• Korean Journal of Weed Science
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• v.16 no.2
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• pp.122-131
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• 1996

The inhibition characteristics of chlorsulfuron [CHL, 2-chloro-N-[{ (4-methoxy-6-methyl-1,3,5-triazin-2-yl)amino}carbonyl]benzenesulfonamide] and imazaquin [IMA, 2-{4,5-dihydro-4-methyl-4-(1-methy-lethyl)-5-oxo-1H-imidazol-2-yl}-3-quinolinecarboxylic acid] on acetolactate synthase(ALS) activity of corn plants were investigated. CHL and IMA rapidly inhibited ALS activity of corn plants in vitro. Their $I_{50}$ values for ALS activity were 100nM and $5{\mu}M$, respectively, indicating that CHL had 50 times more inhibitory effect on ALS activity than IMA. The first applied herbicide had a dominant inhibitory effect on ALS activity when the two herbicides were applied sequentially. Branched-chain amino acids, valine(Val), leucine(Leu), and isoleucine(Ile) showed a feedback inhibition on ALS activity ; Val or Leu had a more inhibitory effect on ALS activity than Ile. Branchedchain amino acids and CHL or IMA exhibited an additive effect on inhibiting ALS activity. This suggests that branched-chain amino acids inhibit ALS activity by a different mechanisms) from that of CHL or IMA. Apparent ALS activity, which was measured on the basis of the conversion of pyruvate to acetolactate, was decreased by the addition of 2-ketobutyrate into the ALS reaction mixture in a concentration-dependent manner. In addition, kinetic studies revealed that CHL acts as a noncompetitive inhibitor, while IMA acts as an uncompetitive inhibitor to ALS with respect to pyruvate.

• Journal of the Korean Chemical Society
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• v.43 no.4
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• pp.461-468
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• 1999

Acetolactate synthase (ALS) catalyzes the first common reaction in the biosynthesis of branched-chain amino acids, valine, leucine, and isoleucine. ALS is the common target of several classes of structurally diverse herbicides, the triazolopyrimidines, the imidazolinones, the sulfonylureas, and pyrimidyl-oxy-benzoates. We examined ihibitory activities of newly synthesized triazolopyrimidine sulfonamide derivatives using partially purified ALS from barley. $IC_{50}$ values for the active derivatives are 0.5nM∼8$\mu$M. Among them TP1 and TP2 are the most potent ALS inhibitors with $IC_{50}$ values of 0.5nM and 1.6nM, respectively. These inhibitors are more potent in the inhibition of barley ALS than commercial herbicides, Metosulam ($IC_{50}$;3.6 nM), Flumetsulam ($IC_{50}$;126 nM), and Cadre ($IC_{50};4 {\mu}M$). The progress curves for inhibition of ALS by TP2 showed that the amount of inhibition increases with time. The inhibition of ALS by TP2 was mixed-type inhibition with respect to pyruvate. Dual inhibition analyses of TP2 versus an imidazolinone, Cadre, and Leu showed parallel and intercepting kinetic pattern, respectively. The results suggest that TP2 binds to ALS competively with Cadre but not with Leu. Chemical modification of cysteinly residues in ALS decreased the sensitivity of ALS to Leu, while the modification did not affect the sensitivity of ALS to TP2 and Cadre.

• The Korean Journal of Pesticide Science
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• v.5 no.3
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• pp.41-46
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• 2001

This study was conducted to develop a high throughput system for screening acetolactate synthase(ALS) inhibitors, and to detect basic mother molecules for developing new novel herbicide candidates. The high throughput screening (HTS) method using 96-well plate and microplate reader was developed. This method is 8 times more effective than basic technique in one cycle per person. Futhermore, considering for less than 1/10 volume of materials required for ALS test and enzyme kinetics with 16 times faster speed compared to those of former procedure, this HTS method has more than 100 times higher efficacy than basic system in a consecutive procedure. We discovered 11 new ALS inhibitors such as 2-oxoglutaric acid, aminooxyacetic acid, azelaic acid, citric acid, cyanuric fluoride, itaconic acid, malonic acid, niclosamide, oxalic acid, glyoxylic acid, and suramin from 107 commercial plant-specific inhibitors using this technique. We hope these results might be useful to discover lead compounds for developing new novel herbicide candidate.

