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BACKGROUND Wheat growers have limited herbicide options to manage Aegilops cylindrica Host (jointed goatgrass), with many relying on mesosulfuron or imazamox in Clearfield™ winter wheat. Both imazamox and mesosulfuron inhibit acetohydroxyacid synthase/acetolactate synthase (AHAS/ALS). In 2015, a suspected imazamox resistant biotype of Ae. cylindrica was found in eastern Washington. RESULTS Imazamox and mesosulfuron were applied to the suspected resistant and susceptible Ae. cylindrica biotypes in increasing application rates to evaluate herbicide dose needed to cause 50% growth reduction (GR50 ). The imazamox resistant biotype had a GR50 of 308.5 g ai ha-1 and was more than 5000 times more resistant to imazamox than a known susceptible biotype with a GR50 of 0.06 g ai ha-1 . The Ae. cylindrica resistant biotype was also resistant to mesosulfuron, with an GR50 of 46.82 g ai ha-1 , which was five times more than the susceptible GR50 of 8.6 g ai ha-1 . Sequencing of the AHAS/ALS gene revealed an Ala122 Thr substitution in the herbicide binding region of the AHAS/ALS gene on the D genome of Ae. cylindrica. The resistance trait was inherited as a dominant trait, and the Ala122 Thr co-segregates with the resistance phenotype. CONCLUSIONS An Ala122 Thr substitution in the AHAS/ALS gene on the D genome of Ae. cylindrica confers resistance to imazamox in Ae. cylindrica. © 2021 Society of Chemical Industry.

Purpose. Provide bioinformatic analysis and comparison of target regions of the acetolactate synthase (als) gene in several members of the Poaceae family and, on the basis of the obtained data, explore the possibility of creating a unified genetic construct for als gene editing using the CRISPR-Cas9 system. Methods. The als gene sequences of various members of the Poaceae family were obtained from the NCBI: Nucleotide database. For comparison, a fragment of the imi-2 gene of wheat of the soft line ‘TealIMI11A’ was used in two regions of the 367–390 and 1729–1749 nucleotide sequences. The Sequence Viewer 3.37.0 tool was used to assess the presence of nucleotide substitutions in the working sequence of the imi-2 gene. The dendrogram was built using the “Blast Tree” tool from the NCBI: Blast: Nucleotide resource. Results. A comparative analysis of the nucleotide sequences of seven different species was carried out: soft wheat (Triticum aestivum L.), common wild oat (Avena fatua L.), barley (Hordeum vulgare L.), Asian rice (Oryza sativa L.), maize (Zea mays L.), aleppo grass (Sorghum halepense Pers.) and Tausch’s goatgrass (Aegilops tauschii Coss.). The dendrogram is based on the gene sequence als, showed that all studied genotypes can be divided into two blocks: the first block included maize and aleppo grass, and the second block, a separate branch includes Asian rice and common wild oat, barley, soft wheat and Tausch’s goatgrass. 367–390 nucleotide sequences of soft wheat showed the highest 100% homology to Asian rice, Tausch’s goatgrass and common wild oat. The lowest homo­logy was for maize and aleppo grass at 83.3%. Evaluation of the nucleotide sequence 1729–1749 showed no complete homology at the 100% level. It was the highest for barley and Tausch’s goatgrass – 95.2%, and the lowest for rice, maize and aleppo grass – 80.9% each. Conclusions. The analysis confirms a significant degree of homology of the als gene sequence for various species of the Poaceae family, which allows us to create a universal genetic vector. However, taking into account the high degree of sequence homology for species such as soft wheat, Tausch’s goatgrass, barley, Asian rice and common wild oat, it can be assumed that the corresponding genetic vector can be used with the greatest efficiency to alter the als gene of these genotypes.

Wild buckwheat (Polygonum convolvulus L.) is a problem weed and ALS-inhibitors (e.g., chlorsulfuron) are commonly used for its management. Recently, a population of wild buckwheat (KSW-R) uncontrolled with ALS-inhibitors was found in a wheat field in Kansas, USA. The objectives of this research were to determine the level and mechanism of resistance to chlorsulfuron and cross resistance to other ALS-inhibitors in the KSW-R population. In response to chlorsulfuron rates ranging from 0 to 16x (x = 18 g ai/ha), the KSW-R wild buckwheat was found &gt, 100-fold more resistant compared to a known ALS-inhibitor susceptible (KSW-S) wild buckwheat. Also, &gt, 90% of KSW-R plants survived field recommended rates of sulfonylurea but not imidazolinone family of ALS-inhibitors. A portion of the ALS gene covering all previously reported mutations known to bestow resistance to ALS-inhibitors was sequenced from both KSW-R and KSW-S plants. The Pro-197-Ser substitution that confers resistance to the sulfonylurea herbicides was found in KSW-R plants. Our results support the evolution of high level of chlorsulfuron resistance as a result of a mutation in the ALS-gene in KSW-R buckwheat. This is the first case of resistance to any herbicides in wild buckwheat in the US.

