Trp_(548)Met mutation of acetolactate synthase in rice confers resistance to a broad spectrum of ALS-inhibiting herbicides

Herbicide resistance in crop plants is valuable for integrated weed management in agriculture. Herbicide resistant rice, in particular, is important to management of weedy rice, a close relative of cultivated rice and a noxious weed prevalent in rice fields that remains challenging to farmers worldwide. Herbicide resistant plants can be obtained through transgenic approach or by mutagenesis of regular plant and screening of mutants with elevated resistance to herbicide. In this study, we conducted ethyl methyl sulfonate mutagenesis(EMS) to elite indica cultivar Huanghuazhan(HHZ) and screened for mutants resistant to imazapic, a herbicide that can inhibit the acetolactate synthase(ALS) in plants. We obtained three mutants of Os ALS gene that have not been reported previously in rice. One of the mutants, with Trp_(548) changed to Met(W_(548)M), was analyzed in more details in this study. This mutation had no negative effect on the plant physiology and morphology as well as rice yield. Compared with the imidazolinone-resistant mutant S_(627)N(Ser_(627) changed to Asn) that has been deployed for Clearfield rice development, W_(548)M mutant showed high levels of resistance to a broad spectrum of five families of ALSinhibiting herbicides, in addition to a higher level of resistance to herbicides of the imidazolinone family.The herbicide-resistance was stably inherited by crossing into other rice lines. Thus, the W_(548)M mutation provides a valuable resource for breeding of herbicide resistant rice and weed management.

Detection of Resistance and Cross-resistance to Four ALS-inhibiting Herbicides of Scirpus juncoides Roxb. in Paddy Field in Heilongjiang Province

The resistance and cross-resistance of Scirpus juncoides Roxb.to ALS-inhibiting herbicides were tested to provide theoretical basis for effectively control Scirpus juncoides Roxb.and rational use of herbicides.To identify the resistance and crossresistance of 19 populations from nine areas in Heilongjiang Province to four ALS-inhibiting herbicides by agar method and the whole-plant assay.The results are as the followings,the two bioassay methods had good correlation,the resistance indexes in the same area were different by two means,but resistant level was the same,similarity up to 89.5%.The resistant rates to pyrazosulfuronethyl,bensulfuron-methyl,penoxsulam and pyribenzoxim were 26.3%,36.8%,36.1%and 21.1%,respectively.Shangzhi populations evolved the highest level of resistance to pyrazosulfuron-methyl with a resistance index of 15.56.Shangzhi,Tonghe,Fangzheng and Jixi populations showed cross-resistance to four ALS-inhibiting herbicides,accounting for 26.3%.The resistance of Scirpus juncoides Roxb.to four ALS-inhibiting herbicides in Heilongjiang paddy field was low to moderate,five populations showed cross-resistance to four ALS-inhibiting herbicides.

Differential Susceptibility of <i>Conyza bonariensis</i>Biotypes to Glyphosate and ALS-Inhibiting Herbicides in Argentina

The aim of this study was to compare control of three stages biotypes (rosette, bolting, and repro- ductive stage) of Conyza bonariensis glyphosate susceptible biotype (S) and tolerant biotype (T) to glyphosate with glyphosate and mixtures of acetolactate synthase (ALS)-inhibiting. For glyphosate, the dose-response curves confirmed that injury of the biotype T relative to biotype S was signifi- cantly lower for both rosette and bolting stages. Resistance index (RI) for this herbicide was approximately 4 for both weed stages. At bolting, for both biotypes doses much higher than the recommended dose were required. For acetolactate synthase (ALS)-inhibiting herbicides, at the rosette stage, control of both biotypes was excellent with doses significantly lower than the recommended rate. All herbicides within this group showed a very low I50 relative to the recommended rate. These results indicate that biotypes difficult to control with glyphosate at the rosette and bolting stage may be controlled using acetolacte synthase (ALS)-inhibiting herbicides. Control for the reproductive stage was poor. A management program based on the combination of glyphosate with pre- and post-emergence acetolactate synthase (ALS) herbicides may be effective to control the weed.

