Sublethal Herbicide Doses Research Articles

Herbicides and their potential to disrupt plant–insect chemical communication

AbstractEcological interactions between plants and insects are of paramount importance for the maintenance of biodiversity and ecosystem functioning. Herbicides have long been considered a threat to plant and insect populations, but global increases in intensive agriculture and availability of herbicide‐resistant crops have intensified concerns about their full impact on biodiversity. Here, we argue that exposure to sublethal herbicide doses has the potential to alter plant–insect interactions as a result of disruptions in their chemical communication. This is because herbicides interfere with biosynthetic pathways and phytohormones involved in the production of several classes of plant volatiles that mediate plant–insect chemical communication. Sublethal herbicide doses can modify the morphological and life‐history plant traits and affect interactions with insects. However, the potential changes in plant volatiles and their consequences for plant–insect chemical communication have not yet received as much attention. We discuss how target‐site (disruptors of primary metabolism) and non‐target‐site (synthetic auxins) herbicides could alter the production of plant volatiles and disrupt plant–insect chemical communication. We suggest research avenues to fill in the current gap in our knowledge that might derive recommendations and applied solutions to minimize herbicides' impacts on plant–insect interactions and biodiversity.

Effects of glyphosate spray-drift on plant flowering.

Recent studies have shown that sub-lethal doses of herbicides may affect plant flowering, however, no study has established a direct relationship between the concentrations of deposited herbicide and plant flowering. Here the aim was to investigate the relationship between herbicide spray drift deposited on non-target plants and plant flowering in a realistic agro-ecosystem setting. The concentrations of the herbicide glyphosate deposited on plants were estimated by measuring the concentration of a dye tracer applied together with the herbicide. The estimated maximal and average deposition of glyphosate within the experimental area corresponded to 30g glyphosate/ha (2.08% of the label rate of 1440g a.i./ha) and 2.4g glyphosate/ha (0.15% label rate), respectively, and the concentrations decreased rapidly with increasing distance from the spraying track. However, there were not a unique relation between distance and deposition, which indicate that heterogeneities of turbulence, wind speed and/or direction can strongly influence the deposition from 1min to another during spraying. The effects of glyphosate on cumulative flower numbers and flowering time were modelled using Gompertz growth models on four non-target species. Glyphosate had a significantly negative effect on the cumulative number of flowers on Trifolium pratense and Lotus corniculatus, whereas there were no significant effects on Trifolium repens, and a positive, but non-significant, effect on number of flowers on Cichorium intybus. Glyphosate did not affect the flowering time of any of the four species significantly. Lack of floral resources is known to be of major importance for pollinator declines. The implications of the presented results for pesticide risk assessment are discussed.

Rapid Reduction of Herbicide Susceptibility in Junglerice by Recurrent Selection with Sublethal Dose of Herbicides and Heat Stress

Global climate change, specifically rising temperature, can alter the molecular physiology of weedy plants. These changes affect herbicide efficacy and weed management. This research aimed to investigate the combined effect of heat stress (HS) and sublethal doses of herbicides (four active ingredients) on adaptive gene expression and efficacy of herbicide on Echinochloa colona (L.) Link (junglerice). Three factors were evaluated; factor A was E. colona generation (G0-original population from susceptible standard; G1 and G2 were progenies of recurrent selection), factor B was herbicide treatment (florpyrauxifen-benzyl, glufosinate-ammonium, imazethapyr, quinclorac and nontreated check) and factor C was HS (30 and 45 °C). The herbicides were applied at 0.125× the recommended dose. Recurrent exposure to HS, combined with sublethal doses of herbicides, favors the selection of plants less susceptible to the herbicide. Upregulation of defense (antioxidant) genes (APX: Ascorbate peroxidase), herbicide detoxification genes (CYP450 family: Cytochrome P450), stress acclimation genes (HSP: Heat shock protein, TPP: Trehalose phosphate phosphatase and TPS: Trehalose phosphate synthase) and genes related to herbicide conjugation (UGT: UDP Glucosyltransferase) was significant. The positive regulation of these genes may promote increased tolerance of E. colona to these herbicides.

Is There a Possibility to Involve the Hormesis Effect on the Soybean with Glyphosate Sub-Lethal Amounts Used to Control Weed Species Amaranthus retroflexus L.?

