Distribution of 14C-glyphosate in legumes parasitised by Orobanche crenata

Absorption and translocation of ¹⁴C-glyphosate was evaluated under controlled conditions in peas ( Pisum sativum L.) and broad beans ( Vicia faba L.) parasitised or not by crenate broomrape Orobanche crenata Forsk.). Absorption increased with time up to 12 days after treatment, and reached about 50% of the ¹⁴C-glyphosate applied. Three days after treatment 70–85% of the total herbicide absorbed had been translocated out of the treated leaflet. There was no consistent differ ence in absorption and translocation between infected and non-infected plants 12 days after treatment. The ¹⁴C-glyphosate concentration in the root system increased with time in broad beans. In peas it remained more or less constant from 3 days after treatment. Generally, pods were stronger sinks for glyphosate than other parts of the plant. Herbicide accumulation in broomrape increased with its growth stage, and the parasite was a much stronger sink for glyphosate than the legume root system.     

Uptake and metabolism of hexazinone in Rubus hispidus L. and Pyrus melanocarpa (Michx.) Willd

Rooted cuttings of Pyrus melanocarpa (Michx.) Willd. had a 3.5-fold greater tolerance to soil-applied hexazinone than those of Rubus hispidus L. in a greenhouse trial. R. hispidus accumulated four times more ¹⁴C-label in the foliage following root uptake of ¹⁴C-hexazinone than P. melano-carpa . Here, the greater uptake and susceptibility of R. hispidus was related to its greater root:fol-iage (weight) ratio compared to the tolerant P. melanocarpa . However, in whole plant metabolism studies employing younger plants there were no differences in ¹⁴C accumulation in the leaves. Here ¹⁴Chexazinone was converted to a number of hydroxylated and/or demethylated triazinone metabolites in both species following vacuum infiltration into leaf disks and root sections, or following root uptake in whole plants. A major difference in metabolism between the two species was the greater formation of the mono demethylated metabolite, B, [3-cyclohexyl-6-methylamino-l-methyl-l,3,5-triazine-2,4-dione] in P. melanocarpa which may also contribute to its greater tolerance. A loss of total ¹⁴Clabel from all plant parts with time may reflect cleavage of the ¹⁴C-ring-labelled herbicide.     

Root exudation of diclofop-methyl and triasulfuron from foliar-treated durum wheat and ryegrass

In this study, we evaluated the release of diclofop-methyl and triasulfuron from the roots of foliar-treated ryegrass and wheat. The study with ¹⁴C-diclofop-methyl indicated a basipetal translocation of foliar-applied herbicide in wheat and ryegrass. No root exudation from ¹⁴C-diclofop-methyl-treated wheat plants was observed, while 20 days after treatment (DAT) 0.2–0.9% of radioactivity absorbed by ryegrass was found exuded in the growing medium. Root exudation was stimulated three to six times by the presence of untreated wheat or ryegrass sharing the growing medium with diclofop-methyl-treated ryegrass. No subsequent uptake of exuded radiolabel by untreated plants (ryegrass or wheat) in the same pot with ¹⁴C-diclofop-methyl-treated ryegrass was observed. The study with ¹⁴C-triasulfuron indicated a basipetal translocation of foliar-applied herbicide in wheat and ryegrass and also into the growing medium. By 20 DAT, 0.5–4.2% of radioactivity absorbed by wheat or ryegrass was found exuded in the growing medium. The presence of untreated plants (wheat or ryegrass) in the same pot as triasulfuron-treated ryegrass or wheat induced exudation seven to 32 times more. The study also revealed a subsequent uptake of exuded compounds by untreated wheat or ryegrass sharing the medium of ¹⁴C-triasulfuron-treated plants. This study has demonstrated for the first time that the root exudation of exogenous compounds can be related to plant arrangement in pots. The implication is that herbicide root exudation and transfer, a form of allelopathy, could be significant in the field. A precise estimation of environmental fate, unexpected ecological side effects and residual activity of herbicides may require quantification of such exudation.     

TRANSLOCATION OF AMINOTRIAZOLE AND DALAPON IN AG ROPY RON REPENS (L.) BEAUV.