Generating broad-spectrum tolerance to ALS-inhibiting herbicides in rice by base editing

Herbicide-tolerant rice varieties generated by genome editing are highly desirable for weed control.We have used a cytosine base editor to create a series of missense mutations in the P171 and/or G628 codons of the acetolactate synthase(ALS)gene to confer herbicide tolerance in rice.The four different missense mutations in the P171 codon,P171S,P171A,P171Yand P171F,exhibited different patterns of tolerance towards five representative herbicides from five chemical families of ALS inhibitors.For example,P171S and P171A had lower levels of tolerance than P171Y and P171F to bispyribac but not to the other herbicides.Interestingly,a novel triple mutant(P171F/G628E/G629S)had the highest tolerance to all five tested herbicides.Field trials showed that both P171F and P171F/G628E/G629S could potentially be used with nicosulfuron.Our work illustrates an effective way of using base editing to generate herbicide tolerance in elite rice varieties.

Evaluation of Pre-Emergence and Post-Emergence Herbicides for Weed Management in Miscanthus sacchariflorus and Miscanthus sinensis

Miscanthus,is a promising bioenergy crop,considered superior to other bioenergy crops because of its higher water and nutrient use efficiency,cold tolerance,and higher production of biomass.Broadleaf weeds and grass weeds,cause major problems in the Miscanthus field.A field experiment was conducted in 2018 and 2019,to assess the effects of pre-emergence(alachlor and napropamide)and post-emergence herbicides(nicosulfuron,dicamba,bentazon,and glufosinate ammonium)on broadleaf and grass weeds in M.sinensis and M.sacchariflorus fields.The weed control efficiency and phytotoxicity of pre-and post-emergence herbicides were evaluated at 30 days after treatment(DAT)and compared to those of the control plots.The results showed wide variations in the susceptibility of the weed species to the treated herbicides.Treatment with nicosulfuron 40 g.a.i.ha^(-1) provided the most effective overall weed control(with 10%visual injury),without affecting the height and biomass of neither Miscanthus species in the field.Post-emergence herbicides such as glufosinate ammonium 400 g.a.i.ha^(-1) and dicamba 482 g.a.i.ha^(-1) were effective and inhibited the growth and density of the majority of weeds to a 100%;however,they showed significant phytotoxicity(toxicity scale of 1-10)to both species of Miscanthus.The application of glufosinate ammonium caused severe injuries to the foliar region(90%visual injury)of both Miscanthus sps.Comparatively,M.sinensis showed a slightly higher tolerance to the herbicides nicosulfuron,bentazon and napropamide with 10%visual injury at the recommended dose than M.sacchariflorus.The present study clearly showed that infestation of broadleaf and grass weeds in Miscanthus fields can cause significant damage to the growth and biomass of Miscanthus and applying pre-emergence and post-emergence herbicides effectively controls the high infestation of these weeds.

Effects of Different Herbicides on Weed Control in Alfalfa Field

[Objectives]The paper was to systematically study the technology of weed control in alfalfa field.[Methods]Reviving alfalfa field and newly sown alfalfa field after emergence were selected,and the effects of different herbicides on weed control and alfalfa yield were discussed.[Results]The optimal herbicides after alfalfa reviving were 5%imazethapyr and 10%imazethapyr,and the optimal dosages were 1.5 and 1.05 L/hm 2,respectively.The optimal herbicides after emergence of newly born alfalfa were 5%imazethapyr and 10%imazethapyr,and the optimal dosages were 1.5 and 0.75 L/hm 2,respectively.[Conclusions]This study will provide a technical support for high quality production of alfalfa.

A nonsynonymous mutation in an acetolactate synthase gene (Gh_D10G1253) is required for tolerance to imidazolinone herbicides in cotton

