Evaluation of allelopathic effects of Parthenium hysterophorus L. methanolic extracts on some selected plants and weeds.

Herbicides made from natural molecules are cost-effective and environmentally friendly alternatives to synthetic chemical herbicides for controlling weeds in the crop field. In this context, an investigation was carried out to ascertain the allelopathic potential of Parthenium hysterophorus L. as well as to identify its phenolic components which are responsible for the allelopathic effect. During the observation, the rate of germination and seedlings' growth of Vigna subterranea (L.) Verdc, Raphanus sativus (L.) Domin, Cucurbita maxima Duchesne., Cucumis sativus L., Solanum lycopersicum L., Capsicum frutescens L., Zea mays L., Abelmoschus esculentus (L.) Moench, Daucus carota L., Digitaria sanguinalis (L.) Scop and Eleusine indica (L.) Gaertn were investigated by using methanol extracts, isolated from leaf, stem and flower of P. hysterophorus. Six concentrations (i.e., 25, 50, 75, 100, and 150 g L-1) of methanol extracts were isolated from P. hysterophorus leaf, stem and flower were compared to the control (distilled water). It was also observed that the concentration of methanol extracts (isolated from P. hysterophorus leaf, stem, and flower) while increased, the rate of seed germination and seedling growth of both selected crops and weeds decreased drastically, indicating that these methanol extracts have allelopathic potential. The allelopathic potential of P. hysterophorus leaf extraction (811) was found higher than the extraction of the stem (1554) and flower (1109), which is confirmed by EC50 values. The principal component analysis (PCA) was also used to re-validate the allelopathic potentiality of these methanol extracts and confirmed that Raphanus sativus, Solanum lycopersicum, Capsicum frutescens, Abelmoschus esculentus, Daucus carota, Digitaria sanguinalis, and Eleusine indica were highly susceptible to allelochemicals of P. hysterophorus. Besides these, the LC-MS analysis also revealed that the P. hysterophorus leaf extract contained 7 phenolic compounds which were responsible for the inhibition of tested crops and weeds through allelopathic effect. The results of the current study revealed that the leaf of P. hysterophorus is a major source of allelopathic potential on crops and weeds and which could be used as a valuable natural herbicide in the future for the sustainability of crop production through controlling weeds.

Determination and Quantification of Phytochemicals from the Leaf Extract of Parthenium hysterophorus L. and Their Physio-Biochemical Responses to Several Crop and Weed Species.

This current investigation was undertaken both in laboratory and glasshouse for documentation and quantification of phytochemicals from different parts of the parthenium (Parthenium hysterophorus L.) plant through LC-MS and HPLC to study their effect on two crops namely, Bambara groundnut (Vigna subterranean L.) and maize (Zea mays L.), and six different types of weed e.g., Digitaria sanguinalis, Eleusine indica, Ageratum conyzoides, Cyperus iria, Euphorbia hirta, and Cyperus difformis. The parthenium methanolic leaf extracts at 25, 50, 75, and 100 g L-1 were sprayed in the test crops and weeds to assess their physiological and biochemical reactions after 6, 24, 48, and 72 h of spraying these compounds (HAS). The LC-MS analysis confirmed seven types of phytochemicals (caffeic acid, ferulic acid, vanillic acid, parthenin, chlorogenic acid, quinic acid, and p-anisic acid) in the parthenium leaf extract that were responsible for the inhibition of tested crops and weeds. From the HPLC analysis, higher amounts in leaf methanol extracts (40,752.52 ppm) than those of the stem (2664.09 ppm) and flower extracts (30,454.33 ppm) were recorded. Parthenium leaf extract at 100 g L-1 had observed higher phytotoxicity on all weed species except C. difformis. However, all crops were found safe under this dose of extraction. Although both crops were also affected to some extent, they could recover from the stress after a few days. The photosynthetic rate, transpiration rate, stomatal conductance, carotenoid and chlorophyll content were decreased due to the application of parthenium leaf extract. However, when parthenium leaf extract was applied at 100 g L-1 for 72 h, the malondialdehyde (MDA) and proline content were increased in all weeds. Enzymatic antioxidant activity (e.g., superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) contents) were also elevated as a result of the sprayed parthenium leaf extract. The negative impact of physiological and biochemical responses as a consequence of the parthenium leaf extract led the weed species to be stressed and finally killed. The current findings show the feasibility of developing bioherbicide from the methanolic extract of parthenium leaf for controlling weeds, which will be cost-effective, sustainable, and environment friendly for crop production during the future changing climate.