Sub-lethal doses of herbicides can promote plant growth and have a positive effect on an organism this is called hormesis. The purpose of this study was to test the effects of sub-lethal doses of glyphosate on soybean (Glycine max (L.) Merr.) (1.8, 3.6, 7.2, 36, 180, and 720 g ha−1) and Amaranthus retroflexus L. (7.2, 36, 180, 720, 1440, and 2880 g ha−1). Different biological parameters, such as phytotoxicity, fresh weight, root length, content of photosynthetic pigments, and shikimate concentration, were measured. Glyphosate in doses of 1440 and 2880 g ha−1 destroyed A. retroflexus plants. A fresh weight of A. retroflexus at a dose of 36 g ha−1 was reduced by 76.31%, while for the soybean it was reduced by 19.26%. At the highest dose, the shikimate concentration was 145% in the soybean, while in A. retroflexus, the concentration increased by 58.80% compared to the control plants. All doses of glyphosate were statistically significantly different in terms of chlorophyll a content, while higher doses in A. retroflexus caused chlorophyll b to decrease. The change in the production of carotenoids was not statistically significant. The results showed that sub-lethal amounts of glyphosate did not lead to stimulation of measured parameters of soybean.

Effects of sublethal herbicides on offspring germination and seedling growth: Redroot pigweed (Amaranthus retroflexus) vs. velvetleaf (Abutilon theophrasti)

The effects of sublethal doses of herbicides on plants cannot be ignored, yet little is known about the effects of sublethal doses of herbicides on the F1 generation of plants. Seed germination and seedling growth of native and invasive plants following the sublethal exposure of parent plants to herbicides were comparatively analyzed in this study. Sublethal atrazine and tribenuron-methyl had carry-over effects on the germination and seedling growth of the F1 generation of invasive redroot pigweed (Amaranthus retroflexus L.) and native velvetleaf (Abutilon theophrasti Medicus), both of which had different responses to the carry-over effects of sublethal herbicide. The germination percentage of the F1 redroot pigweed (decreased) was greater than that of the F1 velvetleaf (increased or not significantly changed) following parental exposure to atrazine or tribenuron-methyl. Atrazine reduced the radical growth of 7-day-old velvetleaf seedlings and decreased the difference in seedling length between velvetleaf and redroot pigweed, while tribenuron-methyl had no significant effects on the growth of 7-day-old velvetleaf seedlings. The herbicide inhibition effect on the germination and growth of F1 velvetleaf and redroot pigweed did not increase as the sublethal dose increased. This study suggests that carry-over effects of sublethal herbicides weaken the growth advantage of the F1 velvetleaf at the seedling stage and may have a more negative influence on progeny population development of native velvetleaf compared with invasive redroot pigweed.

Os herbicidas causam impactos na sobrevivência e desenvolvimento de abelhas?

A polinização é um serviço ecossistêmico chave na manutenção da biodiversidade, essencial no processo reprodutivo das plantas superiores em quase todos os sistemas produtivos terrestres. Para que se possa manter a vasta biodiversidade global, incluindo de espécies cultivadas, é necessária que haja diversidade de polinizadores. No entanto, as abelhas sofrem com as ações antrópicas. Estes insetos vêm desaparecendo de áreas agrícolas devido à introdução de espécies exóticas, às grandes áreas de monocultivo, ao desmatamento para agricultura e pastagem e, principalmente, à fragmentação de habitats e ao uso excessivo ou incorreto de defensivos agrícolas. Esses pesticidas podem afetar não só a longevidade das abelhas, como também sua vitalidade o que pode, ao longo do tempo, influenciar na manutenção das colônias. No caso do uso de defensivos agrícolas, a maioria dos estudos visa avaliar os efeitos dos inseticidas sobre as abelhas, sendo escassos os estudos sobre outras classes de produtos, como os herbicidas. Esta revisão buscou dar foco aos possíveis efeitos, especialmente, de uso de herbicidas na sobrevivência e no desenvolvimento das abelhas. O uso de herbicidas que apresentam toxicidade para os insetos benéficos, como glyphosate, 2,4-D, MSMA e paraquat devem ser continuamente monitorado. As doses subletais dos herbicidas, ou seja, dose menor do que a que provoca a morte, podem provocar alterações comportamentais nos indivíduos, acarretando em sérios prejuízos à manutenção das colônias.