• Weed & Turfgrass Science
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• v.7 no.3
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• pp.191-199
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• 2018

This study was to investigate the occurrence patterns of paddy weeds, their resistance levels to an ALS inhibiting herbicide, and to estimate the areas of resistance in these paddy fields. We used soil collected from 358 paddy fields of Jeonnam province in 2017. Based on their life cycles, weeds were 96% annuals and 4% perennial. Additionally, according to morphological classification, 59% were broad leaves, 28% were sedges and 13% were grasses. Different areas within Jeonnam province contained different numbers and occurrence rates of weed species. However, generally, we observed Lindernia dubia var. dubia, Monochoria vaginalis var. plantaginea, Ludwigia prostrata, L. procumbens, Cyperus difformis, Scirpus juncoides, Eleocharis Kuroguwai, Echinochloa oryzoides, and E. crus-galli var. echinata. We also observed seven weeds resistant to an ALS inhibiting herbicide. They were M. vaginalis, S. juncoides, C. difformis, L. dubia, Ludwigia prostrata, E. oryzoides, and E. crus-galli var. echinata. Although there were differences in the number and occurrence rate of resistant weed species to an ALS inhibiting herbicide among areas in Jeonnam province, the M. vaginalis, C. difformis, and S. juncoides occurred in 23 cities and counties in Jeonnam including Gwangju metropolitan city. Based on the rates (52%) of resistant occurrence to an ALS inhibiting herbicide in Jeonnam province, the area of weed resistant paddy fields was estimated to be 91,543 ha.

• Journal of the Korean Chemical Society
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• v.42 no.3
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• pp.306-314
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• 1998

Acetolactate synthase (ALS) is the common enzyme in the biosynthesis of branched chain amino acids, Val, Leu, and Ile in bacteria, yeast, and higher plants. The enzyme is target site of several classes of structually diverse herbicides, including the sulfonylureas, the imidazolinones, the triazolopyrimidines, and the primidyl-oxy-benzoates. We have synthesized new triazolopyrimidine (TP) derivatives, and determined their inhibitory activities on barley ALS. $lC_{50}$ values for the active compounds were 3.2 nM-0.62 mM, and some of them appeared to be potent inhibitors. The progress curves for inhibition of ALS by TP4, a representative derivative, indicated that the extent of inhibition increased with incubation time. The inhibition of ALS by TP4 showed mixed-type inhibition with respect to pyruvate. Dual inhibition analyses of TP4 versus imidazolinone Cadre and feedback inhibitor Leu suggested that three different classes of inhibitors bind to ALS in a mutually exclusive manner. Chemical modification of tyrosyl residues of ALS decreased sensitivity of ALS to TP4, while modification of tryptophan and cysteine did not affect the sensitivity.

• Journal of the Korean Chemical Society
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• v.41 no.6
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• pp.304-312
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• 1997

Acetolactate synthase(ALS) is the common enzyme in the biosynthetic of valine, leucine, and isoleucine, and is the target of several classes of structually unrelated herbicides, including sulfonylureas, imidazolinones, and triazolopyrimidines. In an effort to develop new and desirable herbicides, we have synthesized 4,6-dimethoxypyrimidine derivatives, and examined their inhibitory activities on pea ALS. The most active compound was found to be K11570 and $IC_{50}$ value for K11570 was 0.2 ${\mu}M.$ The inhibition of pea ALS by K11570 was biphasic, showing increased inhibition with incubation time. The K11570 showed mixed-type inhibition with respect to substrate pyruvate. Dual inhibition analysis of K11570 versus sufonylurea herbicide Ally and feedback inhibitor leucine revealed that three inhibitors were competitive for binding to ALS. The arginine modified enzyme showed decreased inhibition by K11570, sufonylurea Ally, and leucine, in constrast to, tryptophan modification did not affect on the sensitivity of ALS to the inhibitors.