Physiological and biochemical responses of selected weed and crop species to the plant-based bioherbicide WeedLock.

WeedLock is a broad-spectrum plant-based bioherbicide that is currently on the market as a ready-to-use formulation. In this study, we investigated the physiological and biochemical effects of WeedLock (672.75 L ha-1) on Ageratum conyzoides L., Eleusine indica (L.) Gaertn, Zea mays L., and Amaranthus gangeticus L. at four different time points. WeedLock caused significant reductions in chlorophyll pigment content and disrupted photosynthetic processes in all test plants. The greatest inhibition in photosynthesis was recorded in A. conyzoides at 24 h post-treatment with a 74.88% inhibition. Plants treated with WeedLock showed increased malondialdehyde (MDA) and proline production, which is indicative of phytotoxic stress. Remarkably, MDA contents of all treated plants increased by more than 100% in comparison to untreated. The activity of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) was elevated following treatment with WeedLock. Significant increases were observed in the SOD activity of A. conyzoides ranging from 69.66 to 118.24% from 6 to 72 h post-treatment. Our findings confirm that WeedLock disrupts the normal physiological and biochemical processes in plants following exposure and that its mode of action is associated with ROS (reactive oxygen species) production, similar to that of PPO (protoporphyrinogen oxidase) inhibitors, although specific site-of-action of this novel bioherbicide warrants further investigation.

Documentation of Phytotoxic Compounds Existing in Parthenium hysterophorus L. Leaf and Their Phytotoxicity on Eleusine indica (L.) Gaertn. and Digitaria sanguinalis (L.) Scop.

The utilization of the invasive weed, Parthenium hysterophorus L. for producing value-added products is novel research for sustaining our environment. Therefore, the current study aims to document the phytotoxic compounds contained in the leaf of parthenium and to examine the phytotoxic effects of all those phytochemicals on the seed sprouting and growth of Crabgrass Digitaria sanguinalis (L.) Scop. and Goosegrass Eleusine indica (L.) Gaertn. The phytotoxic substances of the methanol extract of the P. hysterophorus leaf were analyzed by LC-ESI-QTOF-MS=MS. From the LC-MS study, many compounds, such as terpenoids, flavonoids, amino acids, pseudo guaianolides, and carbohydrate and phenolic acids, were identified. Among them, seven potential phytotoxic compounds (i.e., caffeic acid, vanillic acid, ferulic acid, chlorogenic acid, quinic acid, anisic acid, and parthenin) were documented, those are responsible for plant growth inhibition. The concentration needed to reach 50% growth inhibition in respect to germination (ECg50), root length (ECr50), and shoot length (ECs50) was estimated and the severity of phytotoxicity of the biochemicals was determined by the pooled values (rank value) of three inhibition parameters. The highest growth inhibition was demarcated by caffeic acid, which was confirmed and indicated by cluster analysis and principal component analysis (PCA). In the case of D. sanguinalis, the germination was reduced by 60.02%, root length was reduced by 76.49%, and shoot length was reduced by 71.14% when the chemical was applied at 800 µM concentration, but in the case of E. indica, 100% reduction of seed germination, root length, and shoot length reduction occurred at the same concentration. The lowest rank value was observed from caffeic acids in both E. indica (rank value 684.7) and D. sanguinalis (909.5) caused by parthenin. It means that caffeic acid showed the highest phytotoxicity. As a result, there is a significant chance that the parthenium weed will be used to create bioherbicides in the future.

Phytochemical Constituents and Allelopathic Potential of Parthenium hysterophorus L. in Comparison to Commercial Herbicides to Control Weeds.