Summary. Tracer studies using single drops of solutions containing 3–amino-1,2,4–triazole-5–¹⁴C (aminotriazole-¹⁴C) or 2,2–dichloropropionic acid-2–¹⁴C (datapon-¹⁴C) revealed that in couch plants (Agropyron repens (L.) Beauv.) growing under field conditions in the autumn and at the stage where the aerial shoots were 40–50 cm long, both compounds moved in both symplast and apoplast. Dalapon was less mobile in the symplast than aminotriazole and only negligible amounts of dalapon were translocated to the rhizomes. The nodes of the treated shoots appeared to act as barriers to translocation, a phenomenon more pronounced for dalapon than for aminotriazole.
Application to a basal green leaf led to a more uniform distribution of the compounds within plants and rhizomes than when the application was made to the youngest fully-expanded leaf.
In couch plants with aerial shoots 10–15 cm long treated in the stubble, the distribution of both aminotriazole and dalapon was mainly restricted to the treated shoots. Even 15 days after application only trace amounts of radioactivity could be found in the rhizomes and untreated shoots.     

Phytotoxic activity of root absorbed glyphosate in corn seedlings (Zea mays L.)

Growth chamber experiments were conducted in order to study the absorption, translocation and activity of glyphosate when applied to roots with aqueous solution avoiding any glyphosate–substrate interaction. Corn seedlings at the first leaf stage were set up in individual graduated cylinders containing different solutions of ¹⁴C-glyphosate (0–30 mg ae kg⁻¹). After 26 h of root exposure, plants were transferred to fresh nutrient solution and grown for the next 5 days. After harvest, plants were separated into seed, root, mesocotyle, coleoptile, cotyledon, first leaf and all new leaves (apex), and quantified ¹⁴C radioactivity contained in each part. Glyphosate uptake was only 11% of the theoretical mass flow into the plant. The amount of glyphosate translocated from roots was positively correlated with plant uptake ( P  < 0.01). Total plant fresh weight presented a logistic response to glyphosate amounts, including a growth stimulant effect (hormesis), when plants absorbed less than 0.6 µg. The treated plants presented a normal pattern of glyphosate allocation, with the apex the principal sink, accumulating more than 38% of mobilized glyphosate. When corn plants absorbed more than 0.6 µg they showed a decrease in growth. The relatively high glyphosate quantities allocated in the new leaves showed the relevance of the symplastic pathway in the translocation process for root absorbed glyphosate.     

Effects of the safeners CGA 154281, oxabetrinil and fenclorim on uptake and degradation of metolachlor in corn (Zea mays L.) seedlings

Summary. The influence of three crop safeners on the uptake and degradation of ¹⁴C-metolachlor was investigated in two corn varieties. Following application of herbicide and safener together to seedling shoots the concentrations of non-metabolized ¹⁴C-metolachlor in the tissues was found to be lower in the tolerant variety LG 9 than in the susceptible variety 211A. The difference between varieties was due to differences in both uptake and degradation of ¹⁴C-metolachlor.
Following shoot application most of the radioactivity was retained in the coleoptile and the mesocotyl. Two hours after application 95% of the herbicide had been degraded in coleoptiles and mesocotyls, whereas approximately 20% of non-metabolized ¹⁴C-metolachlor was present in the enclosed developing shoot leaves. In both corn varieties the safener CGA 154281 caused a substantial lowering of tissue levels of parent ¹⁴C-metolachlor. This was primarily due to an enhanced degradation. Glutalhione- S -transfer-ase (GST) enzyme activity in shoot tissues was found to be enhanced in both varieties by CGA 154281. Oxabetrinil and fenclorim were less effective than CGA 154281 both in reducing tissue levels of non-metabolized ¹⁴C-metolachlor and in enhancing GST activity in either variety.     

SHOOT ZONE UPTAKE OF SOIL-APPLIED HERBICIDES IN PISUM SATIVUM L.