Constitutive redox and phosphoproteome changes in multiple herbicide resistant Avena fatua L. are similar to those of systemic acquired resistance and systemic acquired acclimation

Plants are routinely confronted with numerous biotic and abiotic stressors, and in response have evolved highly effective strategies of systemic acquired resistance (SAR) and systemic acquired acclimation (SAA), respectively. A much more evolutionarily recent abiotic stress is the application of herbicides to control weedy plants, and their intensive use has selected for resistant weed populations that cause substantial crop yield losses and increase production costs. Non-target site resistance (NTSR) to herbicides is rapidly increasing worldwide and is associated with alterations in generalized stress defense networks. This work investigated protein post-translational modifications associated with NTSR in multiple herbicide resistant (MHR) Avena fatua, and their commonalities with those of SAR and SAA. We used proteomic, biochemical, and immunological approaches to compare constitutive protein profiles in MHR and herbicide susceptible (HS) A. fatua populations. Phosphoproteome and redox proteome surveys showed that post-translational modifications of proteins with functions in core cellular processes were reduced in MHR plants, while those involved in xenobiotic and stress response, reactive oxygen species detoxification and redox maintenance, heat shock response, and intracellular signaling were elevated in MHR as compared to HS plants. More specifically, MHR plants contained constitutively elevated levels of three protein kinases including the lectin S-receptor-like serine/threonine-protein kinase LecRK2, a well-characterized component of SAR. Analyses of superoxide dismutase enzyme activity and protein levels did not reveal constitutive differences between MHR and HS plants. The overall results support the idea that herbicide stress is perceived similarly to other abiotic stresses, and that A. fatua NTSR shares analogous features with SAR and SAA. We speculate that MHR A. fatua’s previous exposure to sublethal herbicide doses, as well as earlier evolution under a diversity of abiotic and biotic stressors, has led to a heightened state of stress preparedness that includes NTSR to a number of unrelated herbicides.

Low doses of glyphosate enhance growth, CO2 assimilation, stomatal conductance and transpiration in sugarcane and eucalyptus.

Sublethal doses of herbicides can enhance plant growth and stimulate other process, an effect known as hormesis. The magnitude of hormesis is dependent on the plant species, the herbicide and its dose, plant development stage and environmental parameters. Glyphosate hormesis is well established, but relatively little is known of the mechanism of this phenomenon. The objective of this study was to determine if low doses of glyphosate that cause growth stimulation in sugarcane and eucalyptus concomitantly stimulate CO2 assimilation. Shoot dry weight in both species increased at both 40 and 60 days after application of 6.2 to 20.2 g a.e. ha-1 glyphosate. The level of enhanced shoot dry weight was 11 to 37%, depending on the time after treatment and the species. Concomitantly, CO2 assimilation, stomatal conductance and transpiration were increased by glyphosate doses similar to those that caused growth increases. Glyphosate applied at low doses increased the dry weight of sugarcane and eucalyptus plants in all experiments. This hormetic effect was related to low dose effects on CO2 assimilation rate, stomatal conductance and transpiration rate, indicating that low glyphosate doses enhance photosynthesis of plants. © 2017 Society of Chemical Industry.

Effects of melatonin in rats in the initial third stage of pregnancy exposed to sub-lethal doses of herbicides