Background Herbicide tolerance in crops enables them to survive when lethal doses of herbicides are applied to surrounding weeds.Herbicide-tolerant crops can be developed through transgenic approaches or traditional mutagenesis approaches.At present,no transgenic herbicide tolerant cotton have been commercialized in China due to the genetically-modified organism(GMO)regulation law.We aim to develop a non-transgenic herbicide-tolerant cotton through ethyl methanesulfonate(EMS)mutagenesis,offering an alternative choice for weed management.Results Seeds of an elite cotton cultivar Lumianyan 37(Lu37)were treated with EMS,and a mutant Lu37-1 showed strong tolerance to imidazolinone(IMI)herbicides was identified.A novel nonsynonymous substitution mutation Ser642Asn at acetolactate synthase(ALS)(Gh_D10G1253)in Lu37-1 mutant line was found to be the potential cause to the IMI herbicides tolerance in cotton.The Ser642Asn mutation in ALS did not present among the genomes of natural Gossypium species.Cleaved amplified polymorphic sequence(CAPS)markers were developed to identify the ALS mutant allele.The Arabidopsis overexpressing the mutanted ALS also showed high tolerance to IMI herbicides.Conclusion The nonsynonymous substitution mutation Ser642Asn of the ALS gene Gh_D10G1253 is a novel identi-fied mutation in cotton.This substitution mutation has also been identified in the orthologous ALS genes in other crops.This mutant ALS allele can be used to develop IMI herbicide-tolerant crops via a non-transgenic or transgenic approach.

Limiting Factors and Countermeasures for the Development of Biological Herbicides

This paper mainly analyzes the main limiting factors in the development of biological herbicides,such as narrow herbicidal spectrum,vulnerable to environmental impacts,difficulties in preparing dosage forms,difficult separation and low yield of active substances,biological safety,etc.Moreover,countermeasures and suggestions are put forward,including strengthening the study of biological screening of herbicidal activity,microbial metabolites and their use as lead compounds,dosage forms,fungi complex formulation,fungi and herbicide complex formulation,and fermentation technology.

Review and Analysis: Did the United States Transport, Off-Load and Use Commercial 2,4,5-T Herbicides with Unknown Amounts of Dioxin TCDD on Military Base Grounds in Panama Canal Zone between 1948 and 1999?

The 84 km Panama Canal connecting the Atlantic and Pacific oceans has historically been a strategic waterway for shipping and the location of United States (US) military bases. Since the construction of Lake Gatun reservoir, canal locks and navigation channel through the Isthmus of Panama tropical forests in the 1910s, chemicals, pesticides, and herbicides have been essential for controlling upland and wetland vegetation as well as managing mosquito-borne diseases. Chemicals and pesticides flowed into Lake Gatun via land surface runoff and subsurface drainage either attached to the sediment or in solution during the rainy season. Lake Gatun and the Panama Canal was the drinking water source for most of the civilian and military population living in the Panama Canal Zone. Between 1948 and 1999, US military base commanders had the ability to order, from the Federal Supply Catalog, commercially available herbicide 2,4,5-T with unknown amounts of dioxin TCDD for use on the military base grounds in the Panama Canal Zone. The herbicide 2,4,5-T was transported to Panama Canal Zone ports, including the ports Cristobal on the Caribbean and Balboa on the Pacific, and distributed to the US military bases in Panama by rail or truck. The US National Toxicology Program and the International Agency for the Research on Cancer listed dioxin and TCDD as known human carcinogens. Dioxins are endocrine disrupters and can cause certain chloracne, cancers, developmental and reproductive effects. In 1985, the United States government banned the manufacture of the herbicide 2,4,5-T, with unknown amounts of dioxin TCDD, after it was shown to cause cancer in animals. The objectives of this study are to determine: 1) the fate of dioxin TCDD, a contaminant in the herbicide 2,4,5-T, sprayed on the US military base vegetation in the Panama Canal Zone from 1948 to 1999, 2) the transport of dioxin TCDD rich sediment via soil erosion and overland flow into Lake Gatun and Panama Canal waterways and 3) the human health impacts of dioxin TCDD, a known carcinogen, on US military and Panamanian civilians exposed to dioxin TCDD in the Panama Canal Zone.

Inheritance and molecular characterization of resistance to AHAS- inhibiting herbicides in rapeseed

