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BACKGROUND: Chlorsulfuron resistance and genetic dominance was evaluated in Raphanus raphanistrum genotypes homozygous (122-RR, 376-RR), heterozygous (122-RS, 376-RS) and compound heterozygous (122-R/376-R) for the target-site resistance mutations Ala-122-Tyr and Asp-376-Glu in the AHAS (acetohydroxyacid synthase) gene. RESULTS: At the AHAS level, 122-RR and 122-RS plants exhibited significantly higher I50 values than 376-RR and 376-RS plants, respectively. However, plants of the compound heterozygous genotype (122-R/376-R), showed no difference in AHAS activity compared to the 122-RS genotype but lower activity than the 122-RR genotype, and showed a nearly 400-fold greater I50 value than both genotypes (376-RR and 376-RS) carrying the 376-Glu allele. At the whole-plant level, 100% survival was observed for 122-RR plants at the highest chlorsulfuron dose of 640 g ha-1, yet 376-RR plants showed no survival at 380 g ha-1. Thus, this survival difference resulted in different median lethal dose (LD50) estimates [>640 (122-RR) versus 330 g ha-1(376-RR)]. The effect of chlorsulfuron in reducing aboveground growth of surviving plants also was markedly lower for the homozygous 122-RR (GR50 = 566 g ha-1) than for 376-RR plants (GR50 = 66). Heterozygous plants carrying the 122-Tyr allele (122-RS) exhibited two- and five-fold higher LD50 values than both homozygous and heterozygous plants carrying the 376-Glu allele (376-RR, 376-RS), respectively. Along the difference in plant survival, 122-RS plants also showed four-fold higher GR50 than both 376-RR and 376-RS plants. Survival of plants with the compound heterozygous genotype (122-R/376-R) under increasing chlorsulfuron doses was similar to 122-RR or 122-RS genotypes. However, this compound heterozygous genotype showed two- and six-fold higher LD50 values than 376-RR or 376-RS genotypes, respectively. However, both resistance 122-Tyr and 376-Glu alleles were dominant or nearly dominant over the wild-type susceptible alleles (ALA-122 and ASP-376), and the resistance 122-Tyr allele was dominant over the 376-Glu allele. CONCLUSIONS: These results broaden our understanding of AHAS target-site resistance in R. raphanistrum and strengthens the hypothesis that the AHAS 122-Tyr allele corresponds to a stronger target-site resistance allele than the 376-Glu allele. © 2024 Society of Chemical Industry.

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RESUMO Eleusine indica é uma planta daninha de difícil controle com herbicidas pós-emergentes devido ao curto intervalo entre a emergência e o perfilhamento, em que o controle é mais limitado. Ademais, o surgimento de biótipos resistentes motiva realizar trabalhos desta natureza para encontrar mais opções de controle químico. O objetivo desta investigação foi avaliar a eficiência de herbicidas pós-emergentes no controle do capim-pé-de-galinha em dois estádios fenológicos. O trabalho foi realizado em vasos a céu aberto num delineamento inteiramente casualizado, com onze herbicidas aplicados em dois experimentos: um experimento na espécie com quatro folhas (pré-perfilhamento) e outro na planta com oito folhas (em perfilhamento). Foram atribuídas notas visuais de controle aos 7, 14 e 21 dias após a aplicação, coletando a parte aérea da planta na última avaliação para obter a massa seca. Com esses dados foi realizada a análise de variância e a comparação de médias com o teste Tukey a um nível de significância de 0,05. Dos herbicidas testados, pyroxsulam, mesotrione y nicossulfuron controlaram menos do 80 % da população com quatro folhas. No perfilhamento, pyroxsulam, mesotrione, nicossulfuron y tembotrione mostraram controle menor ou igual a 65 %. Independentemente do estádio fenológico, clethodim, haloxifop, amônio-glufosinato, glyphosate, paraquat, indaziflam e ametrina foram efetivos, com mais de 88 % de controle da espécie.