Specific mutations in the conserved domains of the acetolactate synthase (ALS) gene conduct to different key amino acid substitutions that can confer herbicide resistance in different plant species. This outcome has been widely exploited to produce herbicide-resistant agronomic crops as well as to direct many genome editing studies. Therefore, the ALS gene has become a model sequence target to improve our technological skills for more precise CRISPR/Cas nucleotide base substitution in plants, which is essential for modulation/modification of gene function as opposed to the more general gene knock out obtained by indels in conventional genome editing studies. This review summarizes the main knowledge and experiences attained from the use of the ALS gene as a target in CRISPR/Cas studies. Instituto de Biotecnología Fil: Darqui, Flavia Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Hopp, Horacio Esteban. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina Fil: Lopez Bilbao, Marisa Gisela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Diplotaxis erucoides and Erucaria hispanica are common weeds of the Mediterranean region, they infest various habitats including cultivated fields and roadsides. In several fields across Israel, farmers have reported on poor control of D. erucoides and E. hispanica plants using acetolactate synthase (ALS) inhibitors. Greenhouse experiments were conducted to determine the effect of various ALS inhibitors on plants from two potentially resistant D. erucoides and E. hispanica populations. Additionally, alternative management strategies using auxinic herbicides were studied. Plants from both populations exhibited resistance to all tested ALS inhibitors, up to 20-fold the label field rate, as compared with ALS sensitive populations of D. erucoides and E. hispanica. Sequencing of the ALS gene revealed Trp574 to Leu substitution in ALS-resistant D. erucoides plants, whereas a Pro197 to Ser substitution was detected in ALS-resistant E. hispanica plants. Although high levels of resistance were observed in individuals from both putative resistant populations, sensitive individuals were also detected, suggesting the evolution of resistance in these two populations is still in progress. Auxinic herbicides, 2,4-D, and mecoprop-P, provided excellent control of plants from both ALS-resistant populations. This study documents and confirms the first case of evolution of resistance to ALS inhibitors in D. erucoides and E. hispanica populations.

Pennsylvania smartweed [Persicaria pensylvanica(L.) M. Gómez] is a common weed of rice (Oryza sativaL.) in the midsouthern United States and has recently become a concern for farmers because of reduced tillage systems. Acetolactate synthase (ALS) inhibitors have been extensively used for controlling smartweeds in imidazolinone-resistant and conventional rice. In the present study, we confirmed resistance to commonly used ALS inhibitors in rice and characterized the underlying resistance mechanism in aP. pensylvanicabiotype from southeast Missouri. A dose–response experiment was conducted in the greenhouse using bensulfuron-methyl, imazethapyr, and bispyribac-sodium to determine the resistance index (resistance/susceptibility [R/S]) based on GR50estimates. The target-siteALSgene was amplified from R and S plants, and sequences were analyzed for mutations known to confer ALS-inhibitor resistance. TheP. pensylvanicabiotype in question was found to be resistant to bensulfuron-methyl (R/S=2,330), imazethapyr (R/S=12), and bispyribac-sodium (R/S=6). Sequencing of theALSgene from R plants revealed two previously known mutations (Pro-197-Ser, Ala-122-Ser) conferring resistance to sulfonylureas and imidazolinones. This is the first report of ALS-inhibitor resistance inP. pensylvanica.

Proposed pathogenesis of most neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS), is complex and typically involves interactions between genetic and environmental factors.1,2 The contribution of genetics in neurodegenerative diseases varies from monogenic causes to more multifaceted susceptibility genetic factors. However, identification of even rare single gene causes in these conditions has been crucial to our evolving understanding of neurodegenerative cascades and may ultimately lead to the development of new therapeutic interventions.3

Green foxtail [Setaria viridis (L) Beauv.] and yellow foxtail [Setaria pumila (Poir.) Roem. & Schult.] are among the most abundant and troublesome annual grass weeds in cereal crops in the Northern Plains of the United States and the Prairie Provinces of Canada. Greenhouse and laboratory experiments were conducted to examine the differential responses of both weed species to foliar applications of the new triazolopyrimidine sulfonamide acetolactate synthase-inhibiting herbicide, pyroxsulam, and to determine the mechanism(s) of differential weed control. Foliar applications of pyroxsulam resulted in >90% control of yellow foxtail at rates between 7.5 and 15 g ai ha–1, whereas the same rates resulted in a reduced efficacy on green foxtail (≤81%). The absorption and translocation of [14C]pyroxsulam in green and yellow foxtail were similar and could not explain the differential whole-plant efficacy. Studies with [14C]pyroxsulam revealed a higher percentage of absorbed pyroxsulam was metabolized into an inacti...

