This study was conducted to establish a fluorescence assay system for high efficient mass screening of the herbicides causing rapid membrane peroxidation, based on the fact that peroxide in cellular leakage could be fluorometrically determined through the fuorescent compounds formed after reacting with homovanillic acid (HVA) and peroxidase (HRP). The assay procesure established in this study was as follows. Only single disc (4 mm diameter) excised from cucumber cotyledon is placed on the well containing test solution ($200{\mu}L$) with 96-well microplate. The plate is shaking-incubated for 8 h under light condition. Then after removing the cucumber disc, HVA and HRP are supplied in the medium buffer and incubated for 5 min at room temperature. Fluorescence values are determined at Ex 320 nm/Ex 425 nm. The higher fluorescence values are obtained in the treatment of chemical having higher herbicidal activity. Using this assay with 96-well microplates, a large number of herbicides inducing rapid membrane peroxidation seemed to be screened more efficiently than spectrophotometric microtiter assay reported previously.
To improve the growth inhibitory activity against the shoot and root of rice plant (Oryza sativa L) and barnyard grass (Echinochloa crus-galli), a series of N-substituted phenyl-3,4-dimethylmaleimdes derivatives as substrates were synthesized and then their the inhibitory activities of protoporphyrinogen oxidase (1.3.3.4), protox were measured. The quantitative structure-activity relationships (QSAR) between structures and the inhibitory activities were studied quantitatively using the 2D-QSAR method. And also, molecular sharp similarity between the substrate derivatives and protogen, substrare of protox enzyme were studied. The activities of the two plants indicated that barnyard grass had a higher activity than the rice plant and their correlation relationships have shown in proportion for each. Accordingly, the results of SARs suggest that the electron donating groups as $R_2=Sub.X$ group will bind to phenyl ring because the bigger surface area of negative charged atoms in the substrate molecule derivatives may increase to the higher the activity against barnyard grass. Based on the molecular shape similarity, when the derivatives and protogen, subsbrate of protox enzyme were superimposed by atom fitting, the similarity indices (S) were above 0.8 level but the correlation coefficients (r) between S values and the activities showed not good.
To investigate the physiological and biochemical differences between susceptible and tolerant species of Echinochloa species to the foliar applied cyhalofop-butyl and pyribenzoxim herbicides, herbicidal response, the rate of absorption, translocation and metabolism were studied. Among 148 individuals of Echinochloa species collected from 41 sampling sites in Korea which were classified into 7 geographical regions, based on morphological characteristics of plant and seed type, 46 lines were used for further studies. From them, E. crus-galli var. praticola collected from Hwanggan and E. crus-galli var. crus-galli collected from Namyangju were selected as the most susceptible species to cyhalofop-butyl and pyribenzoxim, respectively. Meanwhile, E. oryzicola(from Cheju) and E. crus-galli var. crus-galli(from Asan) were selected as tolerant species to cyhalofop-butyl and pyribenzoxim, respectively. Application of radio-labelled herbicides on the 1st leaf to both susceptible and tolerant Echinochloa species exhibited that the applied $^{14}C$-Cyhalofop-butyl and $^{14}C$-pyribenzoxim were more easily absorbed and translocated into stem tissues than root. Absorption of $^{14}C$-Cyhalofop-butyl increased rapidly at 1 h after application and reached the maximum at 12 h after application. However, the absorption rate of $^{14}C$-pyribenzoxim was not changed. Two metabolites from cyhalofop-butyl-treated plants and one metabolite from pyribenzoxim-treated plants were separated by TLC. The amount of metabolite 1 in cyhalofop-butyl-treated tolerant species was significantly higher than that in the susceptible one, suggesting differences in detoxification ability between susceptible and tolerant species.
Selective action of root-treated oxyfluorfen [(2-chloro-4-thrifluoromethylphenyl)-3'-ethoxy-4'-nitrophenyl ether] and chlomethoxynil [2, 4-dichlorophenyl-3'-methoxy-4'-nitrophenyl ether] were investigated. Oxyfluorfen showed greater activity to all plant species than chlomethoxynil. $^{14}C$-oxyfluorfen was little metabolized in roots of the plant species and more slowly absorbed than $^{14}C$-chlomethoxynil. These results suggest that herbicidal activity of oxyfluorfen at the site of action is higher than chlomethoxynil. In the tested plants, rice, barnyardgrass, sorghum, and corn were absorbed less of the oxyfluorfen and chlomethoxynil than the broad leaf plant species. However, no clear relationship was observed between a degree of tolerance and absorption and metabolism of both herbicides by the plant species.