ABSTRACT Eleusine indica is difficult to control with post-emergence herbicides due to the short interval between emergence and tillering, which control is more limited. Furthermore, the emergence of resistant biotypes motivates research on this type to find more options for chemical control. The objective of this work was to evaluate the efficiency of post-emergence herbicides in the goosegrass control in two phenological stages. The work was carried out in pots in a completely randomized design with eleven herbicides applied in two experiments: one with plants of four leaves (pre-tillering) and another with plants of eight leaves (in tillering). Visual control notes were assigned at 7, 14, and 21 days after application, collecting the aerial part of the plant in the last evaluation to obtain the dry weight. With these data, variance analysis and means comparison were performed using Tukey's test at a significance level of 0,05. As a principal result, pyroxsulam, mesotrione, and nicosulfuron controlled less than 80 % of the four-leaf population. At tillering, pyroxsulam, mesotrione, nicosulfuron, and tembotrione showed less than or equal to 65 % of control. Regardless of phenological stage, clethodim, haloxyfop, glufosinate ammonium, glyphosate, paraquat, indaziflam, and ametrine were effective, with more than 88 % of control of the species.

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Weedy rice, a pervasive and troublesome weed found across the globe, has often evolved through fertilization of rice cultivars with little importance of crop-weed gene flow. In Argentina, weedy rice has been reported as an important constraint since the early 1970s, and, in the last few years, strains with herbicide-resistance are suspected to evolve. Despite their importance, the origin and genetic composition of Argentinian weedy rice as well its adaptation to agricultural environments has not been explored so far. To study this, we conducted genotyping-by-sequencing on samples of Argentinian weedy and cultivated rice and compared them with published data from weedy, cultivated and wild rice accessions distributed worldwide. In addition, we conducted a phenotypic characterization for weedy-related traits, a herbicide resistance screening and genotyped accessions for known mutations in the acetolactate synthase (ALS) gene, which confers herbicide resistance. Our results revealed large phenotypic variability in Argentinian weedy rice. Most strains were resistant to ALS-inhibiting herbicides with a high frequency of the ALS mutation (A122T) present in Argentinian rice cultivars. Argentinian cultivars belonged to the three major genetic groups of rice: japonica, indica and aus while weeds were mostly aus or aus-indica admixed, resembling weedy rice strains from the Southern Cone region. Phylogenetic analysis supports a single origin for aus-like South American weeds, likely as seed contaminants from the United States, and then admixture with local indica cultivars. Our findings demonstrate that crop to weed introgression can facilitate rapid adaptation to agriculture environments.

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Low glyphosate doses that produce hormesis may alter the susceptibility to herbicides of weeds or enhance their propagation and dispersal. The objective of this work was to evaluate the hormetic effects of glyphosate on the vegetative, phenological and reproductive development in resistant (R) and susceptible (S) Conyza sumatrensis biotypes. The glyphosate resistance level of biotype R was 11.2-fold compared to the S biotype. Glyphosate doses <11.25 g ae ha-1 induced temporary and permanent hormetic effects for the number of leaves, plant height and dry mass accumulation up to 28 d after application in both R and S biotypes. The S biotype required 15-19% fewer thermal units at 1.4 and 2.8 g ae ha-1 glyphosate than untreated plants to reach the bolting stage. Also, this biotype had less thermal units associated with the appearance (1225 vs 1408 units) and opening (1520 vs 1765 units) of the first capitulum than the R biotype. In addition, glyphosate affected reproductive traits of both biotypes compared to their controls, increasing the number of capitulum's and seeds per plant up to 37 and 41% (at 2.8 and 0.7 g ae h-1, respectively) in the S biotype, and by 48 and 114% (both at 5.6 g ae ha-1) in the R biotype. Depending on environmental parameters, glyphosate may or may not cause hormetic effects on the vegetative and phenological development of C. sumatrenis biotypes; however, this herbicide increases the speed and fecundity of reproduction, regardless of the glyphosate susceptibility level, which can alter the population dynamics and glyphosate susceptibility of future generations.