Recent advances in ALS gene discovery have both empowered and challenged clinicians providing evaluation and care for persons with ALS, many of whom seek an answer as to the cause of their condition. In order to study clinician practices and attitudes towards genetic testing, we surveyed members of the Northeast ALS Consortium, an international group of specialist ALS clinicians; responses were received from 80 of 255 (response rate = 31.4%). While 92.3% indicated they offered genetic testing to patients with familial ALS, 57.0% offered testing to patients with ALS and a family history of dementia, and 36.9% offered testing to patients with sporadic ALS, revealing a lack of consensus with respect to the approach to the typical ALS patient encountered in clinical practice. In addition, comparison of clinician and patient attitudes towards genetic testing revealed that clinicians valued the scientific potential of testing, but were less likely to say they would have testing themselves, or to see the value in testing for family members. People with ALS were more likely to see value of testing for themselves and for family members, and less likely to strongly value the scientific potential of testing.

Amaranthus palmeri S. Watson (Amaranthaceae Juss.) is a dioecious noxious weed, native to the Americas, which infests summer crops. It causes high crop losses, and rapidly evolves resistance to herbicides. In Europe, A. palmeri was recorded mostly as a casual alien, but in 2018 it was reported infesting a soybean field in Italy, and the next year two more populations were found in the same area. Experiments were conducted on these three populations to evaluate the resistance to ALS-inhibiting herbicides, to determine the main resistance mechanisms involved and assess the efficacy of alternative herbicides with different sites of action than ALS. The three populations were confirmed cross-resistant to ALS-inhibiting herbicides (thifensulfuron-methyl and imazamox). Gene sequencing identified a Trp to Leu substitution at position 574 of ALS gene in resistant plants, proving that the main resistance mechanism for the three populations is target-site related. The presence of other resistance mechanisms cannot be excluded. Metobromuron, metribuzin and glyphosate are still effective on these populations.

To evaluate the mechanism and levels of herbicide resistance in Sinapis arvensis, a series of experiments were performed. Sequencing of ALS gene in S. arvensis sub-populations was carried out and ALS-based resistance (substitution of Asp by Glu) was detected at position 376 in this weed species. Further, it was reported that combined application of herbicide mesosulfuron + iodosulfuron can cause up to 50% reduction in dry weight (GR50) of heterozygous resistant (FHR3), homozygous resistant (FHR2) and homozygous susceptible (MHS1) sub-populations and it was reported 2409, 603 and 289 g ha -1 , respectively. Further, resistance indices (RI) were reported 8.3 and 2 for FHR3 and FHR2 sub-populations, respectively. Based on LD50, RI values were 8.5 and 4.5 for FHR3 and FHR2, respectively. Growth reduction occurred in the resistant homozygous when compared with the heterozygous sub-population in the presence of mesosulfuron + iodosulfuron.

Recent advances in the genetics of ALS have bolstered hope that a molecular logic for the pathogenesis of the disease is fast approaching. An emerging challenge is the dissection of the common and unique molecular pathways altered by ALS gene mutations. Disease modeling in rodents has yielded many important insights, but as the genetic complexity of the disease grows, additional models with improved speed, cost and genetic tractability will be increasingly necessary. Models such as fruitfly, nematode, and zebrafish have been important for diagramming the molecular pathways that underlie many fundamental biological processes, but have been comparatively underutilized in the study of neurodegeneration. Here we highlight the benefits and opportunities for increased diversity in the models used to study ALS.

© 2018 Elsevier Inc., To identify novel genes associated with ALS, we undertook two lines of investigation. We carried out a genome-wide association study comparing 20,806 ALS cases and 59,804 controls. Independently, we performed a rare variant burden analysis comparing 1,138 index familial ALS cases and 19,494 controls. Through both approaches, we identified kinesin family member 5A (KIF5A) as a novel gene associated with ALS. Interestingly, mutations predominantly in the N-terminal motor domain of KIF5A are causative for two neurodegenerative diseases: hereditary spastic paraplegia (SPG10) and Charcot-Marie-Tooth type 2 (CMT2). In contrast, ALS-associated mutations are primarily located at the C-terminal cargo-binding tail domain and patients harboring loss-of-function mutations displayed an extended survival relative to typical ALS cases. Taken together, these results broaden the phenotype spectrum resulting from mutations in KIF5A and strengthen the role of cytoskeletal defects in the pathogenesis of ALS.