Exposure to the herbicides Paraquat (PQ) and Roundup® may cause cell lesions due to an increase in oxidative stress levels in different biological systems, even in the reproductive system. ObjectiveEvaluate the possible changes in reproductive parameters and hepatic, as well as its prevention by simultaneous application of melatonin. MethodsThirty-five female rats at the age of 3 months were divided into seven groups: three groups exposed to sub-lethal doses of the herbicides PQ (50mg/kg) and Roundup® (500mg/kg) (n=5, G2, G3 and G4); three groups exposed to herbicides and simultaneous treatment with 10mg/kg of Melatonin (n=5, G5, G6 and G7) and control group (n=5, G1) from the first to the seventh day of pregnancy. On the seventh day of pregnancy, the rats were anesthetized and euthanized, followed by laparotomy to remove their reproductive tissues and liver. Body and ovary weights were taken and the number of implantation sites, corpora lutea, preimplantation losses, implantation rates were counted and histopathology of the implantation sites, morphometry of the surface and glandular epithelia of endometrium and hepatic oxidative stress were undertaken. ResultsThe present study shows the decrease in body and ovary weight, decrease in the number of implantation sites, implantation rate, in the total number of corpora lutea and increase of preimplantation percentages were observed when compared to the G1: Fig. 1 and Table 1, (p>0.001 ANOVA/Tukey). The histopathological analysis of the implantation sites showed a disorder of the cytotrophoblast and cell degeneration within the blastocyst cavity in Fig. 4. Morphometry revealed a reduction in surface and glandular epithelia and in the diameter of the endometrial glands (Table 2; p>0.05 ANOVA/Tukey), whereas in liver, serum levels of thiobarbituric acid reactive substances (TBARS) were found to be significantly elevated (Fig. 2; p>0.001; p>0.05 ANOVA/Tukey), and serum level of reduced glutathione (GSH) was significantly lower (Fig. 3; p>0.001 ANOVA/Tukey). However, treatments with melatonin exhibited improvements in reproductive parameters, as well as reduced lesions in the implantation sites (Fig. 4.) and in serum levels TBARS (Fig. 2; p>0.001 ANOVA/Tukey), serum levels GSH (Fig. 3; p>0.001; p>0.05 ANOVA/Tukey). ConclusionsThese results reveal that melatonin is a protective agent against experimentally induced maternal/embryo toxicity with herbicides and favoring normalization of reproductive parameters and hepatic.

Exposure to a glyphosate-based herbicide during pregnancy and lactation induces neurobehavioral alterations in rat offspring

The impact of sub-lethal doses of herbicides on human health and the environment is a matter of controversy. Due to the fact that evidence particularly of the effects of glyphosate on the central nervous system of rat offspring by in utero exposure is scarce, the purpose of the present study was to assess the neurobehavioral effects of chronic exposure to a glyphosate-containing herbicide during pregnancy and lactation. To this end, pregnant Wistar rats were exposed through drinking water to 0.2% or 0.4% of a commercial formulation of glyphosate (corresponding to a concentration of 0.65 or 1.30g/L of glyphosate, respectively) during pregnancy and lactation and neurobehavioral alterations in offspring were analyzed. The postnatal day on which each pup acquired neonatal reflexes (righting, cliff aversion and negative geotaxis) and that on which eyes and auditory canals were fully opened were recorded for the assessment of sensorimotor development. Locomotor activity and anxiety levels were monitored via open field test and plus maze test, respectively, in 45- and 90-day-old offspring. Pups exposed to a glyphosate-based herbicide showed early onset of cliff aversion reflex and early auditory canal opening. A decrease in locomotor activity and in anxiety levels was also observed in the groups exposed to a glyphosate-containing herbicide. Findings from the present study reveal that early exposure to a glyphosate-based herbicide affects the central nervous system in rat offspring probably by altering mechanisms or neurotransmitter systems that regulate locomotor activity and anxiety.

Evolution of glyphosate resistance in a Lolium rigidum population by glyphosate selection at sublethal doses

The majority of the documented cases of field-evolved herbicide-resistant weed biotypes established that single major genes confer glyphosate resistance. However, the contribution of minor genes endowing substantial plant survival at sublethal herbicide doses may be a potential complementary path to herbicide resistance evolution in weed populations under selection. Here, we subjected a number of susceptible individuals of Lolium rigidum to recurrent glyphosate selection to test the potential for sublethal glyphosate doses to additively select for glyphosate resistance. After 3-4 cycles of glyphosate selection in two distinct environments, the progenies of the initially susceptible population were shifted toward glyphosate resistance. The results indicate progressive enrichment of minor gene trait(s) contributing toward plant survival in the glyphosate-selected progenies. After three generations of selection, the estimated LD(50) values were doubled compared with the original population and up to 33% plant survival was obtained in the glyphosate-selected progeny at the recommended glyphosate label rate. This level of resistance probably was the maximum shift achievable with sublethal glyphosate dose selection in this small population. Cross-pollination was a crucial factor enabling the rapid rate of accumulation of minor glyphosate resistance gene trait(s) that are likely to be present at a relatively high frequency in a small susceptible population. The mechanistic basis of the moderate glyphosate resistance level selected by sublethal glyphosate doses remains unknown and warrants future research. Studying the main factors influencing the evolution of resistant weed populations is crucial for understanding, predicting and managing herbicide resistance.