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BACKGROUND: Chloris virgata is a troublesome weed in tropical regions. With the evolution of glyphosate resistance in key grass species, acetyl CoA carboxylase (ACCase) inhibitors have become a commonly used tool in soybean production areas in Brazil. We assessed if suspected resistant populations exhibited cross resistance to the different classes of ACCase inhibitors and investigated the resistance mechanisms in C. virgata. RESULTS: Dose-response experiments revealed resistance to haloxyfop-methyl and pinoxaden, with 432- and 3-fold resistance, respectively, compared to susceptible populations. Due to the lack of genetic resources for C. virgata, we sequenced, assembled, and annotated the genome using short-read Illumina technology. The k-mer analysis estimated a genome size of approximately 336 Mbp, with BUSCO completeness of 97%, and over 36 000 gene models were annotated. We examined if ACCase copy number variation and increased gene expression were involved in the resistance phenotype and found no difference when compared to a susceptible population. A mutation was detected in ACCase that encodes for amino acid position 2027, resulting in a tryptophan-to-cysteine (Trp2027Cys) substitution. We found the resistant population absorbed 11.4% less herbicide and retained 21% more herbicide on the treated leaf compared to the susceptible population. We developed a genotyping assay targeting the resistance-endowing Trp2027Cys substitution for quick resistance diagnosis. CONCLUSION: A Trp2027Cys amino acid substitution in ACCase confers resistance to haloxyfop and pinoxaden in C. virgata. We provide important insights into the evolutionary history of C. virgata and a draft genome as a useful resource to further our understanding of the biology in the genus Chloris. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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BACKGROUND: Eleusine indica (L.) Gaertn. (goosegrass) is a major weed in global cropping systems. It has evolved resistance to glyphosate due to single Pro-106-Ser (P106S) or double Thr-102-Ile + Pro-106-Ser (TIPS) EPSPS target site mutations. Here, experiments were conducted to evaluate the single effect of soybean competition and its combined effect with a glyphosate field dose (1080 g ae ha-1 ) on the growth and fitness of plants carrying these glyphosate resistance endowing target site mutations. RESULTS: TIPS E. indica plants are highly glyphosate-resistant but the double mutation endows a substantial fitness cost. The TIPS fitness penalty increased under the effect of soybean competition resulting in a cost of 95%, 95% and 96% in terms of, respectively, vegetative growth, seed mass and seed number investment. Glyphosate treatment of these glyphosate-resistant TIPS plants showed an increase in growth relative to those without glyphosate. Conversely, for the P106S moderate glyphosate resistance mutation, glyphosate treatment alone reduced survival rate, vegetative growth, aboveground biomass (34%), seed mass (48%) and number (52%) of P106S plants relative to the glyphosate nontreated plants. However, under the combined effects of both soybean competition and the field-recommended glyphosate dose, vegetative growth, aboveground biomass, seed mass and number of P106S and TIPS plants were substantially limited (by ≤99%). CONCLUSION: The ecological environment imposed by intense competition from a soybean crop sets a significant constraint for the landscape-level increase of both the E. indica single and double glyphosate resistance mutations in the agroecosystem and highlights the key role of crop competition in limiting the population growth of weeds, whether they are herbicide-resistant or susceptible. © 2022 Society of Chemical Industry.

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Hybridization between crops and their wild relatives may promote the evolution of de-domesticated (feral) weeds. Wild sunflower (Helianthus annuus L.) is typically found in ruderal environments, but crop-wild hybridization may facilitate the evolution of weedy populations. Using 1 crop-specific mitochondrial marker (CMS-PET1) and 14 nuclear SSR markers, we studied the origin and genetic diversity of a recently discovered weedy population of sunflower (named BRW). Then, using a resurrection approach, we tested for rapid evolution of weedy traits (seed dormancy, herbicide resistance, and competitive ability) by sampling weedy and wild populations 10 years apart (2007 and 2017). All the weedy plants present the CMS-PET1 cytotype, confirming their feral origin. At the nuclear markers, BRW showed higher genetic diversity than the cultivated lines and low differentiation with one wild population, suggesting that wild hybridization increased their genetic diversity. We found support for rapid evolution towards higher seed dormancy, but not for higher competitive ability or herbicide resistance. Our results highlight the importance of seed dormancy during the earliest stages of adaptation and show that crop-wild hybrids can evolve quickly in agricultural environments.