The allelopathic effect of various concentrations (0, 6.25, 12.5, 50 and 100 g L-1) of Parthenium hysterophorus methanol extract on Cyperus iria was investigated under laboratory and glasshouse conditions. No seed germination was recorded in the laboratory when P. hysterophorus extract was applied at 50 g L-1. In the glasshouse, C. iria was mostly injured by P. hysterophorus extract at 100 g L-1. The phytochemical constituents of the methanol extract of P. hysterophorus were analyzed by LC-ESI-QTOF-MS=MS. The results indicated the presence of phenolic compounds, terpenoids, alkaloids, amino acids, fatty acids, piperazines, benzofuran, indole, amines, azoles, sulfonic acid and other unknown compounds in P. hysterophorus methanol extract. A comparative study was also conducted between P. hysterophorus extract (20, 40 and 80 g L-1) with a synthetic herbicide (glyphosate and glufosinate ammonium at 2 L ha-1) as a positive control and no treatment (negative control) on Ageratumconyzoides, Oryzasativa and C. iria. The growth and biomass of test weeds were remarkably inhibited by P. hysterophorus extract. Nevertheless, no significant difference was obtained when P. hysterophorus extract (80 g L-1) and synthetic herbicides (glyphosate and glufosinate ammonium) were applied on A.conyzoides.

Bioherbicides: An Eco-Friendly Tool for Sustainable Weed Management.

Weed management is an arduous undertaking in crop production. Integrated weed management, inclusive of the application of bioherbicides, is an emerging weed control strategy toward sustainable agriculture. In general, bioherbicides are derived either from plants containing phytotoxic allelochemicals or certain disease-carrying microbes that can suppress weed populations. While bioherbicides have exhibited great promise in deterring weed seed germination and growth, only a few in vitro studies have been conducted on the physiological responses they evoke in weeds. This review discusses bioherbicide products that are currently available on the market, bioherbicide impact on weed physiology, and potential factors influencing bioherbicide efficacy. A new promising bioherbicide product is introduced at the end of this paper. When absorbed, phytotoxic plant extracts or metabolites disrupt cell membrane integrity and important biochemical processes in weeds. The phytotoxic impact on weed growth is reflected in low levels of root cell division, nutrient absorption, and growth hormone and pigment synthesis, as well as in the development of reactive oxygen species (ROS), stress-related hormones, and abnormal antioxidant activity. The inconsistency of bioherbicide efficacy is a primary factor restricting their widespread use, which is influenced by factors such as bioactive compound content, weed control spectrum, formulation, and application method.

Physiological and Biochemical Responses of Ageratum conyzoides, Oryza sativa f. spontanea (Weedy Rice) and Cyperus iria to Parthenium hysterophorus Methanol Extract.

The current study was designed to investigate the effect of Parthenium hysterophorus L. methanol extract on Ageratum conyzoides L., Oryza sativa f. spontanea (weedy rice) and Cyperus iria L. in glasshouse condition. Here, Parthenium hysterophorus methanol extract at 20, 40, and 60 g L-1 concentrations was applied on the test species to examine their physiological and biochemical responses at 6, 24, 48 and 72 h after spraying (HAS). The phytotoxicity of P. hysterophorus was strong on A. conyzoides compared to weedy rice and Cyperus iria at different concentrations and exposure times. There was a reduction in photosynthesis rate, stomatal conductance, transpiration, chlorophyll content and carotenoid content when plants were treated with P. hysterophorus extract concentrations. Exposure to P. hysterophorus (60 g L-1) at 24 HAS increased malondialdehyde (MDA) and proline content by 152% and 130%, respectively, in A. conyzoides compared with control. The activities of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD)) were also increased in the presence of P. hysterophorus extract. Present findings confirm that the methanol extract of P. hysterophorus can disrupt the physiological and biochemical mechanism of target weeds and could be used as an alternative to chemical herbicides.

Asp-376-Glu substitution endows target-site resistance to AHAS inhibitors in Limnocharis flava, an invasive weed in tropical rice fields.

Limnocharis flava (L.) Buchenau is a problematic weed in rice fields and water canals of Southeast Asia, and in Malaysia this invasive aquatic weed species has evolved multiple resistance to synthetic auxin herbicide and acetohydroxyacid synthase (AHAS) inhibitors. In this study, it was revealed that, a single nucleotide polymorphism (SNP) at amino acid position 376, where C was substituted to G at the third base of the same codon (GAC to GAG), resulting in Aspartate (Asp) substitution by Glutamate (Glu) was the contributing resistance mechanism in the L. flava population to AHAS inhibitors. In vitro assay further proved that, all the L. flava individuals carrying AHAS resistance mutation exhibited decreased-sensitivity to AHAS inhibitors at the enzyme level. In the bensulfuron-methyl whole-plant bioassay, high resistance indices (RI) of 328- and 437-fold were recorded in the absence and presence of malathion (the P450 inhibitor), respectively. Similarly, translocation and absorption of bensulfuron-methyl in both resistant and susceptible L. flava populations showed no remarkable differences, hence eliminated the possible co-existence of non-target-site resistance mechanism in the resistant L. flava. This study has confirmed another new case of a target-site resistant weed species to AHAS-inhibitors.