The experiment was carried out to find the feasibility of using Oxyfluorfen in the paddy fields by investigating the difference of selective activity of Oxyfluorfen among rice cultivars and major paddy weed species. The dosage of Oxyfluorfen that show selective activity between rice cultivars and weed species ranged from 0.1 to 0.4kg ai/ha. The degree of growth inhibition was in order of whole-plant soaking application > root soaking application > stem bandage application, and in that case $10^{-5}$M Oxyfluorfen was treated after emergence. Especially the growth inhibition of rice cultivars and Cyperus serotinus was low, among others. Photosynthesis was severely inhibited at the Oxyfluorfen level above $10^{-4}$ M in all the tested weeds, but inhibition of respiration was not to be seen. Isolated single cells of two rice cultivars and Cyperus serotinus were tolerant to $10^{-5}$M Oxyfluorfen,but those of Echinochloa crus-galli and Sagittaria pygmaea were susceptible comparatively. The growth inhibition of suspension cultured rice cell induced by the increments of Oxyfluorfen concentration, and the degree of inhibition was higher in C.V. Mushakdanti than in C.V. Aichiasahi.
Antagonistic mode of action of fenoxaprop-P-ethyl [ethyl(R)2-4-{(6-chloro-2-benzoxazolyloxy) phenoxy}propionate] with bentazon was investigated with respect to absorption, translocation, metabolism, and change in target site response of fenoxaprop-P-ethyl using four-leaf stage of rice(Oryza sativa L.) and barnyardgrass [Echinochloa eras-galli (L.) P. Beauv.]. Shoots of rice and barnyardgrass was more sensitive to fenoxaprop-P-ethyl than the roots. More than 90% of fenoxaprop-P-ethyl was absorbed within 6 hours after treatment and 30% of the absorbed was acropetally and basipetally translocated at 24 hours after treatment. Fenoxaprop-P-ethyl was rapidly transformed to its acid form, fenoxaprop(2-[4-(6-chloro-2-benzoxazolyloxy)phenoxy]propionic acid), which was subsequently metabolized to polar conjugates. However, changes in absorption, translocation, and metabolism of fenoxaprop-P-ethyl by bentazon treatment were not found in both species. Background activity of acetyl-CoA carboxylase(ACCase) in rice and barnyardgrass was 26.5 and 23.2nmol/min/mg, respectively. Concentration required to inhibit fifty percent enzyme activity$(I_{50})$ in vitro was 6.5~7.4${\mu}M$ of fenoxaprop-P-ethyl and more than 500${\mu}M$ of bentazon. There were no significant differences in $I_{50}$ value between two treatments of fenoxaprop-P-ethyl alone and its bentazon mixture. However, bentazon reduced ACCase activity in vivo and inhibited electron transport in chloroplast thylakoid. Based on the results obtained, it is concluded that the antagonistic effect of bentazon occurs due not to direct effect on target site of fenoxaprop-P-ethyl, but to indirect involvement in reducing herbicidal activity of fenoxaprop-P-ethyl through physiological disturbances caused by bentazone at whole chloroplast level.
This paper provides the current status of weed science and prospects for the development of weed science based on the research trends presented at the 7th International Weed Science Conference in 2016. Approximately 520 researchers from 59 countries, including Korea, participated in the conference and presented 625 papers in nine research areas. Major research topics were herbicide resistance, weed ecology, weed management in agricultural and non-agricultural lands, herbicide spray technology, and non-chemical weed control. Studies on herbicide resistance presented more than 30% of all papers presented. Particularly, resistance to non-selective herbicides, such as glyphosate and glufosinate-ammonium, and non-target sites of resistance mechanisms were the main subjects of the herbicide resistance research area. Moreover, the conference focused on research concerning herbicide resistant weeds of staple crops of the world (corn, wheat, and rice). Arylex was introduced as a new compound which has a mode of herbicidal action similar to synthetic auxin. Three compounds being developed as HPPD inhibitors were studied for ways to reduce their toxicity and tested as mixed with safeners. Additionally, parasitic weeds, which are not native to Korea, are an expanding research subject in the world. Although 45 years have passed since the first report of herbicide resistance in 1970, herbicide resistance remains a serious problem in most intensive cropping systems of the world and will continue to be a major area of study in the future.