Summary. The effects of localized herbicide placement at different internodes of pea ( Pisum sativum L. cv. Alaska) shoots below the soil surface after plant emergence were studied by removing the soil from around the shoots and replacing with herbicide-treated soil. The first internode proved insensitive to linuron, diuron, atrazine and simazine at 4·5 kg/ha, while treatment of the second and third internodes markedly reduced plant growth 4–6 days after treatment. Separate exposure of the first internode alone, and the second and part of the third together to ¹⁴C-labelled atrazine indicated no difference in herbicide metabolism. However, a two- to threefold increase in ¹⁴C uptake and movement to the foliar parts occurred when the second and part of the third internode was treated, as compared to first internode treatment. Thus the differential sensitivity of the internodes to atrazine, and possibly to the other herbicides, may be because the more mature first internode allows less uptake and subsequent movement to the foliar parts.
Absorption dans la zone des tiges des herbicides appliqués dans le sol chez Pisum sativum L.     

TRANSLOCATION STUDIES WITH ENDOTHAL-14C IN POTAMOGETON NODOSUS POIR.

Summary. Plants of Potamogeton nodosus, a submersed aquatic, were treated with the di-sodium salt of 7-oxabicyclo(2,2,l)heptane-2,3-dicarboxylic acid-¹⁴C (endothal-¹⁴C). Gross radioautographs showed that the ¹⁴C label moved from mature photosynthesizing leaves and accumulated in the apices and developing secondary plants. Similar results were obtained with mid-leaf, stem (internode) and winter-bud applications. No move-ment of radioactivity occurred following root treatment. The results indicate symplastic translocation of endothal-¹⁴C when applied to the leaf, stem or winter bud. It is suggested that endothal can cause the death of plants by direct injury to root tissues subsequent to absorption.
Recherches sur la migration de l'endothal-¹⁴C dans le Potamogeton nodosus Poir.     

STUDIES ON THE TRANSLOCATION IN AGROPYRON REPENS OF 14C-LABELLED 2,2-DICHLOROPROPIONIC ACID

Summary. Previous findings suggested that the translocation in Agropyron repens of 2,2-dichloropropionic acid (dalapon) may be considerably affected by the transpiration rate. This relationship has been studied further in the present investigation using ¹⁴C-labelled material and autoradiography.
It was found that when dalapon was applied to the leaves and the treated plants were placed in darkness a reduction in the transpiration rate of ca. 90% was associated with a marked increase in the amount of dalapon translocation into the roots and tillers. The evidence provided by the autoradiographs was confirmed by a quantitative assay of the ¹⁴C present in the roots. It was also found, however, that translocation was not appreciably affected either by a 50% reduction in the light intensity or when the transpiration rate was reduced by placing the plaints under conditions of high humidity in the light or by the application of white petroleum jelly to the leaves. An alternative hypothesis, namely that the effect of darkness on the movement of the herbicide might be due to changes induced in the normal pattern of assimilate translocation was also investigated but was not supported by the results obtained. Further experimentss, in which various parts of the treated shoot were placed in the dark, suggested that the effect of darkness on the translocation of dalapon is exerted primarily on the treated leaf itself.
Etudes sur to migration dans Agropyron repens de l'acide 2,2-dichloropropionique marqué avec ¹⁴C     

MOVEMENT AND FATE OF FLUORODIFEN-1'-14C IN CUCUMBER SEEDLINGS

Summary. Autoradiographs and liquid scintillation counts of cucumber ( Cucumis sativus L. cv. Ashley) seedlings indicated rapid absorption and translocation of radioactivity from fluorodifen-1'-¹⁴C ( p -nitrophenyl-α,α,α-trifluoro-2-nitro- p -tolyl ether labelled with ¹⁴C in the 1 position of the p -nitrophenyl ring). After 24 h in 1 mg/1 of fluoro-difen-1'-¹⁴C, 25% of the absorbed radioactivity was translocated from the roots.
Cucumber seedlings degraded fluorodifen-1'-¹⁴C via a pathway similar to that reported for peanut seedlings but at a slower rate. The major degradation products were p -nitrophenol and an unidentified polar compound (Unknown I). More rapid absorption and translocation coupled with slower degradation by cucumber as compared with peanut seedlings are factors which may contribute to the susceptibility of cucumber to fluorodifen.     