Correction: Ser-653-Asn substitution in the acetohydroxyacid synthase gene confers resistance in weedy rice to imidazolinone herbicides in Malaysia.

[This corrects the article DOI: 10.1371/journal.pone.0227397.].

Ser-653-Asn substitution in the acetohydroxyacid synthase gene confers resistance in weedy rice to imidazolinone herbicides in Malaysia.

The continuous and sole dependence on imidazolinone (IMI) herbicides for weedy rice control has led to the evolution of herbicide resistance in weedy rice populations across various countries growing IMI herbicide-resistant rice (IMI-rice), including Malaysia. A comprehensive study was conducted to elucidate occurrence, level, and mechanisms endowing resistance to IMI herbicides in putative resistant (R) weedy rice populations collected from three local Malaysian IMI-rice fields. Seed bioassay and whole-plant dose-response experiments were conducted using commercial IMI herbicides. Based on the resistance index (RI) quantification in both experiments, the cross-resistance pattern of R and susceptible (S) weedy rice populations and control rice varieties (IMI-rice variety MR220CL2 and non-IMI-rice variety MR219) to imazapic and imazapyr was determined. A molecular investigation was carried out by comparing the acetohydroxyacid synthase (AHAS) gene sequences of the R and S populations and the MR220CL2 and MR219 varieties. The AHAS gene sequences of R weedy rice were identical to those of MR220CL2, exhibiting a Ser-653-Asn substitution, which was absent in MR219 and S plants. In vitro assays were conducted using analytical grade IMI herbicides of imazapic (99.3%) and imazapyr (99.6%) at seven different concentrations. The results demonstrated that the AHAS enzyme extracted from the R populations and MR220CL2 was less sensitive to IMI herbicides than that from S and MR219, further supporting that IMI herbicide resistance was conferred by target-site mutation. In conclusion, IMI resistance in the selected populations of Malaysian weedy rice could be attributed to a Ser-653-Asn mutation that reduced the sensitivity of the target site to IMI herbicides. To our knowledge, this study is the first to show the resistance mechanism in weedy rice from Malaysian rice fields.

Adsorption-desorption and leaching potential of glyphosate and aminomethylphosphonic acid in acidic Malaysian soil amended with cow dung and rice husk ash.

This study investigates adsorption-desorption and the leaching potential of glyphosate and aminomethylphosphonic acid (AMPA) in control and amended-addition of cow dung or rice husk ash-acidic Malaysian soil with high oxide mineral content. The addition of cow dung or rice husk ash increased the adsorptive removal of AMPA. The isotherm data of glyphosate and AMPA best fitted the Freundlich model. The constant Kf for glyphosate was high in the control soil (544.873 mg g-1) followed by soil with cow dung (482.451 mg g-1) then soil with rice husk ash (418.539 mg g-1). However, for AMPA, soil with cow dung was high (166.636 mg g-1) followed by soil with rice husk ash (137.570 mg g-1) then the control soil (48.446 mg g-1). The 1/n values for both glyphosate and AMPA adsorptions were < 1 indicating their strong affinity for adsorbents. Desorption of both glyphosate and AMPA occurred only in the control soil. The compounds were not detected in soils with added cow dung or rice husk ash. The addition of cow dung or rice husk ash increased glyphosate mobility. However, ground water ubiquity scores for both control and amended soils were < 2.8. This indicated glyphosate is a transitional herbicide; therefore, its leaching potential in the soil is low, despite the addition of cow dung or rice husk ash. Addition of these wastes decreased the mobility and leaching potential of AMPA. The addition of cow dung or rice husk ash could be beneficial in increasing adsorption and enhancing degradation of these compounds.