Rapeseed is a very important oil crop in China; however, its production is challenging due to the absence of effective weed management strategies. This is predominantly because of a shortage of herbicide resistance genes. Acetohydroxyacid synthase （AHAS） herbicides inhibit AHAS, a key enzyme involved in branched-chain amino acid synthesis that is required for plant growth. A rapeseed line designated M342 with AHAS herbicide resistance was developed through seed muta- genesis and was studied to assess the level and mode of inheritance of the resistance and to identify the molecular basis of resistance. M342 possessed a high level of cross-resistance to sulfonylureas （SUs） and imidazolinones （IMIs）. This resistance was due to AHAS insensitivity to these herbicides and was inherited as a dominant trait conferred by a single nuclear-encoded gene. Molecular analysis revealed the presence of a Trp574Leu mutation in M342, and an allele-specific cleaved amplified polymorphic sequence （AS-CAPS） marker was developed and cosegregated with herbicide resistance in the F2, BC1, and BC2 populations. This mutation altered the transcript levels of BnAHAS1 and BnAHAS3 in M342 compared with those in the wild type, but it did not affect the agronomic or quality traits. The simple genetic inheritance of this mutation and the availability of the cleaved amplified polymorphic sequence （CAPS） marker and herbicide resistance gene should facilitate the development of herbicide-resistant rapeseed cultivars for effective weed control in China.

Effect of Cadmium and Herbicides on the Growth, Chlorophyll and Soluble Sugar Content in Rice Seedlings

To investigate the combined effects of Cd^2＋ （ 0,10,100,500 μmol/L） , acetochlor （AC） （0,1.6,4.0,8.0μmol/L）, and bensulfuronmethyl （BSM） （ 0,0.16,0.40,0.80 μmol/L） on the biomass,leaf soluble sugar, total chlorophyll （chl） content, chl a/b ratio, and Cd content in roots and shoots in the rice seedlings, pot culture experiments were conducted with rice （Oryza sativa L. ） cultivar Jinyou 402. The results showed that the Cd and AC combined treatment （Cd＋ AC treatment） significantly inhibited the growth of the roots and shoots. The root dry weight/shoot dry weight （RDW/SDW） ratio, total chlorophyll content and chl a/b ratio decreased by 41%, 50%, 56% , respectively, in comparison with the control, as well as the leaf soluble sugar content, plant dry weight /plant fresh weight （PDW/PFW） ratio increased by 284% and 44% , respectively. Cd content in the roots and shoots had a decreasing tendency with the increasing concentration of AC under the Cd＋AC treatment. The Cd and BSM combined treatment （Cd＋BSM treatment） had a similar result to the Cd＋AC treatment. The results suggest that the toxicity of Cd to rice seedlings has been enhanced due to its combination with herbicides.

Predicting herbicides concentrations in paddy water and runoff to the river basin

This study was conducted to investigate the dissipation pattern and runoff of herbicides to the river basin from the paddy fields. Pesticide paddy field model(PADDY) was applied to predict herbicide concentration in paddy fields. A field study was conducted in a paddy farm of Higashi Hiroshima City, Hiroshima Prefecture, Japan in the year of 2003 paddy season. The herbicides were mefenacet, thiobencarb, and bensulfuron methyl. The sample water was analyzed by using gas chromatography and HPLC after solid phase extraction. Predicted dissipation rate of thiobencarb in paddy water was higher(DT_ 50 = 4 36) than that measured, with a lower k value(-0 069). Two weeks after application no thiobencarb was detected in the drainage channel and down stream. In the down stream, thiobencarb was detected until 3 d after application, with a range of 0 02% to 0 08% of applied herbicide. The predicted dissipation rate(k) and half-life(DT_ 50 ) of mefenacet was not significantly different from that of measured. In the drainage channel, upstream and downstream mefenacet was found during the whole study period. In downstream, the maximum concentration of mefenacet was present 0 61% of applied in the paddy field on 3DAH. The dissipation rate(k) of BSM varied from -0 0860 to -0 1059 to with half-life(DT_ 50 ) 3 5 and 2 84 d. In upstream water, no BSM was detected except trace amounts(0 01 μg/L) at 3 d after application. However, in the drainage channel 8%, 6% and 1 58% of applied BSM was present at 0, 1 and 3 d after application respectively. In the down stream, the highest concentration was 1 06%, shortly after application.