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Sugarcane (Saccharum spp.) is a tropical and sub-tropical, vegetative-propagated crop that contributes to approximately 80% of the sugar and 40% of the world's biofuel production. Modern sugarcane cultivars are highly polyploid and aneuploid hybrids with extremely large genomes (>10 Gigabases), that have originated from artificial crosses between the two species, Saccharum officinarum and S. spontaneum. The genetic complexity and low fertility of sugarcane under natural growing conditions make traditional breeding improvement extremely laborious, costly and time-consuming. This, together with its vegetative propagation, which allows for stable transfer and multiplication of transgenes, make sugarcane a good candidate for crop improvement through genetic engineering. Genetic transformation has the potential to improve economically important properties in sugarcane as well as diversify sugarcane beyond traditional applications, such as sucrose production. Traits such as herbicide, disease and insect resistance, improved tolerance to cold, salt and drought and accumulation of sugar and biomass have been some of the areas of interest as far as the application of transgenic sugarcane is concerned. Although there have been much interest in developing transgenic sugarcane there are only three officially approved varieties for commercialization, all of them expressing insect-resistance and recently released in Brazil. Since the early 1990's, different genetic transformation systems have been successfully developed in sugarcane, including electroporation, Agrobacterium tumefaciens and biobalistics. However, genetic transformation of sugarcane is a very laborious process, which relies heavily on intensive and sophisticated tissue culture and plant generation procedures that must be optimized for each new genotype to be transformed. Therefore, it remains a great technical challenge to develop an efficient transformation protocol for any sugarcane variety that has not been previously transformed. Additionally, once a transgenic event is obtained, molecular studies required for a commercial release by regulatory authorities, which include transgene insertion site, number of transgenes and gene expression levels, are all hindered by the genomic complexity and the lack of a complete sequenced reference genome for this crop. The objective of this review is to summarize current techniques and state of the art in sugarcane transformation and provide information on existing and future sugarcane improvement by genetic engineering.

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The global invasion, and subsequent spread and evolution of weeds provides unique opportunities to address fundamental questions in evolutionary and invasion ecology. Amaranthus palmeri is a widespread glyphosate-resistant (GR) weed in the USA. Since 2015, GR populations of A. palmeri have been confirmed in South America, raising questions about introduction pathways and the importance of pre- vs. post-invasion evolution of GR traits. We used RAD-sequencing genotyping to characterize genetic structure of populations from Brazil, Argentina, Uruguay and the USA. We also quantified gene copy number of the glyphosate target, 5-enolpyruvyl-3-shikimate phosphate synthase (EPSPS), and the presence of an extrachromosomal circular DNA (eccDNA) replicon known to confer glyphosate resistance in USA populations. Populations in Brazil, Argentina and Uruguay were only weakly differentiated (pairwise FST  ≤0.043) in comparison to USA populations (mean pairwise FST  =0.161, range =0.068-0.258), suggesting a single major invasion event. However, elevated EPSPS copy number and the EPSPS replicon were identified in all populations from Brazil and Uruguay, but only in a single Argentinean population. These observations are consistent with independent in situ evolution of glyphosate resistance in Argentina, followed by some limited recent migration of the eccDNA-based mechanism from Brazil to Argentina. Taken together, our results are consistent with an initial introduction of A. palmeri into South America sometime before the 1980s, and local evolution of GR in Argentina, followed by a secondary invasion of GR A. palmeri with the unique eccDNA-based mechanism from the USA into Brazil and Uruguay during the 2010s.

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Weeds are one of the main causes of the decrease in crop yields, with Johnsongrass (Sorghum halepense L.) being one of the most significant. Weeds can be controlled by herbicides, but some have developed resistance. Quantitative PCR is the technique of choice for studying gene expression related to herbicide resistance because of its high sensitivity and specificity, although its quantitative accuracy is highly dependent on the stability of the reference genes. Thus, in this study we evaluated the stability of different reference genes of glyphosate-resistant S. halepense. Nine genes frequently used as reference genes were selected: MDH, ADP, PP2A, EIF4α, ACT, ARI8, DnaJ, Hsp70, and ALS1, and their expression analyzed in susceptible and resistant biotypes at 0, 24 and 72 h post-application of glyphosate. The stability was analyzed with the geNorm, NormFinder, and BestKeeper software programs and using the ΔCt method. RefFinder was used to generate a comprehensive stability ranking. The results showed that PP2A and ARI8 were the most stable genes under the test conditions. EPSPS expression was also verified against the best two and the worst two reference genes. This study provides useful information for gene expression analysis under glyphosate stress and will facilitate resistance mechanism studies in this weed species.