Propanil activity, uptake and metabolism in resistant Echinochloa spp. biotypes

Field resistance of Echinochloa spp. to propanil has been previously reported in Costa Rica, Colombia and Arkansas (USA). In this study, the mechanism of resistance was investigated in three resistant (R) and three susceptible (S) biotypes. The shoot fresh weight reduction in pot-grown plants from a post-emergence spray of propanil at 2.44 kg a.i. ha⁻¹ on biotypes R/S from Costa Rica, Colombia and Arkansas was 35/98%, 25/79% and 20/82% respectively. In vitro chlorophyll fluorescence data from leaf tissue incubated in propanil showed that photosynthesis was inhibited in all biotypes, indicating that the propanil-binding site and enzyme were not altered. After transfer to herbicide-free solution, photosynthesis recovered only in resistant biotypes, indicating that the mechanism of resistance was caused by enhanced metabolism of the herbicide. Simultaneous treatment with fenitrothion, an aryl acylamidase inhibitor, prevented the recovery of photosynthesis in leaf tissue in two resistant biotypes. In contrast, the cytochrome P450 mono-oxygenase inhibitor, 1-aminobenzotriazole, did not prevent recovery from propanil in leaf tissue. Application of ¹⁴C-propanil to the second leaf of intact Echinochloa plants showed that c . 90% of the radioactivity remained in the treated leaf for up to 72 h after application. No major differences in translocation between R and S biotype plants were found. TLC analysis of tissue extracts from the treated leaves showed substantially less radioactivity associated with propanil, present after 72 h in rice or in the three R biotypes, compared with S biotypes.     

EFFECT OF 2,4-D AND PICLORAM ON TRANSLOCATION OF 14C-ASSIMILATES IN VITIS VINIEERA L.

Summary. High concentrations of 2,4-D and picloram interfered with the downward movement of ¹⁴c-assimilates infield-grown vines. The interference in translocation was appreciably greater with picloram than it was with 2,4-D, Although basipetal translocation was retarded, translocation within the treated shoots continued from the vegetative part to the clusters. Translocation of 2,4-D appeared to follow the same route as ¹⁴c-assimilatcs for the most part. Formative effects were absent on untreated grape shoots although the adjacent shoots treated with 2,4-D or picloram on the same cordons were killed; however, formative effects were evident on some of the stump sprouts which developed after the vines were harvested. The malformed leaves on the stump sprouts were twelve or more nodes from the base of the shoots, while ¹⁴c was in the more basal leaves.
Thompson Seedless (Sultanina) rootings treated with 20 000 ppm 2,4-D or picloram transported less ¹⁴C to the roots than did the controls. Treatment with either herbicide resulted in a marked increase in the labelling of the stems.
Effet du 2,4-D et du pichlorame sur la migration de métabolites marqués au ¹⁴C dans Vitis vinifera L.     

THE EFFECT OF SIMAZINE ON THE PHOTOFIXATION OF CO2 AND ON TRANSLOCATION OF ASSIMILATES IN NORWAY SPRUCE (PICEA ABIES)

Summary. A method of exposing seedlings of Norway spruce (Picea abies (L.) Karst.) to ¹⁴CO₂ is described. Within the 1st hr alter ¹⁴CO₂ exposure, no translocation of the ¹⁴C out of the treated branch could be observed. After a 24-hr period, however, the ¹⁴C in dormant seedlings had been translocated basipetally to part of the root system only, with no lateral diffusion of the ¹⁴C-compounds in the stem. About a week after exposure, both symplastic and apoplastic patterns of translocation had caused a more uniform distribution of ¹⁴C. In seedlings at active internode elongation, the translocation patterns were fundamentally identical to those in dormant seedlings, but the active shoot growth had led to a more uniform distribution of the ¹⁴C.
Simazine at 20 ppm had apparently stimulated both the photofixation of ¹⁴CO₂ and the rate of translocation of the ¹⁴C-assimilates. At 30 ppm, however, simazine had blocked the translocation of nutrients to the roots. On the other hand, the ¹⁴CO₂ uptake was not influenced. The simazine incubation had apparently no influence on the synthesis of cationic photosynthate.     

Comparison of chlorophyll fluorescence and whole-plant bioassays of isoproturon

Summary The response to aqueous solutions of isoproturon was examined on intact plants of Matricaria perforata , Apera spica-venti and Sinapis alba in greenhouse tests, comparing visual estimates of shoot phytotoxicity at 30 d after application (DAA) with measurements of fast chlorophyll fluorescence induction at 10 DAA on leaves of the same treated plants. The sensitivity was greatest from pre-emergence application to intact plants of M. perforata and A. spica-venti , whereas post-emergence sprays were relatively more active on S. alba . The use of fluorescence induction was investigated further in Petri-dish experiments on excised M. perforata leaf segments. The Petri-dish bioassay enabled detection of isoproturon at concentrations above 9 × 10⁻⁷ M within 96 h after herbicide exposure. The potential for using chlorophyll fluorescence assays for the determination of isoproturon content in soil extracts is discussed.     