Phytotoxicity of four herbicides on Ceratophyllum demersum,Vallisneria natans and Elodea nuttallii

The physiological effects of 4 herbicides （butachlor, quinclorac, bensulfuron-methyl and atrazine） on 3 submerged macrophytes （Ceratophyllum demersum, Vallisneria natans and Elodea nuttallii） were tested in laboratory. The variables of the relative growth rate and the photosynthetic pigment content showed that all of the tested herbicides affected the growth of the plants obviously, even at the lowest concentration （0.0001 mg/L）. Except for the C. demersum treated with quinclorac at 0.005 and 0.01 mg/L, the relative growth rates of the plants were inhibited significantly （p 〈 0.01）. Statistical analysis of chlorophyll a （Chl-a） contents was carded out with both the t-test and one-way ANOVA to determine the difference between the treatment and control. The results showed that Chl-a contents of the plants in all treatment groups were affected by herbicides significantly, except for the C. demersum treated with bensulfuron-methyl at 0.0005 mg/L. The decrease in Chl-a content was positively correlated to the dosage of the herbicides in most treatment groups. It was suggested that herbicides in water bodies might potentially affect the growth of aquatic macrophytes. Since the Chl-a content of submerged macrophytes responded to the stress of herbicides sensitively and directly, it could be used as a biomaker in environmental monitoring or in the ecological risk assessment of herbicide contamination.

Adsorption and correlation with their thermodynamic properties of triazine herbicides on soils

Adsorption of atrazine, prometryne and prometon was determined on six soils with different physical and chemical properties. The adsorption isotherms of three herbicides could well fit Freundlich equation. On all of six soils, adsorption of herbicides increased in the order: atrazine ≈prometon<prometryne. This order is quite the same to the calculation result of by means of excess thermodynamic properties of triazine. The Freundlich adsorption constants, K f, showed to have good correlation with organic matter(OM%) of soils for each of these herbicides, suggesting that OM is the main factor, which dominates in the adsorption process of these triazine herbicides.

Adsorption of chloroacetanilide herbicides on soil and its components III. Influence of clay acidity, humic acid coating and herbicide structure on acetanilide herbicide adsorption on homoionic clays

Adsorption of chloroacetanilide herbicides on homoionic montmorillonite, soil humic acid, and their mixtures was studied by coupling batch equilibration and FT IR analysis. Adsorption isotherms of acetochlor, alachlor, metolachlor and propachlor on Ca 2+ \|, Mg 2+ \|, Al 3+ \| and Fe 3+ \| saturated clays were well described by the Freundlich equation. Regardless of the type of exchange cations, K f decreased in the order of metolachlor > acetolachlor > alachlor > propachlor on the same clay. FT IR spectra showed that the carbonyl group of the herbicide molecule was involved in binding, probably via H bond with water molecules in the clay interlayer. The type and position of substitutions around the carbonyl group may have affected the electronegativity of oxygen, thus influencing the relative adsorption of these herbicides. For the same herbicide, adsorption on clay increased in the order of Mg 2+ <Ca 2+ <Al 3+ ≤Fe 3+ which coincided with the increasing acidity of homoionic clays. Acidity of cations may have affected the protonation of water, and thus the strength of H bond between the clay water and herbicide. Complexation of clay and humic acid resulted in less adsorption than that expected from independent adsorption by the individual constituents. The effect varied with herbicides, but the greatest decrease in adsorption occurred at a 60:40 clay to humic acid ratio for all the herbicides. Causes for the decreased adsorption need to be characterized to better understand adsorption mechanisms and predict adsorption from soil compositions.

Simultaneous Voltammetric Determination of Three Herbicides in Food and Water Samples with the Aid of Chemometrics

Differential pulse stripping voltammetry method（DPSV） was applied to the determination of three herbicides, ametryn, cyanatryn, and dimethametryn. It was found that their voltammograms overlapped strongly, and it is difficult to determine these compounds individually from their mixtures. With the aid of chemometrics, classical least squares（CLS）, principal component regression（PCR） and partial least squares（PLS）, voltammogram resolution and quantitative analysis of the synthetic mixtures of the three compounds were successfully performed. The proposed method was also applied to the analysis of some real samples with satisfactory results.

Effect of Starch Sources on the Release Rates of Herbicides Encapsulated

The effects of starch sources on the behaviors of starch matrices and on the rates of herbicides released from the matrices were studied for slow release. The starches considered include native corn starch, wheat starch, potato starch and cassava starch. The matrices were prepared through encapsulating 2,4-dichlorophenoxyacetic or 2,4,5 trichlorophenoxyacetic acids as model herbicides with hot-gelatinized starch pastes. The encapsulation was evaluated in terms of herbicide content, swellability, encapsulation efficiency, and release rate. The results show that starch sources play an important role on the matrix behaviors and on release rates. The rate of 2,4-D released follows the order： wheat starch 〈 potaot starch 〈 corn starch 〈 cassava starch. And for the rate of 2,4,5-T, this order is nearly the same only with an exception that the late two kinds of starch are similar. It is also demonstrated that herbicides with different water solubility show different release rates, no matter what type of starch is used as the matrices.