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BACKGROUND: Digitaria insularis is a weed species that has gained considerable importance in Brazil's soybean production areas that rely on glyphosate-resistant cultivars. Herbicide-resistant weed populations of this species have been reported in many regions in Brazil, first in the south, followed by later reports in the north. We hypothesized that the spread of herbicide-resistant D. insularis is facilitated by movement of agricultural machinery from the southern regions of Brazil. RESULTS: Population genomics revealed a weak or no genetic structure (FST  = [0; 0.16]), moderate expected heterozygosity (HE  = 0.15; 0.44) and low inbreeding (FIS  = [-0.1; 0.1]) in D. insularis populations. Our data supported the hypothesis that herbicide resistance gene flow predominantly occurred in a south-to-north direction based on a migration analysis. We also found evidence of local adaptation of resistant populations in the northern soybean-growing regions of Brazil. CONCLUSION: Evidence in our work suggests that gene flow of glyphosate-resistant D. insularis is associated with movement of agricultural machinery, although local selection pressure seems to play an important role in the evolution of herbicide resistance throughout the country. Our results suggest preventive practices such as equipment sanitation should be implemented to limit the spread of herbicide resistant D. insularis. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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Herbicide resistance is broadly recognized as the adaptive evolution of weed populations to the intense selection pressure imposed by the herbicide applications. Here, we tested whether transcriptional gene silencing (TGS) and RNA-directed DNA Methylation (RdDM) pathways modulate resistance to commonly applied herbicides. Using Arabidopsis thaliana wild-type plants exposed to sublethal doses of glyphosate, imazethapyr, and 2,4-D, we found a partial loss of TGS and increased susceptibility to herbicides in six out of 11 tested TGS/RdDM mutants. Mutation in REPRESSOR OF SILENCING 1 (ROS1), that plays an important role in DNA demethylation, leading to strongly increased susceptibility to all applied herbicides, and imazethapyr in particular. Transcriptomic analysis of the imazethapyr-treated wild type and ros1 plants revealed a relation of the herbicide upregulated genes to chemical stimulus, secondary metabolism, stress condition, flavonoid biosynthesis, and epigenetic processes. Hypersensitivity to imazethapyr of the flavonoid biosynthesis component TRANSPARENT TESTA 4 (TT4) mutant plants strongly suggests that ROS1-dependent accumulation of flavonoids is an important mechanism for herbicide stress response in A. thaliana. In summary, our study shows that herbicide treatment affects transcriptional gene silencing pathways and that misregulation of these pathways makes Arabidopsis plants more sensitive to herbicide treatment.

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Since glyphosate was classified as potentially carcinogenic by the International Agency for Research on Cancer, public debate regarding the environmental impact and health risks from its use has intensified. Almost all regulatory agencies throughout the world have concluded that the judicious use of glyphosate does not pose risks to the environment and human health. However, on the last day of 2020 the Mexican government decreed a ban of this herbicide beginning January, 2024. In current Mexican agriculture there are no safer chemical and/or other weed management technologies that allow for the economical substitution of glyphosate for weed control. Many Mexican weed scientists agree that glyphosate use should be reduced, but not banned outright. This decree could have more negative economic and social consequences as well as environmental and human health risks than benefits, which could compromise the country's food and public security. Crop yields are projected by some to decline by up to 40% with this ban, increasing food prices, making food less accessible to low-income consumers. In addition, a black market for the smuggling and illegal sale of glyphosate is possible. The possible environmental, economic and social impacts caused by the glyphosate ban in Mexico are discussed, emphasizing the impact on weed management. © 2021 Society of Chemical Industry.