Comunicación presentada al XVI Congreso de la Sociedad Española de Malherbología, celebrado en la Universidad Pública de Navarra, Pamplona-Iruña, entre los días 25 y 27 de octubre de 2017. Existen bastantes casos de resistencia en Echinochloa spp. descritos a nivel mundial en arroz. En los últimos 2 años se han realizado muestreos periódicos en campos de arroz en Extremadura, en lugares donde se sospecha resistencia. Con estas semillas se han realizado tanto ensayos en invernadero para confirmación de las resistencias, como estudios de mecanismos de resistencia a nivel de sitio de acción (secuenciación de genes ALS y ACCasa). Hasta ahora, de las muestras analizadas, se han encontrado 37 poblaciones con resistencia confirmada (con mutaciones en el sitio de acción) a nivel de ALS y 2 con resistencia a nivel de ACCasa. También se han encontrado poblaciones donde ambas mutaciones (gen ALS y ACCasa) están presentes. Estas poblaciones están distribuidas en distintas zonas arroceras de la región extremeña. El estudio del mecanismo responsable de la resistencia es básico para el diseño de un programa de control integrado. The genus Echinochloa spp. includes the most problematic species in rice worldwide, and in particular in rice fields in Extremadura. The herbicides authorized in Spain at present to control this weed in rice belong mainly to 2 modes of action: ALS inhibitors and ACCase inhibitors. Repeated and continuous use of herbicides with the same mode of action may lead to the appearance of resistance. In the last 2 years, periodic sampling has been carried out in rice fields in Extremadura, in places where resistance is suspected. We have a set of internal assays for confirmation of resistance, such as studies of mechanisms of resistance to a target site-level (ALS and ACCase gene sequencing). So far, 37 populations were resistant (with mutations at the site of action) to ALS inhibitors and 2 to ACCase inhibitors. These populations are distributed in different areas of the Extremadura region. The study of the mechanism responsible for the resistance is basic for the design of an integrated control program. Este trabajo está financiado por el proyecto INIA RTA-2014-00033-C03-01, por la ayuda a Grupos de Investigación de la Junta de Extremadura GR15112 y por el proyecto AGROS (CCESAGROS), fondos FEDER.

Comunicación presentada al XVI Congreso de la Sociedad Española de Malherbología, celebrado en la Universidad Pública de Navarra, Pamplona-Iruña, entre los días 25 y 27 de octubre de 2017. Existen bastantes casos de resistencia en Echinochloa spp. descritos a nivel mundial en arroz. En los últimos 2 años se han realizado muestreos periódicos en campos de arroz en Extremadura, en lugares donde se sospecha resistencia. Con estas semillas se han realizado tanto ensayos en invernadero para confirmación de las resistencias, como estudios de mecanismos de resistencia a nivel de sitio de acción (secuenciación de genes ALS y ACCasa). Hasta ahora, de las muestras analizadas, se han encontrado 37 poblaciones con resistencia confirmada (con mutaciones en el sitio de acción) a nivel de ALS y 2 con resistencia a nivel de ACCasa. También se han encontrado poblaciones donde ambas mutaciones (gen ALS y ACCasa) están presentes. Estas poblaciones están distribuidas en distintas zonas arroceras de la región extremeña. El estudio del mecanismo responsable de la resistencia es básico para el diseño de un programa de control integrado. The genus Echinochloa spp. includes the most problematic species in rice worldwide, and in particular in rice fields in Extremadura. The herbicides authorized in Spain at present to control this weed in rice belong mainly to 2 modes of action: ALS inhibitors and ACCase inhibitors. Repeated and continuous use of herbicides with the same mode of action may lead to the appearance of resistance. In the last 2 years, periodic sampling has been carried out in rice fields in Extremadura, in places where resistance is suspected. We have a set of internal assays for confirmation of resistance, such as studies of mechanisms of resistance to a target site-level (ALS and ACCase gene sequencing). So far, 37 populations were resistant (with mutations at the site of action) to ALS inhibitors and 2 to ACCase inhibitors. These populations are distributed in different areas of the Extremadura region. The study of the mechanism responsible for the resistance is basic for the design of an integrated control program. Este trabajo está financiado por el proyecto INIA RTA-2014-00033-C03-01, por la ayuda a Grupos de Investigación de la Junta de Extremadura GR15112 y por el proyecto AGROS (CCESAGROS), fondos FEDER.