The study researched germination of the plants and growth of Fungus according to concentration of aqueous extracts in order to provide basic data for developing natural agricultural resources by using Persicaria longiseta. The seed germination of Amaranthus spinosus was inhibited at 25% P. longiseta extract, while Agrostis stolonifera ssp. palustris was not affected at all concentrations tested. Especially, the seed germination rate and fresh weight of Trifolium incarnatum at 20% P. blumei extract were higher than those of control plot. The early growth of most receptor plant seedlings was promoted at 25% and 50% of P. blumei extracts, but the radicle growth of all receptor plants was significantly inhibited at > 25% of P. longiseta extract. The response of receptor plants to P. longiseta extract was different according to the plant species and the plant parts. The growth of plant pathogenic fungus in PDA medium showed an increasing inhibition tendency with increasing concentrations of P. longiseta extract. Especially, P. longiseta extract showed the greatest antimicrobial activity against Phytophthora infestans, Phythium graminicola, and Pythium venterpoolii. The content of total phenolic compound in P. longiseta was higher in leaves (1082.3 mg/L) but lower in roots (228.6 mg/L) and stems (207.8 mg/L), which is an allelopathic chemical. As these results are summarized, P. longiseta have competitive advantage because they release phenolic compounds with allelopathic effect and affect on germination, growth and fungi growth on underground flora compared to native plants and they have eligibility for natural herbicide and germicide.
New technologies will have a large impact on the discovery of new herbicide site of action. Genomics, combinatorial chemistry, and bioinformatics help take advantage of serendipity through tile sequencing of huge numbers of genes or the synthesis of large numbers of chemical compounds. There are approximately $10^{30}\;to\;10^{50}$ possible molecules in molecular space of which only a fraction have been synthesized. Combining this potential with having access to 50,000 plant genes in the future elevates tile probability of discovering flew herbicidal site of actions. If 0.1, 1.0 or 10% of total genes in a typical plant are valid for herbicide target, a plant with 50,000 genes would provide about 50, 500, and 5,000 targets, respectively. However, only 11 herbicide targets have been identified and commercialized. The successful design of novel herbicides depends on careful consideration of a number of factors including target enzyme selections and validations, inhibitor designs, and the metabolic fates. Biochemical information can be used to identify enzymes which produce lethal phenotypes. The identification of a lethal target site is an important step to this approach. An examination of the characteristics of known targets provides of crucial insight as to the definition of a lethal target. Recently, antisense RNA suppression of an enzyme translation has been used to determine the genes required for toxicity and offers a strategy for identifying lethal target sites. After the identification of a lethal target, detailed knowledge such as the enzyme kinetics and the protein structure may be used to design potent inhibitors. Various types of inhibitors may be designed for a given enzyme. Strategies for the selection of new enzyme targets giving the desired physiological response upon partial inhibition include identification of chemical leads, lethal mutants and the use of antisense technology. Enzyme inhibitors having agrochemical utility can be categorized into six major groups: ground-state analogues, group specific reagents, affinity labels, suicide substrates, reaction intermediate analogues, and extraneous site inhibitors. In this review, examples of each category, and their advantages and disadvantages, will be discussed. The target identification and construction of a potent inhibitor, in itself, may not lead to develop an effective herbicide. The desired in vivo activity, uptake and translocation, and metabolism of the inhibitor should be studied in detail to assess the full potential of the target. Strategies for delivery of the compound to the target enzyme and avoidance of premature detoxification may include a proherbicidal approach, especially when inhibitors are highly charged or when selective detoxification or activation can be exploited. Utilization of differences in detoxification or activation between weeds and crops may lead to enhance selectivity. Without a full appreciation of each of these facets of herbicide design, the chances for success with the target or enzyme-driven approach are reduced.
A field experiment was conducted in 1981 at the Crop Experiment Station, Suweon, Korea, in machine transplanted paddy rice field, to study the effectiveness of single herbicide, mixture, and systematic application of herbicides on diversity of weed control spectrum. The rice variety planted was Taebaegbyeo, Indica ${\times}$ Japonica cross bred. Experimental field was dominated by Echinochtoa crusgalli, Eleocharis kuroguwai, and Scirpus hotarui, and importance values based on dry weight of these weeds were 89%, 5%, and 3%, respectively. The mixture or systematic treatments of herbicide were generally more effective than single herbicide applications on weed control. Coefficients of similarity based on floristic composition after herbicide application between Perfluidone (5G) and Chloromethoxynil (7G), and between Pertluidone (5G) and Bifenox (7G), and between Perfluidone (5G) and three types of Butachlor (6G) were low, and these sets seemed to be a good mixture herbicide in paddy fields. While, Perfluidone (5G) had low coefficient of similarity with other single herbicides tested. The information on coefficient of similarity could be used as parameter for selecting herbicides to increase the efficiency of herbicidal performance. Simpson's indices from Butachlor (3.5G)/SL-49 (7G), Butachlor (3.5G)/Pyrazolate (6G), and Perfluidone (5G) treatments were high, and these herbicide treatments tended to the weed community type simplified, while the indices from Perfluidone (5G) + Chloromethoxynil (7G), Butachlor (6G) fb Perftuidone (5G), and Butachlor (4G)/Naproanilide (6G) treatments were low, and these herbicide treatments caused to the community type diversified in terms of floristic composition.
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