Metsulfuron-methyl Molecularly Imprinted Stir Bar Sorptive Extraction Coupled with High Performance Liquid Chromatography for Trace Sulfonylurea Herbicides Analysis in Complex Samples

Metsulfuron-methyl molecularly imprinted polymer（MIP）-coated stir bar was prepared for sorptive extraction of sulfonylurea herbicides in complex samples.The MIP-coating was about 21.3 μm thickness with the relative standard deviation（RSD） of 4.4%（n=10）.It was homogeneous and porous with good thermal stability and chemical stability.The extraction capability of the MIP-coating was 2.8 times over that of the non-imprinted polymer（NIP）-coating in hexane.The MIP-coating exhibited selective adsorption ability to the template and its analogues.The extraction conditions,including extraction solvent,desorption solvent,extraction time,desorption time and stirring speed,were optimized.A method for the determination of six sulfonylurea herbicides by MIP-coated stir bar sorptive extraction coupled with high performance liquid chromatography（HPLC） was developed.The linear range was 10―200 μg/L and the detection limits were within a range of 2.0―3.3 μg/L.It was also applied to the analysis of sulfonylurea herbicides in spiked river water,soil and rice samples.

Glutathione-Responsive Carboxymethyl Chitosan Nanoparticles for Controlled Release of Herbicides

Glutathione-responsive carboxymethyl chitosan nanoparticles cross-linked with disulfide bonds were developed for controlled release of herbicides. The nanoparticles were synthesized by selfassembly of amphiphilic carboxymethyl chitosan derivative (CMCS-MUA) in aqueous solution and subsequently producing disulfide cross-linking bonds by ultrasonic treatment. TEM showed that the nanoparticles had a spherical core-shell configuration with a size of about 250 nm. Assessment of stability of the nanoparticles (considering mean diameter, polydispersity, and Zeta potential) was conducted over a period of three months, and the nanoparticles were found to be stable in solution. Herbicide-loaded nanoparticles were prepared using diuron as a model herbicide. In vitro release study revealed that diuron can be released from nanoparticles in a controlled manner depended on the glutathione concentration. Herbicidal activity assays performed with preemergence treatment of target species (Echinochloa crusgalli) showed the effectiveness of diuron- loaded nanoparticles. Assays with nontarget species (Zea mays) showed that the diuronloaded nanoparticles did not affect plant growth. The results indicate that the glutathioneresponsive nanoparticles prepared in this work will be a promising candidate for controlled release of herbicides in agriculture.

Integrated Weed Management Through Tillage and Herbicides for Wheat Production in Rice-Wheat Cropping System in Northwestern Pakistan

Conservational tillage （CT） in combination with broad spectrum herbicide may be more efficient and economical in controlling weeds, reducing cost of cultivation, and enhancing wheat yield. Field experiments were carried out to evaluate the effect of tillage and herbicides on weeds and wheat yield under rice-wheat cropping system （RWCS）. The results indicated that broad spectrum herbicides particularly, carfentrazone ethyl ester＋ isoproturon （Affinity 50 WDG） reduced weeds density by 93 and 95% compared to control and also produced the maximum grain yield （6 818 and 6 996 kg ha-l） and HI （harvest index, 44.7 and 44.6%） in 2007-2008 and 2008-2009 growing seasons, respectively. The maximum weeds infestation and the lowest yield were recorded in control. Tillage methods significantly influenced grain yield and HI, while no significant effect on all other parameters. The overall mean yield was significantly higher in zero tillage （5 575 kg ha-l） and reduced tillage （5 584 kg ha-l） compared to CT （5 479 kg ha-^-1）. Besides increasing wheat yield, Affinity 50 WDG and zero tillage are important weed management practices that can be integrated into wheat production in RWCS. However, in case of perennial weeds or insect pests＇ infestation, CT in combination with Affinity 50 WDG would be more productive than conservation tillage.