BACKGROUND Yellow nutsedge is one of the most problematic sedges in Arkansas rice, requiring the frequent use of halosulfuron (sulfonylurea) for its control. In the summer of 2012, halosulfuron at 53 g ha−1 (labeled field rate) failed to control yellow nutsedge. The level of resistance to halosulfuron was determined in the putative resistant biotype, and its cross-resistance to other acetolactate synthase (ALS) inhibitors from four different herbicide families. ALS enzyme assays and analysis of the ALS gene were used to ascertain the resistance mechanism. RESULTS None of the resistant plants was killed by halosulfuron at a dose of 13 568 g ha−1 (256× the field dose), indicating a high level of resistance. Based on the whole-plant bioassay, the resistant biotype was not controlled by any of the ALS-inhibiting herbicides (imazamox, imazethapyr, penoxsulam, bispyribac, pyrithiobac-sodium, bensulfuron and halosulfuron) tested at the labeled field rate. The ALS enzyme from the resistant biotype was 2540 times less responsive to halosulfuron than the susceptible biotype, and a Trp574-to-Leu substitution was detected by ALS gene sequencing using the Illumina HiSeq. CONCLUSION The results suggest a target-site alteration as the mechanism of resistance in yellow nutsedge, which accounts for the cross-resistance to other ALS-inhibiting herbicide families. © 2014 Society of Chemical Industry

SUMMARYThe lack of control of barnyardgrass in flooded rice cultivated with imidazolinone-resistant rice cultivars is challenging the utilization of this system, which is continuously expanding for new rice areas worldwide. The objectives of the present study were to evaluate the frequency, distribution and mechanisms of imidazolinone resistance in barnyardgrass to establish the best practices to control and prevent this problem. The distribution of resistance was evaluated in 624 populations collected in Southern Brazil. The frequency of imidazolinone-herbicide resistance was 0·81, broadly distributed in all sampled regions. Resistance to quinclorac was also found in 0·19 of the populations, but all of the evaluated populations were susceptible to cyhalofop-butyl. Further studies were conducted in six populations. The enhanced metabolism was assessed with the metabolic inhibitors that reversed the resistance to quinclorac from 0·54 to 1·00 in two populations and the resistance to imazethapyr from 0·15 to 0·41 in three populations. The acetolactase synthase (ALS) enzyme activity also indicated the occurrence of altered target site resistance in two populations caused by the ALS gene mutations Trp574Leu and Ser653Asn, which is a novel finding in this species. The herbicide resistance in barnyardgrass in Southern Brazil presented a complex basis of resistance because it is associated with resistance to multiple herbicides due to multiple mechanisms and with multiple mutations of the ALS gene. This indicates that it is necessary to adopt specific measures to prevent and control the evolution of multiple herbicide resistance in this species.

Populations of water starwort, a winter annual or biennial weed in the pink family (Caryophyllaceae), can no longer be controlled by tribenuron following successive use of this herbicide over several years. Whole-plant bioassays have established that the resistant water starwort populations JS17, JS08, JS16, and JS07 showed high-level (from 203-fold to 565-fold) resistance to tribenuron. In vitro acetolactate synthase (ALS) assays revealed that resistance was due to reduced sensitivity of the ALS enzyme to tribenuron. The half-maximal inhibitory concentration (I50) values for JS17, JS08, JS16, and JS07 were 72, 71, 70, and 76 times greater, respectively, than were those of the susceptible population JS24. This altered ALS sensitivity in the resistant populations was due to a mutation in the ALS gene resulting in a Pro197to Ser substitution (JS17, JS08, and JS16) and a Pro197to Leu substitution (JS07). This study established the first documented case, to our knowledge, of evolved tribenuron resistance in water starwort and concluded that the molecular basis of resistance is due, at least in part, to a target-site modification at Pro197in the ALS gene.

