpenoid gossypol accumulated in greater amounts in clomazone-treated than in control tissues. These results indicate that this herbicide inhibits synthesis of terpenoids after the formation of farnesyl pyrophosphate.

Laboratory assays were conducted to determine the sensitivity of tomato and tobacco cell suspension cultures and tomato and pepper cotyledons to clomazone. A comparison of fresh weight and carotenoid content indicated up to a three-fold difference between the clomazone-tolerant tobacco and clomazone-susceptible tomato cell suspension cultures. In contrast, an approximate 60-fold difference between the tolerant pepper and susceptible tomato cotyledons was observed when total chlorophyll and carotenoid contents were measured. The effect of clomazone and its possible metabolites on in vivo and in vitro extractable IPP isomerase (EC 5.3.3.2) and prenyltransferase (EC 2.5.1.29) activity was investigated. There was no clear inhibitory effect of clomazone or possible clomazone metabolites upon enzyme activity in tomato or tobacco cell suspension cultures or on light or dark grown tomato or pepper cotyledons. No specific enzymatic target site of clomazone was identified in correlation with the reduction in total chlorophyll or carotenoid content.

Cultures of Streptomyces tendae grown on Actinomyces medium containing clomazone contained less geosmin 48 h after inoculation than untreated controls, suggesting a possible inhibition of reactions prior to sesquiterpenoid biosynthesis. However, exposure to clomazone increased geosmin accumulation in S. tendae cultures grown on Hickey-Tresner medium and in cultures of the fungus Penicillium expansum. Additionally, clomazone increased accumulation of a sesquiterpenoid tentatively identified as a eudesmol in S. tendae, indicating possible point(s) of inhibition subsequent to sesquiterpenoid biosynthesis. Geosmin biosynthesis was induced by transferring cultures of S. tendae inoculated on Actinomyces medium to Hickey-Tresner medium 24 h after inoculation. When harvested at 48 h, geosmin biosynthesis-induced-cultures exposed to clomazone contained more geosmin than untreated controls, indicating that response of a single taxon to clomazone can be rapidly altered by changing medium composition.

Experiments were conducted at Lexington and Quicksand, KY, in 1989 and 1990 to determine the effect of preplant incorporated clomazone on weed control, crop injury, and yield of broccoli, cauliflower, green cabbage, red cabbage, and pak choi. Oxyfluorfen and trifluralin were included as standard treatments. Greater than 80% weed control was observed at both locations with 0.8 kg ai ha−1 clomazone, with the exception of smooth pigweed at Lexington. All clomazone treatments caused crop injury 2 WAT. Injury was still evident 8 WAT at clomazone rates of 1.7 and 3.4 kg ha−1, but was minor with rates of 0.8 kg ha−1 or less on broccoli, cauliflower, and red cabbage. Injury was less at Quicksand in both years and seasons over all clomazone rates, most likely due to higher soil organic matter content Yields of broccoli and cauliflower treated with 0.8 kg ha−1 clomazone were similar to yields with oxyfluorfen on a 3% organic matter soil at Lexington in 1989. However, clomazone at 0.8 kg ha−1 at Lexington reduced 1989 spring yields of green cabbage, red cabbage, and pak choi and 1990 spring yields of all cole crops as compared to oxyfluorfen. Clomazone at 0.8 kg ha−1 at Quicksand reduced yield of green cabbage and pak choi in spring 1990 only on a 5.2% organic matter soil. Fall yields of broccoli and cauliflower in both years were not reduced by clomazone at 0.8 kg ha−1 or less at either location. Our studies indicated potential for utilization of clomazone on cole crops in higher organic matter soils, especially if some early season crop injury and occasional yield loss can be tolerated.

Concentrations of clomazone in nutrient solution causing 50% reduction in shoot fresh weights were 325 and 2 μM for soybean and velvetleaf, respectively. Root uptake of 14C–clomazone from nutrient solution and translocation to shoots were similar for the two species. Soybean and velvetleaf contained similar amounts of 14C per plant; however, because velvetleaf had lower tissue weights, concentration of 14C was higher in velvetleaf. Clomazone did not accumulate in roots but was rapidly translocated to shoots of both species. No more than 30% of absorbed clomazone remained in roots at any time in the experiment. Soybean and velvetleaf leaves contained 4.2 and 1.7 nmol clomazone g−1 fresh weight, respectively, and 3.7 and 6.1 nmol clomazone metabolites g−1 fresh weight, respectively. Thus, velvetleaf metabolized clomazone more rapidly than soybean did. Treatment of clomazone metabolites with β-glucosidase demonstrated that metabolism occurred by oxidative cleavage and conjugation of the benzyl moiety in both plant species. One aglycone cochromatographed with 2–chlorobenzylalcohol. The results suggest that reduced absorption or translocation or increased detoxication of clomazone by soybean seedlings is probably not responsible for their tolerance to this herbicide.