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Herbicides play an important role in preventing crop yield losses due to both their weed interference ability and their capacity for increasing soil conservation in no-till systems. Group A herbicides or acetyl-CoA carboxylase (ACCase) are essential tools the selective management of glyphosate resistance in grass weed species. In this review, we describe important aspects of ACCase biology and herbicides targeting this enzyme, along with a discussion on stewardship programs to delay the evolution of herbicide resistance which can evolve either through target site and/or non-target site mechanisms. Sixteen-point mutations have been reported to confer resistance to ACCase inhibitors. Each mutation confers cross resistance to a different group of herbicides. Metabolic resistance can result in resistance to multiple herbicides with different mechanisms of action (MoA), and herbicide detoxification is often conferred by cytochrome P450 monooxigenases and glutathione- S -transferases. Regardless of whether resistance mechanisms are target or non-target site, using herbicides with the same MoA will result in resistance evolution. Therefore, while field surveys and resistance mechanism studies are crucial for designing reactive management strategies, integrated weed management plays a central role in both reactive and proactive mitigation of herbicide resistance evolution.(AU)

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Herbicides play an important role in preventing crop yield losses due to both their weed interference ability and their capacity for increasing soil conservation in no-till systems. Group A herbicides or acetyl-CoA carboxylase (ACCase) are essential tools the selective management of glyphosate resistance in grass weed species. In this review, we describe important aspects of ACCase biology and herbicides targeting this enzyme, along with a discussion on stewardship programs to delay the evolution of herbicide resistance which can evolve either through target site and/or non-target site mechanisms. Sixteen-point mutations have been reported to confer resistance to ACCase inhibitors. Each mutation confers cross resistance to a different group of herbicides. Metabolic resistance can result in resistance to multiple herbicides with different mechanisms of action (MoA), and herbicide detoxification is often conferred by cytochrome P450 monooxigenases and glutathione- S -transferases. Regardless of whether resistance mechanisms are target or non-target site, using herbicides with the same MoA will result in resistance evolution. Therefore, while field surveys and resistance mechanism studies are crucial for designing reactive management strategies, integrated weed management plays a central role in both reactive and proactive mitigation of herbicide resistance evolution.

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Herbicide-resistant oilseed rape (Brassica napus) cultivation in our country entails the risk of gene transfer to related wild species. One of these species is the wild turnip (B. rapa), an important weed of winter crops widely distributed in the Pampas region. Despite hybridization risks, Clearfield ® oilseed rape is available in Argentina. In 2008, a B. rapa population, which was sympatric to an imidazolinone-resistant and a conventional oilseed rape cultivar, was located on a farm in the main cropping area of the country. Herbicide-resistant individuals were found in the progeny of this population in a herbicide screening test. Therefore, a molecular characterization using cleaved amplified polymorphic sequence (CAPS) and simple sequence repeat (SSR) markers was conducted on these plants to determine their hybrid nature and to establish the origin of the imidazolinone resistance trait. The results of this study, along with information of field records, confirmed that the resistant plants were first generation interspecific hybrids. Imidazolinone resistance had been effectively transferred from the herbicide-resistant oilseed rape, even in the particular situation of pollen competition. Oilseed rape resistant cultivars are becoming more common in the country. So, considering that seed loss and crop volunteers are common in these species, it is crucial to avoid the dispersion of new resistant weed biotypes as they reduce the effectiveness of chemical control technologies.

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Herbicide resistant (HR) weeds are of major concern in modern agriculture. This situation is exacerbated by the massive adoption of herbicide-based technologies along with the overuse of a few active ingredients to control weeds over vast areas year after year. Also, many other anthropological, biological, and environmental factors have defined a higher rate of herbicide resistance evolution in numerous weed species around the world. This review focuses on two central points: 1) how these factors have affected the resistance evolution process; and 2) which cultural practices and new approaches would help to achieve an effective integrated weed management. We claim that global climate change is an unnoticed factor that may be acting on the selection of HR weeds, especially those evolving into non-target-site resistance mechanisms. And we present several new tools -such as Gene Drive and RNAi technologies- that may be adopted to cope with herbicide resistance spread, as well as discuss their potential application at field level. This is the first review that integrates agronomic and molecular knowledge of herbicide resistance. It covers not only the genetic basis of the most relevant resistance mechanisms but also the strengths and weaknesses of traditional and forthcoming agricultural practices.