DeutschDie strafrechtlichen Risiken fur Geschaftsleiter von Kapitalgesellschaften sind vielfaltig. Neben spezifischen Straftatbestanden, die diesen Personenkreis unmittelbar adressieren, sowie diversen strafrechtlichen Zurechnungskonstruktionen, mit denen Geschaftsleiter fur Straftaten von Mitarbeitern haftbar gemacht werden konnen, ragt insbesondere der Un- treuetatbestand des § 266 StGB als strafrechtliche Allzweckwaffe heraus. Der Beitrag zeigt auf, wie die Strafsenate des BGH diesen Tatbestand durch kreative Auslegung der Merk- male Vermogensbetreuungspflicht und Vermogensnachteil standig erweitert und als Gene- ralklausel fur nahezu alle Arten wirtschaftlichen Fehlverhaltens passend gemacht ha- ben. EnglishThe German legal system provides a wide range of criminal liabilities for CEOs of corpora- tions. Besides specific criminal offenses dealing with these persons directly, and diverse other criminal law constructions which make managers liable for crimes committed by employees, the criminal offense in Sec. 266 German Criminal Code, „Abuse of Trust“, clearly stands out as a broad spectrum criminal law instrument. This contribution will show how the criminal senates of the German Federal Supreme Court have continuously broadened the scope of this criminal offense by creatively interpreting the „duty to safeguard property interests“ and „causing damage“ in such a way that it now is a general provision against almost all kinds of abusive business behavior.

Wild buckwheat is the most abundant broadleaf weed across the Prairie region of western Canada. Acetolactate synthase (ALS)-inhibiting herbicides are commonly used to control this species and other broadleaf weeds in cereal crops. A field survey in Alberta in 2007 identified a single population that was putatively resistant to ALS-inhibiting herbicides. In herbicide resistance screening in the greenhouse, all F1 progeny tested were resistant to the ALS-inhibiting herbicides thifensulfuron/tribenuron, a sulfonylurea herbicide, or florasulam, a triazolopyrimidine herbicide; dose response of shoot biomass indicated the population was 10- and 20-fold less sensitive to thifensulfuron/tribenuron and florasulam, respectively, than a susceptible control population. ALS gene sequencing of 24 F1 progeny indicated that the Trp574Leu target-site mutation was responsible for conferring ALS-inhibitor resistance in this biotype, the first global report of ALS-inhibitor resistance for this species. Because this mutation typically endows high-level resistance across all five ALS-inhibitor classes, this wild buckwheat biotype may only be controlled by a different site-of-action herbicide.

Cleavers species (false cleavers and catchweed bedstraw) are among the top 10 most abundant weeds across the prairie region of western Canada, and are increasing in relative abundance at the fastest rate since the 1970s. In 2008, two false cleavers populations from Tisdale and Choiceland, Saskatchewan, were suspected of acetolactate synthase (ALS) –inhibitor resistance. Dose-response experiments were conducted with the use of imazethapyr and florasulam, both ALS inhibitors, as well as fluroxypyr, a synthetic auxin. Additionally, a 1,954–base-pair region of theALSgene including sites known to conferALSresistance were sequenced. Both populations were highly resistant to imazethapyr (resistance factors greater than 100), one population (Tisdale) was highly resistant to florasulam (Choiceland population susceptible, although a second, larger screening of 200 individuals indicated low frequency [2%] florasulam resistance), and both populations were susceptible to fluroxypyr. All sequenced Tisdale individuals screened with imazethapyr posessed the Trp574Leu mutation. In contrast, three point mutations were found for Choiceland individuals sequenced: Ser653Asn, Trp574Leu, and Asp376Glu. TheseALStarget-site mutations have not been documented previously in this species.

ALSGene Mutations inApera Spica-VentiConfer Broad-Range Resistance to HerbicidesSeveral biotypes of wind bentgrass in Poland have been identified as being resistant to acetolactate synthase (ALS) inhibitors. We screened these weeds with chlorsulfuron and performed a whole-plant bioassay with a range of doses based on these four herbicides: chlorsulfuron, sulfosulfuron, propoxycarbazone-sodium and mesosulfuron-methyl + iodosulfuron-methyl-sodium mixture. Ten biotypes, diverse in their levels of resistance, were submitted for molecular tests. PCR amplification and sequencing of als domains demonstrated numerous single nucleotide polymorphisms. Nine biotypes showed non-synonymous substitutions in codon Pro197, changing it to Ser or Thr. Mutation in Pro197conferred a high level of resistance to the tested herbicides. Analysis of four biotypes also revealed a substitution in the Ala122codon, changing it to Val. In one biotype this substitution was not accompanied by Pro197mutation and this biotype was resistant to chlorsulfuron and mesosulfuron + iodosulfuron, but not to sulfosulfuron or propoxycarbazone-sodium. Correspondence between mutations and levels of resistan ce to ALS inhibitors may support management of resistant weeds with the existing palette of herbicides.