THE FATE OF PARAQUAT APPLIED TO PLANTS

Summary. Paraquat dichloride was applied to maize, tomato and broad bean plants as droplets of an aqueous solution and analysed al intervals. It was shown to be degraded under some conditions, but the amount of degradation was least under conditions which would have been most favourable for metabolism of the chemical. The analytical results and results from cation-exchange paper chromatography and thin layer chromatography showed that the degradation was caused by photochemical decomposition on the surface of the leaves of the plants, not by metabolism. The degradation was not observed in the dark, and only occurred to a significant extent in daylight in the summer.
Two photochemical degradation products from methyl-labelled paraquat-¹⁴C dichloride, viz., 4-carboxy-l-(methyl-¹⁴C)-pyridinium chloride and methylamine-¹⁴C hydrochloride, were isolated from plants sprayed with the herbicide.
Le devenir du paraquat dans les plantes traitées     

Effects of formulation and environment on absorption and translocation of clopyralid in Cirsium arvense (L.) Scop. and Polygonum convolvulus L

The patterns of absorption and translocation of ¹⁴C clopyralid commercially formulated as the acid, monoethanolamine salt, potassium salt, 2-ethylhexyl ester and 1 -decyl ester were compared in Cirsium arvense and Polygonum convolvulus grown under three environmental regimes. Plants were grown under a 35% or 65% RH regime in silica sand maintained at 33% w/w moisture or under a water stress regime at 65% RH. Approxi mately 26,39,86,93 and 100% of the applied ¹⁴C-activity from the 2-ethylhexyl ester, acid, monoethanolamine salt, 1-decyl ester and potas sium salt, respectively, were recovered 72 h after application to glass cover slips placed in the growth room. However, loss of applied ¹⁴C-activity after application of the five formulations to both plant species was significant only for the 2-ethylhexyl ester. Regardless of environmental regime, the acid was the most readily absorbed formulation. In contrast to the acid, the salts and esters were less readily absorbed. When the data were expressed as a percentage of absorbed radioactivity, there was no significant difference in translocation when the acid, monoethanolamine salt and potassium salt were compared. However, significantly smaller quantities of the absorbed 2-ethylhexyl and 1-decyl ester were exported from the treated leaf. These results indicate that once absorbed, the esters do not readily partition out of the cuticle, whereas the acid and two salts move into the symplast for subsequent translocation. When the results of the three environmental regimes were compared, the absorption of the monoethanolamine and potas sium salts were greatly reduced under low humi dity or water stress, whereas the acid and esters were not affected.     

Optimizing foliar activity of glyphosate on Bidens frondosa and Panicum maximum with different adjuvant types

The influence of non-ionic (X-77) and organosilicone (L-77) adjuvants and of methylated seed oil (MSO) on the uptake, translocation and efficacy of glyphosate was investigated in Bidens frondosa L. and Panicum maximum Jacq. In addition, the physicochemical properties of adjuvants and adjuvant + glyphosate aqueous solutions were determined. Significantly lower surface tension and contact angle values were obtained with aqueous solutions of L-77 alone and with glyphosate. Over a 48-h time course, it was observed that >50% of applied ¹⁴C-glyphosate was absorbed within 15 min in B. frondosa with L-77. At 6 h and thereafter, ¹⁴C glyphosate absorption was significantly higher with MSO compared with X-77 in B. frondosa . In P. maximum , uptake and translocation of ¹⁴C-glyphosate + adjuvant were increased in general up to 48 h after treatment application, except with L-77, which showed no improvement in uptake – instead there was a significant reduction compared with no treatment with L-77. This indicated its antagonistic effect on this grass species. The lower values of ¹⁴C-glyphosate in P. maximum also confirmed that adjuvant effects were species specific. In the efficacy studies, glyphosate formulated with L-77 achieved significantly higher control of B. frondosa , while there was no control of P. maximum with this treatment. This confirmed antagonism in glyphosate absorption into P. maximum by L-77. Furthermore, significantly higher control of tested plants was recorded with MSO in comparison to X-77, which confirms the solubilizing or humectant nature of MSO.     