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Perennial plants which propagate through both seeds and rhizomes are common in agricultural and nonagricultural systems. Due to their multifaceted life cycle, few population models are available for studying such species. We constructed a novel individual-based model to examine the effects of ecological, evolutionary, and anthropogenic factors on the population dynamics of perennial species. To exemplify the application of the model, we presented a case study of an important weed, Sorghum halepense (L.) Pers. (Johnsongrass), in soybean productions in Argentina. The model encompasses a full perennial weed life cycle with both sexual (seeds) and asexual (rhizomes) propagations. The evolution of herbicide resistance was modeled based on either single genes or quantitative effects. Field experiments were conducted in the species' native environment in Argentina to parameterize the model. Simulation results showed that resistance conferred by single-gene mutations was predominantly affected by the initial frequency of resistance alleles and the associated fitness cost. Population dynamics were influenced by evolved resistance, soil tillage, and rhizome fecundity. Despite the pivotal role of rhizomes in driving the population dynamics of Johnsongrass, most herbicides target the aboveground biomass, and chemical solutions to control rhizomes are still very limited. To maintain effective (short-term) and sustainable (long-term) weed management, it is recommended to combine soil tillage with herbicide applications for suppressing the rhizomes and delaying the evolution of resistance. This novel model of seed- and rhizome-propagated plants will also be a useful tool for studying the evolutionary processes of other perennial weeds, cash crops, and invasive species.

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Herbicides inhibiting acetyl-coenzyme A carboxylase (ACCase) are very effective in controlling grass weeds including weedy-rice in paddy rice production systems. The ACCase inhibitor affects the enzyme by blocking fatty acid biosynthesis resulting in plant death. The herbicide resistance in rice is conferred by a single point mutation with an amino acid substitution of the carboxyl transferase domain of the ACCase gene. An assay based on the tetra-primer ARMS-PCR method was developed to detect the SNP G2027T that causes a tryptophan-cysteine substitution in the gene encoding chloroplastic ACCase in rice. The protocol was tested in 453 rice samples from a segregant population for validation of the assay. This technique can be exploited to monitor resistant lines in rice breeding programs to detect homozygous or heterozygous resistant genotypes and homozygous susceptible genotypes. The presence of resistant ACCase allele(s) can be detected with rapidity, simplicity, at low cost and can be used in any molecular biology laboratory with minimal equipment.

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Conyza bonariensis (hairy fleabane) is one of the most problematic and widespread glyphosate-resistant weeds in the world. This highly competitive weed species significantly interferes with crop growth and substantially decreases crop yield. Despite its agricultural importance, the molecular mechanisms of glyphosate resistance are still unknown. The present RNA-Seq study was performed with the goal of identifying differentially expressed candidate transcripts (genes) related to metabolism-based non-target site glyphosate resistance in C. bonariensis. The whole-transcriptome was de novo assembled from glyphosate-resistant and -sensitive biotypes of C. bonariensis from Southern Brazil. The RNA was extracted from untreated and glyphosate-treated plants at several timepoints up to 288 h after treatment in both biotypes. The transcriptome assembly produced 90,124 contigs with an average length of 777 bp and N50 of 1118 bp. In response to glyphosate treatment, differential gene expression analysis was performed on glyphosate-resistant and -sensitive biotypes. A total of 9622 genes were differentially expressed as a response to glyphosate treatment in both biotypes, 4297 (44.6%) being up- and 5325 (55.4%) down-regulated. The resistant biotype presented 1770 up- and 2333 down-regulated genes while the sensitive biotype had 2335 and 2800 up- and down-regulated genes, respectively. Among them, 974 up- and 1290 down-regulated genes were co-expressed in both biotypes. In the present work, we identified 41 new candidate target genes from five families related to herbicide transport and metabolism: 19 ABC transporters, 10 CYP450s, one glutathione S-transferase (GST), five glycosyltransferases (GT), and six genes related to antioxidant enzyme catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD). The candidate genes may participate in metabolic-based glyphosate resistance via oxidation, conjugation, transport, and degradation, plus antioxidation. One or more of these genes might 'rescue' resistant plants from irreversible damage after glyphosate treatment. The 41 target genes we report in the present study may inform further functional genomics studies, including gene editing approaches to elucidate glyphosate-resistance mechanisms in C. bonariensis.