O uso contínuo de herbicidas inibidores da ALS tem levado à evolução de plantas daninhas resistentes mundialmente. Picão-preto é uma das mais importantes plantas daninhas no sistema de produção de soja no Brasil. Recentemente, foi identificado um biótipo de picao-preto resistente (R) aos inibidores da ALS devido à mutação Trp574Leu no gene ALS. Também, foram comparadas características adaptativas entre biótipos de picao-preto suscetível (S) e R aos inibidores da ALS. Especificamente, os objetivos deste trabalho foram: (1) avaliar e comparar o crescimento relativo (RGR) entre os biótipos; (2) analisar características da germinação das sementes dos biótipos R e S sob diferentes condições de temperatura; e (3) avaliar suas habilidades competitivas em um estudo em série de substituição. Os experimentos foram conduzidos na University of Arkansas, EUA, em 2007 e na Universidade Federal do Rio Grande do Sul, Brasil, em 2008. Para os estudos de série de substituição as proporções de plantas dos biótipos foram, respectivamente: 100:0, 75:25, 50:50, 25:75 e 0:100, com uma população total de 150 plantas m-2. Não se verificou diferença no RGR entre os biótipos R e S. A taxa de germinação do biótipo R foi menor que a do biótipo S. Porém, sob baixas condições de temperatura (15 oC), foi observada a situação oposta. Em geral, não há diferença na habilidade competitiva entre os biótipos R e S de picão-preto The continuous use of ALS-inhibiting herbicides has led to the evolution of herbicide-resistant weeds worldwide. Greater beggarticks is one of the most troublesome weeds found in the soybean production system in Brazil. Recently, a greater beggarticks biotype that is resistant (R) to ALS inhibitors due to Trp574Leu mutation in the ALS gene was identified. Also, the adaptive traits between susceptible (S) and R to ALS inhibitors biotypes of greater beggarticks were compared. Specifically, we aimed to: (1) evaluate and compare the relative growth rates (RGR) between the biotypes; (2) analyze the seed germination characteristics of R and S biotypes under different temperature conditions; and (3) evaluate their competitive ability in a replacement series study. The experiments were conducted at the University of Arkansas, USA, in 2007 and at Universidade Federal do Rio Grande do Sul (Federal University of Rio Grande do Sul), Brazil, in 2008. Plant proportions for replacement series studies were respectively 100:0, 75:25, 50:50, 25:75 and 0:100, with a total population of 150 plants m-2. There was no difference in RGR between R and S biotypes. The R-biotype germination rate was lower than that of the S biotype. However, at low temperature conditions (15 oC), the reverse was observed. In general, there is no difference in the competitive ability between R and S greater beggarticks biotypes

Reports arose from major Chinese wheat production regions that flixweed was not controlled by tribenuron after the herbicide was continuously used for several years. Flixweed seeds were collected from wheat fields that had been treated with tribenuron repeatedly over 3 to 15 yr or from road sides and remote hills that had never received tribenuron in Jiangsu, Hebei, Shanxi, Sichuan, Shandong, Shaanxi, and Henan provinces, and Tianjin and Beijing metropolises in China. The response of various biotypes to tribenuron was determined by whole plant experiments in the greenhouse. The experiments demonstrated that 11 of 32 flixweed biotypes were susceptible to tribenuron. The remaining 21 biotypes expressed moderate to high levels of tribenuron resistance with resistance indices ranging from 4 to > 1,500. DNA sequence analysis of acetolactate synthase (ALS) genes of selected biotypes 6, 7, 27, and 29 revealed a point mutation at position 197 of the ALS gene, as numbered relative to the protein sequence ...

Applications of 64 times higher rates of azimsulfuron and halosulfuron-methyl failed to reduce by 50% growth of 10 putative resistant (R) Cyperus difformis (smallflower umbrella-sedge) populations collected from rice fields located in Northern Greece. However, the growth rate of the susceptible (S) population was reduced by 50% by 1/4 of the recommended rate of both herbicides. The als gene sequencing identified point mutations at Pro-197 position, leading to amino acid substitution by Ala or Ser of the ALS enzyme. The in vitro activity of the ALS enzyme indicated that the I 50 values (herbicide concentration required for 50% reduction of the ALS activity) ranged from 10.7 to 55.5 μM azimsulfuron and from 6.7 to 50.6 μM halosulfuron-methyl, whereas the respective values for the S population were 0.09 and 0.11 μM. These results strongly support that cross-resistance of 10 C. difformis populations was due to point mutations of the als gene, which resulted in a less-sensitive ALS enzyme. This study reports the presence of a 1240 bp intron in the C. difformis als gene with the Pro-197 point mutations near the splice junction.

