The activity of glyphosate [N-(phosphonomethyl)glycine], formulated as the isopropylamine salt, on in vitro photosynthesis was investigated. When pH 4.7 glyphosate solutions were titrated to a pH equal to that of the reaction media (pH 7.8), glyphosate additions had no effect on whole chain electron transport between coupled photosystem II (PS II) and photosystem I (PS I) in stroma-free chloroplast thylakoids from peas (Pisum sativum L. ‘Morse's Progress No. 9′). Inhibition did not occur even after a 2-h dark incubation of lamellae in a 5-mM solution of glyphosate. Fluorescence studies failed to detect an effect of glyphosate on PS-II mediated electron transport processes or upon light harvesting properties of PS II even after a 2-h glyphosate/chloroplast preincubation. Glyphosate had no effect on cyclic and noncyclic photophosphorylation even after a 100-min dark incubation of chloroplast membranes in a 5-mM solution of glyphosate. Based on these assays it is concluded that glyphosate has no direct effect on the photochemical reactions of photosynthesis when the pH of the active compound is adjusted to that of the reaction mixture prior to addition to a chloroplast suspension.

Thylakoid membranes isolated from chloroplasts of atrazine [2-chloro-4-(ethylamino)-6-(isopropylamino)-s-triazine]-susceptible (S) and -resistant (R) biotypes of common groundsel (Senecio vulgaris L.) were assayed to determine the effect of diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea], bromacil (5-bromo-3-sec-butyl-6-methyluracil) and several s-triazine herbicides on photosystem II (PSII) activity. Each herbicide inhibited photosynthetic electron transport on the reducing side of PSII in thylakoid membranes from the susceptible biotype. Thylakoids isolated from the resistant biotype were 60 to 3200 times more tolerant to the inhibitors than thylakoids from the susceptible biotype. Electrophoretic analysis revealed no differences in molecular weight of membrane polypeptides of either common groundsel biotype. Three-hour dark incubation did not enhance inhibitor activity in the resistant-type thylakoid membranes. Specific structural or configurational changes associated with the reactive site may account for the differences in Hill reaction inhibition between biotypes. A relationship between herbicide structure and Hill reaction activity is discussed.

Analysis of reciprocal F1, F2, F3, and backcross progeny from crosses between atrazine [2-chloro-4-(ethylamino)-6-(isopropylamino)-s-triazine]-resistant and several susceptible genotypes of Brassica campestris L. indicated that triazine resistance was inherited cytoplasmically from the female parent. A partially dominant chlorotic cotyledon was used as a genetic marker to confirm true hybrids in the reciprocal F1 progenies. Phytotoxicity of atrazine was correlated with loss of the Hill reaction.

The susceptibility of sweet pepper (Capsicum annuum L. ‘Keystone Resistant Giant’) and tolerance of hot pepper (Capsicum chinense L. ‘Bohemian Chili’) to bentazon [3-isopropyl-1H-2,1,3-benzothiadiazin-4(3H)-one 2,2-dioxide] was demonstrated under greenhouse and field conditions. Sweet pepper growth and fruit production were inhibited by foliar applications of bentazon at rates of 0.6 to 6.7 kg ai/ha. Severity of injury increased with application rate. Injury symptoms in sweet pepper were leaf chlorosis, with necrosis and death of shoot apices. Foliar applications of bentazon to hot pepper resulted in little or no injury. Bentazon inhibited Hill reaction activity of isolated chloroplasts from both species to a similar extent, suggesting that selectivity of this compound in hot pepper is not due to resistance at the chloroplast level.

Insensitivity to atrazine [2-chloro-4-(ethylamino)-6-(isopropylamino)-s-triazine] and certain other herbicides was found in plants of horseweed (Conyza canadensis (L.) Crong. ♯4 ERICA) collected from vineyards where atrazine had been applied extensively. There were no differences in the uptake, translocation, or accumulation of radiolabeled atrazine between plants of the susceptible and resistant biotypes. However, atrazine at 1 × 10-4 M caused no inhibition of photosynthetic electron transport in resistant leaves. Photosynthetic characteristics of leaves of susceptible and resistant plants showed different sensitivities to atrazine, but similar sensitivities to diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea] and ioxynil (4-hydroxy-3,5-diiodobenzonitrile). Isolated chloroplast membranes from resistant plants exhibited a resistance factor of 1000 to atrazine but only 10 to 30 to methylthio-s-triazines. DNOC (4,6-dinitro-o-cresol) was found to be more effective in susceptible chloroplasts than in resistant ones. A marked reduction in the galactolipid content, especially digalactosyl diglyceride, was found in resistant leaves. It is suggested that these alterations in lipid content and composition may contribute to atrazine resistance in horseweed.

The effect of chlorotoluron on different slender foxtail populations that survived normal agricultural rates of this herbicide (2.5 to 3.5 kg ai ha–1) was investigated under controlled laboratory conditions. Among five populations tested, two had tolerance for the herbicide. The ED50 values for these biotypes ranged from 0.33 to 2.43 kg ha–1. Assays with radiolabeled 14C-chlorotoluron indicated similar absorption and translocation of the herbicide between susceptible and resistant biotypes. Chlorophyll fluorescence and Hill reaction analysis seem to support the view that the mechanism of resistance of one biotype to the herbicide is due to degradation/detoxification rather than to modification of the target site, although compartmentation processes cannot be ruled out. This resistant biotype had high tolerance to other photosynthetic inhibiting phenylureas, including linuron, chlorbromuron, diuron, monolinuron, and neburon.