A laboratory bioassay was conducted to determine the differential tolerance of cole crops to clomazone as measured by extractable total chlorophyll and carotenoids. Clomazone concentrations causing 50% inhibition (I50) in the biosynthesis of total chlorophyll in broccoli, cauliflower, and green and red cabbage cotyledons were 16, 11, 3, and 11 μM, respectively, while I50 values for carotenoid levels were 20, 10, 4, and 8 μM clomazone, respectively. Therefore, broccoli was the most tolerant to clomazone based upon extractable chlorophyll and carotenoid concentrations. Further laboratory studies were performed to investigate the basis for differential clomazone tolerance in 3-wk-old cole crop seedlings. No differences in total root uptake of 14C-clomazone were observed between these crops after 24 h. There were no differences in rate of metabolism of 14C-clomazone to methanol-soluble metabolites in roots of these crops. Percentage of polar metabolites in roots remained fairly constant over time. There were also no differences between crops in percentage of methanol-soluble 14C-clomazone metabolites formed in shoots between 24 and 96 h. In all crops, levels of 14C-clomazone decreased in a similar manner over time in methanolic extracts of roots and shoots while nonextractable 14C levels increased, indicating a conversion of clomazone to insoluble, nonextractable forms. Differential uptake, translocation, and metabolism do not appear to account for clomazone selectivity differences between cole crop seedlings.

Pronounced differences in the tolerance of tomatoes and bell peppers to clomazone observed in field studies were confirmed in a greenhouse experiment. In greenhouse studies, preemergence clomazone rates causing 50% visible injury on bell pepper and tomato seedlings 10 days after application were 9.4 and 0.1 kg/ha, respectively. Based on growth inhibition, bell peppers were 40-fold more tolerant of clomazone than tomatoes 20 days after clomazone application. In laboratory studies investigating the basis for differential clomazone tolerance, no differences in uptake of 14C-clomazone from nutrient solutions between tomato and bell pepper plants were observed after 24 h. Minor differences were observed in the distribution of 14C label within plants; a higher percentage of 14C was recovered in bell pepper roots than in tomato roots, while the opposite was true for the shoots. Clomazone was metabolized to two products in roots of both bell peppers and tomatoes within 48 h after treatment. Tomato shoots were more active in converting clomazone to these metabolites than were tomato roots. Bell pepper roots converted more clomazone to metabolites than did tomato roots 24 h after treatment. However, by 72 h, differences in clomazone metabolite levels between species were negligible in both roots and shoots. Enzymatic and acid hydrolysis of soluble, polaf clomazone metabolites indicated that these metabolites may be sugar conjugates of clomazone.

Experiments were conducted in 1985 to 1987 to evaluate the effects of conventional and no-tillage systems on the weed control provided by clomazone applied preemergence in soybeans. The persistence of clomazone in soil of the two tillage systems was also determined. Increasing the clomazone rate from 0.8 to 1.4 kg/ha did not increase weed control. Clomazone controlled 80% or more of jimsonweed, velvetleaf, and giant foxtail. Common cocklebur control ranged from about 50 to 70% in no-till and from 80 to 90% in conventional tillage. Generally, soybean pods/plant and yields were lower from clomazone treatments than from handweeded treatments due to inadequate common cocklebur control. Over 40% of the clomazone applied did not reach the soil surface; it was either intercepted by wheat straw, volatilized, or both. Clomazone persisted longer in conventional tillage than in no-tillage in. However, in 1986, clomazone was equally persistent in the two tillage systems. The half-life of clomazone was 34 and 6 days in 1985 in conventional and no-tillage, respectively, and in 1986, 18 and 16 days in conventional and no-tillage, respectively. Significant clomazone concentrations were not found below 10 cm in the soil profile. Corn planted without tillage (no-till) approximately 1 yr after clomazone application was not injured and yields were not reduced due to prior clomazone use.

Field trials were conducted in 1985 and 1986 to determine the effect of incorporation on volatilization of clomazone from soil. Volatilization was detected up to 2 weeks after surface-applied or soil-incorporated treatments of clomazone at 1.1 kg ai/ha. The amount of volatilization detected was greatest following rainfall and varied between years. More clomazone volatilized after surface application than after incorporation, regardless of the climatic conditions present. Clomazone volatilization detected was in the order of no-tillage > minimum tillage > conventional tillage.

Based on chlorophyll content, hydroponically cultured soybean seedlings were 254, 66, and 13 times more tolerant to clomazone than velvetleaf, corn, and smooth pigweed, respectively. Clomazone, at concentrations that inhibited chlorophyll, did not affect fresh weight accumulations of any species except velvetleaf. However, in velvetleaf, fresh weight accumulation was only half as sensitive to clomazone as the leaf chlorophyll content. Uptake of 14C-clomazone from nutrient solution by 72 h after treatment (HAT) (pigweed > velvetleaf > soybean > corn) indicates that differential absorption cannot account for selectivity. Shoot:root ratios of 14C recovered from soybean, corn, velvetleaf, and pigweed by 72 HAT were 0.39, 0.84, 1.67, and 2.37, respectively. The limited acropetal clomazone translocation in soybean seedlings may account to a small degree for soybean tolerance to clomazone. Conversion of clomazone to more polar metabolites was rapid in all four species. There were no significant differences among species in the percentage of 14C activity recovered as clomazone from root tissue by 72 HAT. Of the 14C activity recovered from shoots of soybean, corn, pigweed, and velvetleaf seedlings by 72 HAT, 46, 59, 35, and 54%, respectively, was clomazone. Differences in clomazone uptake, distribution, and metabolism among the four species were either insignificant or poorly correlated to selectivity, and therefore cannot account for the tremendous differences in clomazone sensitivity among these species. These observations indicate, indirectly, that differences at the site of action may account for selectivity.