Multiple cases of ALS inhibitor-resistant weed biotypes are reported for many species, including wild mustard. The physiological extent and molecular basis of resistance to ALS inhibitors was compared in four biotypes of wild mustard from western Canada: a sulfonylurea (SU)-resistant (R) biotype from Manitoba detected in 1992; an SU (ethametsulfuron)-R biotype from Alberta detected in 1993 (metabolism-based resistance); an SU-R biotype from Manitoba detected in 2002; and a SU- and imidazolinone (IMI)-R biotype from Saskatchewan detected in 2002. Herbicide dose-response experiments confirmed that the two Manitoba biotypes were resistant to the SU herbicides ethametsulfuron and tribenuron : thifensulfuron mixture, whereas the Saskatchewan biotype was resistant to both SU herbicides and to imazethapyr, an IMI herbicide. Sequence analysis of theALSgene detected target site mutations in three of the four R biotypes, with amino acid substitutions Pro197(CCT) to Ser (TCT) [Domain A of the gene] in the two SU-R Manitoba biotypes and Trp574(TGG) to Leu (TTG) [Domain B] in the Saskatchewan biotype. The Alberta SU-R biotype had the sameALSnucleotide and amino acid sequence as the susceptible population at these two positions. Two heterozygous individuals [Trp574(Tt/gG)] were detected in the Saskatchewan biotype, and genetic segregation for nucleotide bases and resistance phenotype was consistent with single gene control. Nucleotide variation in neutral regions of theALSgene varied with biotype, with no variation in the two Manitoba biotypes, two variants in the Saskatchewan biotype, and 16 neutral nucleotide polymorphisms (0.9%) in the Alberta biotype. The occurrence of at least three different ALS inhibitor-R biotypes in this important weed species is likely to impact negatively on the use of ALS inhibitors, such as the IMIs, and serves as a warning for strict implementation of herbicide rotations to prevent or delay the evolution and spread of such populations.

The feasibility of using the sex-linked gene for imperfect albinism (S*ALS) to sex chicks during incubation by candling was studied. With this technique, the dark eye of nonalbino embryos can be positively identified. Two trials were performed. In a first trial, 66.5 and 89.5% of the 254 nonalbino and 210 albino chicks produced in four hatches were correctly identified by candling at 7 d of incubation. Of 191 eggs predicted to be nonalbinos, 22 were albinos, resulting in an overall accuracy of 88.5% for identification of nonalbino embryos. In a second trial, the accuracy of the technique from 7 to 10 d of incubation was evaluated. Increased age resulted in a tendency for lower accuracy, but candling at 8, 9, or 10 d of incubation allowed identification of a greater (P < 0.05) proportion of the nonalbino population than at 7 d of incubation. Candling at 8 d of incubation allowed identification of nonalbinos and albinos with an accuracy of 81.3 and 84.9%, respectively, suggesting that the ALS gene could be used to sex chicks during incubation when used in a sex-linked cross. This technique may prove advantageous to the laying industry because of savings of incubator and hatcher space. The males of commercial layer lines are normally killed at hatch. Reducing the number that hatch by eliminating them before 10 d of incubation could diminish animal welfare concerns.

Several studies have suggested an increased frequency of variants in the gene encoding angiogenin (ANG) in patients with amyotrophic lateral sclerosis (ALS). Interestingly, a few ALS patients carrying ANG variants also showed signs of Parkinson disease (PD). Furthermore, relatives of ALS patients have an increased risk to develop PD, and the prevalence of concomitant motor neuron disease in PD is higher than expected based on chance occurrence. We therefore investigated whether ANG variants could predispose to both ALS and PD.We reviewed all previous studies on ANG in ALS and performed sequence experiments on additional samples, which allowed us to analyze data from 6,471 ALS patients and 7,668 controls from 15 centers (13 from Europe and 2 from the USA). We sequenced DNA samples from 3,146 PD patients from 6 centers (5 from Europe and 1 from the USA). Statistical analysis was performed using the variable threshold test, and the Mantel-Haenszel procedure was used to estimate odds ratios.Analysis of sequence data from 17,258 individuals demonstrated a significantly higher frequency of ANG variants in both ALS and PD patients compared to control subjects (p = 9.3 × 10(-6) for ALS and p = 4.3 × 10(-5) for PD). The odds ratio for any ANG variant in patients versus controls was 9.2 for ALS and 6.7 for PD.The data from this multicenter study demonstrate that there is a strong association between PD, ALS, and ANG variants. ANG is a genetic link between ALS and PD.

NeuroscienceAn international consortium has found that a repeating nucleotide sequence on chromosome 9 causes amyotrophic lateral sclerosis (ALS). It's the most important ALS gene discovered to date. Now, the whole field is waiting to discover how the mutation causes ALS—and how, in some people, indistinguishable mutations instead paradoxically trigger frontotemporal dementia.