• Weed & Turfgrass Science
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• v.7 no.3
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• pp.209-218
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• 2018

This study was carried out to investigate the occurrence and distribution of ALS inhibiting herbicide-resistant weeds and to estimate the appeared areas of resistant weeds in the paddy fields of Gyeongnam province of Korea in 2017 and 2018 using a soil assay method. Compared with the 2012 survey, this study showed that the infested ratio of ALS inhibiting herbicide-resistant weeds increased from 1.0% to 66.8% and the infested area increased from 876 ha to 49,008 ha. The infested area of ALS inhibiting herbicide-resistant weeds was estimated in Ulsan-si (8.4%), Hapcheon-gun (8.3%), Haman-gun (7.9%), Goseong-gun (7.9%), Hadong-gun (7.3%), Jinju-si (7.2%), Changnyeong-gun (7.0%), Gimhae-si (6.4%), Miryang-si (5.5%), Busan-si (4.9%), Uiryeong-gun (4.6%), Namhae-gun (4.3%), Geochang-gun (4.2%), Changwon-si (3.8%), Geoje-si (2.9%), Yangsan-si (1.8%), Sancheong-gun (0.9%) and Tongyeong-si (0.4%), and the herbicide resistant weeds was not occurred in Hamyang-gun. The most dominant ALS inhibiting herbicide-resistant weeds in paddy fields were Monochoria vaginalis, followed by Echinochloa oryzicola, Lindernia dubia, Scirpus juncoides, Ludwigia prostrata, Cyperus difformis, Sagittaria trifolia and Rotala indica. ALS inhibiting herbicide-resistant M. vaginalis, L. dubia, and E. oryzoides occurred throughout Gyeongnam province, and ALS inhibiting herbicide-resistant S. trifolia and R. indica were only found in Gimhae-si. Therefore, these results will be utilized to estimate population dynamics of ALS inhibiting herbicide-resistant weeds and provide proper management practices in the paddy fields of Gyeongnam province.

• Korean Journal of Weed Science
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• v.30 no.3
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• pp.291-299
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• 2010

Cyhalofop-butyl and penoxsulam, known for respective acetyl CoA carboxylase (ACCase) and acetolactate synthase (ALS) inhibitor, have been widely using as a post-emergence foliar application for many years in wet-seeded rice field in Korea. Since 2009, control of Echinochloa oryzoides with cyhalofop-butyl and penoxsulam was no longer satisfactory. Greenhouse and experiment in rice field were conducted to confirm E. oryzoides resistance to ACCase and ALS inhibitors and to compare herbicide treatments for control of the resistant E. oryzoides to ACCase and ALS inhibitors. Three resistant E. oryzoides to ACCase and ALS inhibitors accessions were tested for levels of resistance to cyhalofop-butyl and penoxsulam, base on survival rate. Iksan and Kimje accessions were not any affected to the survival by treatment with recommended doses of two herbicides tested. Buan accession displayed an intermediate response of 62 and 72% survival at recommended doses of cyhalofop-butyl and penoxsulam, respectively. Survival rates to herbicides mixed with ACCase and ALS inhibitors at 3.5 leaf stage of E. oryzoides was over 80%. Benzobicyclon+fentrazamide+bensulfuron SC, benzobicyclon+mefenacet+bensulfuron SC, benzobicyclone +cafenstrole+pyrazosulfuron-ethyl GR controlled effectively by 2 leaf stage of resistant E. oryzoides to ACCase and ALS inhibitors. In the field experiment, single treatment of benzobicyclon+ fentrazamide+bensulfuron SC and benzobicyclon+mefenacet+bensulfuron SC failed to control E. oryzoides, but squential treatment of benzobicyclon+thiobencarb SE and benzobicyclon+ mefenacet+ bensulfuron SC controlled effectively it in rice infant seedling culture with machine. Our results suggest that resistant E. oryzoides to ACCase and ALS inhibitors had not developed multiple resistance to herbicides with different modes of action. In particular, cafenstrole, fentrazamide, mefenacet were effective control measures.