AN AUTORADIOGRAPHIC STUDY OF THE MOVEMENT OF 14C-LABELLED ASSIMILATES IN THE DEVELOPING WHEAT PLANT

Summary. ¹⁴CO₂ together with autoradiographic techniques was used to stuy the movement of labelled carbon within wheat plants at different stages in their development. Movement of ¹⁴C about the plain was found to be extensive in the early stages, gradually bccoming more localized until at a stage after ear emergence there was little transference of labelled carbon between tillers within a 24-hour period.
In the early stages of development ¹⁴C from leaves on the main shoot was transported throughout the plant but accumulated in greatest amounts in the meristematic regions. Assimilates produced by the leaves of newly formed tillers were distributed mainly to the meristematic regions of those tillers although labelled carbon was moved in into the rest of the plant. In the later stages of development of each tiller the labelled assimilate was restricted to the tiller itself and to any very young tillers associated with it. After car emergence the pattern of distribution of the labelled carbon from the youngest and second youngest leaves was found to differ. Movement was predominantly towards the ear from the flag leaf and towards the root system from the leaf below the flag.
It was noticed that mature leaves accumulated some labelled material and the possibility that this occurred via the transpiration stream following migration from phloem to xylem, as is the case with some of the case with some of the translocated herbicides, was investigated, but no indication of xylem transference was obtained.
The possible application of these studies to investigations into the relationship between the movement natural assimilates and of translocated herbicides is discussed.     

THE MOVEMENT OF PARAQUAT IN PLANTS

Summary. A qualitative study of the movement of the herbicide paraquat from droplets applied to leaves of tomato plants, using ¹⁴C-methyl-labelled and ¹⁴C-ring-labelled paraquat dichloride and di(methylsulphate), has shown that it moves in the xylem with the transpiration stream. The chemical is as well transported from young leaves as from mature ones, and will move through a steam-ringed petiole. The enhancement of the amount of paraquat transported from the treated leaves which occurs when treated plants are kept in darkness for a period following treatment and then exposed to light, is probably due to the greater movement into the xylem through undamaged tissue which can occur in the dark. Once the chemical has been absorbed into treated leaves, light-induced damage is required for significant movement through the rest of the plant to take place, but the damage then inhibits further entry of paraquat into the xylem. The movement of paraquat in broad bean and maize is essentially similar, though the enhancement of movement by a period of darkness after application is much less marked.
La migration du paraquat dans les plantes     

Biotransformation of [ring-U-14C]atrazine to dealkylated and hydroxylated metabolites in cell-suspension cultures

The biconversion of [¹⁴C]atrazine to deaikyt-ated and hydroxylated products was studied in heteroirophic cell-suspension cultures of carrot ( Daucus caroia L.), Agrostemma githago L. (corn cockle). Digitalis purpurea L. (purple foxglove), soyabean ( Giycine max L. Merr; four different cultivars). Datura stramonium L. (thorn-apple) and wheat ( Tritician aestivum L.). During 48 h of incubation, the herbicide was biotransformed by all species; turnovers yields differed considerably and were between 10.1% and 88.0% of applied ¹⁴C. Differences were also observed among the soyabean cultivars (10.1-73.5%). Hydroxy-atrazine, de-ethyl-, deisopropyl- and de-ethyt-deisopropylatrazine formed in the cultures were identified by thin-layer chromatography (tlc) (co-chromatography with reference compounds); deaikyiated metabolites were also proved by gas chromatography–mass spectrometry (gc–ms). In addition, highly polar transformation products emerged that were not identified. Portions of non-extractable residues were below 5% (one soyabean cultivar: 8.9%). Atrazine was metabolized by the cells, mainly to its dealkylated derivatives and hydroxyatrazine (totals of 9.4-54, 5%), whereas portions of highly polar products were lower (0.1-26.1%). Exceptions were A. githago (26.0 and 33.6%, respectively) and D, purpurea (4.5 and 25.2% respectively). Thus, plants generally contribute to the environmental degradation of atrazine.