Dual action of phosphonate herbicides in plants affected by herbivore—Model study on black bean aphid Aphis fabae rearing on broad bean Vicia faba plants

The interactions between plants, herbicides and herbivore insects were studied as an aspect of possible side effect of the using of phosphonate herbicides. The experimental system was composed of phosphonate herbicides, broad bean Vicia faba (L.) plants and black bean aphid Aphis fabae (Scopoli). Two means of herbicide application, namely standard spraying and direct introduction of the herbicide into stem via glass capillary, were examined. The results obtained for N-2-piridylaminomethylene bisphosphonic acid and its derivatives show 10 times higher inhibition of the plant growth if glass capillary mode was used. When plants were infested by aphids 24 h after the use of herbicide, a significant decrease in plant growth rate was observed in relation to plants treated with herbicides alone. Moreover, the sensitivity of aphids towards glyphosate, N-2-piridylaminomethylene bisphosphonic acid and its 3-methyl derivative introduced to artificial diet indicated that these herbicidal phosphonates possessed also insecticidal activity if applied in a systemic manner. Additionally, olfactometer measurements revealed that aphids preferred intact V. faba leaves over those that had been treated with sublethal doses of herbicides. The results achieved in these experiments indicate that the use of phosphonate herbicides decreases plant resistance and influences the number of aphids accompanied with treated plants. Regarding these facts it can be concluded that the combined effect of herbicide-induced stress and insect herbivory reduced plant fitness and thus should be considered as also a factor enabling the reduction of herbicide doses.

Predicting sublethal effects of herbicides on terrestrial non-crop plant species in the field from greenhouse data

Guidelines provided by OECD and EPPO allow the use of data obtained in greenhouse experiments in the risk assessment for pesticides to non-target terrestrial plants in the field. The present study was undertaken to investigate the predictability of effects on field-grown plants using greenhouse data. In addition, the influence of plant development stage on plant sensitivity and herbicide efficacy, the influence of the surrounding vegetation on individual plant sensitivity and of sublethal herbicide doses on the biomass, recovery and reproduction of non-crop plants was studied. Results show that in the future, it might well be possible to translate results from greenhouse experiments to field situations, given sufficient experimental data. The results also suggest consequences at the population level. Even when only marginal effects on the biomass of non-target plants are expected, their seed production and thereby survival at the population level may be negatively affected.

Effects of sublethal glyphosate rates on fresh market tomato

Two field trials were conducted to determine the effect of low doses of glyphosate on tomato plant growth and marketable yield. The herbicide was applied to the foliage 1 day before transplanting. There was a rapid decline on tomato plant vigour and height as the herbicide dose reached 100 mg/L. Marketable yield was also affected by sublethal herbicide doses, with a 41% yield decrease with 25 mg/L of glyphosate. These results confirm that low glyphosate doses, which could be found in multi-purpose spraying equipment, might severely reduce tomato marketable yield.

Combined effects ofAceria malherbaeand herbicides on field bindweed (Convolvulus arvensis) growth

The effects of a gall mite (Aceria malherbae) and sublethal doses of either 2,4-DB or glyphosate on field bindweed growth were evaluated under laboratory conditions. Mite feeding reduced field bindweed shoot biomass 37 to 48% and root biomass 46 to 50%. 2,4-DB at 0.07 to 0.14 kg ae ha−1or glyphosate at 0.14 to 0.28 kg ai ha−1reduced field bindweed root biomass 25 to 52%. CombiningA. malherbaefeeding with either 2,4-DB or glyphosate application reduced root biomass of field bindweed plants more than mites or either herbicide alone. LiveA. malherbaewere present on field bindweed 3 wk after treatment with either herbicide. Combination ofA. malherbaewith sublethal herbicide doses may allow for field bindweed suppression while reducing potential herbicide injury to crops and maintainingA. malherbaepopulations.