A triazine-resistant (R) biotype of a Graminaceous weed, Brachypodium distachyon (= Trachynia distachya), was found in the lower Galilee area of Israel after repeated treatments of roadsides with simazine [2-chloro-4,6-bis (ethylamino)-s-triazine] and atrazine [2-chloro-4-(ethylamino)-6-(isopropylamino)-s-triazine]. It was not affected at levels up to 3.0 kg/ha of soil-incorporated atrazine or simazine. The (R) biotype was unaffected by atrazine, as ascertained by measuring whole-leaf and plastid fluorescences and the Hill reaction. The triazine-sensitive (S) biotype was inhibited in all these reactions. The data suggest that the (R) biotype has a plastid mode of resistance common to many weed species. The (R) biotype metabolized 14C-(ring)-atrazine more rapidly to both water-soluble and to chloroform-soluble compounds than the (S) biotype did, but that could not explain the immediate resistance at the plastid level. Probably there was evolution towards increased tolerance by detoxification as well as plastid level resistance.

A simple and quick method is described for identifying biotypes of common groundsel (Senecio vulgaris L.), common lambsquarters (Chenopodium album L.), and smooth pigweed (Amaranthus hybridus L.) that are resistant or susceptible to triazine herbicides. Leaf discs (3 mm diam) from the tested plants were vacuum infiltrated with a phosphate medium and exposed to a 3-klux light source. When 1.0 × 10−5M and higher levels of atrazine (2-chloro-4-ethylamino-6-isopropylamino-s-triazine) were present, the leaf discs from susceptible biotypes failed to float to the surface, whereas the leaf discs from resistant biotypes floated within 1 h. The urea herbicide, fluometuron [1,1-dimethyl-3-(α,α,α-trifluoro-m-tolyl)urea] inhibited the floating of leaf discs from both the triazine-susceptible and triazine-resistant biotypes. This method can be used to evaluate large numbers of plants to determine if they are resistant or susceptible to triazines without destroying the plants or preventing seed production.

Triazine-resistant (R) biotypes of common lambsquarters, yellow foxtail, and hairy fleabane were found in the Province of Córdoba (Andalusia, Southern Spain). The former two R biotypes came from atrazine-treated cornfields, whereas the latter came from simazine-treated nontilled olive orchards. The R biotypes of common lambsquarters and yellow foxtail and the R biotype of hairy fleabane survived at doses up to 5 kg ai/ha of soil-applied atrazine or simazine, respectively. Photosynthetic electron transport in R biotypes was unaffected by atrazine and simazine but was inhibited by diuron, as shown by fluorescence induction measurements in whole leaves. In Hill reaction assays, the R biotypes showed high resistance to atrazine and simazine (resistance factors in the range of 350 to 550), medium to high resistance to ametryn, terbumeton, metribuzin, and monolinuron (resistance factors in the range of 80 to 250), slight resistance to diuron and methabenzthiazuron (resistance factors in the range of 1.1 to 15.7), and reverse resistance to swep, ioxynil, and DNOC (resistance factors less than 1). It is concluded that the R biotypes have a chloroplast mode of resistance similar to that previously described for other triazine-resistant weed biotypes.

Biotypes of several annual broadleaved weeds tolerant to 2-chloro-s-triazines have been recorded, including common lambsquarters (Chenopodium album L.). The mechanism of resistance in common lambsquarter was based on the differential inhibition of the Hill reaction in chloroplasts by atrazine [2-chloro-4-(ethylamino)-6-(isopropylamino)-s-triazine]. Chlorophyll fluorescence and electron transport assays were used with isolated chloroplasts of atrazine-tolerant and susceptible biotypes, to determine the effect of atrazine and diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea] on photosystem II (PS II) activity, differential atrazine penetration of the chloroplast envelope, and relative tolerance to chloro, methoxy, and methylthio triazines. Atrazine and diuron inhibited electron transport on the reducing side of PS II in susceptible biotype chloroplasts. In tolerant biotype chloroplasts only diuron inhibited electron transport whereas atrazine had only slight effects. There were no differences in the chloroplast membrane permeability to atrazine in the two biotypes. Chloroplasts of the atrazine-tolerant biotype of common lambsquarters were also tolerant to the other classes of triazines tested.

A downy brome population in a winter wheat field at Córdoba, Spain, survived use rates of chlortoluron (2.5 to 3.5 kg ai ha−1) over 2 consecutive yr, where wheat monoculture and multiple annual chlortoluron applications had been carried out. The resistant (CR) biotype showed a higher ED50 value (7.4 kg ai ha−1; the concentration required for 50% reduction of fresh weight) than the susceptible (S) control (2.2 kg ai ha−1), with a 3.4-fold increase in chlortoluron tolerance. Chlortoluron resistance in the CR downy brome biotype was not caused by altered absorption, translocation, or modification of the herbicide target site but by enhanced detoxification. The inhibition of both the recovery of photosynthetic electron transport and chlortoluron metabolism in the CR biotype due to the presence of the Cyt P450 inhibitor 1-aminobenzotriazole (ABT) indicates that herbicide metabolism catalyzed by Cyt P450 monooxygenases is related to chlortoluron resistance in CR plants. Although both biotypes degraded chlortoluron by N-dealkylation and ring-methyl hydroxylation and seem to share the same ability to form polar conjugates, degradation in the resistant biotype is more efficacious as this biotype metabolizes the parent herbicide faster and to a greater extent than its susceptible counterpart. The ability of the susceptible biotype to ring-hydroxylate chlortoluron, albeit at much slower rate, probably explains its moderate tolerance to chlortoluron observed in the growth assays and its minor photosynthetic electron transport recovery observed in fluorescence measurements.