• Korean Journal of Weed Science
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• v.31 no.2
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• pp.119-133
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• 2011

Sulfonylurea (SU)-resistant weeds include seven annual weeds such as Monochoria vaginalis, Scirpus juncoides and Cyperus difformis, etc., and three perennial weeds of Scirpus planiculmis, Sagittaria pigmaea and Eleocharis acicularis as of 2010 since identification Monochoria korsakowii in the reclaimed rice field in 1998. The Echinochloa oryzoides resistant to acetyl CoA carboxylase (ACCase) and acetolactate synthase (ALS) inhibitors has been confirmed in wet-direct seeding rice field of the southern province, Korea in 2009. In the beginning of occurrence of SU-resistant weeds the M. vaginalis, S. juncoides and C. difformis were rapidly and individually spreaded in different fields, however, theses resistant weeds have been occurring simultaneously in the same filed as time goes by. The resistant biotype by weed species demonstrated about 10- to 1,000-fold resistance, base on $GR_{50}$ (50% growth reduction) values of the SU herbicides tested. And the resistant biotype of E. oryzoides to cyhalofop-butyl, pyriminobac-methyl, and penoxsulam was about 14, 8, and 11 times more resistant than the susceptible biotype base on $GR_{50}$ values. In history of paddy herbicides in Korea, the introduction of SU herbicides including besulfuron-metyl and pyrazosulfuron-ethyl that control many troublesome weeds at low use rates and provide excellent crop safety gave farmers and many workers for herbicide business refreshing jolt. The products and applied area of SU-included herbicides have been rapidly increased, and have accounted for about 69% and 96%, respectively, in Korea. The top ten herbicides by applied area were composed of all SU-included herbicides by 2003. The concentrated and successive treatment of ACCase and ALS inhibitors for control of barnyardgrass in direct-seeded rice led up to the resistance of E. oryzoides. Also, SU-herbicides like pyrazosulfuron-ethyl and imazosulfuron which are effective to barnyardgrass can be bound up with the resistance of E. oryzoides. The ALS activity isolated from the resistant biotype of M. korsakowii to SU-herbicides tested was less sensitive than that of susceptible biotype. The concentration of herbicide required for 50% inhibition of ALS activity ($I_{50}$) of the SU-resistant M. korsakowii was 14- to 76-fold higher as compared to the susceptible biotype. No differences were observed in the rates of [$^{14}C$]bensulfuron uptake and translocation. ALS genes from M. vaginalis resistant and susceptible biotypes against SU-herbicides revealed a single amino acid substitution of proline (CCT), at 197th position based on the M. korsakowii ALS sequence numbering, to serin (TCT) in conserved domain A of the gene. Carfentrazone-ethyl and pyrazolate were used mainly to control SU-resistant M. vaginalis by 2006, the early period, in Korea. However, the alternative herbicides such as benzobicyclone, to be possible to control simultaneously the several resistant weeds, have been developing and using broadly because the several resistant weeds have been occurring simultaneously in the same filed. The top ten herbicides by applied area in Korea have been occupied by products of 3-way mixture type including herbicides with alternative mode of action for the herbicide resistant weeds. Mefenacet, fentrazamide and cafenstrole had excellent controlling effects on the ACCase and ALS inhibitors resistant when they were applied within 2 leaf stage.

• Applied Biological Chemistry
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• v.41 no.1
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• pp.99-103
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• 1998

Triazolopyrimidine sulfonanilide (TP) derivative is one of excellent herbicide compounds. We have synthesized three classes of a new sulfonamide derivative (TPP) as Acetolactate synthase (ALS) inhibitors, in which the benzene ring in TP skeleton was converted to substituted pyrimidyl ring and examined their inhibitory activities on barley for ALS. $I_{50}$ values of the inhibitors ranged from 0.005 to 2 mM. Comparing the $I_{50}$ value of each class of TPP derivatives, the substituents in pyrimidine and triazolopyrimidine ring were found to affect the degree of ALS inhibition. TPP with substituted methyl group in pyrimidine ring showed higher inhibitory activity than that with methoxy group, while the substitution of the cyclopentano group in triazolopyrimidine ring gave very large inhibitory activity than that of methyl group. The present study established that variation of the electron density by substitution at heterocyclic ring is a very important factor for ALS inhibition, but showed no dependence on steric effect by substituents.