Biological control of Chenopodium album L. in Europe

Ascochyta caulina (P. Karst) v.d. Aa and v. Kest is aplant pathogenic fungus which is specific to Chenopodium albumL. It has been suggested as a potential mycoherbicide to this weed,which is important and wide spread in arable crops throughout Europe. Toinvestigate its potential as a biocontrol agent, the fungus has beentested in glasshouse and field experiments. Formulations containingdifferent combinations of A. caulina conidia, the phytotoxinsfrom the fungus and low doses of herbicides have been tested.Significant improvement in the efficacy of the fungus was achieved inglasshouse trials with an aqueous formulation containing PVA(0.1% v/v), Psyllium (0.4% w/v), Sylgard 309(0.1% v/v), nutrients and conidia (5 ×106/ml). The extracellular, hydrophilic phytotoxinsproduced by A. caulina were purified and their structuresdetermined. The main toxin, named ascaulitoxin, was characterised as theN2-β-D-glucopyranoside of the unusual bis-aminoacid2,4,7-triamino-5-hydroxyoctandioic acid. Two other toxins proved to betrans-4-amino-D-proline and the aglycone of ascaulitoxin. Thesetoxins have shown promising herbicidal properties. Field trials haveinvestigated the performance of A. caulina conidia applied atdifferent developmental stages of C. album either as a singletreatment or combined with sub-lethal doses of herbicides or with thefungal phytotoxins. With the available formulation, favourable weatherconditions are needed to obtain infection in the field. The efficacy ofthe strain of A. caulina used so far has proved to beinadequate to justify its development as a bioherbicide. This isprobably due to its low virulence.

Characteristics of seeds and seedlings from weeds treated with sublethal herbicide doses

SummarySize, germination and viability of seeds as well as growth of seedlings derived from three weed species were studied in a pot trial. Fallopin convolvulus (L.) A. Loeve, Galium. spurium L. and Thlaspi arvense L. were treated with MCPA or tribenuron‐methyl at four doses and at five growth stages, from seedling stage to flowering, In G. spurium subnormal doses of tribenuron‐methyl, applied at intermediate growth stages, greatly reduced seed weight, gennination, viability, seedling shoot biomass and root biomass. Germination and viability, as well as the shoot biomass and root biomass of seedlings, were highly correlated with seed weight. In addition, but to a smaller extent, seed weight was reduced in F. convolvidus and T. arvense by tribenuron‐methyl and in G. spurium by MCPA. Germination was reduced in F. convolvulus by MCPA and in T. arvense by tribenuron‐methyl. However, the effects varied greatly depending on the growth stage at application.

The Morphological and Physiological Response of Slender Oat (Avena barbata) to the Herbicides Barban and Difenzoquat

The morphological and physiological response of the slender oat (Avena barbataPott ex Link # AVEBA) to the herbicides barban (4-chloro-2-butynyl 3-chlorophenylcarbamate) and difenzoquat (1,2-dimethyl-3,5-diphenyl-1H-pyrazolium) in conjunction with decreased water availability was determined for seven populations, representing three ecotypes, under greenhouse conditions. Generally, within the range of sublethal herbicide doses, with increasing herbicide rates, phytotoxicity ratings increased, but plant dry weight, tiller height, and number of spikelets were decreased. Also, the number of juvenile tillers decreased, while that of fertile tillers increased. Flag leaf area increased and flowering was delayed. The ratio of number of spikelets to plant dry weight and seed weight was influenced the least. Under the highest rates of difenzoquat, the within-plant variance of spikelet number decreased, indicating that there may have been a more equal partitioning of resources amongst tillers for spikelet production. The general influence of water stress was to amplify the effect of the herbicide. For example, the dry treatment reduced dry weight and tiller height, and delayed flowering. Within a particular herbicide treatment, the effect of the water stress was to cause reduced within-plant variance for days to flowering, flag leaf area, and number of spikelets. Three reactions were observed that could have helped buffer decreases in spikelet production: 1) An increased fraction of the dry weight of the plants was partitioned into the spikelets at the expense of other vegetative matter, 2) the increased leaf area of the primary tiller may have helped counterbalance any reduction in photosynthesis caused by herbicide action, and 3) an increased number of juvenile tillers was converted into fertile tillers resulting in an increased number of mature tillers. These data indicate that the slender oat has a remarkable “phenotypic plasticity,” which enables it to maintain reproductive structures under sublethal herbicide doses.

Yield and protein content of wheat varieties (Kalyansona and Ni-5642) as influenced by sublethal doses of herbicides

A field experiment on two wheat varieties namely NI-5642 and Kalyansona with foliar spray of three herbicides viz 2,4-D, atrataf and taphazine at three concentrations i.e. 250, 500 and 1000 ppm was conducted inrabi season of 1971. From the studies it was observed that all three herbicides at all the concentrations increased the protein content and the maximum increase was to the extent of 56.8 as percentage of control. Of the chemicals used 2,4-D at 500 ppm concentrations was effective in respect of protein content, yield and 1000 grain weight.